Expression of substance P/neurokinin A-encoding preprotachykinin messenger ribonucleic acids in the rat enteric nervous system.
Expression of substance P/neurokinin A-encoding preprotachykinin messenger ribonucleic acids in the rat enteric nervous system.
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DOI：
10.1016/0016-5085(89)90070-x
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发表时间：
1989-08
期刊：
影响因子：
29.4
通讯作者：
C. Sternini;K. Anderson;G. Frantz;J. Krause;N. Brecha
中科院分区：
文献类型：
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作者：
C. Sternini;K. Anderson;G. Frantz;J. Krause;N. Brecha
用35 S或3 H标记的单链核糖核酸（RNA）探针原位杂交组织化学方法研究了大鼠肠神经系统中编码P物质/神经激肽A的前速激肽原mRNA的细胞定位。P物质/神经激肽A编码的mRNA在食管和胃的肌间神经丛内的神经元中表达，在后者中数量更多，并且在分布于肠的肌间神经丛和粘膜下神经丛的神经节细胞中表达。杂交的特异性通过在与正义RNA探针孵育或在杂交前用核糖核酸酶A预处理的切片中缺乏高于背景的特异性信号来证明。核糖核酸印迹杂交分析的RNA提取物从肌肉层肌间神经丛和粘膜下层制剂的十二指肠表现出一个单一的带的杂交在1.3 kb。溶液杂交-核酸酶保护分析显示，在这些RNA提取物中存在多种前速激肽原编码转录本，其丰度水平为γ-mRNA > β-mRNA >> α-mRNA，这与在大鼠脑中观察到的相似。我们的研究结果表明，编码速激肽肽，P物质和神经激肽A的preprotachykinin基因，在肠道神经元，具有区域分布的人口中转录与以前描述的速激肽样免疫反应神经元，这表明特定的mRNA和postperfectionally处理的肽被定位在相同的结构。
The cellular localization of substance P/neurokinin A-encoding preprotachykinin mRNAs in the rat enteric nervous system was studied by means of in situ hybridization histochemistry using35S- or3H-labeled single-stranded ribonucleic acid (RNA) probes which recognize all three preprotachykinin mRNA species, α, β, and γ. Substance P/neurokinin A-encoding mRNAs are expressed in neurons within the myenteric plexus of the esophagus and stomach, being more numerous in the latter, and in ganglion cells distributed to both the myenteric and submucosal plexuses of the intestine. Specificity of the hybridization was demonstrated by the lack of specific signal above background in sections incubated with a sense RNA probe or pretreated with ribonuclease A before hybridization. Ribonucleic acid blot hybridization analysis of RNA extracts from both the muscle layer-myenteric plexus and submucosal layer preparations of the duodenum demonstrated a single band of hybridization at 1.3 kb. Solution hybridization-nuclease protection assays showed multiple preprotachykinin-encoding transcripts in these RNA extracts, with an abundance level of γ-mRNA > β-mRNA >> α-mRNA, which is similar to that observed in the rat brain. Our results indicate that the preprotachykinin gene encoding the tachykinin peptides, substance P and neurokinin A, is transcribed in a population of enteric neurons that have a regional distribution comparable to the previously described tachykinin-like immunoreactive neurons, suggesting that specific mRNAs and the posttranslationally processed peptides are localized in the same structures.