Patch-clamp study reveals that the importance of connexin43-mediated gap junctional communication for ovarian folliculogenesis is strain specific in the mouse.
Genetic ablation of connexin37 (Cx37) or connexin43 (Cx43), the two gap junction proteins expressed by mouse ovarian granulosa cells, has been shown to result in impaired follicle development. We used patch-clamp techniques to evaluate quantitatively the contribution of these connexins to gap junctional intercellular communication (GJIC) among granulosa cells. The coupling conductance derived from a voltage step-induced capacitive current transient was used as a measure of GJIC in cultured granulosa cells. Using this method, we determined that the conductance of wild-type (84.1 +/- 28.6 nS; n = 6) and Cx37-deficient granulosa cells (83.7 +/- 6.4 nS; n = 11) does not differ significantly (P = 0.35), suggesting a limited contribution, if any, of Cx37 to granulosa cell coupling. In contrast, the conductance between granulosa cells of Cx43-deficient mice (2.6 +/- 0.8 nS; n = 5) was not significantly different from that of single, isolated wild-type granulosa cells (2.5 +/- 0.7 nS, n = 5; P = 0.83), indicating that Cx43-deficient granulosa cells were not electrically coupled. A direct measurement of transjunctional conductance between isolated granulosa cell pairs using a dual patch-clamp technique confirmed this conclusion. Interestingly, a partial rescue of folliculogenesis was observed when the Cx43-null mutation in C57BL/6 mice was crossed into the CD1 strain, and capacitive current measurement demonstrated that this rescue was not due to reestablishment of GJIC. These results demonstrate that folliculogenesis is impaired in the absence of GJIC between granulosa cells, but they also indicate that the severity is dependent on genetic background, a phenomenon that cannot be attributed to the expression of additional connexins.
Tong
Dan
D
Dept. of Physiology and Pharmacology, The Univ. of Western Ontario, Dental Science Bldg., Rm. 00073, London, ON, Canada N6A 5C1.
Gittens
Joanne E I
JE
Kidder
Gerald M
GM
Bai
Donglin
D
D000818:Animals; D002450:Cell Communication / Q000502:physiology*; D018031:Connexin 43 / Q000235:genetics / Q000502:physiology*; D025802:Electric Capacitance; D005260:Female; D017629:Gap Junctions / Q000502:physiology*; D006107:Granulosa Cells / Q000166:cytology / Q000502:physiology*; D008564:Membrane Potentials / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009866:Oogenesis / Q000502:physiology*; D018408:Patch-Clamp Techniques
D018031:Connexin 43