RNA and protein synthesis in developing embryos ofSmittia spec. (Chironomidae, Diptera).

Abstract

Embryos of the chironomid midgeSmittia spec. were permeabilized with sodium hypochlorite and octane. Uptake of labeled uridine and amino acids suggested that these compounds are actively transported across the plasma membrane. Before blastoderm formation, uridine was incorporated at low levels into nuclear DNA and mitochondrial RNA. After blastoderm formation, uridine was incorporated rapidly, mostly into cytoplasmic RNA including both ribosomal RNA precursors and poly(A)-containing RNA. Protein synthesis was observed throughout early embryogenesis. By measuring incorporation of labeled amino acids and internal amino acid pool sizes, we found that the rate of protein synthesis increased with development. Experiments with inhibitors of transcription indicated that proteins synthesized before blastoderm formation were translated from maternal mRNA. During blastoderm stages, embryonic mRNAs seemed to replace maternal mRNAs. Proteins synthesized during short incubation periods in vivo were separated by two-dimensional gel electrophoresis. After blastoderm formation, several new proteins were found that could not be detected at earlier stages.