MagnetoWiki - User contributions [en]

SRA Lithium Orotate c-Fos

2026-10-31T19:53:50Z

Houpt: update with rat dob and some times

Induction of c-Fos in rat brain by acute injections of lithium orotate (LiOR), LiCl, or NaCl.

{{Infobox experiment|align=right
| Name = Lithium Orotate c-Fos
| project = Taste Aversion
| code = SRA
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Rashid, Houpt
| species = rat
| date = 2026-10-15
| type = CTA
| drugs = lithium orotate, lithium chloride
| data =
}}

== People ==

Sheikh Rashid

== Agenda ==

=== Prepare Solutions ===

all injections made isotonic with TrisAmino:

* 0.3 M TrisAmino
36.34g / liter, autoclave, pH to 8.0 with 38% HCl

* 0.15 M LiOH
3.6g / liter (no need to autoclave)

* 0.15 M LiCl
6.36 g / liter, autoclave

* LiOr = 75mM LiOrotate, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiOH, 0.47g orotic acid, pH to 8 with 1N HCl, filter

* LiCl = 75mM LiCl, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiCl, pH to 8 with 1N HCl, filter

* NaCl = 75mM NaCl, 150 mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M NaCl, pH to 8 with 1N HCl, filter

Perfusion solutions:

pre-perfusion saline

4% paraformaldehyde

=== Perfusion Day (2026-10-31) ===

Rats (n=9, d.o.ob 2026-7-2, arr. 2026-9-1) injected with one of the following solutions (staggered in pairs by 15-20min):

* 75 mM LiOr/Tris, 6 ml/kg ip. (SRA01, SRA04, SRA07)
* 75 mM LiCl/Tris, 6 ml/kg ip. (SRA02, SRA05, SRA08)
* 75 mM NaCl/Tris, 6 ml/kg ip. (SRA03, SRA06, SRA09)

These are same doses used in [[SNB Lithium Orotate CTA 2]], which showed that LiOr had greater potency in CTA conditioning than LiCl.

One hour after injection, rats are overdosed with sodium pentobarbital and transcardially perfused. Brains were dissected, postfixed for ~1 h, then transferred to 30% sucrose before sectioning.

* 0: inject SRA01,SRA02 (1015)
* 20: inject SRA03, SRA04
* 40: inject SRA05, SRA06
* 60: inject SRA07, SRA08 ; perfuse SRA01, SRA02
* 80: inject SRA09 ; perfuse SRA03, SRA04
* 100: perfuse SRA05, SRA06
* 120: perfuse SRA07, SRA08
* 140: perfuse SRA09

30% sucrose (1500)

=== c-Fos Immunohistochemistry ===

Day 1: section forebrain, start immuno

Day 2: finish immuno

Day 3: mount tissue sections

Repeat for hindbrain...

=== Analysis ===

Digitize sections on microscope to count cells (a couple of days)

SRA Lithium Orotate c-Fos

2026-10-31T13:56:15Z

Houpt: updated with actual pH of 8.0

Induction of c-Fos in rat brain by acute injections of lithium orotate (LiOR), LiCl, or NaCl.

{{Infobox experiment|align=right
| Name = Lithium Orotate c-Fos
| project = Taste Aversion
| code = SRA
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Rashid, Houpt
| species = rat
| date = 2026-10-15
| type = CTA
| drugs = lithium orotate, lithium chloride
| data =
}}

== People ==

Sheikh Rashid

== Agenda ==

=== Prepare Solutions ===

all injections made isotonic with TrisAmino:

* 0.3 M TrisAmino
36.34g / liter, autoclave, pH to 8.0 with 38% HCl

* 0.15 M LiOH
3.6g / liter (no need to autoclave)

* 0.15 M LiCl
6.36 g / liter, autoclave

* LiOr = 75mM LiOrotate, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiOH, 0.47g orotic acid, pH to 8 with 1N HCl, filter

* LiCl = 75mM LiCl, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiCl, pH to 8 with 1N HCl, filter

* NaCl = 75mM NaCl, 150 mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M NaCl, pH to 8 with 1N HCl, filter

Perfusion solutions:

pre-perfusion saline

4% paraformaldehyde

=== Perfusion Day ===

Rats injected with one of the following solutions (staggered in pairs by 15-20min):

* 75 mM LiOr/Tris, 6 ml/kg ip. (SRA01, SRA04, SRA07)
* 75 mM LiCl/Tris, 6 ml/kg ip. (SRA02, SRA05, SRA08)
* 75 mM NaCl/Tris, 6 ml/kg ip. (SRA03, SRA06, SRA09)

These are same doses used in [[SNB Lithium Orotate CTA 2]], which showed that LiOr had greater potency in CTA conditioning than LiCl.

One hour after injection, rats are overdosed with sodium pentobarbital and transcardially perfused. Brains are dissected, and placed into 0.1M PB for storage, then transferred to 30% sucrose overnight before sectioning.

* 0: inject SRA01,SRA02
* 20: inject SRA03, SRA04
* 40: inject SRA05, SRA06
* 60: inject SRA07, SRA08 ; perfuse SRA01, SRA02
* 80: inject SRA09 ; perfuse SRA03, SRA04
* 100: perfuse SRA05, SRA06
* 120: perfuse SRA07, SRA08
* 140: perfuse SRA09

=== c-Fos Immunohistochemistry ===

Day 1: section forebrain, start immuno

Day 2: finish immuno

Day 3: mount tissue sections

Repeat for hindbrain...

=== Analysis ===

Digitize sections on microscope to count cells (a couple of days)

SNB Lithium Orotate CTA 2

2026-10-19T17:50:41Z

Houpt: fix typo in LiCl formula

Continuation of [[SNA Lithium Orotate CTA]].

Test if lithium orotate (LiOr) induces a conditioned taste aversion comparable to LiCl, with a lower dose than SNA experiment

Single pairing of saccharin intake (15 min) after 15-min delay with isomolar doses of LiCl and LiOr to see if LiOr is more potent.

{{Infobox experiment|align=right
| Name = Lithium Orotate and CTA pt2
| project = Taste Aversion
| code = SNB
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-3-13
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SNB
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SNB Raw Data Link]

== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (2026-2-10), n= 16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2025-12-15), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-3-13)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-3-14 thru 2026-3-19)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 900 daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 12: Conditioning Day''' (2026-3-20)

0 min: 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min: rats injected with:

* 75 mM NaCl, 6 ml/kg ip. (SNB01,04,07,10,13)
* 75 mM LiCl, 6 ml/kg ip. (SNB02,05,08,11,14)
* 75 mM LiOr, 6 ml/kg ip. (SNB03,06,09,12,15,16)

all injections made isotonic with TrisAmino:

* 0.3 M TrisAmino
36.34g / liter, autoclave, pH to 7.4 with 38% HCl

* 0.15 M LiOH
3.6g / liter (no need to autoclave)

* 0.15 M LiCl
6.36g / liter, autoclave

* LiOr = 75mM LiOrotate, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiOH, 0.47g orotic acid, pH to 7.4 with LiOH, filter

* LiCl = 75mM LiCl, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiCl, pH to 7.4 with LiOH, filter

* NaCl = 75mM NaCl, 150 mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M NaCl, pH to 7.4 with NaOH, filter

Ad lib water returned after final injection overnight.

'''Days 13 - 22: Preference Tests''' (2026-3-21 through 2026-4-1)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 10 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

SRA Lithium Orotate c-Fos

2026-10-19T17:49:08Z

Houpt: fix typo in LiCl formula

Induction of c-Fos in rat brain by acute injections of lithium orotate (LiOR), LiCl, or NaCl.

{{Infobox experiment|align=right
| Name = Lithium Orotate c-Fos
| project = Taste Aversion
| code = SRA
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Rashid, Houpt
| species = rat
| date = 2026-10-15
| type = CTA
| drugs = lithium orotate, lithium chloride
| data =
}}

== People ==

Sheikh Rashid

== Agenda ==

=== Prepare Solutions ===

all injections made isotonic with TrisAmino:

* 0.3 M TrisAmino
36.34g / liter, autoclave, pH to 7.4 with 38% HCl

* 0.15 M LiOH
3.6g / liter (no need to autoclave)

* 0.15 M LiCl
6.36 g / liter, autoclave

* LiOr = 75mM LiOrotate, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiOH, 0.47g orotic acid, pH to 7.4 with LiOH, filter

* LiCl = 75mM LiCl, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiCl, pH to 7.4 with LiOH, filter

* NaCl = 75mM NaCl, 150 mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M NaCl, pH to 7.4 with NaOH, filter

Perfusion solutions:

pre-perfusion saline

4% paraformaldehyde

=== Perfusion Day ===

Rats injected with one of the following solutions (staggered in pairs by 15-20min):

* 75 mM LiOr/Tris, 6 ml/kg ip. (SRA01, SRA04, SRA07)
* 75 mM LiCl/Tris, 6 ml/kg ip. (SRA02, SRA05, SRA08)
* 75 mM NaCl/Tris, 6 ml/kg ip. (SRA03, SRA06, SRA09)

These are same doses used in [[SNB Lithium Orotate CTA 2]], which showed that LiOr had greater potency in CTA conditioning than LiCl.

One hour after injection, rats are overdosed with sodium pentobarbital and transcardially perfused. Brains are dissected, and placed into 0.1M PB for storage, then transferred to 30% sucrose overnight before sectioning.

* 0: inject SRA01,SRA02
* 20: inject SRA03, SRA04
* 40: inject SRA05, SRA06
* 60: inject SRA07, SRA08 ; perfuse SRA01, SRA02
* 80: inject SRA09 ; perfuse SRA03, SRA04
* 100: perfuse SRA05, SRA06
* 120: perfuse SRA07, SRA08
* 140: perfuse SRA09

=== c-Fos Immunohistochemistry ===

Day 1: section forebrain, start immuno

Day 2: finish immuno

Day 3: mount tissue sections

Repeat for hindbrain...

=== Analysis ===

Digitize sections on microscope to count cells (a couple of days)

SRA Lithium Orotate c-Fos

2026-10-15T16:13:00Z

Houpt: created agenda

Induction of c-Fos in rat brain by acute injections of lithium orotate (LiOR), LiCl, or NaCl.

{{Infobox experiment|align=right
| Name = Lithium Orotate c-Fos
| project = Taste Aversion
| code = SRA
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Rashid, Houpt
| species = rat
| date = 2026-10-15
| type = CTA
| drugs = lithium orotate, lithium chloride
| data =
}}

== People ==

Sheikh Rashid

== Agenda ==

=== Prepare Solutions ===

all injections made isotonic with TrisAmino:

* 0.3 M TrisAmino
36.34g / liter, autoclave, pH to 7.4 with 38% HCl

* 0.15 M LiOH
3.6g / liter (no need to autoclave)

* 0.15 M LiCl
6.45g / liter, autoclave

* LiOr = 75mM LiOrotate, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiOH, 0.47g orotic acid, pH to 7.4 with LiOH, filter

* LiCl = 75mM LiCl, 150mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M LiCl, pH to 7.4 with LiOH, filter

* NaCl = 75mM NaCl, 150 mM TrisAmino
20 ml of 0.3M TrisAmino, 20 ml of 0.15M NaCl, pH to 7.4 with NaOH, filter

Perfusion solutions:

pre-perfusion saline

4% paraformaldehyde

=== Perfusion Day ===

Rats injected with one of the following solutions (staggered in pairs by 15-20min):

* 75 mM LiOr/Tris, 6 ml/kg ip. (SRA01, SRA04, SRA07)
* 75 mM LiCl/Tris, 6 ml/kg ip. (SRA02, SRA05, SRA08)
* 75 mM NaCl/Tris, 6 ml/kg ip. (SRA03, SRA06, SRA09)

These are same doses used in [[SNB Lithium Orotate CTA 2]], which showed that LiOr had greater potency in CTA conditioning than LiCl.

One hour after injection, rats are overdosed with sodium pentobarbital and transcardially perfused. Brains are dissected, and placed into 0.1M PB for storage, then transferred to 30% sucrose overnight before sectioning.

