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Regular Article |
a Departments of Veterinary PathoBiology,
c Pediatrics,
b Pharmacology, Colleges of Veterinary Medicine and Medicine, University of Minnesota, St. Paul, Minnesota 55108
Hormones influence uterine contractility through their effects on intracellular calcium. The regulation of intracellular calcium in uterine smooth muscle is achieved by several mechanisms and includes mobilization from intracellular stores by inositol 1,4,5-trisphosphate and ryanodine-sensitive channels. Cyclic ADP-ribose (cADPR), a metabolite of NAD+, is known to mediate calcium release through ryanodine receptor channels. A cell surface glycoprotein, CD38, catalyzes the synthesis and breakdown of cADPR and thus possesses bifunctional enzymatic activity. The regulation of cADPR synthesis by ADP-ribosyl cyclase (cyclase) or degradation by cADP-ribose hydrolase (hydrolase) by hormones in the myometrium is poorly understood. We investigated the effects of estradiol-17ß on CD38 expression and the synthesis and degradation of cADPR in myometrial smooth muscle obtained from ovariectomized rats. CD38 expression was studied by reverse transcription polymerase chain reaction and Western blot analyses. In uterine microsomal fractions, cyclase and hydrolase activities were measured using nicotinamide guanine dinucleotide and [32P]cADPR as substrates, respectively. Microsomal proteins subfractionated by SDS-PAGE and gel filtration were used to determine the fractions containing cyclase and hydrolase activities. The results demonstrate that cyclase and hydrolase activities are associated with a single protein fraction, similar to CD38 in uteri from both ovariectomized and estradiol-treated rats, and estradiol-17ß causes 1) increased CD38 mRNA and protein expression and 2) significantly enhanced cyclase but not hydrolase activity. The differential regulation of CD38 by estradiol-17ß, resulting in increased cADPR synthesis, would have profound effects on calcium regulation and myometrial contractility.
1 This work was supported by grants from the National Institutes of Health (HL057498 to M.S.K. and DA11806 to T.F.W.), an Academic Health Center Faculty Development Grant, University of Minnesota, and a Fellowship from the Turkish Government (S.D.).
2 Correspondence: Mathur S. Kannan, Department of Veterinary PathoBiology, College of Veterinary Medicine, University of Minnesota, 1971 Commonwealth Ave., St. Paul, MN 55108. FAX: 612 625 5203; kanna001{at}tc.umn.edu
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