Cortisol directly stimulates differentiation of chloride cells in tilapia opercular membrane.
Opercular membranes from freshwater tilapia (Oreochromis mossambicus) were maintained in vitro for 4 days and exposed to several concentrations of cortisol (0, 0.01, 0.1, 1, and 10 micrograms/ml). Chloride cell size, number, and Na(+)-K(+)-ATPase content were examined using a fluorescent mitochondrial dye (dimethylaminostyrylethylpyridiniumiodine), a fluorescent analogue of ouabain (anthroylouabain) that binds specifically to Na(+)-K(+)-ATPase, and a cytological stain specific for plasma and tubular membranes. In the absence of cortisol, chloride cell density of the freshwater tilapia opercular membrane decreased (from initial levels of 6,114 +/- 451 to 18 +/- 9 cells/cm2) and was restored by cortisol in a dose-dependent manner. Chloride cell height (5.5 +/- 0.3 microns initially and 7.8 +/- 0.5 microns after 4 days in vitro) increased twofold (13.1 +/- 0.7 microns) after exposure to 1 microgram/ml cortisol. Initially and after 4 days in control medium, there was no detectable staining with anthroylouabain; exposure to 1 microgram/ml cortisol resulted in the appearance of numerous anthroylouabain-positive chloride cells. Without cortisol, Na(+)-K(+)-ATPase activity of the opercular membrane remained constant through 4 days of culture (0.4-0.6 mumol ADP.mg protein-1.h-1); addition of cortisol caused a dose-dependent increase to a maximum of 1.2 +/- 0.1 mumol Pi.mg protein-1.h-1. In vitro cortisol also maintained the size, density, and appearance of chloride cells from opercular membrane of seawater-adapted tilapia. The results indicate that in vitro cortisol exposure causes morphological and biochemical differentiation of the seawater form of the chloride cell.
Department of Integrative Biology, University of California, Berkeley 94720.
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McCormick
D000818:Animals; D000873:Anthracenes; D002452:Cell Count; D002454:Cell Differentiation; D002712:Chlorides / Q000378:metabolism*; D005260:Female; D005456:Fluorescent Dyes; D005880:Gills / Q000166:cytology* / Q000378:metabolism; D006854:Hydrocortisone / Q000494:pharmacology*; D008297:Male; D009992:Osmium; D010042:Ouabain / Q000031:analogs & derivatives; D010473:Perciformes; D011726:Pyridinium Compounds; D012867:Skin / Q000166:cytology* / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013194:Staining and Labeling; D013268:Stimulation, Chemical
D000873:Anthracenes; D002712:Chlorides; D005456:Fluorescent Dyes; D011726:Pyridinium Compounds; D009992:Osmium; C025042:2-(dimethylaminostyryl)-1-ethylpyridinium; D010042:Ouabain; C013814:anthroylouabain; D000254:Sodium-Potassium-Exchanging ATPase; D006854:Hydrocortisone