Immunolocalization of ion-transport proteins to branchial epithelium mitochondria-rich cells in the mudskipper (Periophthalmodon schlosseri)

Jonathan M. Wilson, David J. Randall, Mark Donowitz, A. Wayne Vogl, Alex K Y Ip

Research output: Contribution to journalArticle

Abstract

The branchial epithelium of the mudskipper Periophthalmodon schlosseri is densely packed with mitochondria-rich (MR) cells. This species of mudskipper is also able to eliminate ammonia against large inward gradients and to tolerate extremely high environmental ammonia concentrations. To test whether these branchial MR cells are the sites of active ammonia elimination, we used an immunological approach to localize ion-transport proteins that have been shown pharmacologically to be involved in the elimination of NH4+ (Na+/NH4+ exchanger and Na+/NH4+-ATPase). We also investigated the role of carbonic anhydrase and boundary-layer pH effects in ammonia elimination by using the carbonic anhydrase inhibitor acetazolamide and by buffering the bath water with Hepes, respectively. In the branchial epithelium, Na+/H+ exchangers (both NHE2- and NHE3-like isoforms), a cystic fibrosis transmembrane regulator (CFTR)-like anion channel, a vacuolar-type H+-ATPase (V-ATPase) and carbonic anhydrase immunoreactivity are associated with the apical crypt region of MR cells. Associated with the MR cell basolateral membrane and tubular system are the Na+/K+-ATPase and a Na+/K+/2Cl- cotransporter. A proportion of the ammonia eliminated by P. schlosseri involves carbonic anhydrase activity and is not dependent on boundary-layer pH effects. The apical CFTR-like anion channel may be serving as a HCO3- channel accounting for the acid-base neutral effects observed with net ammonia efflux inhibition.

Original languageEnglish (US)
Pages (from-to)2297-2310
Number of pages14
JournalJournal of Experimental Biology
Volume203
Issue number15
StatePublished - Aug 2000

Fingerprint

Ion Transport
transport proteins
ion transport
mitochondrion
Ammonia
Carrier Proteins
Mitochondria
mitochondria
ammonia
epithelium
Epithelium
carbonate dehydratase
protein
Carbonic Anhydrases
ion
cystic fibrosis
cells
Cystic Fibrosis
anions
Anions

Keywords

  • Ammonia
  • Carbonic anhydrase
  • CFTR
  • Excretion
  • Gill
  • H-ATPase
  • Mitochondria-rich cell
  • Mudskipper
  • Na/H exchange
  • Na/K-ATPase
  • Na/K/2Cl cotransport
  • Periophthalmodon schlosseri

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Agricultural and Biological Sciences (miscellaneous)

Cite this

Immunolocalization of ion-transport proteins to branchial epithelium mitochondria-rich cells in the mudskipper (Periophthalmodon schlosseri). / Wilson, Jonathan M.; Randall, David J.; Donowitz, Mark; Vogl, A. Wayne; Ip, Alex K Y.

In: Journal of Experimental Biology, Vol. 203, No. 15, 08.2000, p. 2297-2310.

Research output: Contribution to journalArticle

@article{162a8f4a8ec6483296e339cfaf4074c8,
title = "Immunolocalization of ion-transport proteins to branchial epithelium mitochondria-rich cells in the mudskipper (Periophthalmodon schlosseri)",
abstract = "The branchial epithelium of the mudskipper Periophthalmodon schlosseri is densely packed with mitochondria-rich (MR) cells. This species of mudskipper is also able to eliminate ammonia against large inward gradients and to tolerate extremely high environmental ammonia concentrations. To test whether these branchial MR cells are the sites of active ammonia elimination, we used an immunological approach to localize ion-transport proteins that have been shown pharmacologically to be involved in the elimination of NH4+ (Na+/NH4+ exchanger and Na+/NH4+-ATPase). We also investigated the role of carbonic anhydrase and boundary-layer pH effects in ammonia elimination by using the carbonic anhydrase inhibitor acetazolamide and by buffering the bath water with Hepes, respectively. In the branchial epithelium, Na+/H+ exchangers (both NHE2- and NHE3-like isoforms), a cystic fibrosis transmembrane regulator (CFTR)-like anion channel, a vacuolar-type H+-ATPase (V-ATPase) and carbonic anhydrase immunoreactivity are associated with the apical crypt region of MR cells. Associated with the MR cell basolateral membrane and tubular system are the Na+/K+-ATPase and a Na+/K+/2Cl- cotransporter. A proportion of the ammonia eliminated by P. schlosseri involves carbonic anhydrase activity and is not dependent on boundary-layer pH effects. The apical CFTR-like anion channel may be serving as a HCO3- channel accounting for the acid-base neutral effects observed with net ammonia efflux inhibition.",
keywords = "Ammonia, Carbonic anhydrase, CFTR, Excretion, Gill, H-ATPase, Mitochondria-rich cell, Mudskipper, Na/H exchange, Na/K-ATPase, Na/K/2Cl cotransport, Periophthalmodon schlosseri",
author = "Wilson, {Jonathan M.} and Randall, {David J.} and Mark Donowitz and Vogl, {A. Wayne} and Ip, {Alex K Y}",
year = "2000",
month = "8",
language = "English (US)",
volume = "203",
pages = "2297--2310",
journal = "Journal of Experimental Biology",
issn = "0022-0949",
publisher = "Company of Biologists Ltd",
number = "15",

