Somatic cell nuclear transfer(SCNT)has been successfully employed across various mammalian species,yet cloned animals consistently exhibit low pregnancy rates,primarily due to placental abnormalities such as hyperplas...Somatic cell nuclear transfer(SCNT)has been successfully employed across various mammalian species,yet cloned animals consistently exhibit low pregnancy rates,primarily due to placental abnormalities such as hyperplasia and hypertrophy.This study investigated the involvement of the Hippo signaling pathway in aberrant placentaldevelopmentinSCNT-inducedbovine pregnancies.SCNT-derived cattle exhibited placental hypertrophy,including enlarged abdominal circumference and altered placental cotyledon morphology.RNA sequencing analysis indicated significant dysregulation of Hippo signaling pathway genes in SCNT placentas.Coexpression of YAP1 and CCND1 was observed in cloned blastocysts,placental tissues,and bovine placental mesenchymal stem cells(bPMSCs).Manipulation of YAP1expression demonstrated the capacity to regulate bPMSC proliferation.Experimental assays confirmed the direct binding of YAP1 to CCND1,which subsequently promoted CCND1 expression in bPMSCs.Furthermore,inhibition of CDK6,a downstream target of CCND1,attenuated SCNT bPMSC proliferation.This study identified YAP1 as a key regulatory component within the Hippo signaling pathway that drives placental hyperplasia in cloned cattle through up-regulation of CCND1-CDK6 expression,facilitating cell cycle progression.These findings offer potential avenues for enhancing cloning efficiency,with implications for evolutionary biology and the conservation of valuable germplasm resources.展开更多
植物运输抑制响应蛋白1(transport inhibitor protein 1,TIR1)是一种生长素受体,在生长素信号转导过程中发挥重要作用。目前有关马铃薯生长素受体基因的研究尚未见报道。本研究是从前期工作中获得EST序列,利用NCBI中的BLAST工具,对原始...植物运输抑制响应蛋白1(transport inhibitor protein 1,TIR1)是一种生长素受体,在生长素信号转导过程中发挥重要作用。目前有关马铃薯生长素受体基因的研究尚未见报道。本研究是从前期工作中获得EST序列,利用NCBI中的BLAST工具,对原始序列进行同源性分析,选择与原始序列相似度最高的一条同源序列。通过电子克隆技术,得到TIR1基因。采用生物信息学方法对基因序列、编码氨基酸、理化性质、跨膜结构域、亲/疏水性、信号肽、磷酸化位点、N-端糖基化位点、高级结构等进行分析。结果表明,TIR1基因cDNA全长1740 bp,编码580个氨基酸,分子量64.68 kD,理论等电点5.60,亲水性系数0.034,不稳定系数56.67,为不稳定酸性疏水性蛋白,编码蛋白含有59个磷酸化位点,4个N-端糖基化位点,无信号肽、跨膜区域,高级结构主要由α螺旋和无规卷曲序列组成。通过系统进化分析,TIR1蛋白与烟草、番茄和拟南芥等植物中的TIR1蛋白同源性较高。亚细胞定位分析,TIR1基因主要位于细胞核,小部分可能位于线粒体。研究结果为马铃薯TIR1基因的试验克隆及功能验证研究提供了理论依据。展开更多
基金supported by the National Natural Science Foundation of China (32060755)Natural Science Foundation of Inner Mongolia (2024MS03001)+7 种基金Inner Mongolia Autonomous Region Open Competition Projects (2022JBGS0018)Program for Young Talents of Science and Technology in Universities of Inner Mongolia Autonomous Region (NJYT23090)Inner Mongolia Autonomous Region Science and Technology Leading Team (2022LJRC0006)Inner Mongolia Autonomous Region Science and Technology Major Project (2021ZD0009)Major Agricultural Science and Technology Project of the Ministry of Agriculture and Rural Affairs (NK2022130203)Central Government Guides Local Science and Technology Development Funds (2022ZY0212)Inner Mongolia Autonomous Region High-level Talent Support ProgramInner Mongolia University Chief Scientist Program。
文摘Somatic cell nuclear transfer(SCNT)has been successfully employed across various mammalian species,yet cloned animals consistently exhibit low pregnancy rates,primarily due to placental abnormalities such as hyperplasia and hypertrophy.This study investigated the involvement of the Hippo signaling pathway in aberrant placentaldevelopmentinSCNT-inducedbovine pregnancies.SCNT-derived cattle exhibited placental hypertrophy,including enlarged abdominal circumference and altered placental cotyledon morphology.RNA sequencing analysis indicated significant dysregulation of Hippo signaling pathway genes in SCNT placentas.Coexpression of YAP1 and CCND1 was observed in cloned blastocysts,placental tissues,and bovine placental mesenchymal stem cells(bPMSCs).Manipulation of YAP1expression demonstrated the capacity to regulate bPMSC proliferation.Experimental assays confirmed the direct binding of YAP1 to CCND1,which subsequently promoted CCND1 expression in bPMSCs.Furthermore,inhibition of CDK6,a downstream target of CCND1,attenuated SCNT bPMSC proliferation.This study identified YAP1 as a key regulatory component within the Hippo signaling pathway that drives placental hyperplasia in cloned cattle through up-regulation of CCND1-CDK6 expression,facilitating cell cycle progression.These findings offer potential avenues for enhancing cloning efficiency,with implications for evolutionary biology and the conservation of valuable germplasm resources.
