TY - JOUR
T1 - Expression of the β-subunit isoforms of the Na, K-ATpase in rat embryo tissues, inner ear and choroid plexus
AU - González-Martínez, Luis M.
AU - Avila, Julio
AU - Martí, Elisa
AU - Lecuona, Emilia
AU - Martín-Vasallo, Pablo
N1 - Funding Information:
We are grateful to Dr Teresa Alonso for sequence checking of plasmid constructs and to Dr Carlos Hdez Calzadilla for his help and support at all times. This work was supported by grant 93/0831 from Fondo de Investigaciones Sanitarias de la Seguddad Social (FIS) (Spain), and grant 92/08.03.90 from Consejerfa de Educaci6n del Gobierno Aut6nomo de Canarias (Canary Islands, Spain) to PMV. LMGM is recipient of a fellowship from Programa de For-maci6n de Personal Investigador del FIS. LMGM and JA collaborated equally in this work. Both of them are first author.
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 1994
Y1 - 1994
N2 - We report evidence of the apical localization of the two Na,K-ATPase β-subunit isoforms in cells of the inner ear and of the choroid plexus of the rat. To this end, we generated isoform-specific antisera against the human Na,K-ATPase β1 and β2 subunits. These polyclonal rabbit antisera were raised against truncated β-isoform proteins that were made in E coli with pET expression vectors. Deglycosylation of the native antigen with N-endoglycosidase F shows four bands in the β1 isoform and five bands in the β2 iso-form immunoblots. In E15 rat embryos, the β1 isoform was detected in brain, heart and kidney and the β2 isoform only in brain. While β-subunit mRNA expression (Watts AG, Sanchéz-Watts G, Emanuel JR, Levenson R 1991 Proc Natl Acad Sci USA 88, 7425-7429), and immunoblotting and enzymatic activity have been determined (Zlokovic BV, Mackic JB, Wang L, McComb JG, McDonough A 1993 J Biol Chem 268, 8019-8025), very little is known about the specific localization of each β-isoform in the epithelia of choroid plexus and inner ear. Immunocytochemical preparations of 15-day-old whole rat embryos and adult rat brain showed an enhanced staining for the β1 and β2 isoforms in the apical membrane of the ampullary crests of the inner ear's semicircular ducts and in the cuboidal cells of the choroid plexus.
AB - We report evidence of the apical localization of the two Na,K-ATPase β-subunit isoforms in cells of the inner ear and of the choroid plexus of the rat. To this end, we generated isoform-specific antisera against the human Na,K-ATPase β1 and β2 subunits. These polyclonal rabbit antisera were raised against truncated β-isoform proteins that were made in E coli with pET expression vectors. Deglycosylation of the native antigen with N-endoglycosidase F shows four bands in the β1 isoform and five bands in the β2 iso-form immunoblots. In E15 rat embryos, the β1 isoform was detected in brain, heart and kidney and the β2 isoform only in brain. While β-subunit mRNA expression (Watts AG, Sanchéz-Watts G, Emanuel JR, Levenson R 1991 Proc Natl Acad Sci USA 88, 7425-7429), and immunoblotting and enzymatic activity have been determined (Zlokovic BV, Mackic JB, Wang L, McComb JG, McDonough A 1993 J Biol Chem 268, 8019-8025), very little is known about the specific localization of each β-isoform in the epithelia of choroid plexus and inner ear. Immunocytochemical preparations of 15-day-old whole rat embryos and adult rat brain showed an enhanced staining for the β1 and β2 isoforms in the apical membrane of the ampullary crests of the inner ear's semicircular ducts and in the cuboidal cells of the choroid plexus.
KW - K-ATPase
KW - Na
KW - polyclonal rabbit antisera
KW - β-subunits
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U2 - 10.1016/0248-4900(94)90003-5
DO - 10.1016/0248-4900(94)90003-5
M3 - Article
C2 - 7696974
AN - SCOPUS:0028670680
VL - 81
SP - 215
EP - 222
JO - Biologie Cellulaire
JF - Biologie Cellulaire
SN - 0248-4900
IS - 3
ER -