CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Reproduction and Development Laboratory, Lisboa, Portugal.
CBIOS - Faculdade de Medicina Veterinária, Universidade Lusófona de Humanidades e Tecnologias, Lisboa, Portugal.
Reproduction. 2021 Apr;161(4):385-398. doi: 10.1530/REP-20-0435.
This study investigated the role of Notch and Wnt cell signaling interplay in the mouse early embryo, and its effects on fetal development. Developmental kinetics was evaluated in embryos in vitro cultured from the 8-16-cell to the hatched blastocyst stage in the presence of signaling inhibitors of Notch (DAPT) and/or Wnt (DKK1). An embryo subset was evaluated for differential cell count and gene transcription of Notch (receptors Notch1-4, ligands Dll1, Dll4, Jagged1-2, effectors Hes1-2) and Wnt (Wnt3a, Lrp6, Gsk3β, C-myc, Tcf4, β-catenin) components, E-cadherin and pluripotency and differentiation markers (Sox2, Oct4, Klf4, Cdx2), whereas a second subset was evaluated for implantation ability and development to term following transfer into recipients. Notch and Wnt blockades had significant opposing effects on developmental kinetics - Notch blockade retarded while Wnt blockade fastened development. This evidences that Notch and Wnt regulate the pace of embryo kinetics by respectively speeding and braking development. Blockades significantly changed the transcription profile of Sox2, Oct4, Klf4 and Cdx2, and Notch and double blockades significantly changed embryonic cell numbers and cell ratio. The double blockade induced more severe phenotypes than those expected from the cumulative effects of single blockades. Implantation ability was unaffected, but Notch and double blockades significantly decreased fetal development to term. Compared to control embryos, Notch blockade and Wnt blockade embryos originated, respectively, significantly lighter and heavier fetuses. In conclusion, Notch and Wnt signaling interplay in the regulation of the pace of early embryo kinetics, and their actions at this stage have significant carry-over effects on later fetal development to term.
本研究探讨了 Notch 和 Wnt 细胞信号通路相互作用在小鼠早期胚胎中的作用，及其对胎儿发育的影响。在 Notch（DAPT）和/或 Wnt（DKK1）信号抑制剂存在的情况下，将胚胎体外培养从 8-16 细胞期至孵化囊胚期，评估胚胎的发育动力学。评估了一部分胚胎的差异细胞计数和 Notch（受体 Notch1-4、配体 Dll1、Dll4、Jagged1-2、效应物 Hes1-2）和 Wnt（Wnt3a、Lrp6、Gsk3β、C-myc、Tcf4、β-catenin）成分、E-钙粘蛋白和多能性和分化标志物（Sox2、Oct4、Klf4、Cdx2）的基因转录，而另一部分胚胎则评估其植入能力和发育至足月后的情况。Notch 和 Wnt 阻断对发育动力学有显著的相反影响—— Notch 阻断会减缓，而 Wnt 阻断则会加速发育。这表明 Notch 和 Wnt 通过分别加速和制动发育来调节胚胎动力学的速度。阻断显著改变了 Sox2、Oct4、Klf4 和 Cdx2 的转录谱，Notch 和双重阻断显著改变了胚胎细胞数量和细胞比例。双重阻断引起的表型比单一阻断的累积效应更严重。植入能力不受影响，但 Notch 和双重阻断显著降低了胎儿的足月发育。与对照胚胎相比，Notch 阻断和 Wnt 阻断胚胎分别起源于明显较轻和较重的胎儿。总之，Notch 和 Wnt 信号通路在调节早期胚胎动力学速度方面相互作用，它们在这个阶段的作用对后期胎儿发育至足月有显著的后续影响。
