Identification and characterization of Ca2+-activated K plus channels in granulosa cells of the human ovary

Traut, Matthias H.; Berg, Dieter; Berg, Ulrike; Mayerhofer, Artur und Kunz, Lars (2009): Identification and characterization of Ca2+-activated K plus channels in granulosa cells of the human ovary. In: Reproductive Biology and Endocrinology 7:28

Abstract

Background: Granulosa cells (GCs) represent a major endocrine compartment of the ovary producing sex steroid hormones. Recently, we identified in human GCs a Ca(2+)-activated K(+) channel (K(Ca)) of big conductance (BK(Ca)), which is involved in steroidogenesis. This channel is activated by intraovarian signalling molecules (e.g. acetylcholine) via raised intracellular Ca(2+) levels. In this study, we aimed at characterizing 1. expression and functions of K(Ca) channels (including BK(Ca) beta-subunits), and 2. biophysical properties of BK(Ca) channels. Methods: GCs were obtained from in vitro-fertilization patients and cultured. Expression of mRNA was determined by standard RT-PCR and protein expression in human ovarian slices was detected by immunohistochemistry. Progesterone production was measured in cell culture supernatants using ELISAs. Single channels were recorded in the inside-out configuration of the patch-clamp technique. Results: We identified two K(Ca) types in human GCs, the intermediate-(IK) and the small-conductance K(Ca) (SK). Their functionality was concluded from attenuation of human chorionic gonadotropin-stimulated progesterone production by K(Ca) blockers (TRAM-34, apamin). Functional IK channels were also demonstrated by electrophysiological recording of single K(Ca) channels with distinctive features. Both, IK and BK(Ca) channels were found to be simultaneously active in individual GCs. In agreement with functional data, we identified mRNAs encoding IK, SK1, SK2 and SK3 in human GCs and proteins of IK and SK2 in corresponding human ovarian cells. Molecular characterization of the BK(Ca) channel revealed the presence of mRNAs encoding several BK(Ca) beta-subunits (beta2, beta3, beta4) in human GCs. The multitude of beta-subunits detected might contribute to variations in Ca(2+) dependence of individual BK(Ca) channels which we observed in electrophysiological recordings. Conclusion: Functional and molecular studies indicate the presence of active IK and SK channels in human GCs. Considering the already described BK(Ca), they express all three K(Ca) types known. We suggest that the plurality and co-expression of different K(Ca) channels and BK(Ca) beta-subunits might allow differentiated responses to Ca(2+) signals over a wide range caused by various intraovarian signalling molecules (e.g. acetylcholine, ATP, dopamine). The knowledge of ovarian K(Ca) channel properties and functions should help to understand the link between endocrine and paracrine/autocrine control in the human ovary.

• Dokumententyp: Zeitschriftenartikel
• Publikationsform: Publisher's Version
• Fakultät: Biologie > Department Biologie II > Neurobiologie
• Themengebiete: 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften; Biologie
• ISSN: 1477-7827
• Sprache: Englisch