SHH和β-catenin在牦牛蹄缘不同位点毛囊形态发生中的时空表达
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摘要
本研究用胚龄(Ed)45~248牦牛胚胎蹄缘皮肤为实验材料,Ed 55~275黄牛胚胎蹄缘皮肤作为对照,通过石蜡组织切片、HE染色和EnVison免疫组化法,研究牦牛蹄缘不同位点毛囊的形态发生过程中的组织形态学变化,并定性定量检测SHH(Sonic Hedgehog)和β-catenin在牦牛蹄缘不同位点毛囊形态发生中的时空表达,结果如下:
     1.牦牛蹄缘近蹄端毛囊的形态发生约开始于Ed 45,远蹄端为Ed 55,形成成熟毛囊的时间约为Ed 120;近蹄端至远蹄端蹄缘毛囊形态发生的时间、阶段、大小和下陷深度依次呈递减梯度,可见次级毛囊,毛囊均以毛囊群形式存在。
     2. SHH和β-catenin在牦牛蹄缘毛囊的表达时间与位点有关,信号波由近蹄端向远蹄端传递;但在黄牛的表达与位点无关,信号传递方式不同。
     3. SHH与β-catenin在牦牛与黄牛毛囊形态发生中的作用均与毛囊发育阶段相关,但牦牛的信号表达强度远高于黄牛。
     4.在牦牛蹄缘毛囊形态发生过程中,SHH对表皮的作用强于间充质。
     5.在牦牛蹄缘毛囊形态发生过程中,β-catenin对表皮作用强于间充质。
     结果表明,SHH和β-catenin信号在蹄缘毛囊形态发生过程的表皮-间充质相互作用中起着极为重要的作用。据推测,SHH负责发出信号指导细胞分化,决定细胞命运。β-catenin则主管维持细胞分化状态并大量增殖。SHH与β-catenin在毛囊形态发生中既有协同作用,也有各自的分工,提示SHH的表达时间、强度差异可能是决定近蹄端—远蹄端毛囊形态发生时间的关键因素;而β-catenin表达的强度差异可能是决定毛囊的大小梯度的关键因素;在毛囊形态发生过程中SHH与β-catenin对表皮—间充质相互作用调控以表皮为主,间充质为辅。
Ed(embryonic day)45~248 Yak embryos hoof periphery skin and Ed 55~275 the cattle’s were used as test and control material respectively to investigate hair follicle morphogenesis of different hoof periphery site, the paraffin embed tissue section was stained by H.E method for studying the histomorphogenesis changes and EnVision immunohistochemistry for detecting qualitatively and quantitatively temprocal-spatial expression of SHH andβ-catenin during hair follicle morphogenesis,Results is following:
     1. The hair follicle morphogenesis of yak proximal hoof end and distal hoof end initiate from Ed 45 and initiate from Ed 55 respectively, mature hair follicle in Ed 122. From the proximal hoof end to distal hoof end, the time ,the phase,the size and the growing depth of the hair follicle morphogenesis in turn occure degressive gradient. There are secondary hair follicles and hair follicle groups in Yak hoof periphery area.
     2. SHH andβ-catenin expression of hoof periphery hair follicle morphogenesis is related with different hoof periphery site, signaling wave is transmitted from the proximal hoof end to distal hoof end in yak. However, SHH andβ-catenin expression of cattle hoof periphery hair follicle morphogenesis is no related with different hoof periphery site, the signaling transmit pattern is different from that yak work.
     3. SHH andβ-catenin expression of hoof periphery hair follicle morphogenesis in yak and cattle embryos always is coupled with hair follicle morphogenesis phases, whereas the expression intensity of SHH andβ-catenin in yak is much higher than cattle’s.
     4. SHH expression during hair follicle morphogenesis show stronger effect on the hair follicle epidermis than the hair follicle mesenchyme..
     5.β-catenin expression during hair follicle morphogenesis show stronger effect on the hair follicle epidermis than the hair follicle mesenchyme.
     Our results suggest SHH andβ-catenin cooperate each other and respective function during the hoof periphery hair follicle morphogenesis .SHH is responsible for directing differentiation and determining cell fate ,β-catenin regulate differentiated cell proliferation .We speculate on SHH andβ-catenin expression and detail function difference play important role during hair follicle morphogenesis in different hoof periphery site of yak embryos. The epidermis dominate during the hair follicle morphogenesis and the mesenchyme signaling is secondary. Intensity difference ofβ-catenin expression in yak and cattle may act as key factor to determine the hair follicle size gradient.
引文
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