Frequency of Apolipoprotein E (apoE) in Breast Cancer and 载脂蛋白E（APOE）频率在乳腺癌.pptVIP

The effect of ApoE4 on Fear Conditioning Michael A. Manoharan Valerio Dr. Summer Acevedo’s lab Ponce School of Medicine ApoE 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 apoE4 ? Diabetes/Hypertension ? Alzheimer disease /wiki/File:Gfapastr5.jpg /wiki/File:PBB_Protein_APOE.jpg /dynamicImages/chromomap/apoe.jpeg Methods 3 2 1 4 II. PCR I. DNA extraction CfoI cut sites 30 16 19 72 18 48 35 5’-F6 3’-F4 C112R R158C IV. RFLP Analysis III. Digest PCR 95° C 65°C 72°C ApoE Genotyping 3 2 4 30 16 91 18 83 30 16 19 72 18 48 35 30 16 91 18 48 35 C112R R158C Alleles ApoE RFLP’s M1 2/2 3/3 4/4 2/3 2/4 3/4 M2 100bp 75 50 100bp 4 % agarose gel 50 Photos 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. Fear Conditioning Milad 2006 at all ApoE4 effect Fear Recall * * F = 6.70 p0.02 N=20 N=14 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-