1. Manhasset Science Research Paris Thatos
PKC and Memory
Manhasset Science Research
Paris Thatos

2. Need Fig 1. Shows trend in drug related deaths.
Long-term trend in drug-related deaths
Percentage of People Experiencing a Mental Disorder in their Lives
Before I get into my topic, I would first like to introduce two common learned behaviors: drug dependence and chronic fears, or phobias
You can see in this graph, that the trend of drug related deaths has increased drastically since its standing in 1985
In thjs graph, it is shown that about 14.3% of people suffer from some phobia, and in specific numbers, 6.2 million americans suffer from some phobia
Since these two are in essence learned behaviors, it may be possible to suppress them using biochemical means
Fig 1. Shows trend in drug related deaths.
Fig 2. A graph showing % of people experiencing mental disorders
(Gotz, 2004)
(Gallaher, 2007)

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

4. PKC (Protein Kinase C) Has many isoforms (PKMzeta,PKMa)
(Abel, 2004)
Fig 4. Process of PKC interaction in Long term Potentiation
Involved in the stage of maintenance and expression
We believe that memories are stored in the synapses between the neurons, so it would follow that increasing the bond between the neurons would strengthen the memory

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. Bisindolylmaleimide is a Potent and Selective Inhibitor of PKC
Dominique Toullec, The Journal of Biological Chemistry
The levels of PKC and PKA
Toullec, 2005
Fig 6. The concentrations of PKC when exposed to Bisindolylmaleimide, and the concentrations of a PKC isotype

7. Formation of Long-term Olfactory Memory in the cricket Gryllus bimaculatus
Makoto Mizunami, 2005, Chem. Senses
Makoto, 2006
Fig 7. A diagram of the head of the cricket. Brain (In blue) is where PKC is believed to be contained in.

8. PKMzeta maintains spatial, instrumental and classically conditioned Long term memories
Peter Serano, Todd C. Sacktor
PLoS Biology
Fig. 8. The Water Maze in a rat. 1st trial to eigth trial, when the rat has successfully memorized the path to the platform
– December 2008
PKMzeta maintains spatial, instrumemtal and classically conditioned Long term memories
Trained rats to go through a radial arm maze and a water maze, two mazes that train spatial learning
The rats then had the ZIP injected into their bodies
The rats completely forgot how to complete both mazes.
When released, the rat swims around the pool in search of an exit while various parameters are recorded, including the time spent in each quadrant of the pool, the time taken to reach the platform (latency), and total distance traveled. The rat's escape from the water reinforces its desire to quickly find the platform, and on subsequent trials (with the platform in the same position) the rat is able to locate the platform more rapidly. This improvement in performance occurs because the rat has learned where the hidden platform is located relative to the conspicuous visual cues. After enough practice, a capable rat can swim directly from any release point to the platform.
(Morris, 2004)

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
It is hypothesized that with all administeringings of Bisindolymaleide, the formation of memories will be inhibited

10. Grasshopper (Gryllus Bimaculatus) n=120
Statistically analyzed with a T-test (to find statistic significance) followed by a Scheffe post-hoc test (p<o.o5)
Trained to associate one scent with reward and one scent with punishment
Also, trained to run 塗eat maze�
Control group n=30
Reward/Punishment for scent test is water/saline solution
Variable group n=30
Reward/Punishment for scent test is water/saline solution (+5M)
5Min water
Reward/Punishment for scent test is water/saline solution (+5M), also 5Min water supply
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
Scent Testing
Scent Conditioning
(Matsumoto, 2006)

12. Budget Vendor Catalog # page/website Description Quantty Unit $
Total $
EMDbiosciences
203290
Bisindolylmaleimide I 1
$40.00
Carolina
143550
Crickets (Living Adults)
$6.75
143565
House Cricket Food 2
$6.25
$12.50
695145
Sense of Smell Kit
$49.25
199606
Plastic Syringe
$3.85
198697
Distilled Water
721207
Pyrex Beaker 100ml 4
$3.65
$14.60
Total = $133.70

13. Do-ability Materials already stocked in lab: Plastic Syringe Water
Sodium Chloride
Pyrex Beaker
Materials not stocked can be obtained, and protocols are 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,
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):
Serrano P, Yao Y, Sacktor T (2005). "Persistent phosphorylation by protein kinase Mzeta maintains late-phase long-term potentiation". J Neurosci 25 (8):
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):
Cooke SF, Bliss TV (2006). "Plasticity in the human central nervous system". Brain 129 (Pt 7):
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):
Sweatt J (1999). "Toward a molecular explanation for long-term potentiation". Learn Mem 6 (5):
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, p