Expression of 25-hydroxyvitamin D3-1alpha-hydroxylase in pancreatic islets

The Journal of Steroid Biochemistry and Molecular Biology
Rosemary BlandMartin Hewison

Abstract

A number of studies have suggested that Vitamin D has a potential role in the development/treatment of diabetes. These effects may be mediated by circulating levels of 1alpha,25(OH)(2)D(3), but local production of 1alpha,25(OH)(2)D(3), catalysed by the enzyme 25-hydroxyvitamin D(3)-1alpha-hydroxylase (1alpha-OHase), is also likely to be important. RT-PCR analyses demonstrated that both isolated rat islets and MIN6 cells (mouse insulin-secreting cell line, characteristic of beta cells) expressed 1alpha-OHase mRNA. The transcript in both cell types was similar to that seen in HKC-8 cells (a renal cell line, which expresses 1alpha-OHase). Western blot analysis and immunolocalisation identified 1alpha-OHase protein in MIN6 cells and human pancreatic tissue. In addition, suspensions of rat islets were able to convert [3H]-25-hydroxyvitamin D(3) to [3H]-1alpha,25(OH)(2)D(3), demonstrating 1alpha-OHase activity. Both cell systems expressed the Vitamin D receptor and 1alpha,25(OH)(2)D(3) (50nM) evoked a rapid rise in [Ca(2+)](i) in MIN6 cells. This data clearly demonstrates islets are able to produce 1alpha,25(OH)(2)D(3) and respond rapidly to treatment with 1alpha,25(OH)(2)D(3). Therefore, we would postulate that local production of 1...Continue Reading

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