miR-206-3p在小鼠皮肤中的表达及其作用

Expression and distribution of miR-206-3p in mouse skin and its potential roles

目的对小鼠皮肤中miR-206-3p及其靶基因脑源性神经营养因子(Bdnf)进行表达与定位检测,以探讨其在皮肤发育过程中的可能作用。方法手术剪取13.5 dpc(交配后天数)、15.5 dpc、17.5 dpc胎鼠,1 dpp(出生后天数)、4 dpp乳鼠及16周龄雄鼠的背部全层皮肤,用实时荧光定量RT-PCR检测miR-206-3p与Bdnf mRNA的表达水平,western blot检测BDNF蛋白质表达水平,原位杂交与免疫组化分别检测miR-206-3p与BDNF的组织定位。结果皮肤中miR-206-3p在胚胎期持续升高,到17.5 dpc后逐渐降低,于成年鼠时回复至13.5 dpc时水平;BDNF表达趋势与miR-206-3p相反,而Bdnf mRNA在17.5 dpc至成年鼠并无显著改变。miR-206-3p的分布在皮肤发育过程中逐渐区域化,在出生后主要分布于上基部和毛囊外周,与同期BDNF表达区域毗邻但不重叠。结论 miR-206-3p可能通过转录后调控Bdnf参与了小鼠皮肤神经发育过程。

Objective：To investigate the expression and distribution of miR-206-3p and its target gene Bdnf （brain derived neurotrophic factor） in mouse skin and the potential roles during the development of skin. Methods：The full-thickness back skins of BALB/c mice on the 13.5 dpc （day post coition）, 15.5 dpc, 17.5 dpc, 1 dpp （day post partum）, 4 dpp and 16 weeks were collected respectively. The expression levels of miR-206-3p and Bdnf mRNA were measured by real-time RT-PCR, and BDNF protein was detected by western blot. The distributions of miR-206-3p and BDNF in tissue were profiled by in situ hybridization （ISH） and immunohistochemistry （IHC） respectively. Results：The expression level of miR-206-3p increased gradually until 17.5 dpc and subsequently receded to the similar level of the 13.5 dpc during adult stage. The expression of BDNF protein exhibited in the opposite manner of miR-206-3p but dnf mRNA remained stable from the 17.5 dpc until adult stage. ISH results showed that the distribution of miR-206-3p was gradually regionalized during the development of skin in the postnatal days and mainly located in the suprabasal layer and periphery of hair follicles, which was adjacent to the regions of BDNF expression at the same stage in a nonoverlapping manner. Conclusion：The miR-206-3p may be involved in neurodevelopment of mouse skin through the post-transcriptional regulation of Bdnf.