Glucose-dependent Functional Heterogeneity In β-TC-6 Murine Insulinoma
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 33122
Glucose-dependent Functional Heterogeneity In β-TC-6 Murine Insulinoma

Authors: Darren C-W. Tan, Partha Roy

Abstract:

To determine if the murine insulinoma, β-TC-6, is a suitable substitute for primary pancreatic β-cells in the study of β- cell functional heterogeneity, we used three distinct functional assays to ascertain the cell line-s response to glucose or a glucose analog. These assays include: (i) a 2-NBDG uptake assay; (ii) a calcium influx assay, and; (iii) a quinacrine secretion assay. We show that a population of β-TC-6 cells endocytoses the glucose analog, 2- NBDG, at different rates, has non-uniform intracellular calcium ion concentrations and releases quinacrine at different rates when challenged with glucose. We also measured the Km for β-TC-6 glucose uptake to be 46.9 mM and the Vm to be 8.36 x 10-5 mmole/million cells/min. These data suggest that β-TC-6 might be used as an alternative to primary pancreatic β-cells for the study of glucose-dependent β-cell functional heterogeneity.

Keywords: 2-NBDG, Fura-2/AM, functional heterogeneity, quinacrine.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1076632

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 3727

References:


[1] D. E. Ingber, The Pancreas: Biology, Pathobiology, and Disease, 1993, pp. 369-380.
[2] P. E. MacDonald, J. W. Joseph and P. Rorsman, "Glucose-sensing mechanisms in pancreatic beta-cells," Philos Trans R Soc Lond B Biol Sci, vol. 360, 2005, pp. 2211-2225.
[3] F. Matschinsky, Y. Liang, P. Kesavan, L. Wang, P. Froguel, G. Velho, D. Cohen, M. A. Permutt, Y. Tanizawa, T. L. Jetton and et al., "Glucokinase as pancreatic beta cell glucose sensor and diabetes gene," J Clin Invest, vol. 92, 1993, pp. 2092-2098.
[4] T. Aizawa, T. Kaneko, K. Yamauchi, H. Yajima, T. Nishizawa, T. Yada, H. Matsukawa, M. Nagai, S. Yamada, Y. Sato, M. Komatsu, N. Itoh, H. Hidaka, Y. Kajimoto and K. Hashizume, "Size-related and size-unrelated functional heterogeneity among pancreatic islets," Life Sci, vol. 69, 2001, pp. 2627-2639.
[5] H. Heimberg, A. De Vos, A. Vandercammen, E. Van Schaftingen, D. Pipeleers and F. Schuit, "Heterogeneity in glucose sensitivity among pancreatic beta-cells is correlated to differences in glucose phosphorylation rather than glucose transport," EMBO J, vol. 12, 1993, pp. 2873-2879.
[6] D. G. Pipeleers, "Heterogeneity in pancreatic beta-cell population," Diabetes, vol. 41, 1992, pp. 777-781.
[7] W. F. Pralong, C. Bartley and C. B. Wollheim, "Single islet beta-cell stimulation by nutrients: relationship between pyridine nucleotides, cytosolic Ca2+ and secretion," EMBO J, vol. 9, 1990, pp. 53-60.
[8] D. Salomon and P. Meda, "Heterogeneity and contact-dependent regulation of hormone secretion by individual B cells," Exp Cell Res, vol. 162, 1986, pp. 507-520.
[9] P. Smolen, J. Rinzel and A. Sherman, "Why pancreatic islet burst but single beta cells do not: The heterogeneity hypothesis," Biophys J, vol. 64, 1993, pp. 1668-1680.
[10] S. S. Andrali, M. L. Sampley, N. L. Vanderford and S. Ozcan, "Glucose regulation of insulin gene expression in pancreatic beta-cells," Biochem J, vol. 415, 2008, pp. 1-10.
[11] N. Inagaki, K. Yasuda, G. Inoue, Y. Okamoto, H. Yano, Y. Someya, Y. Ohmoto, K. Deguchi, K. Imagawa, H. Imura and et al., "Glucose as regulator of glucose transport activity and glucose-transporter mRNA in hamster beta-cell line," Diabetes, vol. 41, 1992, pp. 592-597.
[12] F. C. Jonkers and J. C. Henquin, "Measurements of cytoplasmic Ca2+ in islet cell clusters show that glucose rapidly recruits beta-cells and gradually increases the individual cell response," Diabetes, vol. 50, 2001, pp. 540-550.
[13] A. Jorns, M. Tiedge and S. Lenzen, "Nutrient-dependent distribution of insulin and glucokinase immunoreactivities in rat pancreatic beta cells," Virchows Arch, vol. 434, 1999, pp. 75-82.
[14] N. Pakhtusova, L. Zaostrovskaya, P. Lindstrom and G. Larsson-Nyren, "Cell-specific Ca(2+) responses in glucose-stimulated single and aggregated beta-cells," Cell Calcium, vol. 34, 2003, pp. 121-129.
[15] K. Yasuda, Y. Yamada, N. Inagaki, H. Yano, Y. Okamoto, K. Tsuji, H. Fukumoto, H. Imura, S. Seino, and Y. Seino, "Expression of GLUT1 and GLUT2 glucose transporter isoforms in rat islets of Langerhans and their regulation by glucose," Diabetes, vol. 41, 1992, pp. 76-81.
[16] V. Poitout, L. E. Stout, M. B. Armstrong, T. F. Walseth, R. L. Sorenson and R. P. Robertson, "Morphological and functional characterization of beta TC-6 cells--an insulin-secreting cell line derived from transgenic mice," Diabetes, vol. 44, 1995, pp. 306-313.
[17] F. Leira, M. C. Louzao, J. M. Vieites, L. M. Botana and M. R. Vieytes, "Fluorescent microplate cell assay to measure uptake and metabolism of glucose in normal human lung fibroblasts," Toxicol In Vitro, vol. 16, 2002, pp. 267-273.
[18] K. Yamada, M. Nakata, N. Horimoto, M. Saito, H. Matsuoka and N. Inagaki, "Measurement of glucose uptake and intracellular calcium concentration in single, living pancreatic beta-cells," J Biol Chem, vol. 275, 2000, pp. 22278-22283.
[19] K. Yoshioka, K. B. Oh, M. Saito, Y. Nemoto and H. Matsuoka, "Evaluation of 2-
[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2- deoxy-D-glucose, a new fluorescent derivative of glucose, for viability assessment of yeast Candida albicans," Appl Microbiol Biotechnol, vol. 46, 1996, pp. 400-404.
[20] C. Zou, Y. Wang and Z. Shen, "2-NBDG as a fluorescent indicator for direct glucose uptake measurement," J Biochem Biophys Methods, vol. 64, 2005, pp. 207-215.
[21] G. G. Holz IV, C. A. Leech, and J. F. Habener, "Actication of a cAMPregulated Ca2+-signaling pathway in pancreatic beta-cells by the insulinotropic hormone glucagon-like peptide-1," J Bio Chem, vol. 270, 1995, pp. 17749-17757.