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Using high throughput or COPAS to study collecting duct development
Author(s) -
Miller R. Lance
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a136-e
Subject(s) - intercalated cell , aquaporin 2 , aquaporin , biology , microbiology and biotechnology , transcription factor , epithelial sodium channel , secretion , chemistry , biochemistry , sodium , gene , water channel , mechanical engineering , organic chemistry , engineering , inlet
The renal collecting duct is composed of principal and intercalated cells. Principal cells play an important role in salt and water homoeostasis and are characterized by the cell‐specific expression of epithelial sodium channels (ENaC), aquaporin type‐2, 3 and 4 water channels (AQP2, 3 and 4), and urea transporter (UTA‐1 and 3). Intercalated cells play a critical role in acid‐base balance and are characterized by the cell‐specific expression of the H+ ATPase B1‐subunit, carbonic anhydrase type‐2 (CAII), anion exchanger 1 and pendrin. Studies involving transgenic mice demonstrate that cell‐specific transcription plays an important role in cell‐specific expression of AQP2 and H+ ATPase B1‐subunit (Zharkikh et. al., 2002 and Miller et. al., 2005). Identification of principal and intercalated cell‐specific transcription factors is critical to our understanding of collecting duct development and maturation. This requires the isolation of large quantities of collecting ducts or principal and intercalated cells, which cannot be achieved using standard methodologies. We describe two novel high throughput methodologies for obtaining large quantities of pure collecting ducts or principal and intercalated cells for genomic and proteomic analysis. These two approaches involve transgenic mice expressing collecting duct‐specific fluorescent protein markers and large particle flow cytometry (COPAS™) (Miller et. al., 2006).