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First published online October 27, 2005
doi: 10.1242/10.1242/jcs.02624

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Differential contributions of connexin37 and connexin43 to oogenesis revealed in chimeric reaggregated mouse ovaries

¹ Departments of Physiology and Pharmacology, Obstetrics and Gynaecology, and Paediatrics, The University of Western Ontario, London, Ontario N6A 5C1, Canada
² Children's Health Research Institute, 800 Commissioners Road East, London, Ontario N6C 2V5, Canada

View larger version (127K): [in a new window]   Fig. 1. Absence of wild-type granulosa cell contamination in ovaries constructed from wild-type oocytes and Cx43-deficient somatic cells. 43+/+oo/43+/+gc (A,C,E) and 43+/+oo/43–/–gc (B,D,F) ovary sections were immunostained for Cx43. Punctate immunoreactivity characteristic of gap junctions is evident between the granulosa cells of 43+/+oo/43+/+gc chimeric ovaries (C,E) but not 43+/+oo/43–/–gc chimeric ovaries (D,F). E and F are magnifications of C and D, respectively. Bar, 50 µm (C,D); 10 µm (E,F).

View larger version (108K): [in a new window]   Fig. 2. Aberrant development in Cx43-deficient ovaries is due to defects in granulosa cells, not oocytes. (A) A 43–/–oo/43+/+gc ovary containing follicles at all developmental stages, including antral stages. (B) A 43+/+oo/43–/–gc ovary at the same magnification as (A) showing a preponderance of small follicles at the primary and early secondary follicle stages. The graphs show mean oocyte (C) and follicle (D) diameters across chimeric ovary combinations. Bars with different letters above them are significantly different (P<0.01). The numbers in parentheses indicate the numbers of oocytes or follicles measured. Bar, 50 µm.

View larger version (100K): [in a new window]   Fig. 3. Aberrant development in Cx37-deficient ovaries is due to defects in oocytes, not granulosa cells. (A) A 37–/–oo/37+/+gc ovary containing only type 4 secondary follicles. (B) A 37+/+oo/37–/–gc ovary showing antral follicles. The graphs show mean oocyte (C) and follicle (D) diameters for chimeric ovary combinations in parallel with grafted wild-type, Cx37 and Cx43 knockout ovaries (BL/6 background). Measurements from 43+/+oo/43–/–gc ovaries on the same background are included in both C and D for comparison. Bars with different letters above them are significantly different (P<0.01). The numbers in parentheses indicate the numbers of oocytes or follicles measured. Bar, 50 µm.

View larger version (134K): [in a new window]   Fig. 4. Cumulus-oocyte complexes from chimeric ovaries composed of either 43+/+oo/43–/–gc or 37–/–oo/37+/+gc do not undergo cumulus expansion. (A) Example of a chimeric follicle with mucified cumulus. (B) Example showing the lack of cumulus expansion. Follicles in A and B were cultured overnight in the same maturation medium and photographed at the same magnification. Bar, 50 µm.

View larger version (109K): [in a new window]   Fig. 5. Cx37 deficiency in granulosa cells does not impair oocyte-granulosa cell gap junctional coupling. 37+/+oo/37+/+gc follicles (A,B), 37–/–oo/37+/+gc follicles (C,D) and 37+/+oo/37–/–gc follicles (E,F) were microinjected with Lucifer Yellow for 10 minutes. Dye transfer from oocytes to granulosa cells was evident when both components were wild-type (B) and when the absence of Cx37 was restricted to the granulosa cells (F). Cx37-deficient oocytes did not pass dye to wild-type granulosa cells (D). Bar, 50 µm.

View larger version (98K): [in a new window]   Fig. 6. Cx43 does not appear to replace Cx37 at the oocyte surface in 37+/+oo/37–/–gc ovaries. 37+/+oo/37+/+gc (A) and 37+/+oo/37–/–gc (B) ovary sections were immunostained for Cx43. Punctate staining for Cx43 was seen between granulosa cells, but very little Cx43 was detected between oocytes and granulosa cells in ovaries of either constitution. Bar, 50 µm.

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