Expression of two isoforms of the vacuolar-type ATPase subunit B in the zebrafish Danio rerio
In the present study we tested the hypothesis that two isoforms of the regulatory subunit B of vacuolar-type ATPase (V-ATPase) are expressed in the zebrafish Danio rerio . The complete coding sequences for both isoforms, vatB1 and vatB2, were cloned and sequenced. BLASTX analysis revealed the greate...
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fthighwire:oai:open-archive.highwire.org:jexbio:206/11/1907 2023-05-15T13:28:08+02:00 Expression of two isoforms of the vacuolar-type ATPase subunit B in the zebrafish Danio rerio Boesch, S. T. Eller, B. Pelster, B. 2003-06-01 00:00:00.0 text/html http://jeb.biologists.org/cgi/content/short/206/11/1907 https://doi.org/10.1242/jeb.00378 en eng Company of Biologists http://jeb.biologists.org/cgi/content/short/206/11/1907 http://dx.doi.org/10.1242/jeb.00378 Copyright (C) 2003, Company of Biologists Research Article TEXT 2003 fthighwire https://doi.org/10.1242/jeb.00378 2015-02-28T16:30:28Z In the present study we tested the hypothesis that two isoforms of the regulatory subunit B of vacuolar-type ATPase (V-ATPase) are expressed in the zebrafish Danio rerio . The complete coding sequences for both isoforms, vatB1 and vatB2, were cloned and sequenced. BLASTX analysis revealed the greatest similarity to amino acid sequences of B subunits from the European eel Anguilla anguilla and rainbow trout Oncorhynchus mykiss . The isoforms were expressed in a bacterial system and the recombinant proteins verified using isoform-specific antibodies directed against vatB isoforms of the eel. The distribution of both isoforms in zebrafish tissues was investigated using reverse transcriptase-polymerase chain reaction and western blot analysis. The results revealed that at the RNA level both isoforms were expressed in all tested organs, i.e. the gills, swimbladder, heart, kidney, liver, spleen, intestine and skeletal muscle. At the protein level, however, there were tissue-specific variations in the levels of the two vatB isoforms expressed. The highest amounts of V-ATPase were detected in total protein preparations from gill, heart and liver tissue. In liver tissue, however, the western blot analysis indicated that vatB1 was not as prominent as vatB2, and immunohistochemistry revealed that antibodies directed against vatB1 yielded a very weak staining in a number of cells, while an antibody directed against vatB1 and vatB2 yielded a strong staining in virtually every cell. Similarly, neurosecretory cells of the small intestine were stained with an antibody directed against vatB1 and vatB2, but not with an antibody specific for vatB1. Therefore we conclude that the differential expression of two isoforms of the V-ATPase subunits, which may serve different functions as in several mammalian species, may also be a common phenomenon in teleost fish. Text Anguilla anguilla European eel HighWire Press (Stanford University) Journal of Experimental Biology 206 11 1907 1915 |
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Research Article Boesch, S. T. Eller, B. Pelster, B. Expression of two isoforms of the vacuolar-type ATPase subunit B in the zebrafish Danio rerio |
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Research Article |
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In the present study we tested the hypothesis that two isoforms of the regulatory subunit B of vacuolar-type ATPase (V-ATPase) are expressed in the zebrafish Danio rerio . The complete coding sequences for both isoforms, vatB1 and vatB2, were cloned and sequenced. BLASTX analysis revealed the greatest similarity to amino acid sequences of B subunits from the European eel Anguilla anguilla and rainbow trout Oncorhynchus mykiss . The isoforms were expressed in a bacterial system and the recombinant proteins verified using isoform-specific antibodies directed against vatB isoforms of the eel. The distribution of both isoforms in zebrafish tissues was investigated using reverse transcriptase-polymerase chain reaction and western blot analysis. The results revealed that at the RNA level both isoforms were expressed in all tested organs, i.e. the gills, swimbladder, heart, kidney, liver, spleen, intestine and skeletal muscle. At the protein level, however, there were tissue-specific variations in the levels of the two vatB isoforms expressed. The highest amounts of V-ATPase were detected in total protein preparations from gill, heart and liver tissue. In liver tissue, however, the western blot analysis indicated that vatB1 was not as prominent as vatB2, and immunohistochemistry revealed that antibodies directed against vatB1 yielded a very weak staining in a number of cells, while an antibody directed against vatB1 and vatB2 yielded a strong staining in virtually every cell. Similarly, neurosecretory cells of the small intestine were stained with an antibody directed against vatB1 and vatB2, but not with an antibody specific for vatB1. Therefore we conclude that the differential expression of two isoforms of the V-ATPase subunits, which may serve different functions as in several mammalian species, may also be a common phenomenon in teleost fish. |
format |
Text |
author |
Boesch, S. T. Eller, B. Pelster, B. |
author_facet |
Boesch, S. T. Eller, B. Pelster, B. |
author_sort |
Boesch, S. T. |
title |
Expression of two isoforms of the vacuolar-type ATPase subunit B in the zebrafish Danio rerio |
title_short |
Expression of two isoforms of the vacuolar-type ATPase subunit B in the zebrafish Danio rerio |
title_full |
Expression of two isoforms of the vacuolar-type ATPase subunit B in the zebrafish Danio rerio |
title_fullStr |
Expression of two isoforms of the vacuolar-type ATPase subunit B in the zebrafish Danio rerio |
title_full_unstemmed |
Expression of two isoforms of the vacuolar-type ATPase subunit B in the zebrafish Danio rerio |
title_sort |
expression of two isoforms of the vacuolar-type atpase subunit b in the zebrafish danio rerio |
publisher |
Company of Biologists |
publishDate |
2003 |
url |
http://jeb.biologists.org/cgi/content/short/206/11/1907 https://doi.org/10.1242/jeb.00378 |
genre |
Anguilla anguilla European eel |
genre_facet |
Anguilla anguilla European eel |
op_relation |
http://jeb.biologists.org/cgi/content/short/206/11/1907 http://dx.doi.org/10.1242/jeb.00378 |
op_rights |
Copyright (C) 2003, Company of Biologists |
op_doi |
https://doi.org/10.1242/jeb.00378 |
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Journal of Experimental Biology |
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206 |
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11 |
container_start_page |
1907 |
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1915 |
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1766402385088872448 |