1. Biochemical Mechanisms for Long-term Information Storage at the Cellular Level From Mechanisms of Memory by J. David Sweatt, Ph.D.

2. Protein Synthesis Dependence of L-LTP Kang et al. (2001) Cell 106:771-783.

3. THE CASE FOR GENE EXPRESSION IN L-LTP EXPERIMENT TYPE and FINDING Block Block of L-LTP with protein synthesis inhibitors Block of L-LTP with RNA synthesis inhibitors Loss of L-LTP with CREB KOs Block of L-LTP with Arc antisense Loss of L-LTP with CaMKIV KO Loss of L-LTP with zif268 KO

4. THE CASE FOR GENE EXPRESSION IN L-LTP EXPERIMENT TYPE and FINDING Measure Increased zif268/krox24 mRNA Increased krox20 Increased expression of fos, jun IEG mRNAs Increased CREB phosphorylation Increased CRE read-out Increased elk-1 phosphorylation Increased Arc/Arg3.1 mRNA expression Increased AMPA receptor protein Increased BDNF message Increased tissue Plasminogen Activator message Increased C/EBP beta (in long-term memory) Increased HOMER Increased MAP Kinase Phosphatase-1 Increased SSAT message Increased MAP2 message Mimic Constitutively active CREB augments L-LTP induction

5. CREB Binding Protein (CBP) cAMP Response Element 5’-TGACGTCA-3’ TATA Sequence CREB Basal Transcription Factor IID (TFIID) Complex ser 133 RNA Helicase A RNA Polymerase II Histone pKAF acetylation Target Gene PO 4 The CREB/CRE Gene Regulation System

6. Figure courtesy of Jennifer Gatchel Brookhaven National Protein Structure Data Bank, Rendered with Rasmol Crystal Structure of CREB bound to the CRE

7. CBPp300 O O P O O Histone acetylation Elk-1 O O P O O CREB O O P O O CBPp300 O O P O O c-fos fosB junB egr-1 etc... Histone Acetyltransferase (HAT) – Regulation of Gene Expression

8. MKP-3/Dual Specificity Phosphatase Upstream Regulatory Elements C/EBP Elk-1 MZF1 Octamer-binding factor 1 NF-  B C HOP -C/EBP  ras-REBP MyT1 c-Rel E2F Wilm's Tumor Suppressor EVI-1 Factor E2F MAZ MKP-3 GC Box 92 total regulatory elements CBF/NFY

9. Components of the LTP Gene Expression System 1. A core signal transduction cascade linking calcium to the transcription factor CREB 2. Modulatory influences that impinge upon this cascade 3. Additional transcription factors besides CREB that may be involved in L-LTP induction 4. Gene targets in L-LTP 5. mRNA targeting and transport 6. Effects of the gene products on synaptic structure

11. Components of the LTP Gene Expression System 1. A core signal transduction cascade linking calcium to the transcription factor CREB 2. Modulatory influences that impinge upon this cascade 3. Additional transcription factors besides CREB that may be involved in L-LTP induction 4. Gene targets in L-LTP 5. mRNA targeting and transport 6. Effects of the gene products on synaptic structure

12. G Protein ERK1/2 MEK1/2 Raf1 R1R1 Grb SOS Ras PKC R2R2 B-Raf PKA R3R3 Rap AC Gene Expression CRE CREB P CBP RSK2 GEF NF1 GAP NO. Ca 2+ Nucleus AChR

13. Components of the LTP Gene Expression System 1. A core signal transduction cascade linking calcium to the transcription factor CREB 2. Modulatory influences that impinge upon this cascade 3. Additional transcription factors besides CREB that may be involved in L-LTP induction 4. Gene targets in L-LTP 5. mRNA targeting and transport 6. Effects of the gene products on synaptic structure

14. Activity-dependent Regulation of Gene Expression

15. Components of the LTP Gene Expression System 1. A core signal transduction cascade linking calcium to the transcription factor CREB 2. Modulatory influences that impinge upon this cascade 3. Additional transcription factors besides CREB that may be involved in L-LTP induction 4. Gene targets in L-LTP 5. mRNA targeting and transport 6. Effects of the gene products on synaptic structure

16. CRESRE zif268 C/EBP krox20 AMPAR HOMER BDNF NT-3 MKP-1 SSAT Arc CBP/CREB CaMKIV + ERK Elk-1 RSK2 Transcriptional Regulation Pathways

17. Components of the LTP Gene Expression System 1. A core signal transduction cascade linking calcium to the transcription factor CREB 2. Modulatory influences that impinge upon this cascade 3. Additional transcription factors besides CREB that may be involved in L-LTP induction 4. Gene targets in L-LTP 5. mRNA targeting and transport 6. Effects of the gene products on synaptic structure

18. Steward and Worley (2001) PNAS 98:7062-7068. Activity-dependent Arc Expression and Dendritic Localization

19. Components of the LTP Gene Expression System 1. A core signal transduction cascade linking calcium to the transcription factor CREB 2. Modulatory influences that impinge upon this cascade 3. Additional transcription factors besides CREB that may be involved in L-LTP induction 4. Gene targets in L-LTP 5. mRNA targeting and transport 6. Effects of the gene products on synaptic structure

20. Hypothetical Mechanisms for Activity-dependent changes in Synaptic Structure

21. Presynaptic Postsynaptic Integrins Ca ++ actin Paxillin FAK (Focal Adhesion Kinase) grb2/sos cas/crk src ERKJNK αα ββ rhorac Dynamic cytoskeletal regulation Cadherins Catenins ? DMPK Myosin Phosphatase Cytoskeletal Myosin II - - α-actinin talin vinculin α-actinin Signaling Mechanisms Used by Integrins and Cadherins

22. Components of the LTP Gene Expression System 1. A core signal transduction cascade linking calcium to the transcription factor CREB 2. Modulatory influences that impinge upon this cascade 3. Additional transcription factors besides CREB that may be involved in L-LTP induction 4. Gene targets in L-LTP 5. mRNA targeting and transport 6. Effects of the gene products on synaptic structure

23. Villarreal et al. (2002). Nat. Neurosci. 5:48-52. L-LTP Decay is an Active Process

24. Trace Eyeblink Delay Eyeblink Morris Watermaze Post training Injection Neurogenesis in the Adult CNS Gould et al. (1999) Nat. Neurosci. 2:260-265.