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Fig. 5. Folliculogenesis in reaggregated ovaries. (A) In ovaries constructed with Cx43-deficient granulosa cells infected with wild-type (WT) Cx43, follicles of all developmental stages were observed (left; white arrow points to an antral follicle), whereas, in ovaries constructed with Cx43-deficient granulosa cells infected with cys-less Cx43 (middle) or control vector (right), only primary (unilaminar) follicles were observed. Scale bars, 50 µm. Notice the different magnifications in the left, middle and right micrographs. (B) Distribution of follicle stages in the three different types of reaggregated ovaries as indicated below each group of bars. Between 22 and 72 follicles from three to four grafted ovaries were categorized for each group. A significant difference (P<0.001 wild type versus cys-less; P<0.01 wild type versus vector control, chi-square) was observed between ovaries expressing wild-type Cx43 and cys-less Cx43 in granulosa cells. Only the ovaries constructed with wild-type Cx43 were seen to contain antral follicles. (C) Expression of wild-type Cx43 and cys-less Cx43 in reaggregated ovaries. Infection was identified by EGFP expression (top row). Cx43 immunostaining (red, bottom row) showed mainly membrane expression of wild-type Cx43 with numerous gap junction-like plaques (left column) and predominately cytoplasmic localization of cys-less Cx43 (middle column). No Cx43 signal was detected in the empty-vector-infected ovaries (right column). Scale bar, 20 µm.
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