Ultrastructural distribution of growth hormone and prolactin mRNAs in normal rat pituitary cells: a comparison between preembedding and postembedding methods
Ultrastructural distribution of growth hormone and prolactin mRNAs in normal rat pituitary cells: a comparison between preembedding and postembedding methods
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正常大鼠垂体细胞中生长激素和催乳素 mRNA 的超微结构分布：包埋前和包埋后方法的比较
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发表时间：
1994
期刊：
影响因子：
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通讯作者：
A. Teramoto
中科院分区：
文献类型：
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作者：
A. Matsuno;T. Kirino;Y. Ohsugi;H. Utsunomiya;S. Takekoshi;R. Osamura;K. Watanabe;A. Teramoto
在电镜水平的原位杂交（ISH）是必要的阐明mRNA在蛋白质合成中的细胞内分布和作用。我们描述了用生物素化寡核苷酸探针检测大鼠生长激素和催乳素mrna的电镜ISH方法，并比较了包埋前和包埋后的方法。预包埋电镜ISH将大鼠生长激素和催乳素mrna定位在粗内质网（RER）的多体上。大鼠生长激素mRNA在内质网呈弥漫性分布，而催乳素mRNA呈散在性、局灶性分布。因此，促乳素mRNA在内质网上可能有一个特定的翻译位点。采用链霉亲和金偶联物后包埋法，在内质网多聚体上识别大鼠生长激素mRNA信号。与预嵌入方法相比，后嵌入方法的杂交信号强度较低，非特异性信号较多。因此，从mRNA的实用性和保存角度来看，预嵌入方法似乎比后嵌入方法更容易和更好。电镜ISH被认为是评估mRNA在细胞内定位和RER上特异性激素合成位点的重要工具。
In situ hybridization (ISH) at the electron microscopic level is essential for elucidating the intracellular distribution and role of mRNA in protein synthesis. We describe our electron microscopic ISH method using biotinylated oligonucleotide probes for rat growth hormone and prolactin mRNAs and compare the preembedding method with the postembedding method. Preembedding electron microscopic ISH localized rat growth hormone and prolactin mRNAs on the polysomes of the rough endoplasmic reticulum (RER). Rat growth hormone mRNA was distributed diffusely on the RER, whereas rat prolactin mRNA was scattered and distributed focally. Thus there might be a specific translational site for prolactin mRNA on the RER. Rat growth hormone mRNA signals were also recognized on the polysomes of the RER, using the postembedding method with streptavidin gold conjugate. The hybridization signal intensity using the postembedding method was lower, and non-specific signals were more frequent, in comparison with the preembedding method. The preembedding method thus appears to be easier and better than the postembedding method from the viewpoint of utility and preservation of mRNA. Electron microscopic ISH is considered to be an important tool for evaluating the intracellular localization of mRNA and the site of specific hormone synthesis on the RER.
通过超微结构原位杂交可视化胚胎肌肉组织中肌球蛋白重链 mRNA 的分布。
影响因子：
2.7
作者：
Pomeroy,ME;Lawrence,JB;Singer,RH;Billings-Gagliardi,S
通讯作者：
Billings-Gagliardi,S