Aberrant regulation of the insulin-like growth factor (IGF)/insulin (IIS)-PI3K-AKT-TOR signaling pathway is linked to major human diseases, and key components of this pathway are targets for therapeutic intervention. Current assays are molecular target- or cell culture-based platforms. Due to the great in vivo complexities inherited in this pathway, there is an unmet need for whole organism based assays. Here we report the development of a zebrafish transgenic line, Tg(igfbp5a:GFP), which faithfully reports the mitotic action of IGF1R-PI3K-Akt-Tor signaling in epithelial cells in real-time. This platform is well suited for high-throughput assays and real-time cell cycle analysis. Using this platform, the dynamics of epithelial cell proliferation in response to low [Ca2+] stress and the distinct roles of Torc1 and Torc2 were elucidated. The availability of Tg(igfbp5a:GFP) line provides a whole organism platform for phenotype-based discovery of novel players and inhibitors in the IIS-PI3K-Akt-Tor signaling pathway.
Large yellow croaker is an important marine fish species for aquaculture in China. Larval rearing of this species is still based on live prey feeding. The aim of this study was to describe the onset and development of the main digestive enzymes during larval development of this species, in order to obtain essential data for the formulation of a compound diet adapted to larvae. Enzymatic assays were conducted from day 1 post hatching to day 40 in larvae fed the following live prey sequence: rotifers, Artemia nauplii, copepods, and then frozen copepods. Assays were done on whole larvae until day 19, then on dissected larvae and on purified brush border membranes of intestine. Pancreatic enzymes, amylase, and trypsin, showed high activities from early stages, even before mouth opening. Mechanisms of pancreatic secretion were achieved from day 19. Enzymes of brush border membranes, alkaline phosphatase, and aminopeptidase N, increased abruptly between day 23 and 25, showing maturation of the intestinal digestive process. These data, together with data describing mouth enlargement during development, suggest that large yellow croaker larvae could be fed successfully with a compound diet from early stages, at least from day 23.
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