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Choi, Yun, Suh, Lim, Cho, Kwon, Cha, Son, Park, and Yoon: Proliferation and Differentiation of Pancreatic β Cells in L-type Calcium Channel α1D Subunit (Cav1.3) Heterozygous Knock Out Mice After Partial Pancreatectomy
Original Article
The Journal of Korean Diabetes Association

The Journal of Korean Diabetes Association 2007; 31(3): 208-219.

Published online: 31 May 2007

DOI: https://doi.org/10.4093/jkda.2007.31.3.208

Proliferation and Differentiation of Pancreatic β Cells in L-type Calcium Channel α1D Subunit (Cav1.3) Heterozygous Knock Out Mice After Partial Pancreatectomy

Yoon-Hee Choi1, Il-Hee Yun3, Sun Hee Suh2, Dong-Jun Lim1, Jae-Hyuung Cho1, Hyuk-Sang Kwon1, Bong-Yun Cha1, Ho-Young Son1, Chung-Gyu Park3, Kun-Ho Yoon1, 2

1Department of Internal Medicine, The Catholic University of Korea, Korea.

2Department of Immunology & Cell Biology Core Laboratory, The Catholic University of Korea, Korea.

3Department of Microbiology and Immunology, Tumor Immunity Medical Research Center, Transplantation Research Institute Seoul National University College of Medicine, Korea.

Received 13 February 2006       Accepted 2 May 2007

Copyright © 2007 Korean Diabetes Association

Abstract

Backgrounds

L-type voltage-dependent calcium channel (LTCC) plays a crucial role in insulin secretion from pancreatic β cells through Ca2+ influx. In the recent report, LTCC Cav1.3 subtype homozygous knock out mice showed impairment of postnatal pancreatic beta cell development as well as insulin secretion.

Methods

We performed 90% partial pancreatectomy in heterozygous Cav1.3 knock out mice to investigate the effect of partial deficiency of Cav1.3 gene on beta cell regeneration in the adult. Glucose homeostasis, metabolic profiles including serum insulin and lipid levels and morphologic changes of pancreatic islets were studied.

Results

90% Partial pancreatectomy induced glucose intolerance only in the heterozygous knock out mice at 8 weeks after surgery. Distribution of islet size was significantly different between two groups after partial pancreatectomy; median value of islet size of heterozygote was larger than that of wild type (642.8 µm2 vs 1459.8 µm2,P < 0.01). The frequency of single beta cell unit, considered as a unit of β cell neogenesis, was much lower in heterozygote than that of wild type (41% vs 23.3%, P < 0.05).

Conclusion

These data suggest that Cav1.3 gene deficiency is specifically associated with impairment of beta cell regeneration, especially neogensis and eventual glucose intolerance in the 90% partial pancreatectomized mice.

Keywords: Beta cell neogenesis, Cav1.3, L-type voltage-dependent calcium channel (LTCC), Partial pancreatectomy

Figures and Tables

Fig. 1
α1D genotyping of animals. PCR of tail DNA showed wild type (+/+) and heterozygote (+/-). C, normal C57BL/6J mouse used as control animal; SM, size marker; NC, negative control of PCR.
jkda-31-208-g001
Fig. 2
Comparison of changes in body weight and fed whole blood glucose levels after partial pancreatectomy between wild type and heterozygote. A, Body weights temporarily decreased after operation and then gradually increased throughout experiment in both groups. B, Whole blood glucose levels were measured at fed state in the morning with portable glucometer weekly. Blood glucose levels increased one week after operation in both groups and maintained in HT and gradually decreased after 3 weeks in WT. WT, wild type; HT, heterozygote. *P < 0.05.
jkda-31-208-g002
Fig. 3
Comparison of baseline IPGTT and ITT between wild type and heterozygote. A, Glucose tolerance test before pancreatectomy showed no significant differences between two groups. WT, wild type, n = 14; HT, heterozygote, n = 39. B, Glucose disappearance rate after insulin injection showed no significant differences between groups. WT, n = 12; HT, n = 11. IPGTT, intraperitoneal glucose tolerance test; ITT, insulin tolerance test.
jkda-31-208-g003
Fig. 4
Comparison of IPGTT and ITT performed 8 weeks after partial pancreatectomy between wild type and heterozygote. A, Blood glucose levels during glucose tolerance test after pancreatectomy in WT and HT groups. Blood glucose level of HT at ninety minute during GTT was significantly higher than that of WT. In each group, n = 5. B, Glucose disappearance rates after insulin injection were not different between groups. In each group, n = 5. WT, wild type; HT, heterozygote. IPGTT, intraperitoneal glucose tolerance test; ITT, insulin tolerance test. *P < 0.05.
jkda-31-208-g004
Fig. 5
Median value of islet size in wild type and heterozygote groups. n = 5 in wild type (WT), n = 6 in heterozygote (HT). *P < 0.05.
jkda-31-208-g005
Fig. 6
Islet size distribution in wild type and heterozygote groups. Frequency of SCU (single cell unit), SI (small islet), MI (medium-sized islet), and LI (large-sized islet) in both groups. Proportion of SCU was less in HT than WT. Size of islets in HT group has a tendency to be larger than that of WT. n = 5 in wild type (WT), n = 6 in heterozygote (HT). *P < 0.05.
jkda-31-208-g006
Fig. 7
Immunohistochemical stain of islet with anti-insulin antibody. Relatively larger sized islets were observed in the heterozygote (HT) (b,d) than wild type (WT) (a, c).
jkda-31-208-g007
Table 1
Biochemical and hormonal parameters of wild type (WT) and heterozygote (HT) of α1D Knock out mice before, 3 days and 8 weeks after partial pancreatectomy
jkda-31-208-i001

Data are mean ± standard error.

Table 2
Total pancreatic weight and beta cell mass in remnant pancreatic tissue of wild type (WT) and heterozygote (HT) of α1D knock out mice before and 3 days, 8 weeks after 90% partial pancreatectomy
jkda-31-208-i002

Data are mean ± standard error.

Table 3
Cross-sectional areas of solitary β cells and islets in wild type (WT) and heterozygote (HT) at 8 weeks after 90% partial pancreatectomy
jkda-31-208-i003

Data are mean ± standard error. SCU, single β cell unit; SI, small sized islet; MI, medium sized islet; LI, large sized islet. *P < 0.05 between wild type and heterozygote.

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