Rainbow Trout (Salmo gairdneri) Growth Hormone: in vitro Translation of Pituitary RNA and Product Analysis

Abstract

We have used RNA blot analysis, in vitro translation, and protein immunoblot analysis to characterize mRNA in the pituitary glands of rapidly growing rainbow trout (Salmo gairdneri). Cell-free translation products of total RNA, analyzed by direct immunoprecipitation or western immunoblot analysis with an antiserum to chum salmon (Oncorphynchus keta) growth hormone (GH), indicate that rainbow trout pregrowth hormone (pre-GH) is a polypeptide of 25 000 Da. RNA blot analysis of total pituitary RNA to its cDNA, using total liver RNA as a competitor, revealed the presence of at least four size classes of pituitary mRNA sequences. One class, with a size smaller than 18S rRNA, is the predominant mRNA component. By in vitro translation and western immunoblot analysis, the pre-GH mRNA activity is associated with this class of mRNA sequences. These results suggest that rainbow trout pre-GH mRNA is in the same size range as that of mammalian GH mRNA.