* 0: inject SRA01,SRA02
* 20: inject SRA03, SRA04
* 40: inject SRA05, SRA06
* 60: inject SRA07, SRA08 ; perfuse SRA01, SRA02
* 80: inject SRA09 ; perfuse SRA03, SRA04
* 100: perfuse SRA05, SRA06
* 120: perfuse SRA07, SRA08
* 140: perfuse SRA09

=== c-Fos Immunohistochemistry ===

Day 1: section forebrain, start immuno

Day 2: finish immuno

Day 3: mount tissue sections

Repeat for hindbrain...

=== Analysis ===

Digitize sections on microscope to count cells (a couple of days)

Conditioned Taste Aversion

2026-10-14T13:11:04Z

Houpt: added some grant text

=Grant Procedure Text=

see Houpt RO1 ResearchStrategy2014 for more text

Conditioned taste aversion (CTA) is a robust form of associative learning in which an animal avoids and reacts aversively to the taste of a food that has been previously paired with illness. CTA is a rich behavioral phenomenon with many of the characteristics of classical conditioning, but with additional unique features such as single-trial learning, long-delay learning, learned safety, taste specificity, and olfactory potentiation [1]. CTA allows integration of gustation with manifold unconditioned stimuli such as ionizing radiation [2], gastrointestinal toxins [3], chemotoxins [4], visual optokinetic stimuli [5], and vestibular perturbation [6,7]. Neurologically, CTA belongs to the class of indirect controls of feeding, by which a distributed forebrain-hindbrain network modulates the direct responses of the brainstem to taste stimulation [1,9]. The robust conditioned change of behavioral responses to tastes in CTA serves as model of a learned control of ingestion.

While CTA has unique adaptations subserving ingestive learning, it presents features of relevance to the general neurobiology of learning. Like fear conditioning, CTA is an amygdala-dependent form of traumatic memory that is rapidly acquired and extremely persistent. Acquisition of CTA is NMDA-receptor mediated, and sensitive to tauopathy. There is considerable overlap of intracellular signaling pathways in CTA with other models of plasticity (i.e. kinases, adaptor proteins, and transcription factors). T unique neuroanatomical and temporal properties of CTA learning permit easier manipulation of intracellular pathways and quantification of the behavioral consequences. Relevant to this proposal, CTA learning is compromised in several mouse and rat models of AD.

== CTA Conditioning and Assessment==
We will employ a standard procedure for conditioning and assessing CTA: The Conditioned Stimulus (CS) is an innately preferred but non-nutritive tastant (0.125% saccharin) presented by 1-bottle access (10min) or intraoral infusion (6 ml/6min). Prior to conditioning, animals are water restricted at 10-min access per day to guarantee ingestion on conditioning day. The Unconditioned Stimulus (US) will be a moderate dose of toxin (e.g. (18-36 mg/kg LiCl, ip, or 7.5 mg/kg apomorphine hydrochloride i.p.) that will allow observation of either potentiation or attenuation of CTA by Li deficiency or supplementation.. The CS-US interval will be 30-min from the start of the CS to the time of US injection, thereby modeling long- delay CTA learning. Depending on the manipulation, the CS-US interval may be probed by administering the treatment during the CS-US interval (e.g. injection of inhibitors 15 min after the CS but before the US). Expression Tests will measure the initial magnitude and rate of extinction of a CTA using 24-h, 2-bottle preference tests of CS vs. water across 14 days post-acquisition. Bar-coded bottles and computer-assisted weighing allows monitoring of up to 96 animals per day. We operationally define a CTA as a significantly lower CS preference (24-h saccharin intake/total intake) compared to controls. When examining short-term effects, intraoral infusions (6 ml/6min) will be used to precisely deliver the CS, 0-6 h after the US [97].

=Scope=
''This article describes some of the fundamental behavioral properties of conditioned taste aversion learning. The article is a work in progress -- in particular, no specific citations are provided yet.''

As a form of associative learning by which an animal avoids a novel taste or food that has been previously paired with a toxin, CTA has been widely used as a marker of aversive effects caused by drugs and treatments. CTA learning is remarkably sensitive, and often reveals a treatment effect even when no other behavioral effect is detectable. Purely exteroceptive sensory stimuli, such as light or auditory cues produced by the MF or the apparatus, are likely to be ignored because CTAs overwhelmingly favor a novel taste as the conditioned stimulus [53]. Likewise, stressful effects of the exposure procedure, such as restraint or cutaneous discomfort, are not sufficient to act as the unconditioned stimulus in CTA learning [39, 53]. Aversive effects that activate interoreceptors or induce nausea in humans, however, are favored to induce CTAs [53].

=Description=
Conditioned taste aversion (CTA) is a form of associative learning in which an animal learns to avoid and reject a food after the taste or flavor of the food is paired with a toxic consequence. For example, an animal encounters a new food with a distinctive (but not necessarily aversive) taste. The animal eats the food, but subsequently gets sick. The animal will associate the taste with the sickness, so that the next time it encounters the food, it will avoid it, or eat less of it. This form of learning has obvious advantages for survival, and animals are extremely good at learning what foods are safe to eat, and what foods are toxic and should be avoided.

The animal can learn a CTA if food is actually toxic, or under experimental conditions in which the food is not toxic, but we administer a separate toxin to the rat after it eats the food. We typically use a sweet tasting solution (e.g. 5% sucrose or 0.1% saccharain) paired with a mildly toxic dose of lithium chloride (0.15M LiCl; 12 ml/kg i.p.). Rats, like most animals, will readily consume a sweet solution the first time they taste it. After letting the rats drink sucrose for a few minutes, we inject the LiCl which makes the rat mildly sick for a couple of hours. When we give the rats sucrose to drink the next day, they drink little if any of the solution. (Control rats get sucrose paired with a non-toxic injection of 0.15 M NaCl; because the rats don’t get sick after the sucrose, they will readily drink it the next day.)

CTA is a form of associative learning, meaning that it involves a learned change in response to a stimulus after the association of the stimulus with another effect. (This is opposed to 1) non-associative forms of learning, such as habituation or sensitization, that rely on repetition of the stimulus to induce a change in response, or 2) declarative or episodic memory (loosely called cognitive or conscious learning.) As a form of classical (Pavlovian) conditioning, CTA learning is almost reflexive in its acqusition and expression, without the requirement for conscious thought.

The components of CTA learning parallel the components of Pavlovian conditioning:
The unconditioned stimulus (US) produces an unconditioned response, but after pairing with an unrealted or neutral conditioned stimulus (CS), the CS will come to elicit a conditioned response (CR) that more or less resembles the UR.

In Pavlovian conditioning:

:US -> UR
::''meat -> salivation''

:CS -> no response
::''bell -> no response''

:CS + US -> UR
::''bell + meat -> salivation''

:CS -> CR
::''bell -> salivation''

In CTA:

:US -> UR
::''LiCl -> toxic effect''

:CS -> ingestion
::''sucrose -> ingestion''

:CS + US -> UR
::''sucrose + LiCl -> toxic effect''

:CS -> rejection
::''sucrose -> rejection''

CTA differs from classical pavlovian conditioning in 2 important ways:

:1. the CS + US must occur almost simultaneously in pavlovian conditioning, with very little delay between the two stimuli (milliseconds to seconds.) In CTA learning, an interval of up to 12 h can be occur between taste and toxin. Thus, CTA learning is “long-delay” learning.

:2. the CR is qualitatively similar to the UR in pavlovian conditioning: the dog will salivate to the bell, just as it salivated to the meat (although it may salivate less to the bell than to the original meat.) In CTA learning, the CR and UR are different: LiCl does not induce a rejection response, it just makes the rat sick: However, when LiCl is paired with sucrose, the subsequent behavioral response of the rat to sucrose is reversed and the rat expresses a CR unrelated to the UR it expresses to LiCl. This second distinction is subtle, but it may turn out to be important.

Some people have proposed finer classification systems, such as conditioned taste avoidance in which the animal only passively avoids or rejects the taste but does not actively reject the food, as measured with active rejection responses during an intraoral infusion (see the work of L. Parker and K.P. Ossenkopf) . However, I am more comfortable refering generically to CTAs in which the animal avoids ingestion of the food, or decreases the amount of food ingested, or actively rejects the food. In all three cases there is a decrease in the rat’s acceptance or preference for the food: it is not clear that the apparent quantitative differences in CTA strength reflect qualitatively different processes.

=Properties of a CTA=
CTA learning has a number of properties that distinguish it from other forms of associative learning. Animals may reject a food for many reasons (e.g. satiety, innate aversion, or non-CTA forms of learning). In order for an animal’s rejection of a food to be labeled a CTA, the following properites of acquisition should be demonstrated:

==1. The CTA is acquired best with novel tastes.==
A rat or other animal learns a CTA best when the toxic effect is paired with a novel taste the rat has never before encountered before. Put another way, if the toxin is paired with a familiar taste the rat has tasted many times before without getting sick, the rat is less likely to learn a CTA (or, the magnitude of the CTA will be attenuated when a familiar taste is paired with a toxin). This is called “learned safety”: the rat has learned that a taste is not associated with illness by non-contingent pre-exposure (i.e. the experience of the taste was not contingent on getting sick afterwards) that a taste is not associated with illness: this makes the rat resistant to CTA against that taste. Thus, If learned safety does not attenuate the animal’s rejection of a food, the animal is not rejecting the food because of a CTA.

==2. The conditioned stimulus is an orosensory stimulus.==
A CTA is the pairing of a toxic effect with a taste, flavor, or oral somatosensory stimulus. Historically, it has been very difficult to associate the effects of a interoceptive toxin with a conditioned stimulus that works through non-oral modalities (e.g. pairing a visual, auditory, or somatic stimulus with a toxic injection of LiCl).

An important consequence of this property is that CTA learning is relatively resistant to contextual conditioning or modulation. Context is a nebulous term refering to multimodal sensory input derived from the environment of the experiment as a whole. Thus the spatial properties, visual characteristics, ambient noise, etc. of the testing situation make up the context; but for CTA learning, the context is far less important than the orosenory stimulation. Thus, if the animal's rejection of a food can be attributed to a learned response to contextual, non-orosensory stimuli, then the behavior is not due to CTA.

(Note that other species may be more sensitive than rats to sensory modalities outside the mouth and nose. For example, birds and primates can easily associate visual cues with intereoceptive stimuli to form visual aversions. Presumably the ability to learn food aversions would evolve to take advantage of the primary senses involved in food detection.)

==3. The toxic unconditioned stimulus is an intereoceptive, not exteroceptive, stimulus.==
Parallel to property number 2, CTA requires that the toxic, negative or aversive stimulus paired with the taste be an interoceptive stimulus. An interoceptive stimulus acts on internal receptors (such as nerves in the gut), while exteroceptive stimuli are detected by receptors on the external surface of the animal (like somatosensation) or that can detect stimuli at a distance (like vision or audition). Thus, a rat finds cutaneous shock, intense light flash, or a loud bang to be aversive stimuli – but the rat finds it very difficult to associate taste with any of these exteroceptive stimuli. Conversely, the rat associates very easily a taste with toxic postingestive effects (such as intragastric copper sulfate that irritates the gut and the sensory vagus.) A convenient consequence of this is that rats do not form CTAs because of the prick of the injection needle!

Although this definition by receptor location sounds clear cut, the interoceptive stimulus is is an ill-defined concept. The essential characteristics of intereoreceptive stimuli that induce CTA have not been rigorously delineated. In addition to literal toxins, internal stimuli as diverse as gastric distention, rotation, or depletion of metabolic fuels can induce CTAs. Other treatments that clearly affect internal state are not sufficient, however, such as restraint stress (which elevates heart rate, glucocorticoid secretion, and gastric acid secretion). It would be interesting to test whether other forms of homestatic stress (e.g. cold stress) or internal afferent stimulation (e.g. intraintestinal capsaicin) are sufficient to induce CTA.

A general rule of thumb is that anything that makes a human feel sick will be sufficient to induce a CTA in a rat. The converse, however, is not true: rats have acquired CTAs to tastes paired with saline, and with drugs of abuse that are usually self-administered, and with other compounds that have little or no negative effect in humans. Jim Smith has an excellent defnition of this property: anything that makes a rat feel “different “ after a meal will be sufficient to induce a CTA.

