【摘要】 目的探讨无翅型小鼠乳房肿瘤病毒整合位点家族[wingless-type mouse mammary tumor virus(MMTV)integration site family,wnt]成员wnt1、wnt2、wnt4、wnt5、wnt11A和wnt11B基因在细粒棘球绦虫(Echinococcus granulosus)原头节和成虫的m RNA差异转录及组织分布。方法用SYBR GreenⅠq RT-PCR方法检测wnt1、wnt2、wnt4、wnt5、wnt11A和wnt11B基因在细粒棘球绦虫原头节和成虫的m RNA相对转录情况。用RNA荧光探针以全量组织荧光原位杂交方法检测6个基因在原头节的组织分布情况。结果 q RT-PCR检测结果显示,wnt1和wnt2基因m RNA在原头节和成虫的相对转录水平分别为1.00、1.49和1.00、2.53,在成虫的相对转录水平分别为原头节的1.49倍(P>0.05)、2.53倍(P<0.05)。wnt4、wnt5、wnt11A和wnt11B m RNA在原头节和成虫的相对转录水平分别为1.00、0.04,1.00、0.03,1.00、0.07和1.00、0.97,在原头节的相对转录水平分别为成虫的25.00倍(P<0.01)、33.33倍(P<0.01)、14.29倍(P<0.01)和1.03倍(P>0.05),原头节和成虫间wnt4、wnt5、wnt11A表达量的差异有统计学意义,而wnt11B的差异则无统计学意义。全量组织荧光原位杂交结果显示,在原头节中,wnt1主要分布于表皮组织,wnt2主要分布于吸盘部位,wnt4主要分布于吸盘和顶突部位,wnt5、wnt11B主要分布于吸盘和表皮,wnt11A主要分布于顶突和小刺。结论 wnt2基因在细粒棘球绦虫成虫的m RNA相对转录水平高于原头节,wnt4、wnt5、wnt11A在原头节的m RNA相对转录水平高于成虫,6个wnt基因均分布于细粒棘球绦虫原头节的前端区域。更多还原

【Abstract】 Objective To investigate the differential mRNA expression and tissue distribution of wnt [wingless-type mouse mammary tumor virus( MMTV) integration site family, wnt ] gene members wnt1, wnt2, wnt4, wnt5,wnt11 A and wnt11 B in protoscoleces and adult worms of Echinococcus granulosus. Methods The mRNA expression of wnt1, wnt2, wnt4, wnt5, wnt11 A and wnt11 B was determined by qRT-PCR. Tissue distribution of wnt1, wnt2,wnt4, wnt5, wnt11 A and wnt11 B in Echinococcus granulosus protoscoleces was determined by the whole-mount in situ hybridization. Results The q RT-PCR results showed that the m RNA expression levels of wnt1 and wnt2 in the adult worms were 1.49(P >0.05) and 2.53 folds(P <0.05) of those in the protoscoleces, respectively. The mRNA expression levels of wnt4, wnt5, wnt11 A and wnt11 B in the protoscoleces were 25.00(P <0.01), 33.33(P <0.01), 14.29(P<0.01) and 1.03 folds(P>0.05) of those in the adult worms, respectively. In brief, there was no significant difference of m RNA expression in wnt2 and wnt11 B between protoscoleces and adult, but there was a significant difference of m RNA expression in wnt1, wnt4, wnt5 and wnt11 A between protoscoleces and adults. Results of the whole-mount in situ hybridization showed that in protoscoleces wnt1 was mainly localized in the epidermaltissue, wnt2 in suckers, wnt4 in suckers and rostellum, wnt5 and wnt11 B in suckers and epidermal tissue, and wnt11 A in rostellum and hooks. Conclusion The m RNA expression of wnt2 in adult E. granulosus was higher than that in protoscoleces, and the m RNA expression of wnt4, wnt5, wnt11 A and wnt11 B in protoscoleces was higher than that in the adult worms. The six wnt gene family members were all distributed in the forward region of protoscoleces.更多还原