J Korean Diabetes Assoc.  2007 May;31(3):208-219. 10.4093/jkda.2007.31.3.208.

Proliferation and Differentiation of Pancreatic beta Cells in L-type Calcium Channel alpha(1D) Subunit (Ca(v)1.3) Heterozygous Knock Out Mice After Partial Pancreatectomy

• Choi YH
• Yun IH
• Suh SH
• Lim DJ
• Cho JH
• Kwon HS
• Cha BY
• Son HY
• Park CG
• Yoon KH

• KMID: 1523009
• DOI: http://doi.org/10.4093/jkda.2007.31.3.208

Abstract

BACKGROUNDS: L-type voltage-dependent calcium channel (LTCC) plays a crucial role in insulin secretion from pancreatic beta cells through Ca2+ influx. In the recent report, LTCC Ca(v)1.3 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 Ca(v)1.3 knock out mice to investigate the effect of partial deficiency of Ca(v)1.3 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 micrometer2 vs 1459.8 micrometer2, P < 0.01). The frequency of single beta cell unit, considered as a unit of beta cell neogenesis, was much lower in heterozygote than that of wild type (41% vs 23.3%, P < 0.05).
CONCLUSION
These data suggest that Ca(v)1.3 gene deficiency is specifically associated with impairment of beta cell regeneration, especially neogensis and eventual glucose intolerance in the 90% partial pancreatectomized mice.

Keyword

Beta cell neogenesis; Ca(v)1.3; L-type voltage-dependent calcium channel (LTCC); Partial pancreatectomy

MeSH Terms

Adult
Animals
Calcium Channels
Calcium Channels, L-Type*
Glucose
Glucose Intolerance
Heterozygote
Homeostasis
Humans
Insulin
Insulin-Secreting Cells*
Islets of Langerhans
Metabolome
Mice*
Pancreatectomy*
Regeneration
Calcium Channels
Calcium Channels, L-Type
Glucose
Insulin

Figure

• 

  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.

• 

  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.

• 

  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.

• 

  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.

• 

  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.

• 

  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.

• 

  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).

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