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The effect of ApoE4 on Fear Conditioning

The effect of ApoE4 on Fear Conditioning. Michael A. Manoharan Valerio Dr. Summer Acevedo’s lab Ponce School of Medicine. ApoE. http://en.wikipedia.org/wiki/File:Gfapastr5.jpg. apoE4  Diabetes/Hypertension  Alzheimer disease. Gene located on chromosome 19q13.2

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The effect of ApoE4 on Fear Conditioning

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  1. The effect of ApoE4 on Fear Conditioning Michael A. Manoharan Valerio Dr. Summer Acevedo’s lab Ponce School of Medicine

  2. ApoE http://en.wikipedia.org/wiki/File:Gfapastr5.jpg • apoE4 •  Diabetes/Hypertension •  Alzheimer disease • Gene located on chromosome 19q13.2 • 299 amino acid residue (34 kDa protein) • Produced by astrocyte cells and is expressed in the brain • Assists in the regulation of lipid metabolism • Major alleles: apoE2, apoE3, apoE4 ”Exon 4” • Risk factor: • apoE2 •  Cardiovascular disease •  Alzheimer disease http://en.wikipedia.org/wiki/File:PBB_Protein_APOE.jpg http://ghr.nlm.nih.gov/dynamicImages/chromomap/apoe.jpeg

  3. Methods I. DNA extraction 1 2 3 4 II. PCR C112R R158C III. Digest CfoI cut sites 30 16 19 72 18 48 35 5’-F6 3’-F4 IV. RFLP Analysis

  4. PCR 95° C 65°C 72°C

  5. ApoEGenotyping C112R R158C Alleles 30 16 91 18 48 35 3 30 16 91 18 83 2 30 16 19 72 18 48 35 4

  6. ApoE RFLP’s 4 % agarose gel M1 2/2 3/3 4/4 2/3 2/4 3/4 M2 100bp 100bp 75 50 50

  7. Photos

  8. Fear Conditioning • This technique is used to study the acquisition and extinction of fear. • The most common technique of Fear conditioning is the Pavlovian conditioning. • The technique used was Associative Learning which is pairing an unconditional stimulus and a condition stimulus so the organism can learn fear. • The Unconditional Stimulus is something that naturally provokes a response. • The Conditioned Stimulus is something that paired with the unconditional stimulus will get a response because naturally it will not create a response.

  9. Fear Conditioning Milad 2006 at all

  10. ApoE4 effect Fear Recall * N=20 N=14 * F = 6.70 p<0.02

  11. Acknowledgements Collaborators: • Summer F. Acevedo, PhD • Tirtsa Porrata-Doria • Jaime Matta, PhD • Julie Dutil, PhD • Project Funded by: • Moffit Cancer Center Grant (60-14599-01-01-54) through Dr. Acevedo,R01 HD050559 • NIH-MBRS Program grant S06 GM008239-20 to PSM through Dr. J. Matta • RCMI High School Supplement Special thanks to the PSM Molecular Biology Core Lab (NCRR-G12RR003050-24) for their assistance and support on the development of this investigation.

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