Ambra1knockdown in zebrafish leads to incomplete development due to severe defects in organogenesis

摘要: AMBRA1 is a positive regulator of the BECN1-dependent program autophagy recently identified in mouse. In this study, we cloned full-length cDNAs ambra1a and ambra1b zebrafish paralogous genes. As mouse, both Ambra1 proteins contain characteristic WD40 repeat region. The transcripts genes are present as maternal RNAs eggs display gradual decline until 8 hpf, being replaced by zygotic mRNAs from 12 hpf onwards. After 24 mainly localized head, suggesting possible role brain development. To check their developmental roles, adopted morpholino knockdown to block either translation (ATGMOs) or splicing (SPLICMOs). Treatment with ATGMOs causes severe embryonic malformations, prelarvae could survive for only 3 4 days b morphants, respectively. SPLICMOs led defects at late stage, indicating importance maternally supplied ambra1 transcripts. Analysis levels Lc3-II, an autophagosome-specific marker, presence lysosome inhibitors evidenced reduction rate autophagosome formation MOs-injected embryos 48 more pronounced case gene. Although some defects, such body growth delay, curved shape hemorrhagic pericardial cavity were occurrence specific phenotypes, major abnormalities development suggests acquisition functions two that required during do not compensate each other following knockdown.