}

TY - JOUR

T1 - Immunolocalization of ion-transport proteins to branchial epithelium mitochondria-rich cells in the mudskipper (Periophthalmodon schlosseri)

AU - Wilson, Jonathan M.

AU - Randall, David J.

AU - Donowitz, Mark

AU - Vogl, A. Wayne

AU - Ip, Alex K Y

PY - 2000/8

Y1 - 2000/8

N2 - The branchial epithelium of the mudskipper Periophthalmodon schlosseri is densely packed with mitochondria-rich (MR) cells. This species of mudskipper is also able to eliminate ammonia against large inward gradients and to tolerate extremely high environmental ammonia concentrations. To test whether these branchial MR cells are the sites of active ammonia elimination, we used an immunological approach to localize ion-transport proteins that have been shown pharmacologically to be involved in the elimination of NH4+ (Na+/NH4+ exchanger and Na+/NH4+-ATPase). We also investigated the role of carbonic anhydrase and boundary-layer pH effects in ammonia elimination by using the carbonic anhydrase inhibitor acetazolamide and by buffering the bath water with Hepes, respectively. In the branchial epithelium, Na+/H+ exchangers (both NHE2- and NHE3-like isoforms), a cystic fibrosis transmembrane regulator (CFTR)-like anion channel, a vacuolar-type H+-ATPase (V-ATPase) and carbonic anhydrase immunoreactivity are associated with the apical crypt region of MR cells. Associated with the MR cell basolateral membrane and tubular system are the Na+/K+-ATPase and a Na+/K+/2Cl- cotransporter. A proportion of the ammonia eliminated by P. schlosseri involves carbonic anhydrase activity and is not dependent on boundary-layer pH effects. The apical CFTR-like anion channel may be serving as a HCO3- channel accounting for the acid-base neutral effects observed with net ammonia efflux inhibition.

AB - The branchial epithelium of the mudskipper Periophthalmodon schlosseri is densely packed with mitochondria-rich (MR) cells. This species of mudskipper is also able to eliminate ammonia against large inward gradients and to tolerate extremely high environmental ammonia concentrations. To test whether these branchial MR cells are the sites of active ammonia elimination, we used an immunological approach to localize ion-transport proteins that have been shown pharmacologically to be involved in the elimination of NH4+ (Na+/NH4+ exchanger and Na+/NH4+-ATPase). We also investigated the role of carbonic anhydrase and boundary-layer pH effects in ammonia elimination by using the carbonic anhydrase inhibitor acetazolamide and by buffering the bath water with Hepes, respectively. In the branchial epithelium, Na+/H+ exchangers (both NHE2- and NHE3-like isoforms), a cystic fibrosis transmembrane regulator (CFTR)-like anion channel, a vacuolar-type H+-ATPase (V-ATPase) and carbonic anhydrase immunoreactivity are associated with the apical crypt region of MR cells. Associated with the MR cell basolateral membrane and tubular system are the Na+/K+-ATPase and a Na+/K+/2Cl- cotransporter. A proportion of the ammonia eliminated by P. schlosseri involves carbonic anhydrase activity and is not dependent on boundary-layer pH effects. The apical CFTR-like anion channel may be serving as a HCO3- channel accounting for the acid-base neutral effects observed with net ammonia efflux inhibition.

KW - Ammonia

KW - Carbonic anhydrase

KW - CFTR

KW - Excretion

KW - Gill

KW - H-ATPase

KW - Mitochondria-rich cell

KW - Mudskipper

KW - Na/H exchange

KW - Na/K-ATPase

KW - Na/K/2Cl cotransport

KW - Periophthalmodon schlosseri

UR - http://www.scopus.com/inward/record.url?scp=0033845015&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033845015&partnerID=8YFLogxK

M3 - Article

C2 - 10887068

AN - SCOPUS:0033845015

VL - 203

SP - 2297

EP - 2310

JO - Journal of Experimental Biology

JF - Journal of Experimental Biology

SN - 0022-0949

IS - 15

ER -