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用生物素化寡核苷酸探针原位杂交法研究雌激素和溴隐亭处理后雌性大鼠垂体细胞中催乳素和生长激素mRNA的超微结构分布变化。

Changes in the ultrastructural distribution of prolactin and growth hormone mRNAs in pituitary cells of female rats after estrogen and bromocriptine treatment, studied using in situ hybridization with biotinylated oligonucleotide probes.

作者信息

Matsuno A, Ohsugi Y, Utsunomiya H, Takekoshi S, Sanno N, Osamura R Y, Watanabe K, Teramoto A, Kirino T

机构信息

Department of Neurosurgery, University of Tokyo Hospital, Japan.

出版信息

Histochem Cell Biol. 1995 Jul;104(1):37-45. doi: 10.1007/BF01464784.

Abstract

The expression and distribution of prolactin (PRL) mRNA and their alterations induced by estrogen and bromocriptine were investigated using non-radioisotopic in situ hybridization (ISH) at the electron microscopic (EM) level. Our EM-ISH studies using biotinylated oligonucleotide probes showed that estrogen induced whirling changes of the rough endoplasmic reticulum (RER) of female rat PRL cells and increased transcription of PRL genes located on the polysomes of the whirling RER. The presence of mammosomatotroph cells in the rat pituitary gland was also verified in our EM-ISH studies. After bromocriptine administration, PRL cells contained many secretory granules due to the inhibition of secretion. Pre- and post-embedding EM-ISH and northern hybridization studies revealed that bromocriptine induced the distorted, vesiculated, and dilated RER, and also the suppressed PRL mRNA expression. The activity of protein kinase C (PKC), which mediates PRL gene expression, tended to be elevated by estrogen and suppressed by bromocriptine. Therefore, it is considered that the ultrastructural and quantitative changes in PRL mRNA expression evoked by estrogen and bromocriptine may be mediated by the intracellular signal transduction system, including PKC.

摘要

采用非放射性原位杂交(ISH)技术,在电子显微镜(EM)水平上研究催乳素(PRL)mRNA的表达、分布及其受雌激素和溴隐亭诱导后的变化。我们使用生物素化寡核苷酸探针进行的EM-ISH研究表明,雌激素可诱导雌性大鼠PRL细胞粗面内质网(RER)发生漩涡状变化,并增加位于漩涡状RER多聚体上的PRL基因的转录。我们的EM-ISH研究还证实了大鼠垂体中存在乳腺生长激素细胞。给予溴隐亭后,由于分泌受到抑制,PRL细胞含有许多分泌颗粒。包埋前和包埋后的EM-ISH及Northern杂交研究显示,溴隐亭可诱导RER扭曲、形成囊泡并扩张,同时也抑制PRL mRNA的表达。介导PRL基因表达的蛋白激酶C(PKC)的活性,倾向于被雌激素升高而被溴隐亭抑制。因此,可以认为雌激素和溴隐亭引起的PRL mRNA表达的超微结构和定量变化可能由包括PKC在内的细胞内信号转导系统介导。

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