Thus, if the rat’s rejection of the food is due to learning due to exteroceptive stimulation, the behavior is not due to CTA

[Properties 2 and 3 are probably true but they are based on only a few early papers: they need to be confirmed in a rigourous (modern?) fashion!]

==4. The CTA can be acquired in a single trial (with sufficiently strong stimuli)==
There are other forms of single-trial learning, such as fear conditioning (pairing a tone with foot-shock.) However, most forms of classical conditioning (such as pavlovian salivation or eyeblink conditioning) require dozens or hundreds of pairings across many days to achieve a robust conditioned response. This makes CTA a convenient model system to study, because the changes in the brain during learning must be relatively large and rapid to persist after only a single trial.

[Note that although CTA can be acquired in a single trial with a strong toxic US, a CTA can also be acquired after multiple pairings of the taste with a weaker stimulus.]

==5. The CTA tolerates a long delay between taste and toxin==
CTA can be aquired even with an interstimulus interval of minutes to hours (as opposed to the upper limit of milliseconds to seconds required by other forms of classical conditioning ). There is at least one report that CTA learning is optimized for a long-delay: in other words, rats learn a CTA better with if there is a delay of 15-30 min (vs a delay of seconds) between the taste and the toxic effect.

This property is quite unique to CTA learning, and has important implications. It makes functional sense: after eating a toxin, the postingestive effects might no be apparent for minutes to hours later, so the animal needs to associate the taste with a delayed effect. However, it might also imply that CTA learning is mediated by unique neural and molecular mechanisms that tolerate long delays (or at least that familiar synaptic mechanisms have been adapted to this particular temporal niche.)

[Note that we may be able to take advantage of the long-delay characteristics: for example, we should be able to intervene between the taste and toxin, e.g. with protein synthesis inhibitors to block gene expression induced by the taste independent of the expression induced by the toxin. At the very least, we can pursue the underlying molecular mechanisms at a leasurely pace, knowing that the taste-induced changes they must persist for a couple of hours…]

These 5 properties define CTA learning as single-trial, long-delay associative learning between taste and toxin. Therefore, in order to establish that a new phenomenon is CTA learning, it is necessary to demonstrate that it possesses these 5 properties. [Of course, additional properties might also be required, for example taste specificity or the ability to extinguish the CTA slowly with unpaired exposures to the taste.]

Importantly for future research, the same properties should be established for non-behavioral correlates of CTA learning, such as physiological or neural responses that parallel the behavioral learning. For example, the taste of sucrose induces c-Fos expression in the NTS of conditioned rats (but not unconditioned rats). Because this neural correlate of CTA expression has met the above 5 properties, we believe it is a true correlate of the associative processes of CTA learning.

Related articles include:
::[[Preferences and Aversions]]
::[[Measuring CTA Expression]]
::[[Conditioned and Unconditioned Stimuli]].

[[Category:Taste Aversion]]

SNF Lithium Orotate Inhibitors 3

2026-09-19T22:02:40Z

Houpt: updated clock times

Continuation of [[SNA Lithium Orotate CTA]] and [[SNB Lithium Orotate CTA 2]] and [[SNC Non-Contingent Lithium Orotate]] [[SND Lithium Orotate Inhibitors]] [[SNE Lithium Orotate Inhibitors 2]]

Test if OAT inhibitor naringin and UMPS inhibitor 6-azauracil decrease effectiveness of lithium orotate (LiOr) in inducing a conditioned taste aversion.

Inject narinign and 6-azauracil 30 min before single pairing of saccharin intake (15 min) after 15-min delay with LiOr.

From Pacholko 2023 PMID 37356352:

naringin MW 580.53 [https://www.sigmaaldrich.com/US/en/product/sigma/71162 Sigma 71162-35g] $50.78: organic anion transporting polypeptide 1A2 (OATP1A2) inhibitor

6-azauracil MW 113.07 [https://www.sigmaaldrich.com/US/en/product/sigma/a1757 Sigma A1757-5g] $99.90: UMPS inhibitor

For injection, combine 100 mg/kg naringin (17mM) + 30 mg/kg 6-azauracil (27mM) dissolved in saline/10% DMSO; in 10 ml/kg i.p. injection.

''Note: Mix with saline on day of injection, otherwise will precipitate overnight.''

* measure 400mg naringin + 120mg 6-azuracil;
* dissolve in '''2ml''' DMSO with heavy vortexing
* slowly add '''38''' ml sterile injectable 0.9% saline, with frequent vortexing (-> clear light yellow solution)
* filter

Use '''5%''' DMSO in injectable 0.9% saline as vehicle control.

''(Note: used only 5% DMSO, while SND used 10% DMSO)''

{{Infobox experiment|align=right
| Name = Lithium Orotate Inhibitors
| project = Taste Aversion
| code = SNF
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-9-12
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SNf
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SNf Raw Data Link]
== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (2026-9-1), n=16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2026-7-2), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-9-13)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-9-14 thru 2026-9-19)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at noon (1200) daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 7: Conditioning Day''' (2026-9-20)

-30 min (~1130): rats weighed and injected with naringin (100mg/kg) + 6-azauracil (30mg/kg) or saline vehicle, 10 ml/kg ip

0 min (~1200): all rats 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min (~1230): rats injected with 75 mM LiOr, 6 ml/kg ip, or Tris-NaCl vehicle.

Thus, four groups:

* Veh-Veh (SNF01,05,09,13)
* NA-Veh (SNF02,06,10,14)
* Veh-LiOr (SNF03,07,11,15)
* NA-LiOr (SNF04,08,12,16)

Ad lib water returned after final injection overnight.

'''Days 8 - 22: Preference Tests''' (2026-9-21 through 2026-10-06)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 14 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

SNE Lithium Orotate Inhibitors 2

2026-09-19T22:01:46Z

Houpt: typo

Continuation of [[SNA Lithium Orotate CTA]] and [[SNB Lithium Orotate CTA 2]] and [[SNC Non-Contingent Lithium Orotate]] [[SND Lithium Orotate Inhibitors]]

Test if OAT inhibitor naringin and UMPS inhibitor 6-azauracil decrease effectiveness of lithium orotate (LiOr) in inducing a conditioned taste aversion.

Inject narinign and 6-azauracil 30 min before single pairing of saccharin intake (15 min) after 15-min delay with LiOr.

From Pacholko 2023 PMID 37356352:

naringin MW 580.53 [https://www.sigmaaldrich.com/US/en/product/sigma/71162 Sigma 71162-35g] $50.78: organic anion transporting polypeptide 1A2 (OATP1A2) inhibitor

6-azauracil MW 113.07 [https://www.sigmaaldrich.com/US/en/product/sigma/a1757 Sigma A1757-5g] $99.90: UMPS inhibitor

For injection, combine 100 mg/kg naringin (17mM) + 30 mg/kg 6-azauracil (27mM) dissolved in saline/10% DMSO; in 10 ml/kg i.p. injection.

''Note: Mix with saline on day of injection, otherwise will precipitate overnight.''

* measure 400mg naringin + 120mg 6-azuracil;
* dissolve in '''2ml''' DMSO with heavy vortexing
* slowly add '''38''' ml sterile injectable 0.9% saline, with frequent vortexing (-> clear light yellow solution)
* filter

Use '''5%''' DMSO in injectable 0.9% saline as vehicle control.

''(Note: used only 5% DMSO, while SND used 10% DMSO)''

{{Infobox experiment|align=right
| Name = Lithium Orotate Inhibitors
| project = Taste Aversion
| code = SNE
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-7-7
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SNE
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SNE Raw Data Link]
== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (2026-6-23), n=16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2026-4-27), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-7-7)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-7-8 thru 2026-7-13)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1000 daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 7: Conditioning Day''' (2026-7-14)

-30 min (~930): rats weighed and injected with naringin (100mg/kg) + 6-azauracil (30mg/kg) or saline vehicle, 10 ml/kg ip

0 min (~1000): all rats 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min (~1030): rats injected with 75 mM LiOr, 6 ml/kg ip, or Tris-NaCl vehicle.

Thus, four groups:

* Veh-Veh (SNE01,05,09,13)
* NA-Veh (SNE02,06,10,14)
* Veh-LiOr (SNE03,07,11,15)
* NA-LiOr (SNE04,08,12,16)

Ad lib water returned after final injection overnight.

'''Days 8 - 22: Preference Tests''' (2026-7-15 through 2026-7-30)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 14 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

SNF Lithium Orotate Inhibitors 3

2026-09-19T22:01:15Z

Houpt: typo

Continuation of [[SNA Lithium Orotate CTA]] and [[SNB Lithium Orotate CTA 2]] and [[SNC Non-Contingent Lithium Orotate]] [[SND Lithium Orotate Inhibitors]] [[SNE Lithium Orotate Inhibitors 2]]

Test if OAT inhibitor naringin and UMPS inhibitor 6-azauracil decrease effectiveness of lithium orotate (LiOr) in inducing a conditioned taste aversion.

Inject narinign and 6-azauracil 30 min before single pairing of saccharin intake (15 min) after 15-min delay with LiOr.

From Pacholko 2023 PMID 37356352:

naringin MW 580.53 [https://www.sigmaaldrich.com/US/en/product/sigma/71162 Sigma 71162-35g] $50.78: organic anion transporting polypeptide 1A2 (OATP1A2) inhibitor

6-azauracil MW 113.07 [https://www.sigmaaldrich.com/US/en/product/sigma/a1757 Sigma A1757-5g] $99.90: UMPS inhibitor

For injection, combine 100 mg/kg naringin (17mM) + 30 mg/kg 6-azauracil (27mM) dissolved in saline/10% DMSO; in 10 ml/kg i.p. injection.

''Note: Mix with saline on day of injection, otherwise will precipitate overnight.''

* measure 400mg naringin + 120mg 6-azuracil;
* dissolve in '''2ml''' DMSO with heavy vortexing
* slowly add '''38''' ml sterile injectable 0.9% saline, with frequent vortexing (-> clear light yellow solution)
* filter

Use '''5%''' DMSO in injectable 0.9% saline as vehicle control.

''(Note: used only 5% DMSO, while SND used 10% DMSO)''

{{Infobox experiment|align=right
| Name = Lithium Orotate Inhibitors
| project = Taste Aversion
| code = SNF
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-9-12
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SNf
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SNf Raw Data Link]
== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (2026-9-1), n=16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2026-7-2), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-9-13)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-9-14 thru 2026-9-19)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1000 daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 7: Conditioning Day''' (2026-9-20)

-30 min (~930): rats weighed and injected with naringin (100mg/kg) + 6-azauracil (30mg/kg) or saline vehicle, 10 ml/kg ip

0 min (~1000): all rats 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min (~1030): rats injected with 75 mM LiOr, 6 ml/kg ip, or Tris-NaCl vehicle.

Thus, four groups:

* Veh-Veh (SNF01,05,09,13)
* NA-Veh (SNF02,06,10,14)
* Veh-LiOr (SNF03,07,11,15)
* NA-LiOr (SNF04,08,12,16)

Ad lib water returned after final injection overnight.

'''Days 8 - 22: Preference Tests''' (2026-9-21 through 2026-10-06)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 14 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

SNF Lithium Orotate Inhibitors 3

2026-09-13T16:30:50Z

Houpt: added dob

Continuation of [[SNA Lithium Orotate CTA]] and [[SNB Lithium Orotate CTA 2]] and [[SNC Non-Contingent Lithium Orotate]] [[SND Lithium Orotate Inhibitors]] [[SNE Lithium Orotate Inhibitors 2]]

Test if OAT inhibitor naringin and UMPS inhibitor 6-azauracil decrease effectiveness of lithium orotate (LiOr) in inducing a conditioned taste aversion.

Inject narinign and 6-azauracil 30 min before single pairing of saccharin intake (15 min) after 15-min delay with LiOr.

From Pacholko 2023 PMID 37356352:

naringin MW 580.53 [https://www.sigmaaldrich.com/US/en/product/sigma/71162 Sigma 71162-35g] $50.78: organic anion transporting polypeptide 1A2 (OATP1A2) inhibitor

6-azauracil MW 113.07 [https://www.sigmaaldrich.com/US/en/product/sigma/a1757 Sigma A1757-5g] $99.90: UMPS inhibitor

For injection, combine 100 mg/kg naringin (17mM) + 30 mg/kg 6-azauracil (27mM) dissolved in saline/10% DMSO; in 10 ml/kg i.p. injection.

