Transcript Slide 1
Marissa Applegate, Shruthi Deivasigamani, Conor Driscoll, Sumeet Jain, Sarah McAlister, Jacquelyn Olwell, Ravi Pancholi, Claire Rhee, David Gabriel Rissman, Adam Rosenstein, Madison Taormina, Justin Zhang Advisors: Dr. Graham Cousens and Zack Vogel Fear Anxiety Directed at a stimulus General uneasiness Dissipates after stimulus Long lasting is removed Construct: Phasic Fear Construct: Sustained Fear Fear Anxiety Phasic Fear Sustained Fear Central Nucleus Bed Nucleus of Stria Terminalis (BNST) Experiment 1: Effects of Strain Differences on the Light Enhanced Startle Response in Lewis and SpragueDawley Rats Experiment 2: Effects of Nicotine Withdrawal on Discrete Cue and Contextual Conditioned Fear Experiment 3: Nicotine Withdrawal-Induced Anxiety in the Elevated Zero Maze Evaluate genetic differences Hypotheses – Lewis vs. Sprague-Dawley Measure Greaterbaseline baseline startle startle response (1,3) Light-enhanced (LES) (2,4,5) Greater percentstartle increase of LES startle response Impact on future research Sustained fear present after LES Lewis Rat H51-58 8 male Sprague-Dawley Rat H59-H65 6 male A. Coulbourn Precision Animal Shocker B. Light C. Fan D. Plexiglass cover for inner chamber E. Odor Emitter F. Amyl Acetate G. Outer Chamber H. Speaker I. Accelerometer 0,0006 Startle Response 0,0005 0,0004 0,0003 Lewis Sprague-Dawley 0,0002 0,0001 0 75 85 95 105 Decibel Level The Lewis rats had a higher baseline startle response when compared to the Sprague-Dawleys. Lewis Sprague Dawley 180 160 160 140 140 120 120 DDD 100 DLD 80 Percent of Baseline Percent of Baseline 180 100 DLD 80 60 60 40 40 20 20 0 2 3 DDD 0 2 At the 95 dB level, the Sprague Dawley rats exhibited a higher LES startle response, and a greater percent increase in startle. The Sprague-Dawley also exhibited a sustained fear. 3 PErcent of Baseline Lewis Sprague-Dawley 250 250 200 200 150 150 DDD DDD DLD DLD 100 100 50 50 0 2 3 0 2 3 At the 105 dB level, the Sprague-Dawley rats still exhibited a higher LES reaction and a greater percent increase. • 43.5% try to quit smoking • Of those, between 70% and 90% fail (6) • Connection between nicotine withdrawal and anxiety • Treatments to limit withdrawal effects Link between withdrawal and sustained fear Withdrawal and Phasic Fear Hypothesis There will be no effect on phasic fear. Sprague-Dawley Rat H1, H2, H3, H8, H9 5 male High Dosage of Nicotine (.5mg/kg) Sprague-Dawley Rat H4, H11, H12, H13, H14 5 male Low Dosage of Nicotine (.25mg/kg) Injected every day for three weeks Tested two hours after injection Sprague-Dawley Rat H5, H6, H7, H10 4 male Control (Saline) Percent Potentiation of Contextual Fear Contextual Fear in Sprague-Dawley Rats 1400 300 Mean Percent Potentiation Mean Startle Amplitude 1200 350 250 200 50 0 0 1000 800 200 Control Low Dose 600 150 High Dose 400 100 Control Pre-conditioning Baseline Low Dose High Dose Context Re-exposure Conclusion: Robust contextual fear potentiated startle with no difference between groups Olfactory Fear Olfactory Conditioning Fear Potentiation in Sprague-Dawley Rats 1400 25 20 1200 1000 800 600 400 Percent Potentiation Mean Startle Amplitude 15 10 5 Control 0 Low Dose High Dose -5 -10 -15 200 -20 Control Low Dose High Dose 0 Odor Odor+30 Odor+60 Conclusion: No olfactory fear potentiated startle in all groups Odor+90 Elevated zero-maze Nicotine withdrawal leads to anxiety Hypothesis: