Expression profiling and immunolocalization of Na

["Maduni\u0107 I", "Breljak D", "Karaica D", "Koepsell H", "Saboli\u0107 I"]
Pflugers Archiv : European journal of physiology 2017
Open on PubMed

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.

10 Figures Extracted
Fig. 1
Fig. 1 PMC
Relative expression of mSglt1 mRNA in various organs/tissues of wild-type mice estimated by end-point RT-PCR and qRT-PCR. a End-point RT-PCR. The ...
Fig. 2
Fig. 2 PMC
Expression of mSglt1 mRNA ( a ) and mSglt1 protein ( b , c ) in the gastrointestinal tract of wild-type mice. a The expression levels of mSglt1 ...
Fig. 3
Fig. 3 PMC
Immunostaining of the nephron segments in various kidney zones of male wild-type mice with mSglt1-Ab. a Cortex. No staining was observed in glomerul...
Fig. 4
Fig. 4 PMC
Double immunostaining of kidney cortex of wild-type mice with mSglt1-Ab (red fluorescence) and Na/K-ATPase-Ab (green fluorescence) confirms the locali...
Fig. 5
Fig. 5 PMC
Immunolocalization of mSglt1 protein in salivary glands from mSglt1 +/+ and mSglt1 −/− mice. Parotid gland. In mSglt1 +/+ mice (main image...
Fig. 6
Fig. 6 PMC
Immunolocation of Sglt1 in the tongue of mSglt1 +/+ and mSglt1 −/− mice; co-staining with Na/K-ATPase-Ab. a In mSglt1 +/+ mice, the plas...
Fig. 7
Fig. 7 PMC
Immunolocation of Sglt1 in pancreas and liver with mSglt1-Ab in mSglt1 +/+ and mSglt1 −/− mice and co-staining with the Na + /K + -ATPase. Pan...
Fig. 8
Fig. 8 PMC
Immunolocalization of mSglt1 protein in prostate and periurethral glands. a Prostate. In mSglt1 +/+ mice (Ab), mSglt1-Ab stained the luminal mem...
Fig. 9
Fig. 9 PMC
Immunolocalization of mSglt1 protein in tissue cryosections of eyes from mSglt1 +/+ and mSglt1 −/− mice. In mSglt1 +/+ mice, mSglt1-Ab str...
Fig. 10
Fig. 10 PMC
Sex-related expression of mSglt1 mRNA and mSglt1 protein in the kidneys of wild-type mice. a In qRT-PCR, relative expression of mSglt1 mRNA in f...