The mineralocorticoid receptor contributes to barrier function of a model fish gill epithelium

Author:

Kolosov Dennis1ORCID,Kelly Scott P.1ORCID

Affiliation:

1. Department of Biology, York University, Toronto, Ontario Canada

Abstract

Cortisol-induced epithelial tightening of a primary cultured rainbow trout gill epithelium model occurs in association with reduced paracellular permeability and increased abundance of select barrier-forming TJ proteins. Corticosteroid receptor (CR) pharmacological blocker studies have suggested that to produce this tightening effect, cortisol acts on the mineralocorticoid receptor (MR) as well as glucocorticoid receptors (GRs). This study considered how cortisol influenced model gill epithelium permeability and TJ properties by transcriptional knockdown of the gene encoding MR (mr-KD) using double-stranded RNA. Following mr-KD a significant reduction in MR protein abundance was observed in the epithelium. The mr-KD epithelium demonstrated reduced transepithelial resistance (TER) and increased the paracellular flux of [3H] polyethylene glycol (MW 400 kDa, PEG-400). Concurrently, mRNA abundance of gr2 and 11βhsd increased, indicating a possible compensatory response to mr-KD. Transcript abundance of claudin (cldn) -6, -8d, 23a and -28b decreased while cldn-20a increased in mr-KD preparations. Cortisol-induced epithelial tightening was enhanced in mr-KD preparations, suggesting that alterations in CRs and TJ composition augmented model epithelium barrier function in response to lowered MR abundance. Cortisol treatment significantly increased the transcript and protein abundance of TJ proteins such as cldn/Cldn -8d, and -28b. However, in mr-KD preparations, Cldn-28b protein abundance did not significantly alter in response to cortisol treatment, while Cldn-8d significantly elevated. Data suggest that mr-KD compromises normal barrier function of a primary cultured rainbow trout gill epithelium both in the presence and absence of cortisol and that Cldn-28b protein abundance may be modulated by cortisol via the MR only.

Funder

Natural Sciences and Engineering Research Council of Canada

Government of Canada

York University

Publisher

The Company of Biologists

Subject

Insect Science,Molecular Biology,Animal Science and Zoology,Aquatic Science,Physiology,Ecology, Evolution, Behavior and Systematics

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