Effect of cortisol on permeability and tight junction protein transcript abundance in primary cultured gill epithelia from stenohaline goldfish and euryhaline trout

Abstract

Primary cultured gill epithelia from goldfish and rainbow trout were used to investigate a role for cortisol in the regulation of paracellular permeability and tight junction (TJ) protein transcript abundance in representative stenohaline versus euryhaline freshwater (FW) fish gills. Glucocorticoid and mineralocorticoid receptors are expressed in cultured goldfish gill preparations and cortisol treatment (100, 500 and 1000 ng/mL) dose-dependently elevated transepithelial resistance (TER) and reduced paracellular [(3)H]PEG-4000 flux across cultured goldfish gill epithelia. Despite these dose-dependent 'tightening' effects of cortisol, the response of goldfish TJ protein transcripts (i.e. occludin, claudin b, c, d, e, h, 7, 8d and 12, and ZO-1) were surprisingly small, with only claudin c and h, and ZO-1 transcript levels significantly decreasing at a dose of 1000 ng/mL. Extending the duration of cortisol exposure from 24 to 48 or 96 h (at 500 ng/mL) did little to alter this phenomenon. By comparison, exposing primary cultured trout gill epithelia (i.e. a euryhaline fish gill model) to 500 ng/mL cortisol resulted in a qualitatively similar, but quantitatively stronger epithelial 'tightening' response. Furthermore, transcript abundance of orthologous trout TJ proteins (i.e. occludin, and claudin 30, 28b, 3a, 7, 8d and 12) significantly elevated as would be expected in a 'tighter' epithelium. Taken together, data suggest a conservative role for cortisol in the endocrine regulation of paracellular permeability across the goldfish gill that may relate to stenohalinity.