1. Inherited Disorders of Human Memory: Mental Retardation Syndromes From Mechanisms of Memory by J. David Sweatt, Ph.D.

2. ERK1/2 MEK1/2 Raf1 R1R1 Grb SOS Ras PKC R2R2 B-Raf PKA R3R3 Rap AC Gene Expression Mnk1/2 eIF4E Protein Synthesis MAPs Spine Structure CRE CREB P CBP RSK2 GEF NF1 GAP NO. Ca 2+ G Protein NF1 Ca 2+ Nucleus R 4

3. ERK1/2 MEK1/2 Raf1 R1R1 Grb SOS Ras PKC R2R2 B-Raf PKA R3R3 Rap AC Gene Expression Mnk1/2 eIF4E Protein Synthesis MAPs Spine Structure CRE CREB P CBP RSK2 GEF NF1 GAP Neurofibromatosis MR NO. Ca 2+ G Protein NF1 Ca 2+ Nucleus R 4

4. PO 4 ras GDPGTP GTP Hydrolysis GAPs GTPase Activating Proteins e.g. NF1, SynGAP GEFs Guanine Nucleotide Exchange Factor Proteins e.g. SOS, cAMP GEF, Ca 2+ /DAG GEF, ras GRF InactiveActive + + Activation of ras Weeber and Sweatt. Neuron 33:845-848.

5. K-ras N-ras Farnesyl Transferase Inhibitor Costa et al (2002) Nature 415:526-530. Ras-dependent Spatial Learning in Nf1 +/- Animals

6. Ras-dependent LTP deficits in Nf1 +/- animals Costa et al (2002) Nature 415:526-530.

7. ERK1/2 MEK1/2 Raf1 R1R1 Grb SOS Ras PKC R2R2 B-Raf PKA R3R3 Rap AC Gene Expression Mnk1/2 eIF4E Protein Synthesis MAPs Spine Structure CRE CREB P CBP RSK2 GEF NF1 GAP Neurofibromatosis MR NO. Ca 2+ G Protein NF1 Ca 2+ Nucleus R 4

8. ERK1/2 MEK1/2 Raf1 R1R1 Grb SOS Ras PKC R2R2 B-Raf PKA R3R3 Rap AC Gene Expression Mnk1/2 eIF4E Protein Synthesis MAPs Spine Structure CRE CREB P CBP RSK2 GEF NF1 GAP Neurofibromatosis MR NO. Ca 2+ G Protein NF1 Ca 2+ Nucleus R 4 Coffin-Lowry Syndrome

9. ERK1/2 MEK1/2 Raf1 R1R1 Grb SOS Ras PKC R2R2 B-Raf PKA R3R3 Rap AC Gene Expression Mnk1/2 eIF4E Protein Synthesis MAPs Spine Structure CRE CREB P CBP RSK2 GEF NF1 GAP Neurofibromatosis MR NO. Ca 2+ G Protein NF1 Ca 2+ Nucleus R 4 Coffin-Lowry Syndrome Rubinstein-Taybi Syndrome

10. Current Model of Fragile X Mental Retardation Coding Region Regulatory Region CGG Expansion in Regulatory Region Point Mutation in Coding Region Disruption Of FMR1 Gene Loss of FMR1 Protein (FMRP) FMR1/FXR Interaction domain Ribosome Interaction Domain RGG Box KH Domain KH Domain RGG Box = Arginine & Glycine-rich domain KH domain = Ribonucleoprotein K homology domain FMRP = 63K RNA binding protein that binds to poly (G) and poly (U) structures FMR1 Gene Gene Structure FMRP Structure

11. ERK1/2 MEK1/2 Raf1 R1R1 Grb SOS Ras PKC R2R2 B-Raf PKA R3R3 Rap AC Gene Expression Mnk1/2 eIF4E Protein Synthesis MAPs Spine Structure CRE CREB P CBP RSK2 GEF NF1 GAP Neurofibromatosis MR NO. Ca 2+ G Protein NF1 Ca 2+ Nucleus R 4 Coffin-Lowry Syndrome Rubinstein-Taybi Syndrome Fragile X Syndrome

12. Enhanced LTP in FMR2 knockout mice Gu et al. (2002) J. Neurosci. 22:2753-2763.

13. Angelman Syndrome- Clinical Features Severe MR Absent speech Happy disposition Seizures

14. Ube3a codes for an E6-AP Ubiquitin Ligase Ube3a Ubiquitin Ligase Maternal Imprinting of Ube3a Expression of E6-AP in the Hippocampus and Cerebellum

15. Ube3a Deletion, MutationUniparental Disomy Mismethylation

16. The Ubiquitination Pathway Step 1 Step 2 Complex Formation Step 3 Step 4 Step 5 E1 Charging E2 Charging E2—E3 Transfer Target Poly- Ubiquitination E1 Ub ATP E2 Ligase Ub E3 Ligase Target Protein E2 Ub E3 Target Protein Ub Weeber and Sweatt (2000) Recent Res. Devel. Neurochem. 3:289-299.

