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东亚三角涡虫DjPLCβ4基因克隆及在胚胎发育中的组织特异性表达
Molecular Cloning and Expression Pattern of the DjPLCβ4 Gene in Dugesia japonica During Embryonic Development
甄辉, 赵博生*
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DOI:10.11984/j.issn.1000-7083.20190414
作者单位:山东理工大学发育与进化生物学研究所, 山东淄博 255000
中文关键字:东亚三角涡虫;DjPLCβ4基因;整体原位杂交技术;胚胎发育
英文关键字:Dugesia japonica; DjPLCβ4 gene; whole-mount in situ hybridization; embryonic development
中文摘要:磷脂酶C的β亚型(PLCβ)在突触传递和可塑性的调节中发挥着重要作用。东亚三角涡虫Dugesia japonica作为首次出现三胚层的两侧对称动物,是研究发育和进化的重要模式生物。为了研究DjPLCβ4基因在东亚三角涡虫胚胎发育不同阶段的表达,通过RT-PCR和RACE技术克隆并验证了DjPLCβ4 cDNA的功能与表达图谱。结果显示,DjPLCβ4基因全长3 245 bp,编码969个氨基酸。系统进化树显示,东亚三角涡虫与其他物种的PLCβ4亚型聚群,并位于两侧对称动物的基部,可能是PLCβ4的原始类型。通过整体原位杂交技术研究DjPLCβ4基因在胚胎中的时空表达,DjPLCβ4基因最开始表达于胚胎发育的第3阶段,阳性信号主要位于胚胎咽及胚带;随着发育,胚胎孵化出幼虫,DjPLCβ4基因主要表达于神经原基和中枢神经系统。综上所述,东亚三角涡虫DjPLCβ4基因参与胚胎神经原基的形成和分化。
英文摘要:The β isoform of phospholipase C (PLCβ) is crucial for the modulation of synaptic transmission and plasticity. As a bilaterally symmetrical animal with the earliest emergence of three germ layers, the planarian (Dugesia japonica) is an important model organism for studying the mechanisms of development and evolution. To explore the expression the DjPLCβ4 gene during the embryonic development, the DjPLCβ4 cDNA of D. japonica was obtained by RACE and RT-PCR. The expression pattern of DjPLCβ4 gene was detected by whole-mount in situ hybridization. The results demonstrated that the DjPLCβ4 gene was 3 245 bp in length and encoding 969 amino acids. The result of phylogenetic analysis showed that the planarian DjPLCβ4 might be the archetype of the bilateral species. DjPLCβ4 gene expressed in the embryonic pharynx and germ band at the stage 3 of embryonic development. Along with the embryonic development, the number of DjPLCβ4-positive embryonic cells was increased and mainly distributed in the neurons blastema, the nervous system regions and the brain of the fully hatched larvae and intact animals. These results suggested that the DjPLCβ4 gene participates in the formation and differentiation of the neural primordium during the embryonic development of planarian.
2020,39(3): 249-257 收稿日期:2019-12-03
分类号:Q959.151;Q78
基金项目:国家自然科学基金项目(31970430)
作者简介:甄辉,博士研究生,研究方向:发育生物学,E-mail:zhenhui1013@126.com
*通信作者:赵博生,教授,博导,E-mail:zhaobosheng@sdut.edu.cn
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