title: Cortisol alters carbonic anhydrase-mediated renal sulfate secretion.
abstract: Active transepithelial sulfate secretion rate by winter flounder renal proximal tubule epithelium in primary culture (fPTC) is dependent on intracellular carbonic anhydrase (CA) and enhanced by cortisol. To further evaluate this relationship, a partial cDNA clone (327 bp) of carbonic anhydrase II (CAII) with high sequence similarity to CAII from numerous species including fish, chicken, and human was obtained from fPTCs. The majority of CA activity and CAII protein was present in the cytosol of fPTCs; however, significant amounts of both (in addition to SDS-resistant CA activity, i.e., CAIV-like isoform) were present in concentrated plasma membranes. CAII from concentrated membranes migrated differently than purified CAII on nondenaturing PAGE gels, suggesting that CAII associates with another membrane component. Treatment of fPTCs with the cell-soluble CA inhibitor methazolamide (100 microM) caused a 58% reduction in active transepithelial SO4(2-) secretion. fPTCs that were previously cultured under high-cortisol concentrations, when subjected to 5 days of low physiological levels of cortisol, had decreased CA activity (28%), CAII protein abundance (65%), and net active SO4(2-) secretion (28%), with no effect on epithelial differentiation. Methazolamide and low-cortisol treatment in combination inhibited net active SO4(2-) secretion 56%, which was not different than the effect of methazolamide treatment alone. These data indicate that cortisol directly increases renal CA activity, CAII protein abundance, and CA-dependent SO4(2-) secretion in the marine teleost renal proximal tubule.
authors.affiliation: Department of Physiology and Neurobiology, U-4156, Univiversity of Connecticut, 3107 Horsebarn Hill Rd., Storrs, Connecticut 06269-4156, USA.
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Pelis
authors.forename: James E
authors.initials: JE
authors.lastname: Goldmeyer
authors.forename: Joseph
authors.initials: J
authors.lastname: Crivello
authors.forename: J Larry
authors.initials: JL
authors.lastname: Renfro
mesh_terms: D000138:Acidosis / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport / Q000187:drug effects; D024402:Carbonic Anhydrase II / Q000235:genetics / Q000378:metabolism*; D005432:Flounder; D006854:Hydrocortisone / Q000494:pharmacology*; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism*; D008969:Molecular Sequence Data; D009924:Organ Culture Techniques; D012333:RNA, Messenger / Q000032:analysis; D013431:Sulfates / Q000378:metabolism*
chemical_list: D012333:RNA, Messenger; D013431:Sulfates; D024402:Carbonic Anhydrase II; D006854:Hydrocortisone