A gap junctionally transmitted epithelial cell signal regulates endocytic yolk uptake in Oncopeltus fasciatus

Abstract

For endocytic uptake of vitellogenins, developing oocytes of Oncopeltus require a soluble, diffusible molecular signal from their surrounding epithelial cells, and this signal must be transmitted through open gap junctions. Hormonal stimulation triggering synthesis and processing of vitellogenins into mature insect yolk spheres has been intensely studied, and follicle epithelial cells are known in several insects to contribute to the blood products which are endocytosed along with vitellogenins synthesized in the fat bodies. However, there has been little evidence that direct gap junctional communication is a requirement for endocytic activity by oocytes. In untreated control follicles, both electrical and dye coupling occur, and follicles incubated in vitro in physiological salt solution containing small amounts of blood and fluorescent dye produce fluorescently labeled nascent yolk spheres. Labeled yolk spheres were visible in both sectioned material, and, with (Laser) Confocal Scanning, in living material. Dye coupling was abolished by treatment with either 1 mM octanol, 0.5 mM ethyl methanesulfonate (EMS), or cytoplasmic acidification, with coupling coefficients also being affected as each of these gap junction antagonists down-regulated the connexons. With each of these treatments, after gap junctions were down-regulated, receptor-mediated endocytic uptake of blood-born vitellogenins came to a halt. Furthermore, Oncopeltus follicles with endocytic activity blocked in this manner could be rescued by microinjection of the soluble fraction of lysed epithelial cell cytoplasm, confirming that the process depended upon a molecular signal from the epithelial cells.