QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

This Article Figures Only Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Reid, S. D. Articles by Goss, G. G. Search for Related Content PubMed PubMed Citation Articles by Reid, S. D. Articles by Goss, G. G. Social Bookmarking                         What's this?

The Journal of Experimental Biology 206, 551-559 (2003)
doi: 10.1242/jeb.00109

Localization and characterization of phenamil-sensitive Na⁺ influx in isolated rainbow trout gill epithelial cells

Scott D. Reid^1,2,*, G. S. Hawkings², F. Galvez² and G. G. Goss²

¹ Dept of Biology, Okanagan University College, Kelowna, British Columbia, VIV 1V7, Canada
² Dept of Biological Sciences, University of Alberta, Edmonton, Alberta, T5G 2E9, Canada

^* Author for correspondence (e-mail: sdreid{at}ouc.bc.ca)

Accepted 28 October 2002

Percoll density-gradient separation, combined with peanut lectin agglutinin (PNA) binding and magnetic bead separation, was used to separate dispersed fish gill cells into sub-populations. Functional characterization of each of the sub-populations was performed to determine which displayed acid-activated phenamil- and bafilomycin-sensitive Na⁺ uptake. Analysis of the mechanism(s) of ²²Na⁺ influx was performed in control and acid-activated (addition of 10 mmoll^-1 proprionic acid) cells using a variety of Na⁺ transport inhibitors (ouabain, phenamil, HOE-694 and bumetanide) and a V-type ATPase inhibitor (bafilomycin). We found that cells migrating to a 1.03-1.05 g ml^-1 Percoll interface [pavement cells (PVCs)] possessed the lowest rates of Na⁺ uptake and that influx was unchanged during either bafilomycin (10 nmoll^-1) treatment or internal acidification with addition of proprionic acid (10 mmoll^-1). Mitochondria-rich (MR) cells that migrated to the 1.05-1.09 g ml^-1 interface of the Percoll gradient demonstrated acidification-activated bafilomycin and phenamil-sensitive Na⁺ influx. Further separation of the MR fraction into PNA⁺ and PNA^- fractions using magnetic separation demonstrated that only the PNA^- cells (-MR cells) demonstrated phenamil-and bafilomycin-sensitive acid-activated ²²Na⁺ uptake. We confirm the coupling of a V-type H⁺-ATPase with phenamil-sensitive Na⁺ uptake activity and conclude that high-density -MR cells function in branchial Na⁺ uptake in freshwater fish.

Key words: mitochondria-rich cells, MR, chloride cells, CC, pavement cells, PVC, peanut lectin agglutinin, PNA, transport, fish, Na⁺ influx, density gradient

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				S. F. Perry, B. Vulesevic, M. Grosell, and M. Bayaa Evidence that SLC26 anion transporters mediate branchial chloride uptake in adult zebrafish (Danio rerio) Am J Physiol Regulatory Integrative Comp Physiol, October 1, 2009; 297(4): R988 - R997. [Abstract] [Full Text] [PDF]

				P.-P. Hwang Ion uptake and acid secretion in zebrafish (Danio rerio) J. Exp. Biol., June 1, 2009; 212(11): 1745 - 1752. [Abstract] [Full Text] [PDF]

				S. K. Parks, M. Tresguerres, and G. G. Goss Cellular mechanisms of Cl- transport in trout gill mitochondrion-rich cells Am J Physiol Regulatory Integrative Comp Physiol, April 1, 2009; 296(4): R1161 - R1169. [Abstract] [Full Text] [PDF]

				D. Mitrovic and S. F. Perry The effects of thermally induced gill remodeling on ionocyte distribution and branchial chloride fluxes in goldfish (Carassius auratus) J. Exp. Biol., March 15, 2009; 212(6): 843 - 852. [Abstract] [Full Text] [PDF]

				J. Hiroi, S. Yasumasu, S. D. McCormick, P.-P. Hwang, and T. Kaneko Evidence for an apical Na-Cl cotransporter involved in ion uptake in a teleost fish J. Exp. Biol., August 15, 2008; 211(16): 2584 - 2599. [Abstract] [Full Text] [PDF]

