背景:胎盘外泌体是妊娠期间胎盘细胞分泌的一种细胞间通讯介质,外泌体的发生和释放受许多分泌糖蛋白的调节。CREG1是一种分泌型糖蛋白,广泛表达于机体的各种器官和组织中,抑制细胞增殖并增强细胞分化。本研究的目的是通过靶向IGF2R并通过外泌体转运参与调节器官分化,探讨CREG1在早期妊娠奶牛胎盘滋养层细胞增殖和分化过程中调节外泌体生成的作用。
方法:首先采用分子生物学方法研究了CREG1、IGF2R和外泌体标记蛋白在妊娠奶牛胎盘发育早期的表达规律。随后,研究了CREG1通过靶向IGF2R对牛胎盘滋养层(BTC)来源的外泌体形成和释放的影响。Further,研究了CREG1对基因表达模式的改变以及外泌体向受体细胞的转运以及参与调节类器官分化的影响。
结果:奶牛胎盘滋养层细胞的早期进化过程中,CREG1、IGF2R和外泌体标记蛋白的表达随妊娠月数的增加而增加。Creg1的过表达增强了来自BTC的外泌体的发生和释放,而抑制Igf2r基因的表达不仅抑制了外泌体的发生,而且还抑制了CREG1蛋白过表达诱导的外泌体的发生和释放。有趣的是,IGF2R可通过反向分泌调节CREG1的表达。更重要的是,滋养细胞来源的外泌体的发生和释放受CREG1与IGF2R结合的调控,随后与Rab11绑定。CREG1不仅能促进供体细胞外泌体的形成和释放,还随着外泌体向受体细胞的转运而调节基因表达模式的变化,并参与调节胎盘的早期发育。
结论:我们的研究证实,CREG1通过靶向IGF2R参与早期妊娠奶牛胎盘滋养层细胞增殖和分化过程中外泌体的发生和释放,并通过外泌体转运参与类器官分化的调节。
BACKGROUND: Placental exosomes are a kind of intercellular communication media secreted by placental cells during pregnancy, exosomogenesis and release are regulated by many secretory glycoproteins. CREG1 is a kind of secreted glycoprotein widely expressed in various organs and tissues of the body, which inhibits cell proliferation and enhances cell differentiation. The aim of this study was to explore the role of CREG1 in regulating exosomogenesis during the proliferation and differentiation of placental trophoblast cells in early pregnant dairy cows by targeting IGF2R and participating in regulating organoid differentiation via exosomes transport.
METHODS: Molecular biological methods were firstly used to investigate the expression patterns of CREG1,
IGF2R and exosomal marker proteins in early placental development of pregnant dairy cows. Subsequently, the effects of CREG1 on the formation and release of bovine placental trophoblast (BTCs) derived exosomes by targeting IGF2R were investigated. Further, the effects of CREG1 on the change of gene expression patterns along with the transport of exosomes to recipient cells and participate in regulating the differentiation of organoids were explored.
RESULTS: The expression of CREG1,
IGF2R and exosomal marker proteins increased with the increase of pregnancy months during the early evolution of placental trophoblast cells in dairy cows. Overexpression of Creg1 enhanced the genesis and release of exosomes derived from BTCs, while knocking down the expression of
Igf2r gene not only inhibited the genesis of exosomes, but also inhibited the genesis and release of exosomes induced by overexpression of CREG1 protein. Interestingly,
IGF2R can regulate the expression of CREG1 through reverse secretion. What\'s more, the occurrence and release of trophoblast-derived exosomes are regulated by CREG1 binding to
IGF2R, which subsequently binds to Rab11. CREG1 can not only promote the formation and release of exosomes in donor cells, but also regulate the change of gene expression patterns along with the transport of exosomes to recipient cells and participate in regulating the early development of placenta.
CONCLUSIONS: Our study confirmed that CREG1 is involved in the exosomogenesis and release of exosomes during the proliferation and differentiation of placental trophoblast cells in early pregnant dairy cows by targeting IGF2R, and is involved in the regulation of organoid differentiation through exosome transport.