''Note: Mix with saline on day of injection, otherwise will precipitate overnight.''

* measure 400mg naringin + 120mg 6-azuracil;
* dissolve in '''2ml''' DMSO with heavy vortexing
* slowly add '''38''' ml sterile injectable 0.9% saline, with frequent vortexing (-> clear light yellow solution)
* filter

Use '''5%''' DMSO in injectable 0.9% saline as vehicle control.

''(Note: used only 5% DMSO, while SND used 10% DMSO)''

{{Infobox experiment|align=right
| Name = Lithium Orotate Inhibitors
| project = Taste Aversion
| code = SNF
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-9-12
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SNf
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SNf Raw Data Link]
== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (2026-9-1), n=16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2026-7-2), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-9-13)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-9-14 thru 2026-9-19)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1000 daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 7: Conditioning Day''' (2026-9-20)

-30 min (~930): rats weighed and injected with naringin (100mg/kg) + 6-azauracil (30mg/kg) or saline vehicle, 10 ml/kg ip

0 min (~1000): all rats 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min (~1030): 6 rats injected with 75 mM LiOr, 6 ml/kg ip, or Tris-NaCl vehicle.

Thus, four groups:

* Veh-Veh (SNF01,05,09,13)
* NA-Veh (SNF02,06,10,14)
* Veh-LiOr (SNF03,07,11,15)
* NA-LiOr (SNF04,08,12,16)

Ad lib water returned after final injection overnight.

'''Days 8 - 22: Preference Tests''' (2026-9-21 through 2026-10-06)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 14 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

SNF Lithium Orotate Inhibitors 3

2026-09-12T17:01:28Z

Houpt: created page

Continuation of [[SNA Lithium Orotate CTA]] and [[SNB Lithium Orotate CTA 2]] and [[SNC Non-Contingent Lithium Orotate]] [[SND Lithium Orotate Inhibitors]] [[SNE Lithium Orotate Inhibitors 2]]

Test if OAT inhibitor naringin and UMPS inhibitor 6-azauracil decrease effectiveness of lithium orotate (LiOr) in inducing a conditioned taste aversion.

Inject narinign and 6-azauracil 30 min before single pairing of saccharin intake (15 min) after 15-min delay with LiOr.

From Pacholko 2023 PMID 37356352:

naringin MW 580.53 [https://www.sigmaaldrich.com/US/en/product/sigma/71162 Sigma 71162-35g] $50.78: organic anion transporting polypeptide 1A2 (OATP1A2) inhibitor

6-azauracil MW 113.07 [https://www.sigmaaldrich.com/US/en/product/sigma/a1757 Sigma A1757-5g] $99.90: UMPS inhibitor

For injection, combine 100 mg/kg naringin (17mM) + 30 mg/kg 6-azauracil (27mM) dissolved in saline/10% DMSO; in 10 ml/kg i.p. injection.

''Note: Mix with saline on day of injection, otherwise will precipitate overnight.''

* measure 400mg naringin + 120mg 6-azuracil;
* dissolve in '''2ml''' DMSO with heavy vortexing
* slowly add '''38''' ml sterile injectable 0.9% saline, with frequent vortexing (-> clear light yellow solution)
* filter

Use '''5%''' DMSO in injectable 0.9% saline as vehicle control.

''(Note: used only 5% DMSO, while SND used 10% DMSO)''

{{Infobox experiment|align=right
| Name = Lithium Orotate Inhibitors
| project = Taste Aversion
| code = SNF
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-9-12
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SNf
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SNf Raw Data Link]
== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (2026-XXX), n=16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2026-XXXX), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-9-13)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-9-14 thru 2026-9-19)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1000 daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 7: Conditioning Day''' (2026-9-20)

-30 min (~930): rats weighed and injected with naringin (100mg/kg) + 6-azauracil (30mg/kg) or saline vehicle, 10 ml/kg ip

0 min (~1000): all rats 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min (~1030): 6 rats injected with 75 mM LiOr, 6 ml/kg ip, or Tris-NaCl vehicle.

Thus, four groups:

* Veh-Veh (SNF01,05,09,13)
* NA-Veh (SNF02,06,10,14)
* Veh-LiOr (SNF03,07,11,15)
* NA-LiOr (SNF04,08,12,16)

Ad lib water returned after final injection overnight.

'''Days 8 - 22: Preference Tests''' (2026-9-21 through 2026-10-06)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 14 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

Experiment Metadata

2026-09-04T16:55:15Z

Houpt: created page

Notes on experiment metadata expected to be provided and imbedded in datafiles or ancillary config files.

NOB Novel Object Recognition Pilot

2026-08-28T23:30:47Z

Houpt: /* Training (2026-7-30) */

Pilot to test [[Novel Object Recognition]] (NOR) set-up.

(describe arena setup)

{{Infobox experiment|align=right
| Name = NOB Novel Object Recognition Pilot
| project = Memory
| code = SNB
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Abreu-Price, Houpt
| species = rat
| date = 2026-6-22
| type = CTA
| drugs = NA
| data = NA
}}

== People ==

Julia Abreu-Price

==Agenda==

NOB01-06, Adult male Sprague Dawley rats (Charles River), DOB 2026-4-12, arrived 2026-6-9, housed in BRF169.

All arena tests run in BRF115.

===1-hour Retention Test===

====Habituation phase (2026-6-22,23,24)====

For each rat, once per day for 3 consecutive days:

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* bring rat cage from housing rm BRF169 to test room BRF115, weigh rat, place in arena for 3 minutes, return to home cage.

====Training (2026-6-25)====

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* place 2 identical toys (i.e., 2 x kong or 2 x jack) in corners 20cm away from each wall
* bring rat cage from housing rm BRF169, weigh rat
* start video recording on iMac, place rat in arena for 3 (THREE) minutes
* return to home cage and BRF 169.
*wash toys in soapy water and rinse
* repeat for each rat, alternating toys:

* 2 x Kong: NOB01,NOB03,NOB05
* 2 x Jack: NOB2,NOB4,NOB6

===Testing (2026-6-25)===

At 60 min (ONE HOUR) after training, repeat test with 2 different toys:

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* place 1 kong and 1 jack in corners 20cm away from each wall
* bring rat cage from housing rm BRF169 (don't weigh rat again)
* start video recording on iMac, place rat in arena for 3 (THREE) minutes
* return to home cage and BRF 169.
* repeat for each rat, alternating position of jack and kong.

<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top>
AID Left Right
NB01 kong* jack
NB02 kong jack*
NB03 jack kong*
NB04 jack* kong
NB05 kong* jack
NB06 jack* kong
</tab>

''<nowiki>*</nowiki> indicates familiar object''

===24-hour Retention Test===

====Habituation phase (2026-7-27,28,29)====

* All 6 rats cages brought on cart from housing rm BRF169 to test room BRF115, weighed, placed in arena for 3 minutes, returned to home cage.

====Training (2026-7-30)====

At 24 hours after training:

* All 6 rats cages brought on cart from housing rm BRF169 to test room BRF115.
* place 2 identical toys (i.e., 2 x shroom or 2 x ball) in corners 20cm away from each wall
* rat weighed, placed in arena for 3 minutes.

* 2 x shroom: NB01, 03, 05
* 2 x ball: NB02,04,06

===Testing (2026-7-32)===

* All 6 rats cages brought on cart from housing rm BRF169 to test room BRF115.
* place 1 of each toys (shroom and ball) in corners 20cm away from each wall
* rat weighed, placed in arena for 3 minutes.

<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top>
AID Left Right
NB01 shroom* ball
NB02 ball* shroom
NB03 shroom* ball
NB04 ball* shroom
NB05 shroom* ball
NB06 ball* shroom
</tab>

''<nowiki>*</nowiki> indicates familiar object''

==Analysis==

===Yolo Detection (2026-7-9)===

yolo-detect.py run on NOB Training and Test movies, using pose model from 2026-7-6.

[[Category:Memory]]

NOB Novel Object Recognition Pilot

2026-08-28T23:30:19Z

Houpt: moved first table to correct place

Pilot to test [[Novel Object Recognition]] (NOR) set-up.

(describe arena setup)

{{Infobox experiment|align=right
| Name = NOB Novel Object Recognition Pilot
| project = Memory
| code = SNB
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Abreu-Price, Houpt
| species = rat
| date = 2026-6-22
| type = CTA
| drugs = NA
| data = NA
}}

== People ==

Julia Abreu-Price

==Agenda==

NOB01-06, Adult male Sprague Dawley rats (Charles River), DOB 2026-4-12, arrived 2026-6-9, housed in BRF169.

All arena tests run in BRF115.

===1-hour Retention Test===

====Habituation phase (2026-6-22,23,24)====

For each rat, once per day for 3 consecutive days:

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* bring rat cage from housing rm BRF169 to test room BRF115, weigh rat, place in arena for 3 minutes, return to home cage.

====Training (2026-6-25)====

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* place 2 identical toys (i.e., 2 x kong or 2 x jack) in corners 20cm away from each wall
* bring rat cage from housing rm BRF169, weigh rat
* start video recording on iMac, place rat in arena for 3 (THREE) minutes
* return to home cage and BRF 169.
*wash toys in soapy water and rinse
* repeat for each rat, alternating toys:

* 2 x Kong: NOB01,NOB03,NOB05
* 2 x Jack: NOB2,NOB4,NOB6

===Testing (2026-6-25)===

At 60 min (ONE HOUR) after training, repeat test with 2 different toys:

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* place 1 kong and 1 jack in corners 20cm away from each wall
* bring rat cage from housing rm BRF169 (don't weigh rat again)
* start video recording on iMac, place rat in arena for 3 (THREE) minutes
* return to home cage and BRF 169.
* repeat for each rat, alternating position of jack and kong.

<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top>
AID Left Right
NB01 kong* jack
NB02 kong jack*
NB03 jack kong*
NB04 jack* kong
NB05 kong* jack
NB06 jack* kong
</tab>

''<nowiki>*</nowiki> indicates familiar object''

===24-hour Retention Test===

====Habituation phase (2026-7-27,28,29)====

* All 6 rats cages brought on cart from housing rm BRF169 to test room BRF115, weighed, placed in arena for 3 minutes, returned to home cage.

====Training (2026-7-30)====

* All 6 rats cages brought on cart from housing rm BRF169 to test room BRF115.
* place 2 identical toys (i.e., 2 x shroom or 2 x ball) in corners 20cm away from each wall
* rat weighed, placed in arena for 3 minutes.

* 2 x shroom: NB01, 03, 05
* 2 x ball: NB02,04,06

===Testing (2026-7-32)===

* All 6 rats cages brought on cart from housing rm BRF169 to test room BRF115.
* place 1 of each toys (shroom and ball) in corners 20cm away from each wall
* rat weighed, placed in arena for 3 minutes.

<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top>
AID Left Right
NB01 shroom* ball
NB02 ball* shroom
NB03 shroom* ball
NB04 ball* shroom
NB05 shroom* ball
NB06 ball* shroom
</tab>

''<nowiki>*</nowiki> indicates familiar object''

==Analysis==

===Yolo Detection (2026-7-9)===

yolo-detect.py run on NOB Training and Test movies, using pose model from 2026-7-6.

[[Category:Memory]]

NOB Novel Object Recognition Pilot

2026-08-28T23:29:15Z

Houpt: added 24-h test, tables of objects

Pilot to test [[Novel Object Recognition]] (NOR) set-up.

(describe arena setup)

{{Infobox experiment|align=right
| Name = NOB Novel Object Recognition Pilot
| project = Memory
| code = SNB
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Abreu-Price, Houpt
| species = rat
| date = 2026-6-22
| type = CTA
| drugs = NA
| data = NA
}}

== People ==

Julia Abreu-Price

==Agenda==

NOB01-06, Adult male Sprague Dawley rats (Charles River), DOB 2026-4-12, arrived 2026-6-9, housed in BRF169.

All arena tests run in BRF115.

