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Myc proteins control cell proliferation, cell cycle progression, and apoptosis, and play important roles in cancer as well in establishment of pluripotency. Here we investigated the control of  myc  gene expression by the Pou5f1/Oct4 pluripotency factor in the early zebrafish embryo. We analyzed the expression of all known zebrafish Myc family members,  myca ,  mycb ,  mych ,  mycl1a, mycl1b , and  mycn , by whole mount  in situ  hybridization during blastula and gastrula stages in wildtype and maternal plus zygotic  pou5f1  mutant (MZ spg ) embryos, as well as by quantitative PCR and in time series microarray data. We found that the broad blastula and gastrula stage  mych  expression, as well as late gastrula stage  mycl1b  expression, both depend on Pou5f1 activity. We analyzed ChIP-Seq data and found that both Pou5f1 and Sox2 bind to  mych  and  mycl1b  control regions. The regulation of  mych  by Pou5f1 appears to be direct transcriptional activation, as overexpression of a Pou5f1 activator fusion protein in MZ spg  embryos induced strong  mych  expression even when translation of zygotically expressed mRNAs was suppressed. We further showed that MZ spg  embryos develop enhanced apoptosis already during early gastrula stages, when apoptosis was not be detected in wildtype embryos. However, Mych knockdown alone did not induce early apoptosis, suggesting potentially redundant action of several early expressed  myc  genes, or combination of several pathways affected in MZ spg.  Experimental  mych  overexpression in MZ spg  embryos did significantly, but not completely suppress the apoptosis phenotype. Similarly, p53 knockdown only partially suppressed apoptosis in MZ spg  gastrula embryos. However, combined knockdown of p53 and overexpression of Mych completely rescued the MZ spg  apoptosis phenotype. These results reveal that Mych has anti-apoptotic activity in the early zebrafish embryo, and that p53-dependent and Myc pathways are likely to act in parallel to control apoptosis at these stages.

Pou5f1/Oct4 Promotes Cell Survival via Direct Activation of mych Expression during Zebrafish Gastrulation