1. Genetic Effects of Stress in Vervet Monkey Olivera Grujic Dr. Eleazar Eskin’s Lab, UCLA Dr. Nelson Freimer’s Lab,UCLA SoCalBSI, 2008

2. Project Importance Evaluate biomedical bases of inter-individual differences in response to stressor. Stress Related Diseases: Depression Post Traumatic Stress Disorder Response to stress has genetic component!

3. Challenging of Studying Genetic Factors of Stress Complex trait Experiments in humans Need: Model organism that reacts to stress!

4. Vervet Research Colony Inbred pedigree 1000 members Small number brought to the Carribean (300- 400 years ago) 57 wild-caught brought from St. Kitts to UCLA (1975-1989) Colony moved from UCLA to Wake Forest (in January 2008) African Vervets (Chlorocebus aethiops sabaeus)

5. Same stressor Controlled environment Good quality of tissue Simultaneous effect in multiple organ systems Highly informative pedigree Vervet genetic map suitable for QTL mapping Advantages of Researching Vervet over Human Population Freimer NB, et al. A quantitative trait locus for variation in dopamine metabolism mapped in a primate model using reference sequences from related species. Proc Natl Acad Sci U S A. 2007 Oct 2;104(40):15811-6. Epub 2007 Sep 20. Jasinska AJ, et al. A genetic linkage map of vervet monkey. (2007) Mamm Genome 18:347–360.

6. Goal: Use samples to determine effects of stress in terms of: Moving can be stressful. Vervet Colony exposed to a major stressor - all of them were moved under the same conditions, at the same time (in controlled way)! Data Collection Before move: –Blood samples from ~380 individuals –Brain tissue from 12 individuals After move: –Blood samples from ~340 individuals –Brain tissue from 4 individuals gene expression profiles interindividual differences

7. Challenges 1. No vervet genome and no vervet microarrays 2. Not much known about gene expression in primate brain 3. Mostly collecting blood data 4. Available expression data only before move

8. First Task  Assess quality of DNA probes  Identify inter-species sequence differences * Vervet BAC end sequences submitted to NCBI in batches

9. Probe Comparison Workflow Illumina BeadStudio Output File Extract and Add Headings Count Frequency Parse Convert to Compare Using BLAST 341,172 Vervet Sequences 22184 Probe SequencesVervet Database BLAST Output File Top Hit for Each Probe Probes per Nucleotides Matched

10. Results Probe Comparison LengthFreq 17 nucleotides 28% 18 nucleotides 15% 19 nucleotides 8% 16 nucleotides 23% 50197 49225 48186 47160 46146 45121 44104 4370 4281 4155 4057 3945 3844 3743 3643 3564 3444 3341 3245 3147 3035 2939 2851 2761 2667 25104 2494 23208 22322 21638 201198 191662 183382 176286 165100 151046 140 15 nucleotides 5% 20 nucleotides 5%

11. Probe Alignments

12. Second Task  Characterize regional gene expression in vervet brain  Characterize group of genes with low gene expression variability between brain and blood  Approach:  Cluster expression data from blood and following brain tissues: Head of Caudate Cereballar Vermis Hippocampus Frontal Pole Dorsolateral Prefrontal Cortex Orbital Frontal Cortex Pulvinar Occipital Pole Obtain a list of genes where more than 75% of variability is due to inter-individual differences!

13. Pulvinar Head of Caudate Cerrebalar Vermus Hippocampus Frontal Pole DLPFC Orbital Frontal Occipital pole Blood

14. Results: Clustering Individuals Tissue Type

15. Samples Genes Cluster1 Cluster2 Blood Tissue

16. GO Analysis on Cluster 1

17. GO Analysis on Cluster 2

18. Future Work Third Task: brain to blood mapping Fourth Task: compare pre-move and post-move expression data

19. Acknowledgments SoCalBSI Dr. Jamil Momand Dr. Sandra Sharp Dr. Nancy Warter-Perez Dr. Wendie Johnston Dr. Beverly Krilowicz Dr. Silvia Heubach Dr. Jennifer Faust Ronnie Cheng SoCalBSI 2008 Interns  Funded by: UCLA Dr. Eleazar Eskin Dr. Nelson Freimer Dr. Ania Jasinska My Labmates