title: Short-term regulation of murine colonic NBCe1-B (electrogenic Na+/HCO3(-) cotransporter) membrane expression and activity by protein kinase C.
abstract: The colonic mucosa actively secretes HCO3(-), and several lines of evidence point to an important role of Na+/HCO3(-) cotransport (NBC) as a basolateral HCO3(-) import pathway. We could recently demonstrate that the predominant NBC isoform in murine colonic crypts is electrogenic NBCe1-B, and that secretagogues cause NBCe1 exocytosis, which likely represents a component of NBC activation. Since protein kinase C (PKC) plays a key role in the regulation of ion transport by trafficking events, we asked whether it is also involved in the observed NBC activity increase. Crypts were isolated from murine proximal colon to assess PKC activation as well as NBC function and membrane abundance using fluorometric pHi measurements and cell surface biotinylation, respectively. PKC isoform translocation and phosphorylation occurred in response to PMA-, as well as secretagogue stimulation. The conventional and novel PKC inhibitors Gö6976 or Gö6850 did not alter NBC function or surface expression by themselves, but stimulation with forskolin (10(-5) M) or carbachol (10(-4) M) in their presence led to a significant decrease in NBC-mediated proton flux, and biotinylated NBCe1. Our data thus indicate that secretagogues lead to PKC translocation and phosphorylation in murine colonic crypts, and that PKC is necessary for the increase in NBC transport rate and membrane abundance caused by cholinergic and cAMP-dependent stimuli.
authors.affiliation: Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: May
authors.affiliation: Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
authors.forename: Haoyang
authors.initials: H
authors.lastname: Yu
authors.affiliation: Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Riederer
authors.affiliation: Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Manns
authors.affiliation: Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
authors.forename: Ursula
authors.initials: U
authors.lastname: Seidler
authors.affiliation: Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Bachmann
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D018679:Cholinergic Agonists / Q000494:pharmacology; D003106:Colon / Q000166:cytology / Q000378:metabolism*; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D015398:Signal Transduction; D027982:Sodium-Bicarbonate Symporters / Q000235:genetics* / Q000378:metabolism
chemical_list: D018679:Cholinergic Agonists; C498899:Slc4a4 protein, mouse; D027982:Sodium-Bicarbonate Symporters; D011493:Protein Kinase C