Related Experiment Videos
A fluorescent compound for glucose uptake measurements in isolated rat cardiomyocytes
S W Ball1, J R Bailey, J M Stewart
1Department of Biology, Mount Allison University, Sackville, Canada.
Abstract:
A focus of current diabetes research is the development of insulinomimetic compounds for oral treatment of diabetes and its associated cardiac complications. Screening compounds for their potential insulinomimetic effects usually involves the use of radioactive isotopes. The focus of this study was to investigate a nonradioactive fluorescent compound for its use in screening insulinomimetic compounds. The indicator 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG) has been used by some workers to measure glucose uptake in Escherichia coli and Candida albicans. We propose that 2-NBDG will also be a suitable indicator for mammalian cell lines, in particular rat cardiomyocytes. We found that the indicator could give a reliable reproducible standard curve following appropriate dilution and is taken up by isolated cardiomyocytes. The insulinomimetic compounds vanadyl sulfate and sodium molybdate showed rates of glucose uptake similar to that of insulin. Furthermore, the rate of uptake measured for insulin using this technique (0.04 +/- 0.003 nmol x min(-1) x 10(6) cells(-1) is comparable with previous literature using 2-deoxyglucose uptake measurements on isolated myocytes (0.040 nmol x min(-1) x 10(6) cells(-1), demonstrating the validity of this fluorescent compound for glucose uptake studies.
Insights
This study validates a nonradioactive fluorescent compound, 2-NBDG, for measuring glucose uptake in rat heart cells. This method offers a reliable alternative for screening insulin-mimicking diabetes drugs.
Area of Science:
- Biochemistry
- Pharmacology
- Cardiovascular Research
Background:
- Diabetes research focuses on oral insulinomimetic compounds for treatment.
- Current screening methods often rely on radioactive isotopes.
- Developing nonradioactive alternatives is crucial for efficient drug discovery.
Purpose of the Study:
- To evaluate the fluorescent compound 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG) as a nonradioactive indicator for glucose uptake.
- To assess the suitability of 2-NBDG for use in mammalian cell lines, specifically rat cardiomyocytes.
- To validate 2-NBDG as a tool for screening insulinomimetic compounds.
Main Methods:
- Investigated the use of 2-NBDG, a fluorescent glucose analog.
- Established a reproducible standard curve for 2-NBDG following appropriate dilution.
- Measured glucose uptake in isolated rat cardiomyocytes using 2-NBDG.
- Compared glucose uptake rates induced by insulinomimetic compounds (vanadyl sulfate, sodium molybdate) and insulin with established literature values.
Main Results:
- 2-NBDG demonstrated reliable and reproducible standard curve generation.
- Isolated cardiomyocytes effectively took up 2-NBDG.
- Insulinomimetic compounds showed glucose uptake rates comparable to insulin.
- Measured insulin-induced glucose uptake rates using 2-NBDG align with previous studies using 2-deoxyglucose.
Conclusions:
- 2-NBDG is a valid and reliable nonradioactive fluorescent indicator for measuring glucose uptake in rat cardiomyocytes.
- This method provides a suitable alternative to radioactive assays for screening insulinomimetic compounds.
- The findings support the use of 2-NBDG in diabetes research for evaluating potential oral treatments and their effects on cardiac cells.