论文部分内容阅读
为了探讨miR-202在鱼类胚胎发育中的功能,采用实时定量反转录PCR技术和整体原位杂交技术检测了miR-202在斑马鱼胚胎发育阶段的表达。结果发现miR-202是母源性分子并在斑马鱼胚胎发育过程中持续表达,尤其在早期胚胎发育阶段表达水平较高。在此基础上,采用基因沉默技术在斑马鱼受精卵中显微注射miR-202的反义锁核苷酸,实时荧光定量反转录PCR技术和整体原位杂交技术结果显示miR-202反义锁核苷酸可以显著下调斑马鱼胚胎中miR-202表达水平,同时发现反义抑制miR-202后胚胎发育停滞在4 hpf时左右。共同注射miR-202前体可以部分挽救反义抑制miR-202后导致的胚胎发育停滞。本研究证明miR-202在斑马鱼胚胎发育过程中起着重要的调控作用,其功能是斑马鱼胚胎早期发育所必需的。为进一步探索miR-202在鱼类胚胎发育过程的功能奠定了基础。
In order to investigate the function of miR-202 in fish embryo development, the expression of miR-202 in embryonic development of zebrafish was detected by real-time quantitative reverse transcription-polymerase chain reaction (PCR) and in situ hybridization. The results showed that miR-202 is a maternal molecule and expressed continuously during zebrafish embryogenesis, especially during early embryo development. On the basis of this, microinjection of antisense oligonucleotides of miR-202 into zebrafish zygotes by gene silencing technique, real-time fluorescence quantitative reverse transcription PCR and whole-mount in situ hybridization showed that miR-202 antisense Locked nucleotides significantly down-regulated the expression of miR-202 in zebrafish embryos, and found that antisense inhibition of miR-202 arrested embryonic development at around 4 hpf. Co-injection of miR-202 precursor can partially rescue the embryonic developmental arrest caused by antisense inhibition of miR-202. This study demonstrated that miR-202 plays an important regulatory role in the zebrafish embryogenesis, and its function is essential for the early development of zebrafish embryos. Which laid the foundation for further exploring the function of miR-202 in fish embryonic development.