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Cell-specific qRT-PCR of renal epithelial cells reveals a novel innate immune signature in murine collecting duct
- Source :
- American Journal of Physiology-Renal Physiology. 315:F812-F823
- Publication Year :
- 2018
- Publisher :
- American Physiological Society, 2018.
-
Abstract
- 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.
- Subjects :
- 0301 basic medicine
Vacuolar Proton-Translocating ATPases
Physiology
Urinary system
Mice, Transgenic
Biology
Kidney
Polymerase Chain Reaction
Microbiology
03 medical and health sciences
medicine
Animals
Intercalated Cell
Kidney Tubules, Collecting
Cell specific
Aquaporin 2
Innate immune system
Epithelial Cells
Immunity, Innate
Up-Regulation
030104 developmental biology
Real-time polymerase chain reaction
medicine.anatomical_structure
Immunology
Culture negative
Duct (anatomy)
Research Article
Subjects
Details
- ISSN :
- 15221466 and 1931857X
- Volume :
- 315
- Database :
- OpenAIRE
- Journal :
- American Journal of Physiology-Renal Physiology
- Accession number :
- edsair.doi.dedup.....d8f417c5fe7b4c1d9c3a21770c58c7c3