Cargando…

Expression profiling and immunolocalization of Na(+)-d-glucose-cotransporter 1 in mice employing knockout mice as specificity control indicate novel locations and differences between mice and rats

The expression and localization of sodium-d-glucose cotransporter SGLT1 (SLC5A1), which is involved in small intestinal glucose absorption and renal glucose reabsorption, is of high biomedical relevance because SGLT1 inhibitors are currently tested for antidiabetic therapy. In human and rat organs,...

Descripción completa

Detalles Bibliográficos
Autores principales: Madunić, Ivana Vrhovac, Breljak, Davorka, Karaica, Dean, Koepsell, Hermann, Sabolić, Ivan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5691098/
https://www.ncbi.nlm.nih.gov/pubmed/28842746
http://dx.doi.org/10.1007/s00424-017-2056-1
_version_ 1783279727015886848
author Madunić, Ivana Vrhovac
Breljak, Davorka
Karaica, Dean
Koepsell, Hermann
Sabolić, Ivan
author_facet Madunić, Ivana Vrhovac
Breljak, Davorka
Karaica, Dean
Koepsell, Hermann
Sabolić, Ivan
author_sort Madunić, Ivana Vrhovac
collection PubMed
description The expression and localization of sodium-d-glucose cotransporter SGLT1 (SLC5A1), which is involved in small intestinal glucose absorption and renal glucose reabsorption, is of high biomedical relevance because SGLT1 inhibitors are currently tested for antidiabetic therapy. In human and rat organs, detailed expression profiling of SGLT1/Sglt1 mRNA and immunolocalization of the transporter protein has been performed. Using polyspecific antibodies and preabsorption with antigenic peptide as specificity control, in several organs, different immunolocalizations of SGLT1/Sglt1 between human and rat were obtained. Because the preabsorption control does not exclude cross-reactivity with similar epitopes, some localizations remained ambiguous. In the present study, we performed an immunocytochemical localization of Sglt1 in various organs of mice. Specificities of the immunoreactions were evaluated using antibody preabsorption with the Sglt1 peptide and the respective organs of Sglt1 knockout mice. Because staining in some locations was abolished after antibody preabsorption but remained in the knockout mice, missing staining in knockout mice was used as specificity criterion. The immunolocalization in mouse was identical or similar to rat in many organs, including small intestine, liver, and kidney. However, the male-dominant renal Sglt1 protein expression in mice differed from the female-dominant expression in rats, and localization in lung, heart, and brain observed in rats was not detected in mice. In mice, several novel locations of Sglt1, e.g., in eyes, tongue epithelial cells, pancreatic ducts, prostate, and periurethral glands were detected. Using end-point and quantitative RT-PCR in various organs, different Sglt1 expression in mice and rats was confirmed. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00424-017-2056-1) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-5691098
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Springer Berlin Heidelberg
record_format MEDLINE/PubMed
spelling pubmed-56910982017-11-30 Expression profiling and immunolocalization of Na(+)-d-glucose-cotransporter 1 in mice employing knockout mice as specificity control indicate novel locations and differences between mice and rats Madunić, Ivana Vrhovac Breljak, Davorka Karaica, Dean Koepsell, Hermann Sabolić, Ivan Pflugers Arch Ion Channels, Receptors and Transporters The expression and localization of sodium-d-glucose cotransporter SGLT1 (SLC5A1), which is involved in small intestinal glucose absorption and renal glucose reabsorption, is of high biomedical relevance because SGLT1 inhibitors are currently tested for antidiabetic therapy. In human and rat organs, detailed expression profiling of SGLT1/Sglt1 mRNA and immunolocalization of the transporter protein has been performed. Using polyspecific antibodies and preabsorption with antigenic peptide as specificity control, in several organs, different immunolocalizations of SGLT1/Sglt1 between human and rat were obtained. Because the preabsorption control does not exclude cross-reactivity with similar epitopes, some localizations remained ambiguous. In the present study, we performed an immunocytochemical localization of Sglt1 in various organs of mice. Specificities of the immunoreactions were evaluated using antibody preabsorption with the Sglt1 peptide and the respective organs of Sglt1 knockout mice. Because staining in some locations was abolished after antibody preabsorption but remained in the knockout mice, missing staining in knockout mice was used as specificity criterion. The immunolocalization in mouse was identical or similar to rat in many organs, including small intestine, liver, and kidney. However, the male-dominant renal Sglt1 protein expression in mice differed from the female-dominant expression in rats, and localization in lung, heart, and brain observed in rats was not detected in mice. In mice, several novel locations of Sglt1, e.g., in eyes, tongue epithelial cells, pancreatic ducts, prostate, and periurethral glands were detected. Using end-point and quantitative RT-PCR in various organs, different Sglt1 expression in mice and rats was confirmed. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00424-017-2056-1) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2017-08-26 2017 /pmc/articles/PMC5691098/ /pubmed/28842746 http://dx.doi.org/10.1007/s00424-017-2056-1 Text en © The Author(s) 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Ion Channels, Receptors and Transporters
Madunić, Ivana Vrhovac
Breljak, Davorka
Karaica, Dean
Koepsell, Hermann
Sabolić, Ivan
Expression profiling and immunolocalization of Na(+)-d-glucose-cotransporter 1 in mice employing knockout mice as specificity control indicate novel locations and differences between mice and rats
title Expression profiling and immunolocalization of Na(+)-d-glucose-cotransporter 1 in mice employing knockout mice as specificity control indicate novel locations and differences between mice and rats
title_full Expression profiling and immunolocalization of Na(+)-d-glucose-cotransporter 1 in mice employing knockout mice as specificity control indicate novel locations and differences between mice and rats
title_fullStr Expression profiling and immunolocalization of Na(+)-d-glucose-cotransporter 1 in mice employing knockout mice as specificity control indicate novel locations and differences between mice and rats
title_full_unstemmed Expression profiling and immunolocalization of Na(+)-d-glucose-cotransporter 1 in mice employing knockout mice as specificity control indicate novel locations and differences between mice and rats
title_short Expression profiling and immunolocalization of Na(+)-d-glucose-cotransporter 1 in mice employing knockout mice as specificity control indicate novel locations and differences between mice and rats
title_sort expression profiling and immunolocalization of na(+)-d-glucose-cotransporter 1 in mice employing knockout mice as specificity control indicate novel locations and differences between mice and rats
topic Ion Channels, Receptors and Transporters
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5691098/
https://www.ncbi.nlm.nih.gov/pubmed/28842746
http://dx.doi.org/10.1007/s00424-017-2056-1
work_keys_str_mv AT madunicivanavrhovac expressionprofilingandimmunolocalizationofnadglucosecotransporter1inmiceemployingknockoutmiceasspecificitycontrolindicatenovellocationsanddifferencesbetweenmiceandrats
AT breljakdavorka expressionprofilingandimmunolocalizationofnadglucosecotransporter1inmiceemployingknockoutmiceasspecificitycontrolindicatenovellocationsanddifferencesbetweenmiceandrats
AT karaicadean expressionprofilingandimmunolocalizationofnadglucosecotransporter1inmiceemployingknockoutmiceasspecificitycontrolindicatenovellocationsanddifferencesbetweenmiceandrats
AT koepsellhermann expressionprofilingandimmunolocalizationofnadglucosecotransporter1inmiceemployingknockoutmiceasspecificitycontrolindicatenovellocationsanddifferencesbetweenmiceandrats
AT sabolicivan expressionprofilingandimmunolocalizationofnadglucosecotransporter1inmiceemployingknockoutmiceasspecificitycontrolindicatenovellocationsanddifferencesbetweenmiceandrats