Abstract
BackgroundIn the Drosophila ovary, germ-line and soma cells are interconnected via gap junctions. The main gap-junction proteins in invertebrates are members of the innexin family. In order to reveal the role that innexins play in cell-cell communication during oogenesis, we investigated the localization of innexins 1, 2, 3 and 4 using immunohistochemistry, and analyzed follicle development following channel blockade.ResultsWe found innexin 1 predominantly localized to the baso-lateral domain of follicle cells, whereas innexin 2 is positioned apico-laterally as well as apically between follicle cells and germ-line cells. Innexin 3 was observed laterally in follicle cells and also in nurse cells, and innexin 4 was detected in the oolemma up to stage 8 and in nurse-cell membranes up to stage 12. In order to test whether innexins form channels suitable for intercellular communication, we microinjected innexin antibodies in combination with a fluorescent tracer into the oocyte of stage-10 follicles. We found that dye-coupling between oocyte and follicle cells was largely reduced by innexin-2 antibodies directed against the intracellular C-terminus as well as against the intracellular loop. Analyzing in vitro, between stages 10 and 14, the developmental capacities of follicles following microinjections of innexin-2 antibodies revealed defects in follicle-cell differentiation, nurse-cell regression, oocyte growth and choriogenesis.ConclusionOur results suggest that all analyzed innexins are involved in the formation of gap junctions in the ovary. While innexins 2 and 3 are colocalized between soma cells, innexins 2 and 4 are colocalized between soma and germ-line cells. Innexin 2 is participating in cell-cell communication via hemichannels residing in the oolemma. It is obvious that gap-junctional communication between germ-line and soma cells is essential for several processes during oogenesis.
Highlights
In the Drosophila ovary, germ-line and soma cells are interconnected via gap junctions
Our results suggest that all analyzed innexins are involved in the formation of gap junctions in the ovary
It is obvious that gap-junctional communication between germ-line and soma cells is essential for several processes during oogenesis
Summary
In the Drosophila ovary, germ-line and soma cells are interconnected via gap junctions. In order to reveal the role that innexins play in cell-cell communication during oogenesis, we investigated the localization of innexins 1, 2, 3 and 4 using immunohistochemistry, and analyzed follicle development following channel blockade. During Drosophila oogenesis, germ-line and soma cells are known to interact intimately with each other, for example concerning follicle organization, vitellogenesis and eggshell production [1,2,3]. We investigated by immunoblots and immunohistochemistry the occurrence and detailed localization of innexins 1, 2, 3 and 4 throughout oogenesis. Our results suggest that all analyzed innexins are involved in the formation of ovarian gap junctions
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