1. The Effects of Bisindolymaleimide on Long Term Memory in Gryllus Bimaculatus Manhasset Science Research Paris Thatos

2. Need Long-term trend in drug-related deaths (Gotz, 2004) Percentage of People Experiencing a Mental Disorder in their Lives (Gallaher, 2007) Fig 1. Shows trend in drug related deaths. Fig 2. A graph showing % of people experiencing mental disorders

3. Long Term Potentiation (Kanehisa, 1999) Fig. 3 The Process of Long Term Potentiation The complex chemical processes that form long-term memories (Cook, 2006)

4. PKC (Protein Kinase C) Has many isoforms (PKMzeta,PKMa) (Abel, 2004) Fig 4. Process of PKC interaction in Long term Potentiation

5. Olfactory Memory Capacity of the Cricket Gryllus Bimaculatus Yukihisa Matsumoto, Biology Letters Yukihisa Matsumoto Fig 5. Olfactory Preferences of crickets trained to associate Scent A with water reward and Scent B with saline punishment

6. Place Memory in Crickets Jan Wessnitzer, Biology Letters Fig 6. Mean time (seconds) taken for cricket to find cool spot Wessnitzer, 2008

7. Bisindolylmaleimide is a Potent and Selective Inhibitor of PKC Dominique Toullec, The Journal of Biological Chemistry Toullec, 2005 Fig 7. The concentrations of PKC when exposed to Bisindolylmaleimide, and the concentrations of a PKC isotype The levels of PKC and PKA

8. Formation of Long-term Olfactory Memory in the cricket Gryllus bimaculatus Makoto Mizunami, 2005, Chem. Senses Makoto, 2006 Fig 8. A diagram of the head of the cricket. Brain (In blue) is where PKC is believed to be contained in.

9. Purpose/Hypothesis The purpose of this experiment is to see whether Bisindolymaleide disrupts long term memories in Gryllus Bimaculatus when treated with it pre- and post- conditioning H (a) - It is hypothesized that with all administeringings of Bisindolymaleide, the number of retained associations will be significantly lower, and it will take more time to complete the heat maze

10. Crickets (Gryllus Bimaculatus) n=120 Statistically analyzed with a one-way ANOVA (to find statistic significance) followed by a Scheffe post-hoc test (p<0.05) Trained to associate one scent with reward and one scent with punishment Also, trained to run heat maze Control group n=30 Reward/Punishme nt for scent test is water/saline solution Given water after each heat maze run Variable group n=30 Reward/Punishment for scent test is water/saline solution (+  ) Given water after each heat maze run Variable group n=30 Reward/Punishment for scent test is water/saline solution Given Water (+5  M)  after each heat maze run Variable group n=30 Reward/Punishment for scent test is water/saline solution (+5  M), Given Water (+5  M)  after each heat maze run Time spent on each scent is recorded for all groups, and time spent finding cool spot in heat maze also recorded

11. Protocol Gryllus Bimaclatus scent conditioning Obtaining specific concentration of Bisindolylmaleimide Exposure to scents to test whether negative or positive connotations were preserved Recording of time spent on top of each conditioned scent Gryllus Bimaculatus Heat Maze training Recording of time spent for each heat maze run Scent ConditioningScent Testing (Matsumoto, 2006)

12. Budget VendorCatalog #page/websiteDescriptionQuantityUnit $Total $ EMDbiosciences203290http://www.emdbiosciences.comBisindolylmaleimide1$40.00 Carolina143550http://carolina.com/Crickets (Living Adults)1$6.75 Carolina143565http://carolina.com/House Cricket Food2$6.25$12.50 Carolina695145http://carolina.com/Sense of Smell Kit1$49.25 Carolina199606http://carolina.com/Plastic Syringe1$3.85 Carolina198697http://carolina.com/Distilled Water1$6.75 Carolina721207http://carolina.com/Pyrex Beaker 100ml4$3.65$14.60 Walmart-http://walmart.com/Plastic Bin5$10$50 Walmart-http://walmart.com/Electrical Skillet1$80 Empire Metal--Brass Cylinder1 - - Home Depot-Rubber Tubing1$7 Home Depot--Hose Section1$10 $280.70Total

13. Do-ability Materials already stocked in lab: Plastic Syringe Water Sodium Chloride Pyrex Beaker Materials not stocked can be obtained, and protocols are straightforward, and not particularly complex

14. Bibliography Olton, D.S., & Samuelson, R.J. (1976). Remembrance of places passed: Spatial memory in rats. Journal of Experimental Psychology: Animal Behavior Processes, 2, 97-116. Ling D, Benardo L, Serrano P, Blace N, Kelly M, Crary J, Sacktor T (2002). "Protein kinase Mzeta is necessary and sufficient for LTP maintenance". Nat. Neurosci. 5 (4): 295-6.Protein kinase Mzeta is necessary and sufficient for LTP maintenance Serrano P, Yao Y, Sacktor T (2005). "Persistent phosphorylation by protein kinase Mzeta maintains late-phase long-term potentiation". J Neurosci 25 (8): 1979-84.Persistent phosphorylation by protein kinase Mzeta maintains late-phase long-term potentiation Pastalkova E, Serrano P, Pinkhasova D, Wallace E, Fenton A, Sacktor T (2006). "Storage of spatial information by the maintenance mechanism of LTP". Science 313 (5790): 1141-4Storage of spatial information by the maintenance mechanism of LTP Cooke SF, Bliss TV (2006). "Plasticity in the human central nervous system". Brain 129 (Pt 7): 1659-73 Boron, Walter F. (2005). Medical Physiology: A Cellular And Molecular Approach. Elsevier/Saunder Clugnet, MC; LeDoux JE (01 Aug 1990). "Synaptic plasticity in fear conditioning circuits: induction of LTP in the lateral nucleus of the amygdala by stimulation of the medial geniculate body." (PDF). J Neurosci 10 (8): 2818-24.Synaptic plasticity in fear conditioning circuits: induction of LTP in the lateral nucleus of the amygdala by stimulation of the medial geniculate body. Sweatt J (1999). "Toward a molecular explanation for long-term potentiation". Learn Mem 6 (5): 399-416.Toward a molecular explanation for long-term potentiation Toullec, Bisindolylmaleimide is a Potent and Selective Inhibitor of PKC, Biology Letters, p15711 – 15781 Olfactory Memory Capacity of the Cricket Gryllus Bimaculatus, Matsumoto, Biology Letters, volume 4, p608 - 610 Jan Wessnitzer, Place Memory in Crickets, Biology Letters, Volume 275, January 2008, pg 915 - 921