DME Sodium Butyrate and Flavor Conditioning 2

Test if systemic Sodium Butyrate (NaB), an HDAC inhibitor, enhances preference learning

Replication of [DMC Sodium Butyrate and Flavor Conditioning] with following modifications:

1. To prevent formation of side preferencs, rats were trained with 2 choice water and 0.05% saccharin prior to conditioning

2. To prevent ceiling effect of preferences being too strong, CS+ used only 4% glucose

3. n=9 male, n=9 female, counterbalanced

Dose of NaB based on Kwon and Houpt, Brain Research, 1333 (2010) 36–47, PMID: 20346924 PMCID: PMC2871962

• 0.3 M Na Butyrate = 3.3g/100ml water
• 0.3 M NaCl = 18g/liter water (osmotic control)

Data

Raw Data Link

People

Durba Mukherjee

CFP Conditioning

Agenda

Rats arrive. (July 7, 2020)

n=18 adult male and female sprague-dawley rats (Charles River CD), individually housed in BRF 169.

Days 0: Food and water removed. (July 10, 2020)

Days 1-7: Food and water restricted. (July 11 - July 17, 2020)

Weigh rats daily. water for 30 min, food 3-4 pellets to maintain at 90% body weight.

30 min water at 1100am daily, pellet ration at 1130am

Day 8: Conditioning Day 1 (July 18, 2020)

0 min: 30 min access to CS+ 005% Kool-Aid + 0.05% saccharin + 4% glucose (counterbalanced cherry, grape) at 1135am - 1155am

30 min: remove CS+, inject with 12ml/kg of 0.3M sodium butyrate or 12ml/kg of 0.3M NaCl, 1200noon-1213pm

Give daily pellet ration at 1215am

• Rats 1-9 : female, Rats 10-18: male
• Rats 1,2,5,6,9,10,13,14,17,18: Cherry CS+
• Rats 3,4,7,8,11,12,15,16: Grape CS+
• Odd rats: NaB injection; Even rats: NaCL injection

Day 9: Conditioning Day 2 (July 19, 2020)

0 min: 30 min access to CS- 005% Kool-Aid + 0.05% saccharin (counterbalanced cherry, grape) at 1106-1136am

CS- bottles not weighed, so estimated volumetrically from fill lines.

30 min: remove CS-, inject with 12ml/kg of 0.3M sodium butyrate or 12ml/kg of 0.3M NaCl, 1138am-1150am

return food and water ad lib at 12noon

• Rats 1-9 : female, Rats 10-18: male
• Rats 1,2,5,6,9,10,13,14,17,18: Grape CS-
• Rats 3,4,7,8,11,12,15,16: Cherry CS-.
• Odd rats: NaB injection; Even rats: NaCL injection.

Days 10-18: 2 bottle preference tests (July 20 - 28, 2020)

Each rat given adlib 24-h access to of CS+/sac vs CS-/sac. Position of bottle alternated each day.

Notes:

• Rat DME 16 did not drink from CS+/glucose on conditioning day 1
• CS- bottles not weighed in conditioning day 2, so estimated volumetrically from fill lines

CTA Positive Control

Agenda

Day 0: (August 30, 2020) Water removed, ad lib food access.

Day 1-9: (August 31-Sept 8, 2020) the rats were placed on a water deprivation schedule under which they received daily water access in one 30-min drinking session. Rats had ad lib food access throughout.

Day 10: Conditioning day (Sept 9, 2020): rats were divided into 4 different groups (NaCl/NaCl, NaCl/NaB, LiCl/NaCl and LiCl/NaB)

0-10 min: 10-min access to a bottle containing 75 mM NaCl solution (half-isotonic, 4.5g NaCl/liter).

30 min: (20 minutes after the end of NaCl) intake:

• LiCl/NaCl and LiCl/NaB groups were injected with LiCl (0.15 M, 3 ml/kg, i.p.)

• NaCl/NaCl and NaCl/NaB groups were injected with saline (0.15 M NaCl, 3 ml/kg, i.p.).

40 min: (10 minutes after LiCl or saline injections)

• LiCl/NaB and NaCl/NaB groups were injected with NaB (0.3 M, 12 ml/kg, i.p.)
• LiCl/NaCl and NaCl/NaCl groups were injected with iso-osmotic saline (0.3 M NaCl, 12 ml/kg, i.p.).

3 h: (3 hours after conditioning) rats received ad libitum access to water overnight.

Day 11 (Sept 10, 2020): CTA acquisition and extinction was measured with 48-h, 2-bottle preference tests. Rats were given 24-h free access to both 75 mM NaCl and water bottles, alternating position for 6 days.