As nicotine dosage increases and causes a higher withdrawal affect, anxiety, locomotor activity, and time spent in the closed sections of the maze will increase. Walled Nicotine Injections 2 hour waiting period 15 minute testing period Rat placed in maze Time Line Spent Crosses in Open vs. Dosage vs Dosage 250 450 Open (Seconds) Crosses Time Spent Line in 400 200 350 300 150 250 200 100 150 100 50 50 0 0.0 mg/kg 0.25 mg/kg 0.5 mg/kg 0.0 mg/kg Dosage Nicotine 0.25of mg/kg 0.5 mg/kg 0 Dosage of Nicotine Nicotine dosage had no effect on the amount of time the rats spent in Nicotine the open dosage sectionshad of the maze.locomotor activity. no effect 500 450 Time Spent in Open (seconds) 400 350 300 250 200 150 100 50 0 10:00 10:30 11:00 11:30 12:00 12:30 13:00 13:30 14:00 Time of Day 14:30 15:00 15:30 16:00 16:30 17:00 Surprising Data Need more conclusive data Future experiments Dr. Graham Cousens , who is getting married today! Dr. Miyamoto, our fearless leader Myrna Papier, our indispensible organizer Zack Vogel, our epic counselor All the NJGSS Staff And our gracious benefactors: John and Laura Overdeck Bayer Healthcare The Crimmins Family Charitable Foundation NJGSS Alumni and Parents 1984 – 2011 Bristol-Meyer Squibb Roche Kinder Morgan Bain Capital Childrens Charity GlastoSmithKline Novartis Reference Sources 1. Ramos A, Kangerski AL, Basso PF, Da Silva Santos JE, Assreuy J, Vendruscolo LF, Takahashi RN. Evaluation of Lewis and SHR rat strains as a genetic model for the study of anxiety and pain. Behavioral Brain Research. 2002 Feb; [Internet]. [cited 2011 Jul 27] 129(1-2): 113-123. Available from: http://www.ncbi.nlm.nih.gov.ezproxy.drew.edu/pubmed/11809502 2. Steiner MA, Lecourt H, Rakotoariniaina A, Jenck F. Favoured genetic background for testing anxiolytics in the fearpotentiated and light-enhanced startle paradigms in the rat. Behavioural Brain Research. 2011 Aug; [Internet]. [cited 2011 Jul 27] 221(1): 34-42. Available from: http://www.ncbi.nlm.nih.gov.ezproxy.drew.edu/pubmed?term=Favoured%20genetic%20background%20for%20testing%20anxi olytics%20in%20the%20fear-potentiated%20and 3. Freet CS, Tesche JD, Tompers DM, Riegel KE, Grigson PS. Lewis rats are more sensitive than Fischer rats to successive negative contrast, but less sensitive to the anxiolytic and appetite-stimulating effects of chlordiazepoxide. Pharmacology, Biochemistry, and Behavior. 2006 Oct; [Internet]. [cited 2011 Jul 27] 85(2): 378-384. Available from: http://www.ncbi.nlm.nih.gov.ezproxy.drew.edu/pubmed/17049372 4. Davis M, Walker DL, Miles L, Grillon C. Phasic vs sustained fear in rats and humans: role of the extended amygdala in fear vs anxiety. Neuropsychopharmacology. 2010 Jan; [Internet]. [cited 2011 Jul 27] 35(1): 105-135. Available from: http://www.ncbi.nlm.nih.gov/pubmed/19693004 5. Jonkman S, Risbrough VB, Geyer MA, Markou A. Spontaneous nicotine withdrawal potentiates the effects of stress in rats. Neuropsychopharmacology. 2008 Aug; [Internet]. [cited 2011 Jul 27] 33(9): 2131-2138. Available from: http://www.ncbi.nlm.nih.gov/pubmed/18033237 6. (Bronars C, Saul J. Increasing Reach of Tobacco Cessation Quitlines: A Review of the Literature and Promising Practices [home page on the Internet]. Phoenix (AZ): North American Quitline Consortium; 2009. [Introduction; cited 2011 July 27]. 18 p. Available from: http://www.naquitline.org/resource/resmgr/issue_papers....)