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Multiple transcripts of Ca²⁺ channel ₁-subunits and a novel spliced variant of the _1C-subunit in rat ductus arteriosus

Departments of ¹Pediatrics, ²Physiology, and ³Neuroanatomy, Yokohama City University, Yokohama; ⁴Consolidated Research Institute for Advanced Science and Medical Care, Waseda University; ⁵Department of Pharmacology, School of Medicine, Faculty of Medicine, Toho University, Tokyo; ⁶Laboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan; and ⁷Cardiovascular Research Institute, Departments of Cell Biology and Molecular Medicine and Medicine (Cardiology), New Jersey Medical School, Newark, New Jersey

Submitted 3 February 2004 ; accepted in final form 27 October 2005

Voltage-dependent Ca²⁺ channels (VDCCs), which consist of multiple subtypes, regulate vascular tone in developing arterial smooth muscle, including the ductus arteriosus (DA). First, we examined the expression of VDCC subunits in the Wistar rat DA during development. Among ₁-subunits, _1C and _1G were the most predominant isoforms. Maternal administration of vitamin A significantly increased _1C- and _1G-transcripts. Second, we examined the effect of VDCC subunits on proliferation of DA smooth muscle cells. We found that 1 µM nitrendipine (an L-type Ca²⁺ channel blocker) and kurtoxin (a T-type Ca²⁺ channel blocker) significantly decreased [³H]thymidine incorporation and that 3 µM efonidipine (an L- and T-type Ca²⁺ channel blocker) further decreased [³H]thymidine incorporation, suggesting that L- and T-type Ca²⁺ channels are involved in smooth muscle cell proliferation in the DA. Third, we found that a novel alternatively spliced variant of the _1C-isoform was highly expressed in the neointimal cushion of the DA, where proliferating and migrating smooth muscle cells are abundant. The basic channel properties of the spliced variant did not differ from those of the conventional _1C-subunit. We conclude that multiple VDCC subunits were identified in the DA, and, in particular, _1C- and _1G-subunits were predominant in the DA. A novel spliced variant of the _1C-subunit gene may play a distinct role in neointimal cushion formation in the DA.

alternative spliced; development; gene expression; fetal circulation

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