Project description:Transcriptional profiling of miRNAs from rat brain tissues comparing controls (Sham) with ischemic rats (tMCAO) and neuroprotected rats (RLIP) Internal normalization: ischemic core vs. periischemic and ANOVA comparison across three experimental conditions: Sham, tMCAO and RLIP
Project description:Purpose: we performed comparative RNA-seq analyses to identify molecular network characteristics of kidney injury at different ischemia times Methods: Unilateral clamping of the left renal pedicle plus contralateral nephrectomy was performed. Briefly, mice were anesthetized, right kidney was excised. The left kidney was clamped for indicated minutes (0min for sham-24h group, 16min for uIRIx16min-24h group, 18minfor uIRIx18min-24h group, 20min for uIRIx20min-24h group, 22min for uIRIx22min-24h group, 24min for uIRIx24min-24h group, 26min for uIRIx26min-24h group, 28min for uIRIx28min-24h group, 30min for uIRIx30min-24h group). Mice were placed on 37°C heating platform while operation, ischemia and before awakening. At 24h after ischemia, mice were anesthetized and blood was taken to detect creatinine, kidney was harvest for PAS staining and mRNA seq. Results: After sequence, the clean reads rate of all samples were ≥98%.The quality of the assembled transcriptome is good enough for functional annotation and further analysis. Conclusions: We performed RNA-sequencing (RNA-Seq) analyses to identify molecular network characteristics of kidney injury at different ischemia time.
Project description:This experiment was designed to study the alteration in expression of mRNA is kidney following recovery from transient acute renal failure. The model used was a 52 min. bilateral renal artery clamping, followed by reperfusion, which resulted in a transient loss of renal function, followed by a functional recovery. All tissue in this study was harvested 35 days post-surgery, when renal function was restored, and renal structure largelyy normal. For controls, sham-operated animals were used. An N of 6 post-ischemia reperfusion animals were used with 6 sham-operated controls. For hybridization studies, RNA from one of six post-ischemic acute renal failure animals were compared with with RNA from kidney of six sham-operated control animals. Each ARF vs. sham-operated comparison was performed twice, alternating the cy3 and cy5 label between the two hybridizations for each pair. A second set of hybridizations was carried out using sham vs. sham hybridizations. This was done to get a quantitative analysis of the variation of the biological and microarry platform. Keywords: parallel sample
