4-3-3 and Hippo pathway protiens upregulation during adipogenesis.

LIM, G.; UHART, M; DEL VELIZ, S.; BUSTOS, D. M.

Mendoza

Congreso; LVI Annual SAIB on line meeting.; 2020

SAIB

3T3-L1 cells undergo a complex roadmap of signals to differentiate into fat cells. The exact mechanism for the coordination of these signals remains elusive, although 14-3-3 proteins could be key players. In our laboratory, we examined how the combination of different adipogenic differentiation drugs affects the expression of the Hippo kinase pathway genes, as well as the expression of specific 14-3-3 paralogs. We achieve adipocyte differentiation in vitro by adding an adipogenic differentiation medium (ADM) that includes Dulbecco´s modified Eagle´s medium, 10% fetal bovine serum, synthetic drugs (dexamethasone, IBMX, rosiglitazone), and peptide hormones (insulin). We performed qPCR experiments to measure the gene expression of 14-3-3 and the most important proteins of the Hippo pathway, on days 3 and 7 of adipogenic differentiation. We have determined that the conditions which most promoted adipogenic differentiation (evidenced as a larger number and size of lipid droplets) showed higher levels of Hippo pathway proteins, and both 14-3-3 gamma and beta isoforms on day 7. These effects were especially evident when IBMX was replaced by GLP-1 in the ADM. These results confirm previous qPCR data, obtained under similar experimental conditions. The main question is whether such increased expression is related to the effects of differentiation inducers (glucocorticoids, thiazolidinediones, incretins, or insulin) during early or late adipogenesis. We also would like to determine if the differentiation and the observed increased expression correlate with the activation of the Hippo pathway