Differential expression and localisation of connexin-37 and connexin-43 in follicles of different stages in the 4-week-old mouse ovary

Abstract

Mammalian female fertility is critically dependent on timely coordinated intercellular communication between follicle cells. Gap junctions are small, membrane-bound, aqueous channels that couple adjacent cells ionically and gap junction proteins, connexin-37 (Cx37) and connexin-43 (Cx43) both participate in sustaining proper growth and maturation of the oocyte. The purpose of the present study was to localise Cx37 and Cx43 in sections of ovarian mouse follicles of different developmental stage, and compare their relative expression using immunofluorescence and confocal microscopy. Cx37 exclusively localised to oocytes of all follicle stages, and the estimated number of Cx37 gap junctions per μm oocyte circumference in follicles of stage 2 to stage 6 increased in parallel to oocyte and follicle growth, reaching a maximum in the pre-antral stage. Acetylated α-tubulin and Cx37 double labelling revealed that the majority of Cx37 gap junctions, irrespective of follicle stage, were located on the outer surface of the oocyte cytoskeleton. Cx43 was detected in granulosa cells of all follicle stages, beginning with distinct spots in resting primordial follicles. The estimated number of Cx43 gap junctions per granulosa cell increased significantly with follicle development, in particular in the transition from pre-antral to the antral stage. A subpopulation of Cx43 in the outer part of the zona pellucida was identified and it is suggested these Cx43 gap junctions connect cumulus trans-zonal projections. Together, these data suggests specific roles for Cx37 and Cx43 in folliculogenesis and offers new insights in patterns of intercellular communication of the mouse follicle.