Cultured gill epithelia from freshwater tilapia (Oreochromis niloticus)

Effect of cortisol and homologous serum supplements from stressed and unstressed fish

S. P. Kelly, C. M. Wood

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Procedures for the preparation and culture of branchial epithelia from dispersed gill cells of freshwater tilapia (Oreochromis niloticus) are described. Epithelia were cultured on permeable supports (terephthalate membranes, "filters") and bathed on both the apical and basolateral side with isotonic media containing 6% fetal bovine serum (FBS). When the apical medium was replaced with freshwater (pseudo in vivo asymmetrical culture conditions), transepithelial resistance (TER) increased markedly, transepithelial potential became negative, and paracellular permeability decreased. The physiological effects of cortisol and 10% homologous (tilapia) serum were investigated. Tilapia serum (TS) was prepared from unstressed and stressed fish and therefore allowed comparison between the effects of homologous serum derived from fish in differing physiological states. Under both symmetrical and asymmetrical culture conditions, cortisol significantly elevated TER across cultured tilapia gill epithelia, indicative of a significant increase in epithelial "tightness." Cortisol reduced transepithelial Na+ and Cl- movement and paracellular permeability. The glucocorticoid agonist dexamethasone elicited a similar response, which was inhibited by the glucocorticoid antagonist (receptor blocker) RU486. Cortisol did not stimulate active ion transport across epithelia under either symmetrical or asymmetrical culture conditions. In epithelia supplemented with TS from stressed fish, physiological changes in cultured preparations were consistent with those observed in FBS + cortisol-supplemented epithelia. Differences between the physiological status of epithelia supplemented with TS from unstressed and stressed fish could be abolished with RU486. Using TS as a medium supplement did not stimulate active ion transport under asymmetrical culture conditions, although Na+-K+-ATPase activity increased in TS-supplemented epithelia relative to FBS-supplemented preparations.

Original languageEnglish
Pages (from-to)29-42
Number of pages14
JournalJournal of Membrane Biology
Volume190
Issue number1
DOIs
StatePublished - Nov 1 2002

Fingerprint

Tilapia
Cichlids
Fresh Water
Hydrocortisone
Fishes
Epithelium
Serum
Ion Transport
Permeability
Glucocorticoid Receptors
Dexamethasone
Glucocorticoids

Keywords

  • Corticosteroids
  • Cultured gill epithelia
  • Ion transport
  • Na-K-ATPase
  • Permeability
  • RU486

ASJC Scopus subject areas

  • Biophysics
  • Physiology
  • Cell Biology

Cite this

Cultured gill epithelia from freshwater tilapia (Oreochromis niloticus) : Effect of cortisol and homologous serum supplements from stressed and unstressed fish. / Kelly, S. P.; Wood, C. M.

In: Journal of Membrane Biology, Vol. 190, No. 1, 01.11.2002, p. 29-42.

Research output: Contribution to journalArticle

@article{d66e4a7855b648e4924e06af6a319842,
title = "Cultured gill epithelia from freshwater tilapia (Oreochromis niloticus): Effect of cortisol and homologous serum supplements from stressed and unstressed fish",
abstract = "Procedures for the preparation and culture of branchial epithelia from dispersed gill cells of freshwater tilapia (Oreochromis niloticus) are described. Epithelia were cultured on permeable supports (terephthalate membranes, {"}filters{"}) and bathed on both the apical and basolateral side with isotonic media containing 6{\%} fetal bovine serum (FBS). When the apical medium was replaced with freshwater (pseudo in vivo asymmetrical culture conditions), transepithelial resistance (TER) increased markedly, transepithelial potential became negative, and paracellular permeability decreased. The physiological effects of cortisol and 10{\%} homologous (tilapia) serum were investigated. Tilapia serum (TS) was prepared from unstressed and stressed fish and therefore allowed comparison between the effects of homologous serum derived from fish in differing physiological states. Under both symmetrical and asymmetrical culture conditions, cortisol significantly elevated TER across cultured tilapia gill epithelia, indicative of a significant increase in epithelial {"}tightness.{"} Cortisol reduced transepithelial Na+ and Cl- movement and paracellular permeability. The glucocorticoid agonist dexamethasone elicited a similar response, which was inhibited by the glucocorticoid antagonist (receptor blocker) RU486. Cortisol did not stimulate active ion transport across epithelia under either symmetrical or asymmetrical culture conditions. In epithelia supplemented with TS from stressed fish, physiological changes in cultured preparations were consistent with those observed in FBS + cortisol-supplemented epithelia. Differences between the physiological status of epithelia supplemented with TS from unstressed and stressed fish could be abolished with RU486. Using TS as a medium supplement did not stimulate active ion transport under asymmetrical culture conditions, although Na+-K+-ATPase activity increased in TS-supplemented epithelia relative to FBS-supplemented preparations.",
keywords = "Corticosteroids, Cultured gill epithelia, Ion transport, Na-K-ATPase, Permeability, RU486",
author = "Kelly, {S. P.} and Wood, {C. M.}",
year = "2002",
month = "11",
day = "1",
doi = "10.1007/s00232-002-1020-x",
language = "English",
volume = "190",
pages = "29--42",
journal = "Journal of Membrane Biology",
issn = "0022-2631",
publisher = "Springer New York",
number = "1",

