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L- and T-Type Voltage-Gated Ca²⁺ Channels in Human Granulosa Cells: Functional Characterization and Cholinergic Regulation

Department of Human and General Physiology (D.P., G.A.) and Interdepartment Center Luigi Galvani for the Study of Biophysics, Bioinformatics, and Biocomplexity (G.A.), University of Bologna, I-40127 Bologna, Italy; and Società Italiana Studi di Medicina della Riproduzione, Reproductive Medicine Unit (M.C.M., A.P.F., L.G.), I-40138 Bologna, Italy

Address all correspondence and requests for reprints to: Dr. Giorgio Aicardi, Department of Human and General Physiology, University of Bologna, Via S. Donato 19/2, I-40127 Bologna, Italy. E-mail: aicardi{at}biocfarm.unibo.it.

Using the whole-cell configuration of the patch-clamp technique, we have characterized two types of ionic currents through voltage-dependent Ca²⁺ channels in human granulosa cells. One is long-lasting, activates at approximately –20 mV, reaches the peak at approximately +20 mV, has an inactivation time constant of 132.5 ± 5.6 msec at 20 mV, and is sensitive to dihydropyridines. The other is transient, activates at approximately –40 mV, peaks at approximately –10 mV, has an inactivation time constant of 38.8 ± 1.8 msec at –10 mV, displays a voltage-dependent inactivation, and is sensitive to 100 µM Ni²⁺, but not to dihydropyridines. Biophysical and pharmacological properties of these currents indicate that they are gated through L- and T-type calcium channels, respectively. The cholinergic receptor agonist carbachol (50 µM) reduces the amplitude of the currents through both L-type (–34.7 ± 6.4%; n = 10) and T-type (–52.6 ± 7.4%; n = 8) channels, suggesting a possible role of these channels in the cholinergic regulation of human ovarian functions.

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