Intracellular calcium ion response to glucose in β-cells of calbindin-D28k nullmutant mice and in βHC13 cells overexpressing calbindin-D28k

Abstract

This article describes studies on the glucose-induced responses of intracellular Ca2+ concentration ([Ca2+]i), insulin release, and redistribution of calbindin-D28k, a calcium-binding regulatory protein, in β-cells of pancreatic islets of calbindin-D28k knockout (KO) and wild-type mice (C57BL6) as well as in βHC-13 control cells and βHC-13 CaBP40 cells (β-cell line overexpressing
calbindin-D28k). Upon increasing the glucose concentration from 2.8 to 30 mM, islets of KO mice showed a significantly greater increase in [Ca2+]i (mean increase in [Ca2+]i, i.e., Δ[Ca2+], was 296 nM) compared with wild-type mice (Δ[Ca2+]i=97 nM). βHC-13 CaBP40 cells showed little change in [Ca2+]i upon elevation of glucose from 5.5 to 32.7 mM, whereas βHC-13 control cells exhibited significant increases in [Ca2+]i (Δ[Ca2+]i=510 nM). Similarly, upon addition of 30 mM glucose, the rate of insulin release increased from 25.2 (basal rate) to 145.2 pg/mL/min in βHC-13 control cells, whereas
in βHC-13 CaBP40 cells the rate of insulin release was only 27.5 pg/mL/min in high glucose. Thus, levels of calbindin-D28k in β-cells affect both [Ca2+]i and insulin secretion in response to glucose. The three-dimensional reconstruct of confocal immunofluorescent images showed
that glucose caused redistribution of calbindin-D28k resulting in co-localization in the region of L-type voltage-dependent calcium channels (VDCC). This colocalization may be
an important regulatory function concerning Ca2+ influx via L-type VDCC and exocytosis of insulin granules.