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Publication : Cell-specific qRT-PCR of renal epithelial cells reveals a novel innate immune signature in murine collecting duct.

First Author  Saxena V Year  2018
Journal  Am J Physiol Renal Physiol Volume  315
Issue  4 Pages  F812-F823
PubMed ID  28468965 Mgi Jnum  J:283906
Mgi Id  MGI:6368419 Doi  10.1152/ajprenal.00512.2016
Citation  Saxena V, et al. (2018) Cell-specific qRT-PCR of renal epithelial cells reveals a novel innate immune signature in murine collecting duct. Am J Physiol Renal Physiol 315(4):F812-F823
abstractText  The urinary tract is usually culture negative despite its close proximity to microbial flora. The precise mechanism by which the kidneys and urinary tract defends against infection is not well understood. The initial kidney cells to encounter ascending pathogens are the collecting tubule cells that consist of principal cells (PCs) that express aquaporin 2 (AQP2) and intercalated cells (ICs) that express vacuolar H(+)-ATPase (V-ATPase, B1 subunit). We have previously shown that ICs are involved with the human renal innate immune defense. Here we generated two reporter mice, VATPase B1-cre(+)tdT(+) mice to fluorescently label ICs and AQP2-cre(+)tdT(+) mice to fluorescently label PCs, and then performed flow sorting to enrich PCs and ICs for analysis. Isolated ICs and PCs along with proximal tubular cells were used to measure antimicrobial peptide (AMP) mRNA expression. ICs and PCs were significantly enriched for AMPs. Isolated ICs responded to uropathogenic Escherichia coli (UPEC) challenge in vitro and had higher RNase4 gene expression than control while both ICs and PCs responded to UPEC challenge in vivo by upregulating Defb1 mRNA expression. To our knowledge, this is the first report of isolating murine collecting tubule cells and performing targeted analysis for multiple classes of AMPs.
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