In vitro culture ofDrosophila ovarian follicles: The influence of different media on development, RNA synthesis, protein synthesis and potassium uptake

Abstract

Contradictory electrophysiological results were recently obtained inDrosophila ovarian follicles kept in different salines or complete media during measurements. Therefore, I checked follicles maintained in various solutions using morphological, physiological and biochemical criteria. Defined complete media were the best for supporting development from stage 10 to stage 14 (end of oogenesis). Supplementation of the solutions with insect pupal haemolymph had negative effects. For the maintenance of RNA synthesis, complete media were again superior to simple salines. Total protein synthesis was not very sensitive to the culture conditions during short incubation periods, but electrophoretic protein patterns were slightly less complex in the salines than in complete media. Furthermore, some major proteins (e.g. chorion proteins) synthesized during longterm culture failed to appear in the salines. In view of extrafollicular electrical currents and intracellular electrical potentials, potassium uptake experiments were conducted with several inhibitors, using rubidium-86 as a probe. Both potassium concentration and osmolarity were found to exert strong influences on total potassium uptake of the follicles. In the tested media and salines differing amounts of potassium were taken up via (Na⁺, K⁺)-ATPase, via other K⁺-pump(s) or passively. The possible influences of several parameters on the outcome of earlier in vitro experiments withDrosophila follicles are discussed. For further in vitro studies the chemically defined complete R-14 medium seems to be the most suitable.

Keywords: Culture media; Drosophila oogenesis; Intrafollicular transport; Ionic current patterns; Membrane potentials.