View source for TW Innate Odor Aversions in 14-3-3 FKO mice

Innate odor aversion in 14-3-3 FKO mice expressed as locomotor activity in response to [[TMT]] or [[Rose odor|2PE]] in [[Odor Arena]], scored by [[Tracker]].

Adam Kimbrough, Charles E. Houpt, Yuying Wu, Dr. Thomas A. Houpt, Dr. Yi Zhou, Dr. Michael Meredith

== Procedure ==

Male 14-3-3 FKO mice (n=6) and wildtype C57 mice (n=6), singly housed with ad lib access to food and water. Habituated to [[Odor Arena]] (without stimulus dish) for 5 min per day for 4 days. 

===Trials 1 - 3: response to water, TMT, or 2PE ===

All mice exposed to an odor stimuli for 5 min, 1 stimulus/day, for 3 consective days.

Odor stimuli are either 5 ul distilled water, 5 ul TMT (1% in water), or 1.25 ul 2PE (neat). Stimuli pipetted onto filter paper (#1 Whatman, 25mm diameter disc) and placed in small glass petri dish. Stimuli were prepared prior to bringing mouse home cage into test room. Stimulus was placed in random corner of the 29cm x 29cm arena immediately prior to mouse being placed in center of arena. Each stimulus was presented once, in random order for each mouse.

Locomotor activity video was captured with a Logitech Quick Pro webcam and an Apple iMac computer into Quicktime Player at 640x480 pixel at 15 fps. Video was processed by [[Tracker]] program. The position of the mouse, defined as the center of the mouse body as detected by Tracker to within ~2 mm, was recorded 3 times per second. From the position data, several variables were calculated: total distance traveled during the 5 min test, distance from the stimulus, etc.

Between mice, the arena was rinsed with soapy water (Alconox) and 70% isopropanol. Stimulus filter paper and pipette tips were disposed into sealed container of bleach.

===Trial 4: contrasting response to water vs. TMT===

All mice exposed to H20 stimulus for 5 min, then odor stimulus was replaced with TMT for 5 min to evaluate contrasting responses to water & TMT back to back. 

==Stimuli==

[[TMT|'''TMT''']] (ConTech, Inc) received April 1, 2013

* Product code 3000000368
* Lot number 11125
* Manufactured May 5, 2011
* Expiry May 2014

[[Rose odor|'''2PE''']] (Sigma-Aldrich) received March, 2013

* Item #77861
* Lot# BCBH9821V

=== Solution Prep===

made 50% TMT in water: 50 ul TMT + 50 ul water

made 1% TMT in water: 10 ul TMT + 990 ul water

aliquoted 1000 ul 2PE

aliquoted 50 ml distilled water

==Random Trial Schedule==


<tab sep=tab class="wikitable" border = "1" cellspacing="3" cellpadding = "3" head = top>
Mouse	Genotype	Trial 1	Trial 2	Trial 3
TW01	Tg	2PE, br	TMT, tl	H20, tr
TW02	Tg	H20, bl	TMT, br	2PE, tl
TW03	Tg	2PE, br	H20, tl	TMT, br
TW04	Tg	TMT, bl	2PE, tl	H20, tl
TW05	Tg	TMT, br	H20, br	2PE, tr
TW06	Tg	H20, br	2PE, bl	TMT, tr
TW07	wt	H20, tl	TMT, bl	2PE, tr
TW08	wt	TMT, tl	H20, br	2PE, br
TW09	wt	TMT, br	H20, tl	2PE, br
TW10	wt	H20, bl	TMT, br	2PE, br
TW11	wt	H20, tr	TMT, bl	2PE, tl
TW12	wt	H20, tr	2PE, tl	TMT, bl
</tab>

''tl, tr, bl, br = corner of odor arena.''

[[Category: CTA Experiments]]