ABSTRACT: Unraveling the Specific Ischemic Core and Penumbra Transcriptome in Permanent Middle Cerebral Artery Occlusion Model Mice Brain Treated with Neuropeptide PACAP38
Project description:Permanent middle cerebral artery occlusion (PMCAO) mouse model has been established and used by our group with a focus on systematically investigating effects of the neuropeptide pituitary adenylate-cyclase activating polypeptide (PACAP38) on the ischemic brain. Using an intracerebroventrically PACAP38 (1 pmol) injection over a control saline (0.9% sodium chloride, NaCl) treatment, we have been successful in generating a vast inventory of gene expression data in both whole hemispheres and specific brain regions of infract/ischemic core (IC) and penumbra (P) with high-throughput Agilent whole genome 4 × 44 K oligo DNA microarray chips. In this study, we first perform a specific analysis of the gene expression in the IC and P compared to the healthy control regions to know the gene profile status of the progression of the IC. We will use this data together with generated biological function and network analysis from our previous studies, and present new insight into the potential mechanism behind PACAP38 neuroprotective function in the ischemic brain. Three mice each in PMCAO groups for ischemic core and penumbra regions in ischemic brain over corresponding controls were used, respectively, that exhibited neurological grades G1 and G2 for the subsequent downstream analysis. Ischemic core and penumbra regions and corresponding healthy core and penumbra were carefully removed with a sterile scalpel, and placed in 2 ml Eppendorf tubes. Samples were then quickly immersed in liquid nitrogen and stored in -80 ºC prior to further analysis. Whole genome DNA microarray analysis was performed on a 4x44K, Agilent DNA Microarray chip.
Project description:Permanent middle cerebral artery occlusion (PMCAO) mouse model has been established and used by our group with a focus on systematically investigating effects of the neuropeptide pituitary adenylate-cyclase activating polypeptide (PACAP38) on the ischemic brain. Using an intracerebroventrically PACAP38 (1 pmol) injection over a control saline (0.9% sodium chloride, NaCl) treatment, we have been successful in generating a vast inventory of gene expression data in both whole hemispheres and specific brain regions of infract/ischemic core (IC) and penumbra (P) with high-throughput Agilent whole genome 4 × 44 K oligo DNA microarray chips. In this study, we first perform a specific analysis of the gene expression in the IC and P compared to the healthy control regions to know the gene profile status of the progression of the IC. We will use this data together with generated biological function and network analysis from our previous studies, and present new insight into the potential mechanism behind PACAP38 neuroprotective function in the ischemic brain.
Project description:To analyze gene expression we isolated total mRNA from the core, periinfarct and contralateral cortex of 60 sprague-dawley rats after 24 hours or 3 days of permanent middle cerebral artery occlusion (pMCAo). Also ipsilateral and contralateral cortex was obtained from sham-operated and healthy animals. A total of 44 microarrays were performed with RNA pooled from 3 independent animals. For each experimental condition the total n number is of 12. Microarray analysis of different areas of the rat brain (core, periinfarct and contralateral cortex) after 24h and 3days of permanent ischemia
Project description:profiling gene transcription in a mouse model of permanent focal cerebral ischemia that was induced by middle cerebral artery occlusion (MCAO)
Project description:Bioinformatics Analysis of the Permanent Middle Cerebral Artery Occlusion (PMCAO) Mouse Model Brain Regions Gene Profiles for Unraveling Progression of the Ischemic Core
Project description:The mammalian brain consists of heterogeneous cell populations with distinct responses to injury. Here we used single nucleus RNA sequencing (snRNAseq) to analyze cell type specific transcriptional responses of the rodent brain to ischemic stroke, 48h after the induction of middle cerebral artery occlusion.
Project description:To investigate the effect of stroke on the transcriptome of intestinal epithelial cells, mice underwent permanent middle cerebral artery occlusion (pMCAO) or sham surgery for 24 hours
Project description:To investigate the effect of stroke on the trasnscriptome of intestinal muscularis macrophages, mice underwent permanent middle cerebral artery occlusion (pMCAO) or sham surgery for 24 hours
Project description:To investigate the effect of stroke on the trasnscriptome of intestinal lamina propria macrophages, mice underwent permanent middle cerebral artery occlusion (pMCAO) or sham surgery for 24 hours
