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First published online 30 October 2007
doi: 10.1242/jcs.011775

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In vivo analysis of undocked connexin43 gap junction hemichannels in ovarian granulosa cells

¹ Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON N6A 5C1, Canada
² Department of Obstetrics and Gynecology, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON N6A 5C1, Canada
³ Department of Paediatrics, Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON N6A 5C1, Canada
⁴ Children's Health Research Institute, 800 Commissioners Road East, London, ON, Canada

^* Author for correspondence (e-mail: gerald.kidder{at}schulich.uwo.ca)

Accepted 3 September 2007

Connexin43 (Cx43, encoded by Gja1) is required for ovarian follicle development in the mouse. It is strongly expressed in granulosa cells, in which it forms intercellular gap junction channels that couple the cells metabolically. However, recent evidence indicates that undocked gap junction hemichannels can also have physiological roles such as mediating the release of small messenger molecules, including ATP. In this study, the presence of undocked Cx43 hemichannels in granulosa cells was revealed by dye uptake induced either by mechanical stimulation or by the reduction of extracellular divalent cations, both of which are known triggers for hemichannel opening. ATP release was also detected, and could be abolished by connexin-channel blockers. None of these putative hemichannel-mediated activities were detected in Cx43-deficient granulosa cells. Therefore, we hypothesized that hemichannels account for the essential role of Cx43 in folliculogenesis. To test this, a Cx43 mutant lacking the conserved cysteines on the extracellular loops (cys-less Cx43), reported to form hemichannels but not intercellular channels, was retrovirally expressed in Cx43-deficient granulosa cells. The infected cells were then combined with wild-type oocytes to make reaggregated ovaries, which were grafted into host kidneys. Although re-introduction of wild-type Cx43 rescued folliculogenesis, introduction of cys-less Cx43 did not. Therefore, although Cx43 gap junction hemichannels might play a role in ovarian folliculogenesis, their contribution does not supplant the need for intercellular gap junction channels.

Key words: Oogenesis, Folliculogenesis, Undocked connexons

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				D. Tong, T. Y. Li, K. E. Naus, D. Bai, and G. M. Kidder In vivo analysis of undocked connexin43 gap junction hemichannels in ovarian granulosa cells Development, December 1, 2007; 134(23): e1 - e1. [Full Text]