Deep Sequencing in Microdissected Renal Tubules Identifies Nephron Segment–Specific Transcriptomes

摘要: The function of each renal tubule segment depends on the genes expressed therein. High-throughput methods used for global profiling gene expression in unique cell types have shown low sensitivity and high false positivity, thereby limiting usefulness these transcriptomic research. However, deep sequencing RNA species (RNA-seq) achieves highly sensitive quantitative by RNAs a massive, parallel manner. Here, we RNA-seq coupled with classic microdissection to comprehensively profile 14 segments from proximal through inner medullary collecting duct rat kidneys. Polyadenylated mRNAs were captured oligo-dT primers processed into adapter–ligated cDNA libraries that sequenced using an Illumina platform. Transcriptomes identified median depth 8261 microdissected samples (105 replicates total) glomeruli (5 replicates). Manual allowed degree sample purity, which was evidenced observed distributions well established cell–specific markers. main product this work is extensive database along nephron provided as publicly accessible webpage (https://helixweb.nih.gov/ESBL/Database/NephronRNAseq/index.html). data also provide genome-wide maps alternative exon usage polyadenylation sites kidney. We illustrate use transcription factor mapping metabolic pathways.