A Comprehensive Map of mRNAs and Their Isoforms across All 14 Renal Tubule Segments of Mouse.
basic_science · Level V
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- Record sourced from PubMed, PMID 33769951.
- Also identified by DOI 10.1681/ASN.2020101406 and PMC identifier 8017530.
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Abstract
The repertoire of protein expression along the renal tubule depends both on regulation of transcription and regulation of alternative splicing that can generate multiple proteins from a single gene. A full-length, small-sample RNA-seq protocol profiled transcriptomes for all 14 renal tubule segments microdissected from mouse kidneys. This study identified >34,000 transcripts, including 3709 that were expressed in a segment-specific manner. All data are provided as an online resource (https://esbl.nhlbi.nih.gov/MRECA/Nephron/). Many of the genes expressed in unique patterns along the renal tubule were solute carriers, transcription factors, or G protein-coupled receptors that account for segment-specific function. Mapping the distribution of transcripts associated with Wnk-SPAK-PKA signaling, renin-angiotensin-aldosterone signaling, and cystic diseases of the kidney illustrated the applications of the online resource. The method allowed full-length mapping of RNA-seq reads, which facilitated comprehensive, unbiased characterization of alternative exon usage along the renal tubule, including known isoforms of <i>Cldn10</i>, <i>Kcnj1</i> (ROMK), <i>Slc12a1</i> (NKCC2), <i>Wnk1</i>, <i>Stk39</i> (SPAK), and <i>Slc14a2</i> (UT-A urea transporter). It also identified many novel isoforms with segment-specific distribution. These included variants associated with altered protein structure (<i>Slc9a8</i>, <i>Khk</i>, <i>Tsc22d1</i>, and <i>Scoc</i>), and variants that may affect untranslated, regulatory regions of transcripts (<i>Pth1r</i>, <i>Pkar1a</i>, and <i>Dab2</i>). Full-length, unbiased sequencing of transcripts identified gene-expression patterns along the mouse renal tubule. The data, provided as an online resource, include both quantitative and qualitative differences in transcripts. Identification of alternative splicing along the renal tubule may prove critical to understanding renal physiology and pathophysiology.