SLC2A2 gene expression in kidney of diabetic rats is regulated by HNF-1α and HNF-3β

Freitas, Helayne Soares; Schaan, Beatriz D.; David-Silva, Aline; Sabino-Silva, Robinson; Okamoto, Maristela Mitiko; Alves-Wagner, Ana Bárbara; Mori, Rosana Cristina Tieko; Machado, Ubiratan Fabres

doi:10.1016/j.mce.2009.02.014

Cited by 36 publications

(31 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…[4][5][6][7][8][22][23][24] So far, in vitro functional analyses on cultured PT cells 8 or on brush border membrane vesicles 25 from diabetic kidneys are performed with radioactive substrates. Also, the in vitro effectiveness of new antidiabetes drugs such as dapagliflozin and sergliflozin-A, which target SGLT2 activity, were investigated for their abilities to inhibit [ 14 C]AMG transport.…”

Section: Discussionmentioning

confidence: 99%

A Fluorescence Method for Measurement of Glucose Transport in Kidney Cells

Blodgett

Kothinti

Kamyshko

et al. 2011

Diabetes Technology & Therapeutics

View full text Add to dashboard Cite

Background: Diabetes may alter renal glucose reabsorption by sodium (Na + )-dependent glucose transporters (SGLTs). Radiolabeled substrates are commonly used for in vitro measurements of SGLT activity in kidney cells. We optimized a method to measure glucose uptake using a fluorescent substrate, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG). Methods: Uptake buffers for 2-NBDG were the same as for 14 C-labeled a-methyl-d-glucopyranoside ([ 14 C]AMG). Cell lysis buffer was optimized for fluorescence of 2-NBDG and Hoechst DNA stain. Uptake was performed on cultures of primary mouse kidney cells (PMKCs), the LLC-PK 1 proximal tubule cell line, or COS-7 cells transiently overexpressing mouse SGLT1 or SGLT2 by incubating cells at 378C in buffer containing 50-200 mM 2-NBDG. Microscopy was performed to visualize uptake in intact cells, while a fluorescence microplate reader was used to measure intracellular concentration of 2-NBDG ([2-NBDG] i ) in cell homogenates. Results: Fluorescent cells were observed in cultures of PMKCs and LLC-PK 1 cells exposed to 2-NBDG in the presence or absence of Na

show abstract

Section: Discussionmentioning

confidence: 99%

A Fluorescence Method for Measurement of Glucose Transport in Kidney Cells

Blodgett

Kothinti

Kamyshko

et al. 2011

Diabetes Technology & Therapeutics

View full text Add to dashboard Cite

show abstract

“…Upregulation of GLUT2 expression has been reported in renal proximal tubules in diabetic rats (29,34,70) and has been linked to transcriptional activity of both HNF-1␣ and HNF-3␤ (42). In contrast, GLUT1 appears to be downregulated in cortical tubules in diabetes (29,34).…”

Section: Hyperglycemia Enhances the Reabsorption Of Glucose In The Prmentioning

confidence: 98%

The proximal tubule in the pathophysiology of the diabetic kidney

Vallon

2011

American Journal of Physiology-Regulatory, Integrative and Comp

308

306

View full text Add to dashboard Cite

Diabetic nephropathy is a leading cause of end-stage renal disease. A better understanding of the molecular mechanism involved in the early changes of the diabetic kidney may permit the development of new strategies to prevent diabetic nephropathy. This review focuses on the proximal tubule in the early diabetic kidney, particularly on its exposure and response to high glucose levels, albuminuria, and other factors in the diabetic glomerular filtrate, the hyperreabsorption of glucose, the unique molecular signature of the tubular growth phenotype, including aspects of senescence, and the resulting cellular and functional consequences. The latter includes the local release of proinflammatory chemokines and changes in proximal tubular salt and fluid reabsorption, which form the basis for the strong tubular control of glomerular filtration in the early diabetic kidney, including glomerular hyperfiltration and odd responses like the salt paradox. Importantly, these early proximal tubular changes can set the stage for oxidative stress, inflammation, hypoxia, and tubulointerstitial fibrosis, and thereby for the progression of diabetic renal disease.

show abstract

“…Four oligonucleotides were designed to contain the wild type or the c.-223C>T (-223WT and -223AV respectively) or c.-185T>C (-185WT and -185AV) variants (Supplementary Table 3). The constructs were labelled with P 32 as described previously (21). The sense probe and complementary labelled probes were annealed to produce double-stranded DNA.…”

Section: Electrophoretic Mobility Shift Assays (Emsas)mentioning

confidence: 99%

A homozygous point mutation in the GH1 promoter (c.-223C>T) leads to reduced GH1 expression in siblings with isolated GH deficiency (IGHD)

Madeira¹,

Jorge

Martin³

et al. 2016

European Journal of Endocrinology

View full text Add to dashboard Cite

Context: Mutations in the GH1 promoter are a rare cause of isolated growth hormone deficiency (IGHD). Objective: To identify the molecular aetiology of a family with IGHD. Design: DNA sequencing, electromobility shift (EMSA) and luciferase reporter assays. Setting: University Hospital. Patients: Three siblings (2M) born to consanguineous parents presented with IGHD with normal pituitary on MRI. Methods: The GH1 proximal promoter, locus control region, five exons and four introns as well as GHRHR gene were sequenced in genomic DNA by Sanger method. DNA-protein interaction was evaluated by EMSA in nuclear extracts of GH3 pituitary cells. Dual-luciferase reporter assays were performed in cells transiently transfected with plasmids containing four different combinations of GH1 allelic variants (AV). Results: The patients harboured two homozygous variants (c.-185T>C and c.-223C>T) in the GH1 promoter within a highly conserved region and predicted binding sites for POU1F1/SP1 and SP1 respectively. The parents and brother were carriers and these variants were absent in 100 controls. EMSA demonstrated absent binding of GH3 nuclear extract to the c.-223C>T variant and normal binding of both POU1F1 protein and GH3 nuclear extract to the c.-185T>C variant. In contrast to GH1 promoter with AV only at c.-185, the GH1 promoter containing the AV only at c.-223 and at both positions drove significantly less expression of luciferase compared with the promoter containing either positions wild type in luciferase reporter assays. Conclusion: To our knowledge, c.-223C>T is the first homozygous point mutation in the GH1 promoter that leads to short stature due to IGHD.

show abstract

SLC2A2 gene expression in kidney of diabetic rats is regulated by HNF-1α and HNF-3β

Cited by 36 publications

References 27 publications

A Fluorescence Method for Measurement of Glucose Transport in Kidney Cells

A Fluorescence Method for Measurement of Glucose Transport in Kidney Cells

The proximal tubule in the pathophysiology of the diabetic kidney

A homozygous point mutation in the GH1 promoter (c.-223C>T) leads to reduced GH1 expression in siblings with isolated GH deficiency (IGHD)

Contact Info

Product

Resources

About