}

TY - JOUR

T1 - Cultured gill epithelia from freshwater tilapia (Oreochromis niloticus)

T2 - Effect of cortisol and homologous serum supplements from stressed and unstressed fish

AU - Kelly, S. P.

AU - Wood, C. M.

PY - 2002/11/1

Y1 - 2002/11/1

N2 - Procedures for the preparation and culture of branchial epithelia from dispersed gill cells of freshwater tilapia (Oreochromis niloticus) are described. Epithelia were cultured on permeable supports (terephthalate membranes, "filters") and bathed on both the apical and basolateral side with isotonic media containing 6% fetal bovine serum (FBS). When the apical medium was replaced with freshwater (pseudo in vivo asymmetrical culture conditions), transepithelial resistance (TER) increased markedly, transepithelial potential became negative, and paracellular permeability decreased. The physiological effects of cortisol and 10% homologous (tilapia) serum were investigated. Tilapia serum (TS) was prepared from unstressed and stressed fish and therefore allowed comparison between the effects of homologous serum derived from fish in differing physiological states. Under both symmetrical and asymmetrical culture conditions, cortisol significantly elevated TER across cultured tilapia gill epithelia, indicative of a significant increase in epithelial "tightness." Cortisol reduced transepithelial Na+ and Cl- movement and paracellular permeability. The glucocorticoid agonist dexamethasone elicited a similar response, which was inhibited by the glucocorticoid antagonist (receptor blocker) RU486. Cortisol did not stimulate active ion transport across epithelia under either symmetrical or asymmetrical culture conditions. In epithelia supplemented with TS from stressed fish, physiological changes in cultured preparations were consistent with those observed in FBS + cortisol-supplemented epithelia. Differences between the physiological status of epithelia supplemented with TS from unstressed and stressed fish could be abolished with RU486. Using TS as a medium supplement did not stimulate active ion transport under asymmetrical culture conditions, although Na+-K+-ATPase activity increased in TS-supplemented epithelia relative to FBS-supplemented preparations.

AB - Procedures for the preparation and culture of branchial epithelia from dispersed gill cells of freshwater tilapia (Oreochromis niloticus) are described. Epithelia were cultured on permeable supports (terephthalate membranes, "filters") and bathed on both the apical and basolateral side with isotonic media containing 6% fetal bovine serum (FBS). When the apical medium was replaced with freshwater (pseudo in vivo asymmetrical culture conditions), transepithelial resistance (TER) increased markedly, transepithelial potential became negative, and paracellular permeability decreased. The physiological effects of cortisol and 10% homologous (tilapia) serum were investigated. Tilapia serum (TS) was prepared from unstressed and stressed fish and therefore allowed comparison between the effects of homologous serum derived from fish in differing physiological states. Under both symmetrical and asymmetrical culture conditions, cortisol significantly elevated TER across cultured tilapia gill epithelia, indicative of a significant increase in epithelial "tightness." Cortisol reduced transepithelial Na+ and Cl- movement and paracellular permeability. The glucocorticoid agonist dexamethasone elicited a similar response, which was inhibited by the glucocorticoid antagonist (receptor blocker) RU486. Cortisol did not stimulate active ion transport across epithelia under either symmetrical or asymmetrical culture conditions. In epithelia supplemented with TS from stressed fish, physiological changes in cultured preparations were consistent with those observed in FBS + cortisol-supplemented epithelia. Differences between the physiological status of epithelia supplemented with TS from unstressed and stressed fish could be abolished with RU486. Using TS as a medium supplement did not stimulate active ion transport under asymmetrical culture conditions, although Na+-K+-ATPase activity increased in TS-supplemented epithelia relative to FBS-supplemented preparations.

KW - Corticosteroids

KW - Cultured gill epithelia

KW - Ion transport

KW - Na-K-ATPase

KW - Permeability

KW - RU486

UR - http://www.scopus.com/inward/record.url?scp=0041530337&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0041530337&partnerID=8YFLogxK

U2 - 10.1007/s00232-002-1020-x

DO - 10.1007/s00232-002-1020-x

M3 - Article

VL - 190

SP - 29

EP - 42

JO - Journal of Membrane Biology

JF - Journal of Membrane Biology

SN - 0022-2631

IS - 1

ER -