Document Detail

Localization and characterization of phenamil-sensitive Na+ influx in isolated rainbow trout gill epithelial cells.
MedLine Citation:
PMID:  12502776     Owner:  NLM     Status:  MEDLINE    
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 (22)Na(+) 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 (alpha-MR cells) demonstrated phenamil-and bafilomycin-sensitive acid-activated (22)Na(+) uptake. We confirm the coupling of a V-type H(+)-ATPase with phenamil-sensitive Na(+) uptake activity and conclude that high-density alpha-MR cells function in branchial Na(+) uptake in freshwater fish.
Scott D Reid; G S Hawkings; F Galvez; G G Goss
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of experimental biology     Volume:  206     ISSN:  0022-0949     ISO Abbreviation:  J. Exp. Biol.     Publication Date:  2003 Feb 
Date Detail:
Created Date:  2002-12-27     Completed Date:  2003-10-03     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0243705     Medline TA:  J Exp Biol     Country:  England    
Other Details:
Languages:  eng     Pagination:  551-9     Citation Subset:  IM    
Dept of Biology, Okanagan University College, Kelowna, British Columbia, VIV 1V7, Canada.
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MeSH Terms
Amiloride / analogs & derivatives*,  pharmacology*
Anti-Bacterial Agents / pharmacology
Biological Transport / drug effects
Bumetanide / pharmacology
Epithelial Cells / drug effects,  physiology*
Gills / cytology,  drug effects,  physiology*
Guanidines / pharmacology
Hydrogen-Ion Concentration / drug effects
Mitochondria / drug effects,  metabolism
Models, Biological
Oncorhynchus mykiss / physiology*
Ouabain / pharmacology
Peanut Agglutinin / pharmacology
Sodium / antagonists & inhibitors,  metabolism*
Sodium Radioisotopes
Sulfones / pharmacology
Reg. No./Substance:
0/Anti-Bacterial Agents; 0/Guanidines; 0/Macrolides; 0/Peanut Agglutinin; 0/Sodium Radioisotopes; 0/Sulfones; 141923-47-9/3-methylsulfonyl-4-piperidinobenzoyl guanidine; 2038-35-9/phenylamil; 2609-46-3/Amiloride; 28395-03-1/Bumetanide; 630-60-4/Ouabain; 7440-23-5/Sodium; 88899-55-2/bafilomycin A1

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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