Macaca mulatta spleen sample exposed to SHIV 89.6 strain
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ABSTRACT: Splenic tissue was isolated from four adult male Indian-origin Rhesus monkeys serologically positive for non-pathogenic SHIV 89.6 and from matched uninfected four adult male Indian-origin Rhesus monkeys respectively. The corresponding RNA was processed by cDNA microarray analysis. Keywords: SIV infection
Project description:Splenic tissue was isolated from four adult male Indian-origin Rhesus monkeys serologically positive for non-pathogenic SHIV 89.6 and from matched uninfected four adult male Indian-origin Rhesus monkeys respectively. The corresponding RNA was processed by cDNA microarray analysis. Sample RNA extracted from the tissue samples. Reference RNA was prepared from a pool of Rhesus monkeys' RNA. Microarray hybridization was carried out by labeling Reference RNA as Cy3 and sample RNA as Cy5. The sample was incubated at 55C for 16hr.
Project description:Objective: The dietary xanthophylls, lutein and zeaxanthin, accumulate in primate brain and may be beneficial for cognition. Brain xanthophyll content varies greatly among individuals and genetic factors are likely to be significant contributors. Subspecies of rhesus macaques originating from different geographic locations differ genetically, but the effect of origin on gene expression and carotenoid status has not been determined. The study objective was to determine whether xanthophyll status and expression of carotenoid-related genes, as well as genes with known variants between subspecies, differ between the brains of adult rhesus monkeys of Indian and Chinese origin. Methods: Next generation RNA sequencing was used to determine differentially expressed carotenoid-related genes and genes with known variants among rhesus monkey subspecies in the prefrontal cortex, cerebellum, and striatum of Indian-origin monkeys (n=3) versus Chinese-origin monkeys (n=3). Serum and brain xanthophylls were determined using HPLC. FastQC was performed on raw sequenced reads to determine the quality of each read. Reads were mapped to the Rhesus Macaque reference genome and differences in gene expression (FPKM) were determined using TopHat and Cuffdiff, respectively. Findings from RNAseq were validated using RT-PCR. Results: Indian-origin monkeys had higher xanthophyll levels in brain tissue compared to Chinese-origin monkeys despite consuming similar amounts of dietary carotenoids. In a region-specific manner, 4 genes related to carotenoid and fatty acid metabolism (BCO2, RPE65, ELOVL4, FADS2) and 4 genes involved in the immune response (CD4, CD74, CXCL12 LTBR) were differentially expressed between Indian- and Chinese-origin monkeys. Expression of all four genes involved in carotenoid and fatty acid metabolism were correlated with brain xanthophyll concentration in a region-specific manner. Conclusions: These results indicate that origin is related to differences in both gene expression and xanthophyll content in the brain. Findings from this study may have important implications regarding genetic diversity, lutein status, and cognition in primates.