===1-hour Retention Test===

====Habituation phase (2026-6-22,23,24)====

For each rat, once per day for 3 consecutive days:

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* bring rat cage from housing rm BRF169 to test room BRF115, weigh rat, place in arena for 3 minutes, return to home cage.

====Training (2026-6-25)====

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* place 2 identical toys (i.e., 2 x kong or 2 x jack) in corners 20cm away from each wall
* bring rat cage from housing rm BRF169, weigh rat
* start video recording on iMac, place rat in arena for 3 (THREE) minutes
* return to home cage and BRF 169.
*wash toys in soapy water and rinse
* repeat for each rat, alternating toys:

* 2 x Kong: NOB01,NOB03,NOB05
* 2 x Jack: NOB2,NOB4,NOB6

===Testing (2026-6-25)===

At 60 min (ONE HOUR) after training, repeat test with 2 different toys:

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* place 1 kong and 1 jack in corners 20cm away from each wall
* bring rat cage from housing rm BRF169 (don't weigh rat again)
* start video recording on iMac, place rat in arena for 3 (THREE) minutes
* return to home cage and BRF 169.
* repeat for each rat, alternating position of jack and kong.

===24-hour Retention Test===

<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top>
AID Left Right
NB01 kong* jack
NB02 kong jack*
NB03 jack kong*
NB04 jack* kong
NB05 kong* jack
NB06 jack* kong
</tab>

''<nowiki>*</nowiki> indicates familiar object''

====Habituation phase (2026-7-27,28,29)====

* All 6 rats cages brought on cart from housing rm BRF169 to test room BRF115, weighed, placed in arena for 3 minutes, returned to home cage.

====Training (2026-7-30)====

* All 6 rats cages brought on cart from housing rm BRF169 to test room BRF115.
* place 2 identical toys (i.e., 2 x shroom or 2 x ball) in corners 20cm away from each wall
* rat weighed, placed in arena for 3 minutes.

* 2 x shroom: NB01, 03, 05
* 2 x ball: NB02,04,06

===Testing (2026-7-32)===

* All 6 rats cages brought on cart from housing rm BRF169 to test room BRF115.
* place 1 of each toys (shroom and ball) in corners 20cm away from each wall
* rat weighed, placed in arena for 3 minutes.

<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top>
AID Left Right
NB01 shroom* ball
NB02 ball* shroom
NB03 shroom* ball
NB04 ball* shroom
NB05 shroom* ball
NB06 ball* shroom
</tab>

''<nowiki>*</nowiki> indicates familiar object''

==Analysis==

===Yolo Detection (2026-7-9)===

yolo-detect.py run on NOB Training and Test movies, using pose model from 2026-7-6.

[[Category:Memory]]

Novel Object Recognition

2026-07-28T18:19:58Z

Houpt: added stub for walk DSL

==Useful Papers==

see [https://houptlab.org/courses/nor/ NOR papers] on lab website.

==Rat Setup==

===Arena===

external size: 79.2 cm square (not including handles), with 112cm diagonal -- so will fit on a 48" round table.

interior size: 70cm square, with 35cm high walls.

[[File:RatNORArenaSchematic.png|200px|thumb|right|Schematic of arena used for NOR test]]

Place 36x36" sterilization wrap from Schein for substrate. [https://www.henryschein.com/us-en/dental/p/infection-control-products/instrument-sterilization-processing/csr-wrap/1002016 CSR Wrap 36x36" 100/Bx 1002016EZ | Henry Schein Inc. - 849]

Constructed from [https://8020.net 8020] materials.

===Objects===

By having 4 unique toys, should be able to do 2 tests on each rat: a short-term test (e.g. 1 hour post-training) and a long-term test (e.g. 24 hours post-training), with 2 different pairs of toys.

All the toys are approximately the same size, and all are made of similar red rubber (the SHITEC mushroom is a slightly lighter shade of red than the Kongs).

Kong Classic Dog Toy (large) vs. Kong Jump'N Jack Dog Toy (medium).

''Note that Jump'N Jack has been discontinued.''

Kong Stuff-A-Ball (large0 vs. SHITEC Dog Chew Toy

[[File:Kongs4NOR.jpg|200px|thumb|right|Kongs used for rat NOR test]]

[[File:MoreToys4NOR.jpg|200px|thumb|right|More Toys used for rat NOR test]]

==Video Detection==

[[File:Label_Studio_rat_between_toys.png|200px|thumb|right|Annotation in Label_Studio of rat between two dog toys (Kong and Jack)]]

[[File:Label_Studio_rat_head_nose_interaural.png|200px|thumb|right|Annotation in Label Studio of rat's head and key points at nose and interaural point (poll)]]

==Walk DSL for Analysis==

Novel Object Recognition

2026-07-28T18:19:21Z

Houpt: small edits

Novel Object Recognition

2026-07-28T18:16:29Z

Houpt: added more toys

==Useful Papers==

see [https://houptlab.org/courses/nor/ NOR papers] on lab website.

==Rat Setup==

===Arena===

external size: 79.2 cm square (not including handles), with 112cm diagonal -- so will fit on a 48" round table.

interior size: 70cm square, with 35cm high walls.

[[File:RatNORArenaSchematic.png|200px|thumb|right|Schematic of arena used for NOR test]]

Place 36x36" sterilization wrap from Schein for substrate. [https://www.henryschein.com/us-en/dental/p/infection-control-products/instrument-sterilization-processing/csr-wrap/1002016 CSR Wrap 36x36" 100/Bx 1002016EZ | Henry Schein Inc. - 849]

Constructed from [8020.net 8020] materials.

===Objects===

By having 4 unique toys, should be able to do 2 tests on each rat: a short-term test (e.g. 1 hour post-training) and a long-term test (e.g. 24 hours post-training), with 2 different pairs of toys.

Kong Classic Dog Toy (large) vs. Kong Jump'N Jack Dog Toy (medium).

''Note that Jump'N Jack has been discontinued.''

Kong Stuff-A-Ball (large0 vs. SHITEC Dog Chew Toy

[[File:Kongs4NOR.jpg|200px|thumb|right|Kongs used for rat NOR test]]

[[File:MoreKongs4NOR.jpg|200px|thumb|right|More Kongs used for rat NOR test]]

==Video Detection==

[[File:Label_Studio_rat_between_toys.png|200px|thumb|right|Annotation in Label_Studio of rat between two dog toys (Kong and Jack)]]

[[File:Label_Studio_rat_head_nose_interaural.png|200px|thumb|right|Annotation in Label Studio of rat's head and key points at nose and interaural point (poll)]]

File:MoreToys4NOR.jpg

2026-07-28T18:13:52Z

Houpt: Two more dog toys for use in NOR tests

== Summary ==
Two more dog toys for use in NOR tests

SND Lithium Orotate Inhibitors

2026-07-14T13:59:48Z

Houpt: note on precipitation, update weighing

Continuation of [[SNA Lithium Orotate CTA]] and [[SNB Lithium Orotate CTA 2]] and [[SNC Non-Contingent Lithium Orotate]]

Test if OAT inhibitor naringin and UMPS inhibitor 6-azauracil decrease effectiveness of lithium orotate (LiOr) in inducing a conditioned taste aversion.

Inject narinign and 6-azauracil 30 min before single pairing of saccharin intake (15 min) after 15-min delay with LiOr.

From Pacholko 2023 PMID 37356352:

naringin MW 580.53 [https://www.sigmaaldrich.com/US/en/product/sigma/71162 Sigma 71162-35g] $50.78: organic anion transporting polypeptide 1A2 (OATP1A2) inhibitor

6-azauracil MW 113.07 [https://www.sigmaaldrich.com/US/en/product/sigma/a1757 Sigma A1757-5g] $99.90: UMPS inhibitor

For injection, combine 100 mg/kg naringin (17mM) + 30 mg/kg 6-azauracil (27mM) dissolved in saline/10% DMSO; in 10 ml/kg i.p. injection.

''Note: Mix with saline on day of injection, otherwise will precipitate overnight.''

* measure 400mg naringin + 120mg 6-azuracil;
* dissolve in 4ml DMSO with heavy vortexing
* slowly add 36 ml sterile injectable 0.9% saline, with frequent vortexing (-> clear light yellow solution)
* filter

Use 10% DMSO in injectable 0.9% saline as vehicle control.

{{Infobox experiment|align=right
| Name = Lithium Orotate Inhibitors
| project = Taste Aversion
| code = SND
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-6-2
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SND
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SND Raw Data Link]
== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (2026-5-26), n=16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2026-3-30), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-6-2)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-6-3 thru 2026-6-8)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1000 daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 7: Conditioning Day''' (2026-6-9)

-30 min (~930): rats weighed and injected with naringin (100mg/kg) + 6-azauracil (30mg/kg) or saline vehicle, 10 ml/kg ip

0 min (~1000): all rats 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min (~1030): 6 rats injected with 75 mM LiOr, 6 ml/kg ip, or Tris-NaCl vehicle.

Thus, four groups:

* Veh-Veh (SND01,05,09,13)
* NA-Veh (SND02,06,10,14)
* Veh-LiOr (SND03,07,11,15)
* NA-LiOr (SND04,08,12,16)

Ad lib water returned after final injection overnight.

'''Days 8 - 22: Preference Tests''' (2026-6-10 through 2026-6-25)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 14 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

SNE Lithium Orotate Inhibitors 2

2026-07-14T13:59:07Z

Houpt:

Continuation of [[SNA Lithium Orotate CTA]] and [[SNB Lithium Orotate CTA 2]] and [[SNC Non-Contingent Lithium Orotate]] [[SND Lithium Orotate Inhibitors]]

Test if OAT inhibitor naringin and UMPS inhibitor 6-azauracil decrease effectiveness of lithium orotate (LiOr) in inducing a conditioned taste aversion.

Inject narinign and 6-azauracil 30 min before single pairing of saccharin intake (15 min) after 15-min delay with LiOr.

From Pacholko 2023 PMID 37356352:

naringin MW 580.53 [https://www.sigmaaldrich.com/US/en/product/sigma/71162 Sigma 71162-35g] $50.78: organic anion transporting polypeptide 1A2 (OATP1A2) inhibitor

6-azauracil MW 113.07 [https://www.sigmaaldrich.com/US/en/product/sigma/a1757 Sigma A1757-5g] $99.90: UMPS inhibitor

For injection, combine 100 mg/kg naringin (17mM) + 30 mg/kg 6-azauracil (27mM) dissolved in saline/10% DMSO; in 10 ml/kg i.p. injection.

''Note: Mix with saline on day of injection, otherwise will precipitate overnight.''

* measure 400mg naringin + 120mg 6-azuracil;
* dissolve in '''2ml''' DMSO with heavy vortexing
* slowly add '''38''' ml sterile injectable 0.9% saline, with frequent vortexing (-> clear light yellow solution)
* filter

Use '''5%''' DMSO in injectable 0.9% saline as vehicle control.

''(Note: used only 5% DMSO, while SND used 10% DMSO)''

{{Infobox experiment|align=right
| Name = Lithium Orotate Inhibitors
| project = Taste Aversion
| code = SNE
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-7-7
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SNE
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SNE Raw Data Link]
== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (2026-6-23), n=16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2026-4-27), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-7-7)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-7-8 thru 2026-7-13)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1000 daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 7: Conditioning Day''' (2026-7-14)

-30 min (~930): rats weighed and injected with naringin (100mg/kg) + 6-azauracil (30mg/kg) or saline vehicle, 10 ml/kg ip

0 min (~1000): all rats 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min (~1030): 6 rats injected with 75 mM LiOr, 6 ml/kg ip, or Tris-NaCl vehicle.

Thus, four groups:

* Veh-Veh (SNE01,05,09,13)
* NA-Veh (SNE02,06,10,14)
* Veh-LiOr (SNE03,07,11,15)
* NA-LiOr (SNE04,08,12,16)

Ad lib water returned after final injection overnight.

'''Days 8 - 22: Preference Tests''' (2026-7-15 through 2026-7-30)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 14 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

SNE Lithium Orotate Inhibitors 2

2026-07-14T13:58:18Z

Houpt: updated DMSO, weighing, some dates

Continuation of [[SNA Lithium Orotate CTA]] and [[SNB Lithium Orotate CTA 2]] and [[SNC Non-Contingent Lithium Orotate]] [[SND Lithium Orotate Inhibitors]]

Test if OAT inhibitor naringin and UMPS inhibitor 6-azauracil decrease effectiveness of lithium orotate (LiOr) in inducing a conditioned taste aversion.

