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 juncti...

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Published in:American Journal of Physiology: Cell Physiology Vol. 290; no. 1; p. C290
Main Authors: Tong, Dan, Gittens, Joanne E I, Kidder, Gerald M, Bai, Donglin
Format: Journal Article
Language:English
Published: United States 01-01-2006
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Abstract 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.
AbstractList 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.
Author Kidder, Gerald M
Gittens, Joanne E I
Tong, Dan
Bai, Donglin
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  fullname: Kidder, Gerald M
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  givenname: Donglin
  surname: Bai
  fullname: Bai, Donglin
BackLink https://www.ncbi.nlm.nih.gov/pubmed/16135542$$D View this record in MEDLINE/PubMed
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Snippet Genetic ablation of connexin37 (Cx37) or connexin43 (Cx43), the two gap junction proteins expressed by mouse ovarian granulosa cells, has been shown to result...
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StartPage C290
SubjectTerms Animals
Cell Communication - physiology
Connexin 43 - genetics
Connexin 43 - physiology
Electric Capacitance
Female
Gap Junctions - physiology
Granulosa Cells - cytology
Granulosa Cells - physiology
Membrane Potentials - physiology
Mice
Mice, Inbred C57BL
Mice, Knockout
Oogenesis - physiology
Patch-Clamp Techniques
Title Patch-clamp study reveals that the importance of connexin43-mediated gap junctional communication for ovarian folliculogenesis is strain specific in the mouse
URI https://www.ncbi.nlm.nih.gov/pubmed/16135542
Volume 290
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