Using RNA-Seq to evaluate the aged female Sprague-Dawley rat cortex transcriptome after repeated bouts of sleep deprivation induced by the gentle-handling method.
Elliott-AS; Turner-RC; O'Callaghan-JP; Rosen-CL; Huber-JD; Miller-DB
Neuroscience 2012: 42nd Annual Meeting of the Society for Neuroscience, October 13-17, 2012, New Orleans, Louisiana. Washington, DC: Society for Neuroscience, 2012 Oct; :486.26/VV14
Sleep deprivation (SD) has been linked to cardiovascular and cerebrovascular disease, as well as the metabolic syndrome. Animal models of long-term SD show dramatic changes in gene expression in the cerebral cortex. Unfortunately, most SD models induce activity of the hypothalamus-pituitary-axis (HPA), indicating stress. This study utilizes the "gentle-handling" method to examine effects of repeated bouts of SD on the transcriptome of the aged female Sprague-Dawley rat cortex. Rats were sleep deprived for approximately half of their normal "active period" for 6 consecutive days. Duration of inactivity was monitored during the daily non-deprivation period via locomotor chambers (Accuscan) and increased over the days of SD in SD group compared to controls. Adrenal and thymus weights were measured as a biological indicator of corticosterone release and did not differ between groups. Transcriptome analysis of cortex tissue was conducted with Next Gen RNA sequencing. Using RNA-Seq, over 25 million short sequencing reads were generated per sample for 12 samples (6 SD and 6 controls). These reads were aligned to the rat reference genome allowing 29,516 Ensembl genes to be searched with over 15,000 genes detected. To determine differential expression, ‘digital mRNA counting was applied based on reads that map to exons. Data was analyzed through a candidate gene approach and Ingenuity Pathway Analysis (IPA) with threshold parameters set at > 1.5 fold change and < .05 p-values. In accordance with previous reports, results showed an upregulation of genes encoding for proteins involved in the unfolded protein response, such as the heat shock 70kDa protein 5 (HSPA5) and growth arrest and DNA-damage-inducible alpha (GADD45a) and related to the cytoskeleton, such as collagen type III alpha 1 (Col3a1) and collagen type I alpha 1 and 2 (Col1a1, Col1a2). Further analysis of the transcriptome of rat cortex of SD rat will promote insight into the molecular consequences of sleep deprivation.
Sleep-deprivation; Cardiovascular-system-disease; Cardiovascular-system-disorders; Cardiovascular-system; Cardiovascular-disease; Cerebrovascular-system; Cerebrovascular-system-disorders; Metabolic-disorders; Animals; Genes; Models; Stress; Laboratory-animals; Proteins;
Author Keywords: SLEEP DEPRIVATION
Healthcare and Social Assistance; Transportation, Warehousing and Utilities
Neuroscience 2012: 42nd Annual Meeting of the Society for Neuroscience, October 13-17, 2012, New Orleans, Louisiana