摘要
研究cPouV和cNanog基因在鸡早期胚胎中的时空表达规律,探索鸡胚早期发育过程中胚胎干细胞的增殖分化规律,为进一步研究2基因在鸡胚早期发育中的作用奠定基础。本试验分别构建了pMD-18T-cPouV和pMD-18T-cNanog重组质粒,制备了地高辛(DIG)标记cPouV和cNanog基因的cDNA探针,采用原位杂交的方法检测cPouV和cNanog在鸡早期胚胎中的表达模式。结果表明:在原条发生前,cPouV和cNanog基因均匀表达于明区。原条期,cPouV基因在原条及其周围表达,cNanog基因则表达于原条周围,原条上未见表达。随着神经板的形成,2基因大量表达于神经板、神经管和脑部周围。待鸡胚发育至14HH阶段,2基因主要集中表达于头部和心等血管富集处,在外周血管中也能检测到其表达信号。综上表明:随着胚胎的发育,能够表达cPouV和cNanog基因的多能性干细胞逐渐由中央明区向外周迁移。
Spatio-temporal expression of cNanogand cPouVgenes in the early chicken embryos was studied to provide basic information for studing functions of those two genes on chicken embryonic stem cells(CES)self-renewal and differentiation.Digoxigenin(DIG)-labeled cPouVand cNanogprobes were produced by cDNA synthesization from the plasmids of pMD-18T-cPouVand pMD-18T-cNanog respectively according to a standard protocol.The expression patterns of cPouVand cNanogin the early chicken embryos were detected using in situ hybridization with the synthesized probes.The results showed that at pre-primitive streak stage,cPouVand cNanog genes were uniformly expressed in the pellucid area of embryos.At primitive streak stage,cPouV genes were strongly expressed in primitive streak and surrounding area.Hybridization signals of cNanoggenes were detected only in surrounding area of primitive streak,no signals were detected inside streak.As the neural plate formed,two genes were strongly expressed in the neural plate, neuraltube and the anterior hindbrain/posterior midbrain.Later,at stage 14HH and subsequently,two genes were strongly expressed in the head,the heart and peripheral vessels.As chickenembryo development,the pluripotent stem cells which can express cPouVand cNanoggenes migrated from pellucid area to peripheral area.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2014年第5期733-739,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(31072101
31201871)
广东省自然科学基金(S201101004876
S2012010010526)
