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鸡血管活性肠肽RNA探针制备及其在鸡肠Remak神经的原位杂交反应

DOI: 10.7685/j.issn.1000-2030.2007.02.023, PP. 111-115

Keywords: 鸡肠Remak神经,血管活性肠肽(VIP),原位杂交,RNA探针

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Abstract:

应用RT-PCR方法从鸡脑中扩增血管活性肠肽(VIP)基因片段,将其连接于pGM-Teasy。分别利用pGM-Teasy中T7和SP6启动子及其RNA聚合酶,以线性化的VIP/pGM-Teasy为模板转录合成正、反义DIG标记RNA探针。通过原位杂交方法将合成的探针用于探查VIP-mRNA在鸡肠Remak神经(INR)中的分布情况。结果表明,INR的神经元胞体中有VIP-mRNA的转录,且这种神经元数量众多;原位杂交阳性神经元胞体大部分是大型神经元细胞,呈圆形或椭圆形;阳性神经元胞体在INR神经节中呈层状或成群分布。INR的神经纤维呈弱阳性。在节间束也有少量的阳性细胞分布。本研究从基因水平证明INR中有VIP神经元存在。

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