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

Autor: Hermann Koepsell, Ivan Sabolić, Davorka Breljak, Ivana Vrhovac Madunić, Dean Karaica
Rok vydání: 2017
Předmět:
Male
0301 basic medicine
Pathology
medicine.medical_specialty
Mouse
Physiology
Clinical Biochemistry
Immunocytochemistry
030204 cardiovascular system & hematology
SGLT1
Epitope
Mice
03 medical and health sciences
Sodium-Glucose Transporter 1
sodium-d-glucose cotransporter
mouse
tissue distribution
localization
immunocytochemistry
nonspecific antibody binding
mRNA expression
sex differences
0302 clinical medicine
Nonspecific antibody binding
Physiology (medical)
Sex differences
medicine
Animals
Receptor
Mice
Knockout
Kidney
SLC5A1
biology
Gene Expression Profiling
digestive
oral
and skin physiology
Immunohistochemistry
Molecular biology
Small intestine
Rats
Renal glucose reabsorption
Mice
Inbred C57BL
Tissue distribution
030104 developmental biology
medicine.anatomical_structure
Localization
Knockout mouse
biology.protein
Female
Ion Channels
Receptors and Transporters
Sodium-d-glucose cotransporter
Zdroj: Pflugers Archiv
ISSN: 1432-2013
0031-6768
DOI: 10.1007/s00424-017-2056-1
Popis: 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.
Databáze: OpenAIRE
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