17. Selective Deficit in Context-dependent Fear conditioning in Ube3a Maternal Deficient Mice Jiang et al (1998) Neuron 21:799-811.

18. Impairment of hippocampal LTP in Ube3a Maternal Deficient mice Jiang et al (1998) Neuron 21:799-811.

19. AS mouse model LTP

20. Saturating HFS-induced LTP

21. NMDAR Independent LTP

22. Pathogenesis of Angelman Syndrome E6-AP P53 HHR23A E6-AP MCM7 ?

23. Adenylyl Cyclase ATP CaMCa ++ PLC Ras B-Raf Rap1 Raf-1 Ras MEK ERK PKC CaMKII PKA DAG cAMP Ca ++ Channel NMDA Receptor

24. A Total Kinase Concentrations are Normal

25. A Increased P-Thr 286 CaMKII in AS Mice P-PKC P-PKA P-ERK p42 P-CaMKII 0 20 40 60 80 100 120 140 160 180 * Total  CaMKII P-Thr 286  CaMKII WT m-/p+ Immunoreactivity (% of Control)

26. Reduced Post-Hoc CaMKII Phosphorylation

27. Thr 286 Regulatory Subunit Ca 2+ CAM  CaMKII Catalytic Subunit

28.  CaMKII Catalytic Subunit Autonomously Active P Thr 286 Ca 2+ CAM Regulatory Subunit

29. Hippocampal CaMKII Activity

30. Thr 286Thr 305 Regulatory Subunit Ca 2+ CAM  CaMKII Catalytic Subunit

31.  CaMKII Catalytic Subunit P P Thr 286 Thr 305 Ca 2+ CAM Regulatory Subunit Inactive

32. Wildtypem-/p+ 0 25 50 75 100 125 150 * Immunoreactivity (% of Control)  CaMKII Total  CaMKII Thr 305/306 CaMKII Hippocampal HomogenatesPurified CaMKII WTm-/p+WTm-/p+ Control Peptide Block Ctrl +Ca 2+ /CaM Ctrl +Ca 2+ /CaM Ctrl +Ca 2+ /CaM  CaMKII  

33. Wildtypem-/p+ 0 25 50 75 100 125 150 * Immunoreactivity (% of Control)  CaMKII Total  CaMKII Thr 305/306 CaMKII Hippocampal HomogenatesPurified CaMKII WTm-/p+WTm-/p+ Control Peptide Block Ctrl +Ca 2+ /CaM Ctrl +Ca 2+ /CaM Ctrl +Ca 2+ /CaM  CaMKII  

34.  100 Hz/1s No  CaMKII  CaMKII T305D Mice are Impaired in LTP Inhibited Wild type 60 80 100 120 140 160 180 200 220 -100102030405060 Time (min) WT T305D CaMKII Elgersma et al.

35. T305D Mutants Have Impaired Fear-Conditioned Learning Elgersma et al.

36.  Cam KII Catalytic Subunit P P Thr 286 Thr 305 Ca 2+ CAM Regulatory Subunit Inactive

37. NR1 NR2B CaMKII P PP1 PP2A ? E6-AP P

38. AS Mouse Model-Conclusions Aberrant hyper-autophosphorylation of CaMKII in Angelman mouse hippocampus. Altered phosphatase activity (PP1 and/or PP2A). Alterations in CaMKII autophosphorylation can explain the physiologic and behavioral deficits in our model. A role for CaMKII activity in human learning.

39. rho PAK, ROCK LIMK-1 (Williams Syndrome) Actin Depolymerization Factor (ADF) / cofilin Actin Cytoskeleton—Loss of LIMK-1 causes increased actin turnover Altered Dendritic Spine Augmented LTP, Learning Impairments rac PKC Direct phosphorylation (inhibitory) ADF / Cofilin promotes Actin depolymerization Williams Syndrome

40. Rho PAK3 (p21 Activated Kinase) JNK p38 Cytoskeletonraf-1LTD disruption? Dbl ( Diffuse B-cell Lymphoma) Rho GEF6 Rho GAP Rho GDI GEFs + _ Nonsyndromic X-Linked Mental Retardation

41. Mental Retardation Syndromes Disruptions of memory-related signal transduction systems. Impact of basic research on clinically relevant studies, and vice-versa. Identifying new avenues of treatment.

42. ERK1/2 MEK1/2 Raf1 R1R1 Grb SOS Ras PKC R2R2 B-Raf PKA R3R3 Rap AC Gene Expression Mnk1/2 eIF4E Protein Synthesis MAPs Spine Structure CRE CREB P CBP RSK2 GEF NF1 GAP Neurofibromatosis MR NO. Ca 2+ G Protein NF1 Ca 2+ Nucleus R 4 Coffin-Lowry Syndrome Rubinstein-Taybi Syndrome Fragile X Syndrome