Inject narinign and 6-azauracil 30 min before single pairing of saccharin intake (15 min) after 15-min delay with LiOr.

From Pacholko 2023 PMID 37356352:

naringin MW 580.53 [https://www.sigmaaldrich.com/US/en/product/sigma/71162 Sigma 71162-35g] $50.78: organic anion transporting polypeptide 1A2 (OATP1A2) inhibitor

6-azauracil MW 113.07 [https://www.sigmaaldrich.com/US/en/product/sigma/a1757 Sigma A1757-5g] $99.90: UMPS inhibitor

For injection, combine 100 mg/kg naringin (17mM) + 30 mg/kg 6-azauracil (27mM) dissolved in saline/10% DMSO; in 10 ml/kg i.p. injection.

* measure 400mg naringin + 120mg 6-azuracil;
* dissolve in '''2ml''' DMSO with heavy vortexing
* slowly add '''38''' ml sterile injectable 0.9% saline, with frequent vortexing (-> clear light yellow solution)
* filter

Use '''5%''' DMSO in injectable 0.9% saline as vehicle control.

''(Note: used only 5% DMSO, while SND used 10% DMSO)''

{{Infobox experiment|align=right
| Name = Lithium Orotate Inhibitors
| project = Taste Aversion
| code = SNE
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-7-7
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SNE
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SNE Raw Data Link]
== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (2026-6-23), n=16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2026-4-27), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-7-7)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-7-8 thru 2026-7-13)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1000 daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 7: Conditioning Day''' (2026-7-14)

-30 min (~930): rats weighed and injected with naringin (100mg/kg) + 6-azauracil (30mg/kg) or saline vehicle, 10 ml/kg ip

0 min (~1000): all rats 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min (~1030): 6 rats injected with 75 mM LiOr, 6 ml/kg ip, or Tris-NaCl vehicle.

Thus, four groups:

* Veh-Veh (SNE01,05,09,13)
* NA-Veh (SNE02,06,10,14)
* Veh-LiOr (SNE03,07,11,15)
* NA-LiOr (SNE04,08,12,16)

Ad lib water returned after final injection overnight.

'''Days 8 - 22: Preference Tests''' (2026-7-15 through 2026-7-30)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 14 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

SNE Lithium Orotate Inhibitors 2

2026-07-12T22:52:25Z

Houpt: /* Agenda */ added dob

Continuation of [[SNA Lithium Orotate CTA]] and [[SNB Lithium Orotate CTA 2]] and [[SNC Non-Contingent Lithium Orotate]] [[SND Lithium Orotate Inhibitors]]

Test if OAT inhibitor naringin and UMPS inhibitor 6-azauracil decrease effectiveness of lithium orotate (LiOr) in inducing a conditioned taste aversion.

Inject narinign and 6-azauracil 30 min before single pairing of saccharin intake (15 min) after 15-min delay with LiOr.

From Pacholko 2023 PMID 37356352:

naringin MW 580.53 [https://www.sigmaaldrich.com/US/en/product/sigma/71162 Sigma 71162-35g] $50.78: organic anion transporting polypeptide 1A2 (OATP1A2) inhibitor

6-azauracil MW 113.07 [https://www.sigmaaldrich.com/US/en/product/sigma/a1757 Sigma A1757-5g] $99.90: UMPS inhibitor

For injection, combine 100 mg/kg naringin (17mM) + 30 mg/kg 6-azauracil (27mM) dissolved in saline/10% DMSO; in 10 ml/kg i.p. injection.

* measure 400mg naringin + 120mg 6-azuracil;
* dissolve in 4ml DMSO with heavy vortexing
* slowly add 36 ml sterile injectable 0.9% saline, with frequent vortexing (-> clear light yellow solution)
* filter

Use 10% DMSO in injectable 0.9% saline as vehicle control.

{{Infobox experiment|align=right
| Name = Lithium Orotate Inhibitors
| project = Taste Aversion
| code = SNE
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-7-7
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SNE
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SNE Raw Data Link]
== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (2026-6-23), n=16 adult male Sprague Dawley rats (Charles River CD, d.o.b 2026-4-27), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-7-7)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-7-8 thru 2026-6-13)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1000 daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 7: Conditioning Day''' (2026-7-14)

-30 min (~930): rats injected with naringin (100mg/kg) + 6-azauracil (30mg/kg) or saline vehicle, 10 ml/kg ip

0 min (~1000): all rats 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min (~1030): 6 rats injected with 75 mM LiOr, 6 ml/kg ip, or Tris-NaCl vehicle.

Thus, four groups:

* Veh-Veh (SNE01,05,09,13)
* NA-Veh (SNE02,06,10,14)
* Veh-LiOr (SNE03,07,11,15)
* NA-LiOr (SNE04,08,12,16)

Ad lib water returned after final injection overnight.

'''Days 8 - 22: Preference Tests''' (2026-7-15 through 2026-7-30)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 14 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

Lithium Orotate Dissociation

2026-07-12T22:43:48Z

Houpt: formatting tweak

It is something of an open question as to whether lithium orotate (LiOr) dissociates in solution (free Li+ and the base orotate), or whether it remains as an metalorganic complex.

This should be approachable by measuring 3 characteristics of LiOr solutions:

1. conductivity -- higher conductance implies free ions

2. osmolality -- higher osmolality implies dissociation

3. free Li+ concentration -- Li-selective electrode will detect Li+ if dissociated, but not in LiOr complex.

==Conductivity==

From Aron 2025 et al:

Conductivity measurements
To measure the conductivity of Li salts, the salts were dissolved in water
to achieve Li concentrations of 4.3 mEq l−1, 430 μEq l−1, 43 μEq l−1 or
21.5 μEq l−1 in each case. Conductivity was measured using an ST300C
conductivity meter (OHAUS, 83033964) equipped with a STCON7
electrode (OHAUS, 30080693) calibrated with potassium chloride
conductivity standards. For each lithium salt, three independent solu-
tion replicates (n = 3) were prepared. Conductivity values are reported
in μS per cm at 25 °C.

==Free Li+ Concentration==

[https://www.ntsensors.com/lithium-ion-selective-electrode/ NT Sensors Li ISE]

==Osmolality==

[https://faculty.smcm.edu/wihatch/courses/436web/436resources/osmoWescore5520.html Wescor Vapro 5520 osmometer]

==Test Solutions==

Make mixtures of LiCl, Li2CO3, and LiOr and several "physiological" solutions. Measure free Li+ with electrode. For Tris and plasma, also measure osmolality.

===Tris===

Tris-LiCl, Tris-LiOH, Tris-LiOH-Orotate (across concentration and pH ranges)

===Simulated Gastric Fluid (SGF)===

Li+ electrode cannot function at low pH (~2.0) of SGF, so need to neutralize mixtures with sodium bicarbonate (NaCH03). Assay free Li+ when added before or after neutralization.

Measure at 37° C, across concentration ranges:

SGF-LiCl -> + NaHC03, SGF-Tris-LiOr -> + NaHC03, SGF -> + NaHC03 + LiCl, SGF -> + NaHC03 + Tris-LiOr

===Rat plasma===

Blood taken from terminal cardiac puncture, heparinized, plasma separated and frozen at -20° C.

Measure at 37° C, across concentration ranges:

plasma-LiCl, plasma-Li2CO3, plasma-LiOr

===Artificial Urine (AU)===

Artificial urine prepared according to DIN EN 1616:1999, from Pickering Laboratories (Mountain View, USA).

Measure at 37° C, across concentration ranges:

AU-LiCl, AU-Li2CO3, AU-LiOr

Lithium Orotate Dissociation

2026-07-12T22:42:40Z

Houpt: added para on Aron paper

It is something of an open question as to whether lithium orotate (LiOr) dissociates in solution (free Li+ and the base orotate), or whether it remains as an metalorganic complex.

This should be approachable by measuring 3 characteristics of LiOr solutions:

1. conductivity -- higher conductance implies free ions

2. osmolality -- higher osmolality implies dissociation

3. free Li+ concentration -- Li-selective electrode will detect Li+ if dissociated, but not in LiOr complex.

==Conductivity==

From Aron 2025 et al:

Conductivity measurements
To measure the conductivity of Li salts, the salts were dissolved in water
to achieve Li concentrations of 4.3 mEq l−1, 430 μEq l−1, 43 μEq l−1 or
21.5 μEq l−1 in each case. Conductivity was measured using an ST300C
conductivity meter (OHAUS, 83033964) equipped with a STCON7
electrode (OHAUS, 30080693) calibrated with potassium chloride
conductivity standards. For each lithium salt, three independent solu-
tion replicates (n = 3) were prepared. Conductivity values are reported
in μS per cm at 25 °C.

==Free Li+ Concentration==

[https://www.ntsensors.com/lithium-ion-selective-electrode/ NT Sensors Li ISE]

==Osmolality==

[https://faculty.smcm.edu/wihatch/courses/436web/436resources/osmoWescore5520.html Wescor Vapro 5520 osmometer]

==Test Solutions==

Make mixtures of LiCl, Li2CO3, and LiOr and several "physiological" solutions. Measure free Li+ with electrode. For Tris and plasma, also measure osmolality.

===1. Tris===

Tris-LiCl, Tris-LiOH, Tris-LiOH-Orotate (across concentration and pH ranges)

===2. Simulated Gastric Fluid (SGF)===

Li+ electrode cannot function at low pH (~2.0) of SGF, so need to neutralize mixtures with sodium bicarbonate (NaCH03). Assay free Li+ when added before or after neutralization.

Measure at 37° C, across concentration ranges:

SGF-LiCl -> + NaHC03, SGF-Tris-LiOr -> + NaHC03, SGF -> + NaHC03 + LiCl, SGF -> + NaHC03 + Tris-LiOr

===3. Rat plasma===

Blood taken from terminal cardiac puncture, heparinized, plasma separated and frozen at -20° C.

Measure at 37° C, across concentration ranges:

plasma-LiCl, plasma-Li2CO3, plasma-LiOr

===4. Artificial Urine (AU)===

Artificial urine prepared according to DIN EN 1616:1999, from Pickering Laboratories (Mountain View, USA).

Measure at 37° C, across concentration ranges:

AU-LiCl, AU-Li2CO3, AU-LiOr

NOB Novel Object Recognition Pilot

2026-07-09T15:20:31Z

Houpt: added yolo detection note

Novel Object Recognition

2026-07-09T15:06:03Z

Houpt: added link to lab website

Novel Object Recognition

2026-07-09T14:43:13Z

Houpt: typo

==Rat Setup==

===Arena===

external size: 79.2 cm square (not including handles), with 112cm diagonal -- so will fit on a 48" round table.

interior size: 70cm square, with 35cm high walls.

[[File:RatNORArenaSchematic.png|200px|thumb|left|Schematic of arena used for NOR test]]

Place 36x36" sterilization wrap from Schein for substrate. [https://www.henryschein.com/us-en/dental/p/infection-control-products/instrument-sterilization-processing/csr-wrap/1002016 CSR Wrap 36x36" 100/Bx 1002016EZ | Henry Schein Inc. - 849]

Constructed from [8020.net 8020] materials.

===Objects===

Kong Classic Dog Toy (large) vs. Kong Jump'N Jack Dog Toy (medium).

''Note that Jump'N Jack has been discontinued.''

[[File:Kongs4NOR.jpg|200px|thumb|left|Kongs used for rat NOR test]]

==Video Detection==

[[File:Label_Studio_rat_between_toys.png|200px|thumb|left|Annotation in Label_Studio of rat between two dog toys (Kong and Jack)]]

[[File:Label_Studio_rat_head_nose_interaural.png|200px|thumb|left|Annotation in Label Studio of rat's head and key points at nose and interaural point (poll)]]

Novel Object Recognition

2026-07-09T14:42:51Z

Houpt: addedd some label-studio images

==Rat Setup==

===Arena===

external size: 79.2 cm square (not including handles), with 112cm diagonal -- so will fit on a 48" round table.

interior size: 70cm square, with 35cm high walls.

