反义抑制miR-202引起斑马鱼早期胚胎发育异常研究

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。

miR-202 inhibition leads to early termination of zebrafish embryonic development

To learn the role of miR-202 in the zebrafish embryonic development, miR-202 expression was detected using qRT-PCR and WISH, the results indicated that miR-202 is a maternal molecule and expressed continuously during the zebrafish embryonic development, especially at the early stages. Knockdown of miR-202 was performed by microinjecting the locked nucleic acid modified antisense oligo of miR-202. The results showed that antisense oligo of miR-202 decreased miR-202 level significantly, accompanied with an early termination of the zebrafish embryonic development at around 4 hpf. And miR-202 inhibition induced early termination could be partly rescued by co-injection of pre-miR-202, indicating that miR-202 is required for the early zebrafish embryonic development. The present study for the first time unveils the essential role of miR-202 for the zebrafish embryonic development.