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Abstract
Noxa functions in apoptosis and immune system of vertebrates, but its activities in embryo development remain unclear. In this study, we have studied the role of zebrafish Noxa (zNoxa) by using zNoxa-specifc morpholino knockdown and overexpression approaches in developing zebrafish embryos. Expression pattern analysis indicates that zNoxa transcript is of maternal origin, which displays a uniform distribution in early embryonic development until shield stage, and the zygote zNoxa transcription is initiated from this stage and mainly localized in YSL of the embryos. The zNoxa expression alterations result in strong embryonic development defects, demonstrating that zNoxa regulates apoptosis from 75% epiboly stage of development onward, in which zNoxa firstly induces the expression of zBik, and then cooperates with zBik to regulate apoptosis. Moreover, zNoxa knockdown also causes a reduction in number of mitotic cells before 8 h.p.f., suggesting that zNoxa also promotes mitosis before 75% epiboly stage. The effect of zNoxa on mitosis is mediated by zWnt4b in early embryos, whereas zMcl1a and zMcl1b suppress the ability of zNoxa to regulate mitosis and apoptosis at different developmental stages. In addition, mammalian mouse Noxa (mNoxa) mRNA was demonstrated to rescue the arrest of mitosis when zNoxa was knocked down, suggesting that mouse and zebrafish Noxa might have similar dual functions. Therefore, the current findings indicate that Noxa is a novel regulator of early mitosis before 75% epiboly stage when it translates into a key mediator of apoptosis in subsequent embryogenesis.
Highlights
ZNoxa and zPuma, have been observed to express before mid-gastrula stage.[8]
To determine the biological function of zebrafish Noxa (zNoxa) in early embryonic development, we examined its effects in early embryos through blocking translation by morpholino (MO) microinjection or through overexpression by synthetic mRNA microinjection
Noxa was originally isolated from an adult T-cell leukemia line as a phorbol 12-myristate-13-acetate inducible protein (PMAIP), and was considered as a canonical tumor suppressor,[18] but its function was not reported in regulating embryonic development except proapoptotic function
Summary
ZNoxa and zPuma, have been observed to express before mid-gastrula stage.[8] their exact functions have remained unclear in early embryonic development of zebrafish. We have characterized zebrafish Noxa (zNoxa) and provided evidence that zNoxa is essential in early embryonic development of zebrafish, and regulates mitosis through zWnt4b and apoptosis through zBik at different development stages. ZMcl1a and zMcl1b have been revealed to suppress the ability of zNoxa to control mitosis and apoptosis. MNoxa can regulate mitosis in zebrafish embryos. The current findings indicate that zNoxa is a novel regulator of mitosis during early embryogenesis except its mediator role for apoptosis
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