[[File:RatNORArenaSchematic.png|200px|thumb|left|Schematic of arena used for NOR test]]

Place 36x36" sterilization wrap from Schein for substrate. [https://www.henryschein.com/us-en/dental/p/infection-control-products/instrument-sterilization-processing/csr-wrap/1002016 CSR Wrap 36x36" 100/Bx 1002016EZ | Henry Schein Inc. - 849]

Constructed from [8020.net 8020] materials.

===Objects===

Kong Classic Dog Toy (large) vs. Kong Jump'N Jack Dog Toy (medium).

''Note that Jump'N Jack has been discontinued.''

[[File:Kongs4NOR.jpg|200px|thumb|left|Kongs used for rat NOR test]]

==Video Detection==

[[File:File:Label_Studio_rat_between_toys.png|200px|thumb|left|Annotation in Label_Studio of rat between two dog toys (Kong and Jack)]]

[[File:File:Label_Studio_rat_head_nose_interaural.png|200px|thumb|left|Annotation in Label Studio of rat's head and key points at nose and interaural point (poll)]]

File:Label Studio rat head nose interaural.png

2026-07-09T14:00:00Z

Houpt: Annotation in Label Studio of rat's head and key points at nose and interaural point (poll)

== Summary ==
Annotation in Label Studio of rat's head and key points at nose and interaural point (poll)

File:Label Studio rat between toys.png

2026-07-09T13:57:44Z

Houpt: Annotation in Label_Studio of rat between two dog toys (Kong and Jack)

== Summary ==
Annotation in Label_Studio of rat between two dog toys (Kong and Jack)

Novel Object Recognition

2026-07-09T13:56:23Z

Houpt: added arena schematic

NOB Novel Object Recognition Pilot

2026-07-09T13:54:17Z

Houpt: fixed training/test times

Lithium Salts

2026-07-07T23:13:43Z

Houpt: fixed Na mg/L

MW of lithium: 6.94 (weighted average of Li6 and Li7 isotopes)

For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day).

Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl.

We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li2CO3) and lithium orotate (LiH3N2O4; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:

<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top>
Salt MW % Li g/L for 0.15M mg Li/L dose ml 0.15M/kg mg salt/kg mg Li/kg
NaCl 58.44 (0.39 Na) 8.8 (3448 Na) 12 105.19 (41.4 Na)
NaOr 178.08 (0.13 Na) 26.71 (3448 Na) 12 320.54 (41.4 Na)
LiOH 23.95 0.29 3.6 1041 na na na
LiCl 42.39 0.16 6.35 1041 12 76.30 12.5
LiCl 42.39 0.16 6.35 1041 20 127.17 41.6
Li2CO3 73.89 0.19 11.08 2082 12 66.54 12.5
LiOr 162.00 0.04 24.31 1041 12 291.79 12.5
</tab>

''note that 0.15M Li2CO3 is 0.45 Osm; 0.1M Li2CO3 is isotonic''

== LiOr Preparation ==

From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was adjusted to 0.9% and pH to 7.4. Doses were administered as 0.1 ml/10g BW (10 ml/kg), with the highest dose of 15 mg/kg Li+ => 375 mg/kg LiOr => 230 mM (NB: not sure how they got this much to dissolve in water)

[https://www.sigmaaldrich.com/US/en/product/sigma/o2750 orotic acid, Sigma Aldrich O2750-10G ($62.01)] CAS 65-86-1, MW 156.10

[https://www.sigmaaldrich.com/US/en/product/sigald/545856 lithium hydroxide, powder, reagent grade >= 98%, 545856-100G ($45.45)] CAS 1310-65-2, MW 23.95

sodium hydroxide, CAS 1310-73-2, MW 40

* Make 500 ml of 0.15M LiOH (1.80 g LiOH / 500ml dH20).

* Make 500 ml of 0.15M NaOH (3.00 g NaOH / 500ml dH20).

* Make 20 ml of 0.15M LiOr by adding 0.47 g orotic acid to 20 ml of 0.15M LiOH, vortex: partially dissolved after about 2 min of vortexing -- but soon precipiates out

* So dissolve in 0.3M TrisAmino...

* Make 20 ml of 0.15M NaOr by adding 0.47 g orotic acid to 20 ml of 0.15M NaOH, vortex: did NOT dissolve -- milky white suspension! Left shaking overnight...

Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).

== Milliequivalents ==
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.

== Osmolarity ==

NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).

LiCO should also dissociate completely (citation needed).

LiOr, however is more tightly bound complex (citation needed). So may need to mix up LiOr in 0.15 NaCl to preserve osmolarity.

Lithium Salts

2026-07-07T23:11:51Z

Houpt: fixed mg/L calculation

MW of lithium: 6.94 (weighted average of Li6 and Li7 isotopes)

For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day).

Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl.

We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li2CO3) and lithium orotate (LiH3N2O4; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:

<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top>
Salt MW % Li g/L for 0.15M mg Li/L dose ml 0.15M/kg mg salt/kg mg Li/kg
NaCl 58.44 (0.39 Na) 8.8 (3 Na) 12 105.19 (41.4 Na)
NaOr 178.08 (0.13 Na) 26.71 (3 Na) 12 320.54 (41.4 Na)
LiOH 23.95 0.29 3.6 1041 na na na
LiCl 42.39 0.16 6.35 1041 12 76.30 12.5
LiCl 42.39 0.16 6.35 1041 20 127.17 41.6
Li2CO3 73.89 0.19 11.08 2082 12 66.54 12.5
LiOr 162.00 0.04 24.31 1041 12 291.79 12.5
</tab>

''note that 0.15M Li2CO3 is 0.45 Osm; 0.1M Li2CO3 is isotonic''

== LiOr Preparation ==

From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was adjusted to 0.9% and pH to 7.4. Doses were administered as 0.1 ml/10g BW (10 ml/kg), with the highest dose of 15 mg/kg Li+ => 375 mg/kg LiOr => 230 mM (NB: not sure how they got this much to dissolve in water)

[https://www.sigmaaldrich.com/US/en/product/sigma/o2750 orotic acid, Sigma Aldrich O2750-10G ($62.01)] CAS 65-86-1, MW 156.10

[https://www.sigmaaldrich.com/US/en/product/sigald/545856 lithium hydroxide, powder, reagent grade >= 98%, 545856-100G ($45.45)] CAS 1310-65-2, MW 23.95

sodium hydroxide, CAS 1310-73-2, MW 40

* Make 500 ml of 0.15M LiOH (1.80 g LiOH / 500ml dH20).

* Make 500 ml of 0.15M NaOH (3.00 g NaOH / 500ml dH20).

* Make 20 ml of 0.15M LiOr by adding 0.47 g orotic acid to 20 ml of 0.15M LiOH, vortex: partially dissolved after about 2 min of vortexing -- but soon precipiates out

* So dissolve in 0.3M TrisAmino...

* Make 20 ml of 0.15M NaOr by adding 0.47 g orotic acid to 20 ml of 0.15M NaOH, vortex: did NOT dissolve -- milky white suspension! Left shaking overnight...

Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).

== Milliequivalents ==
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.

== Osmolarity ==

NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).

LiCO should also dissociate completely (citation needed).

LiOr, however is more tightly bound complex (citation needed). So may need to mix up LiOr in 0.15 NaCl to preserve osmolarity.

Lithium Salts

2026-07-07T23:06:25Z

Houpt: add mg/L to match lithium ISE readings

MW of lithium: 6.94 (weighted average of Li6 and Li7 isotopes)

For conditioned taste aversion experiments in rats, we use a "standard" dose of LiCl of 76 mg/kg, administered as 12 ml/kg of 0.15 M LiCl. The rat dose is based on early work of Nachman and Asche 1973 PMID 4697023, who found this to be the minimum dose to induce a maximum CTA (i.e. after 1 pairingof sucrose with 76 mg/kg LiCl, rats showed nearly zero preference for sucrose the next day).

Mice appear somewhat resistant, and require either 20 ml/kg (127 mg/kg) or 40 ml/kg (254 mg/kg) LiCl.

We are also interested in the effects of other lithium salts, in particular lithium carbonate (Li2CO3) and lithium orotate (LiH3N2O4; LiOr). This table helps convert molarity and dose of lithium of the different salts for equivalent doses of Li:

<tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top>
Salt MW % Li g/L for 0.15M mg Li/L dose ml 0.15M/kg mg salt/kg mg Li/kg
NaCl 58.44 (0.39 Na) 8.8 (3 Na) 12 105.19 (41.4 Na)
NaOr 178.08 (0.13 Na) 26.71 (3 Na) 12 320.54 (41.4 Na)
LiOH 23.95 0.29 3.6 1044 na na na
LiCl 42.39 0.16 6.35 1016 12 76.30 12.5
LiCl 42.39 0.16 6.35 1016 20 127.17 41.6
Li2CO3 73.89 0.19 11.08 2105 12 66.54 12.5
LiOr 162.00 0.04 24.31 972 12 291.79 12.5
</tab>

''note that 0.15M Li2CO3 is 0.45 Osm; 0.1M Li2CO3 is isotonic''

== LiOr Preparation ==

From Pacholko 2023 PMID 37356352: LiOr was synthesized by combining lithium hydroxide (Sigma-Aldrich), and orotic acid (Sigma-Aldrich) in a 1:1 M ratio in distilled water; the NaCl concentration was adjusted to 0.9% and pH to 7.4. Doses were administered as 0.1 ml/10g BW (10 ml/kg), with the highest dose of 15 mg/kg Li+ => 375 mg/kg LiOr => 230 mM (NB: not sure how they got this much to dissolve in water)

[https://www.sigmaaldrich.com/US/en/product/sigma/o2750 orotic acid, Sigma Aldrich O2750-10G ($62.01)] CAS 65-86-1, MW 156.10

[https://www.sigmaaldrich.com/US/en/product/sigald/545856 lithium hydroxide, powder, reagent grade >= 98%, 545856-100G ($45.45)] CAS 1310-65-2, MW 23.95

sodium hydroxide, CAS 1310-73-2, MW 40

* Make 500 ml of 0.15M LiOH (1.80 g LiOH / 500ml dH20).

* Make 500 ml of 0.15M NaOH (3.00 g NaOH / 500ml dH20).

* Make 20 ml of 0.15M LiOr by adding 0.47 g orotic acid to 20 ml of 0.15M LiOH, vortex: partially dissolved after about 2 min of vortexing -- but soon precipiates out

* So dissolve in 0.3M TrisAmino...

* Make 20 ml of 0.15M NaOr by adding 0.47 g orotic acid to 20 ml of 0.15M NaOH, vortex: did NOT dissolve -- milky white suspension! Left shaking overnight...

Also from Pacholko 2023: in mice, 100 mg/kg naringin (inhibitor of Organic anion transporting polypeptides
(OATPs) OATP1A2/Oatp1a1/Oatp1a4) and/or 30 mg/kg 6-azauracil (inhibitor of uridine monophosphate synthase (UMPS), which decarboxylases orotic acid) delivered via IP injection (both dissolved in isotonic saline).

== Milliequivalents ==
Clincial literature often refers to milliequivalents (mEq) of lithium salts. A milliequivalent is the millimoles of positive charge, so for monovalent ions like Na+ and Li+, 1 mEq = 1 mmol.

== Osmolarity ==

NaCl and LiCl at low concentrations dissociate almost completely, so 0.15M NaCl or 0.15M LiCl are essentially 0.3 OsM (Citation needed).

LiCO should also dissociate completely (citation needed).

LiOr, however is more tightly bound complex (citation needed). So may need to mix up LiOr in 0.15 NaCl to preserve osmolarity.

SNE Lithium Orotate Inhibitors 2

2026-07-07T17:06:46Z

Houpt: created

Continuation of [[SNA Lithium Orotate CTA]] and [[SNB Lithium Orotate CTA 2]] and [[SNC Non-Contingent Lithium Orotate]] [[SND Lithium Orotate Inhibitors]]

Test if OAT inhibitor naringin and UMPS inhibitor 6-azauracil decrease effectiveness of lithium orotate (LiOr) in inducing a conditioned taste aversion.

