Cathepsin B differential expression and enzyme processing and activity during Fundulus heteroclitus embryogenesis

Abstract

The role of lysosomal proteases such as cathepsin B (Ctsb) and one of the paralogs of cathepsin L (Ctsla) during yolk metabolism in fish oocytes is well established. However, the function of Ctsb during embryogenesis, particularly in marine teleosts, has been poorly documented. In this study, the spatio-temporal expression of Ctsb and Ctsla, their enzymatic activities, and the processing of the Ctsb and its cellular localization, was investigated in developing embryos of the killifish (Fundulus heteroclitus). Both fhctsb and fhctsla transcript levels, as well as cathepsin B- and L-like activities, gradually increased in embryos from the 2–4 cell stage up to 7 days post-fertilization. During the morula to gastrula transition an increase of the active FhCtsb single chain form was followed by a rise in cathepsin B activity, which were apparently regulated by post-transcriptional mechanisms. During neurulation, a 8-fold increase in cathepsin B activity was accompanied by a more moderate increase in cathepsin L activity, which was 6-fold enhanced by 7 dpf. These increased catalytic activities were well-correlated to changes in the electrophoretic pattern of yolk proteins and a strong expression of fhctsb and its protein product in the yolk syncytial layer. The increase of cathepsin B activity was further correlated with an increment of the relative amount of the FhCtsb single and double chain forms, both active forms of FhCtsb. These results suggest that FhCtsb may be involved in the mechanisms underlying the onset of gastrulation in F. heteroclitus embryos, and may play complementary roles with FhCtsla during yolk metabolism