Autoradiographic studies of protein and polysaccharide synthesis during vitellogenesis in Drosophila

Abstract

Quantitative light- and electron-microscopic autoradiography was used to evaluate metabolic processes that occur during late developmental stages (10-14) of oogenesis in Drosophila melanogaster. Major differences in radiolabelling patterns were found after in vivo (10-45 min) uptake of [3H]-monosaccharides and [3H]-L-lysine. Several different methods of data analysis were required to facilitate interpretation of these patterns. [3H]-L-lysine produced extensive cytoplasmic labelling at all developmental stages. In addition, about 15% of alpha yolk spheres were intensely labelled at stage 10, reflecting the incorporation of radiolabelled vitellogenins synthesized during the incubation period. Subsequent stages showed low silver grain density over alpha yolk spheres until stage 14, when a burst of [3H]-L-lysine incorporation by most alpha spheres was observed, possibly indicative of a maturation process for embryogenesis. [3H]-D-glucose and [3H]-D-galactose (10 min, in vivo) both induced intense labelling of the beta yolk spheres in a manner suggesting in situ assembly beginning at early stage 13. Inasmuch as the polysaccharide of beta yolk spheres has the properties of glycogen (e.g., rosette structure digested by alpha-amylase) and the radiolabelled monosaccharides were introduced intra-abdominally, it is evident that transport systems as well as enzymes utilizing glucose and galactose for glycogenesis must be readily available. It is notable that wide-spread labelling of egg chambers was elicited by [3H]-D-glucose and [3H]-D-galactose (e.g., nurse cells, follicle cells, chorion, vitelline membrane), but the labelling induced by [3H]-N-acetylmannosamine was restricted mainly to the endochorion. A possible role of microtubules in distribution and assembly of yolk spheres was inferred when colchicine, admixed to the culture medium (2-5 ppm), produced abnormal distribution and diminution in number of both alpha and beta yolk spheres. In addition to revealing previously unknown metabolic events of vitellogenesis, the results provide additional criteria for stage characterization as well as a means to specifically label certain macromolecules for purposes of isolation.