Inject narinign and 6-azauracil 30 min before single pairing of saccharin intake (15 min) after 15-min delay with LiOr.

From Pacholko 2023 PMID 37356352:

naringin MW 580.53 [https://www.sigmaaldrich.com/US/en/product/sigma/71162 Sigma 71162-35g] $50.78: organic anion transporting polypeptide 1A2 (OATP1A2) inhibitor

6-azauracil MW 113.07 [https://www.sigmaaldrich.com/US/en/product/sigma/a1757 Sigma A1757-5g] $99.90: UMPS inhibitor

For injection, combine 100 mg/kg naringin (17mM) + 30 mg/kg 6-azauracil (27mM) dissolved in saline/10% DMSO; in 10 ml/kg i.p. injection.

* measure 400mg naringin + 120mg 6-azuracil;
* dissolve in 4ml DMSO with heavy vortexing
* slowly add 36 ml sterile injectable 0.9% saline, with frequent vortexing (-> clear light yellow solution)
* filter

Use 10% DMSO in injectable 0.9% saline as vehicle control.

{{Infobox experiment|align=right
| Name = Lithium Orotate Inhibitors
| project = Taste Aversion
| code = SNE
| protocol = 20220000006
| funding = Chair's Fund
| investigators = Nabi, Houpt
| species = rat
| date = 2026-7-7
| type = CTA
| drugs = lithium orotate, lithium chloride
| data = https://houptlab.org/bartab/expt/?id=SNE
}}

== Data ==

[https://houptlab.org/bartab/expt/?id=SNE Raw Data Link]
== People ==

Soheila Nabi

==CTA Conditioning==

===Agenda===

Rats arrive (XXXX), n=16 adult male Sprague Dawley rats (Charles River CD, d.o.b XXX), individually housed in BRF 169.

 

'''Day 0: Water removed''' (2026-7-7)

make labels for bottles.

 

'''Days 1 - 6: Water Restriction''' (2026-7-8 thru 2026-6-13)

Weigh rats daily. ad lib food.

30 min, 30 min, 30 min, 30 min, 30 min, 15 min water at 1000 daily.

1 water bottle, 1 empty bottle to accustom to 2-bottle choice

 

'''Day 7: Conditioning Day''' (2026-7-14)

-30 min (~930): rats injected with naringin (100mg/kg) + 6-azauracil (30mg/kg) or saline vehicle, 10 ml/kg ip

0 min (~1000): all rats 15 min access to 0.125% saccharin (5 g sodium saccharin/4L water)

30 min (~1030): 6 rats injected with 75 mM LiOr, 6 ml/kg ip, or Tris-NaCl vehicle.

Thus, four groups:

* Veh-Veh (SNE01,05,09,13)
* NA-Veh (SNE02,06,10,14)
* Veh-LiOr (SNE03,07,11,15)
* NA-LiOr (SNE04,08,12,16)

Ad lib water returned after final injection overnight.

'''Days 8 - 22: Preference Tests''' (2026-7-15 through 2026-7-30)

Rats CTA acquisition and extinction was measured with 24-h, 2-bottle preference tests. Rats were given 24-h free access to both 0.125% saccharin and water bottles, alternating position for 14 days.

[[Category:Taste Aversion]]
[[Category:CTA Experiments]]

Lithium Orotate Dissociation

2026-06-28T18:51:04Z

Houpt:

It is something of an open question as to whether lithium orotate (LiOr) dissociates in solution (free Li+ and the base orotate), or whether it remains as an metalorganic complex.

This should be approachable by measuring 3 characteristics of LiOr solutions:

1. conductance -- higher conductance implies free ions

2. osmolality -- higher osmolality implies dissociation

3. free Li+ concentration -- Li-selective electrode will detect Li+ if dissociated, but not in LiOr complex.

==Free Li+ Concentration==

[https://www.ntsensors.com/lithium-ion-selective-electrode/ NT Sensors Li ISE]

==Osmolality==

[https://faculty.smcm.edu/wihatch/courses/436web/436resources/osmoWescore5520.html Wescor Vapro 5520 osmometer]

==Test Solutions==

Make mixtures of LiCl, Li2CO3, and LiOr and several "physiological" solutions. Measure free Li+ with electrode. For Tris and plasma, also measure osmolality.

===1. Tris===

Tris-LiCl, Tris-LiOH, Tris-LiOH-Orotate (across concentration and pH ranges)

===2. Simulated Gastric Fluid (SGF)===

Li+ electrode cannot function at low pH (~2.0) of SGF, so need to neutralize mixtures with sodium bicarbonate (NaCH03). Assay free Li+ when added before or after neutralization.

Measure at 37° C, across concentration ranges:

SGF-LiCl -> + NaHC03, SGF-Tris-LiOr -> + NaHC03, SGF -> + NaHC03 + LiCl, SGF -> + NaHC03 + Tris-LiOr

===3. Rat plasma===

Blood taken from terminal cardiac puncture, heparinized, plasma separated and frozen at -20° C.

Measure at 37° C, across concentration ranges:

plasma-LiCl, plasma-Li2CO3, plasma-LiOr

===4. Artificial Urine (AU)===

Artificial urine prepared according to DIN EN 1616:1999, from Pickering Laboratories (Mountain View, USA).

Measure at 37° C, across concentration ranges:

AU-LiCl, AU-Li2CO3, AU-LiOr

Lithium Orotate Dissociation

2026-06-28T01:07:40Z

Houpt: add some potential test solutions

It is something of an open question as to whether lithium orotate (LiOr) dissociates in solution (free Li+ and the base orotate), or whether it remains as an metalorganic complex.

This should be approachable by measuring 3 characteristics of LiOr solutions:

1. conductance -- higher conductance implies free ions

2. osmolality -- higher osmolality implies dissociation

3. free Li+ concentration -- Li-selective electrode will detect Li+ if dissociated, but not in LiOr complex.

==Free Li+ Concentration==

[https://www.ntsensors.com/lithium-ion-selective-electrode/ NT Sensors Li ISE]

==Osmolality==

[https://faculty.smcm.edu/wihatch/courses/436web/436resources/osmoWescore5520.html Wescor Vapro 5520 osmometer]

==Test Solutions==

Make mixtures of LiCl, Li2CO3, and LiOr and several "physiological" solutions. Measure free Li+ with electrode. For Tris and plasma, also measure osmolality.

===1. Tris===

Tris-LiCl, Tris-LiOH, Tris-LiOH-Orotate (across concentration and pH ranges)

===2. Simulated Gastric Fluid (SGF)===

Li+ electrode cannot function at low pH (~2.0) of SGF, so need to neutralize mixtures with sodium bicarbonate (NaCH03). Assay free Li+ when added before or after neutralization.

Measure at 37° C, across concentration ranges:

SGF-LiCl -> + NaHC03, SGF-Tris-LiOr -> + NaHC03, SGF -> + NaHC03 + LiCl, SGF -> + NaHC03 + Tris-LiOr

===3. Rat plasma===

Blood taken from terminal cardiac puncture, heparinized, plasma separated and frozen at -20° C.

Measure at 37° C, across concentration ranges:

plasma-LiCl, plasma-Li2CO3, plasma-LiOr

===4. Artificial Urine (AU)===

Artificial urine prepared according to DIN EN 1616:1999, from Pickering Laboratories (Mountain View, USA).

Measure at 37° C, across concentration ranges:

AU-LiCl, AU-Li2CO3, AU-LiOr

===5. Artificial Cerebrospinal Fluid (aCSF)===

NOB Novel Object Recognition Pilot

2026-06-27T13:38:05Z

Houpt: added experiment box

NOB Novel Object Recognition Pilot

2026-06-25T14:05:07Z

Houpt: /* Training (2026-6-25) */

Pilot to test [[Novel Object Recognition]] (NOR) set-up.

(describe arena setup)

==Agenda==

NOB01-11, Adult male Sprague Dawley rats (Charles River), DOB 2026-4-12, arrived 2026-6-9.

All arena tests run in BRF115.

===Habituation phase (2026-6-22,23,24)===

For each rat, once per day for 3 consecutive days:

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* bring rat cage from housing rm BRF169, weigh rat, place in arena for 3 minutes, return to home cage.

===Training (2026-6-25)===

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* place 2 identical toys (i.e. 2 x kong or 2 x jack) in corners 20cm away from each wall
* bring rat cage from housing rm BRF169, weigh rat
* start video recording on iMac, place rat in arena for 5 (FIVE) minutes
* return to home cage and BRF 169.
*wash toys in soapy water and rinse
* repeat for each rat, alternating toys:

* 2 x Kong: NOB01,03,05
* 2 x Jack: NOB2,NOB4,NOB6

===Testing (2026-6-25)===

At 60 min (ONE HOUR) after training, repeat test with 2 different toys:

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* place 1 kong and 1 jack in corners 20cm away from each wall
* bring rat cage from housing rm BRF169 (don't weigh rat again)
* start video recording on iMac, place rat in arena for 5 (FIVE) minutes
* return to home cage and BRF 169.
* repeat for each rat, alternating position of jack and kong.

==Analysis==

NOB Novel Object Recognition Pilot

2026-06-25T13:35:26Z

Houpt:

Pilot to test [[Novel Object Recognition]] (NOR) set-up.

(describe arena setup)

==Agenda==

NOB01-11, Adult male Sprague Dawley rats (Charles River), DOB 2026-4-12, arrived 2026-6-9.

All arena tests run in BRF115.

===Habituation phase (2026-6-22,23,24)===

For each rat, once per day for 3 consecutive days:

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* bring rat cage from housing rm BRF169, weigh rat, place in arena for 3 minutes, return to home cage.

===Training (2026-6-25)===

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* place 2 identical toys (i.e. 2 x kong or 2 x jack) in corners 20cm away from each wall
* bring rat cage from housing rm BRF169, weigh rat
* start video recording on iMac, place rat in arena for 5 (FIVE) minutes
* return to home cage and BRF 169.
* repeat for each rat, alternating toys:

* 2 x Kong: NOB01,03,05
* 2 x Jack: NOB2,NOB4,NOB6

===Testing (2026-6-25)===

At 60 min (ONE HOUR) after training, repeat test with 2 different toys:

* wipe down sides of arena with Windex multisurface disinfectant cleaner
* place new wrap under arena
* place 1 kong and 1 jack in corners 20cm away from each wall
* bring rat cage from housing rm BRF169 (don't weigh rat again)
* start video recording on iMac, place rat in arena for 5 (FIVE) minutes
* return to home cage and BRF 169.
* repeat for each rat, alternating position of jack and kong.

==Analysis==

NOB Novel Object Recognition Pilot

2026-06-22T16:55:51Z

Houpt: details

NOB Novel Object Recognition Pilot

2026-06-22T14:17:52Z

Houpt:

Pilot to test [[Novel Object Recognition]] (NOR) set-up.

(describe arena setup)

==Habituation phase==

* wipe down sides with Windex multisurface
* new wrap under arena
* weigh rats, place in arena for 3 minutes, return to home cage.

==Training==

==Testing==

NOB Novel Object Recognition Pilot

2026-06-22T14:16:06Z

Houpt: created page

Pilot to test Novel Object Recognition (NOR) set-up.

(describe arena setup)

==Habituation phase==

* wipe down sides with Windex multisurface
* new wrap under arena
* weigh rats, place in arena for 3 minutes, return to home cage.

==Training==

==Testing==

Lithium Orotate Dissociation

2026-06-20T19:48:14Z

Houpt:

Lithium Orotate Dissociation

2026-06-20T19:47:48Z

Houpt: added osmometer

Lithium Orotate Dissociation

2026-06-20T19:44:37Z

Houpt: add li electrode link

Lithium Orotate Dissociation

2026-06-20T19:41:09Z

Houpt: created page

Label-Studio

2026-06-11T13:05:40Z

Houpt:

https://labelstud.io

== Installation ==
installation to python virtual environment:

python3 -m venv labelstudio-env
source labelstudio-env/bin/activate
pip3 install -U label-studio

run by typing ''label-studio'', it will launch Chrome, and navigate to http://localhost:8080/user/login/

create local account:

username: lab@houptlab.org, pw: novelobject

create project