				G. Ivanis, A. J. Esbaugh, and S. F. Perry Branchial expression and localization of SLC9A2 and SLC9A3 sodium/hydrogen exchangers and their possible role in acid-base regulation in freshwater rainbow trout (Oncorhynchus mykiss) J. Exp. Biol., August 1, 2008; 211(15): 2467 - 2477. [Abstract] [Full Text] [PDF]

				D. H. Evans Teleost fish osmoregulation: what have we learned since August Krogh, Homer Smith, and Ancel Keys Am J Physiol Regulatory Integrative Comp Physiol, August 1, 2008; 295(2): R704 - R713. [Abstract] [Full Text] [PDF]

				J.-L. Horng, L.-Y. Lin, C.-J. Huang, F. Katoh, T. Kaneko, and P.-P. Hwang Knockdown of V-ATPase subunit A (atp6v1a) impairs acid secretion and ion balance in zebrafish (Danio rerio) Am J Physiol Regulatory Integrative Comp Physiol, May 1, 2007; 292(5): R2068 - R2076. [Abstract] [Full Text] [PDF]

				S. K. Parks, M. Tresguerres, and G. G. Goss Interactions between Na+ channels and Na+-HCO3- cotransporters in the freshwater fish gill MR cell: a model for transepithelial Na+ uptake Am J Physiol Cell Physiol, February 1, 2007; 292(2): C935 - C944. [Abstract] [Full Text] [PDF]

				M. Esaki, K. Hoshijima, S. Kobayashi, H. Fukuda, K. Kawakami, and S. Hirose Visualization in zebrafish larvae of Na+ uptake in mitochondria-rich cells whose differentiation is dependent on foxi3a Am J Physiol Regulatory Integrative Comp Physiol, January 1, 2007; 292(1): R470 - R480. [Abstract] [Full Text] [PDF]

				F. Galvez, D. Wong, and C. M. Wood Cadmium and calcium uptake in isolated mitochondria-rich cell populations from the gills of the freshwater rainbow trout Am J Physiol Regulatory Integrative Comp Physiol, July 1, 2006; 291(1): R170 - R176. [Abstract] [Full Text] [PDF]

				J. S. Bystriansky, J. G. Richards, P. M. Schulte, and J. S. Ballantyne Reciprocal expression of gill Na+/K+-ATPase {alpha}-subunit isoforms {alpha}1a and {alpha}1b during seawater acclimation of three salmonid fishes that vary in their salinity tolerance J. Exp. Biol., May 15, 2006; 209(10): 1848 - 1858. [Abstract] [Full Text] [PDF]

				L.-Y. Lin, J.-L. Horng, J. G. Kunkel, and P.-P. Hwang Proton pump-rich cell secretes acid in skin of zebrafish larvae Am J Physiol Cell Physiol, February 1, 2006; 290(2): C371 - C378. [Abstract] [Full Text] [PDF]

				J. Burke and R. D. Handy Sodium-sensitive and -insensitive copper accumulation by isolated intestinal cells of rainbow trout Oncorhynchus mykiss J. Exp. Biol., January 15, 2005; 208(2): 391 - 407. [Abstract] [Full Text] [PDF]

				D. H. Evans, P. M. Piermarini, and K. P. Choe The Multifunctional Fish Gill: Dominant Site of Gas Exchange, Osmoregulation, Acid-Base Regulation, and Excretion of Nitrogenous Waste Physiol Rev, January 1, 2005; 85(1): 97 - 177. [Abstract] [Full Text] [PDF]

				G. R. Scott, J. G. Richards, B. Forbush, P. Isenring, and P. M. Schulte Changes in gene expression in gills of the euryhaline killifish Fundulus heteroclitus after abrupt salinity transfer Am J Physiol Cell Physiol, August 1, 2004; 287(2): C300 - C309. [Abstract] [Full Text] [PDF]

				L. B. Kirschner The mechanism of sodium chloride uptake in hyperregulating aquatic animals J. Exp. Biol., April 1, 2004; 207(9): 1439 - 1452. [Abstract] [Full Text] [PDF]

				G. S. Hawkings, F. Galvez, and G. G. Goss Seawater acclimation causes independent alterations in Na+/K+- and H+-ATPase activity in isolated mitochondria-rich cell subtypes of the rainbow trout gill J. Exp. Biol., February 22, 2004; 207(6): 905 - 912. [Abstract] [Full Text] [PDF]