Application of biotinylated oligonucleotide probes to the detection of pituitary hormone mRNA using Northern blot analysis,in situ hybridization at the light- and electron-microscope levels

Application of biotinylated oligonucleotide probes to the detection of pituitary hormone mRNA using Northern blot analysis,in situ hybridization at the light- and electron-microscope levels
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生物素化寡核苷酸探针在光镜和电镜水平的Northern印迹分析、原位杂交检测垂体激素mRNA中的应用

DOI:
10.1007/bf02388634
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发表时间:
2004
期刊:
The Histochemical Journal
影响因子:
--
通讯作者:
R. Lloyd
R. Lloyd
中科院分区:
--
文献类型:
--
作者:
A. Matsuno;A. Teramoto;S. Takekoshi;H. Utsunomiya;Y. Ohsugi;S. Kishikawa;R. Osamura;T. Kirino;R. Lloyd

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摘要放射性同位素原位杂交(ISH)在正常垂体和垂体腺瘤中证实了垂体激素mRNAs的表达。最近的研究表明,与放射性同位素方法相比,非放射性同位素ISH具有几个优点。利用生物素标记的寡核苷酸探针ISH,我们已经能够在大鼠正常脑垂体腺和人垂体腺瘤的石蜡或冰冻切片中定位各种垂体激素mRNAs。在对照研究中,我们使用带有正义探针的ISH,没有探针的ISH,用核糖核酸酶预处理,用探针进行β-肌动蛋白和Northern杂交的ISH。用生物素化的探针,通过Northern杂交检测大鼠生长激素和催乳素的基因转录本。同样的生物素化探针不仅用于光学显微镜ISH,还用于大鼠生长激素和催乳素mRNAs在粗面内质网多聚体上的电子显微镜演示。生物素化寡核苷酸探针适用于Northern杂交和ISH三种杂交方法,在光镜和电子显微镜水平上均可用于垂体内分泌功能的分析。
SummaryThere have been many reports on radioisotopicin situ hybridization (ISH) studies for the demonstration of pituitary hormone mRNAs in normal pituitary gland and pituitary adenomas. Recent studies have revealed that non-radioisotopic ISH has several advantages over the radioisotopic method. Using ISH with biotinylated oligonucleotide probes, we have been able to localize various pituitary hormone mRNAs in paraffin wax or frozen sections of rat normal pituitary gland and human pituitary adenomas. For control studies we used ISH with sense probes, ISH without probes, pretreatment with ribonuclease, ISH with a probe for β-actin and Northern blot hybridization. Using biotinylated probes, gene transcripts of rat growth hormone and prolactin were detected by Northern blot hybridization. The same biotinylated probes were used not only for light microscope ISH but also for the electron microscopical demonstration of rat growth hormone and prolactin mRNAs on the polysomes of the rough endoplasmic reticula. It is emphasized that biotinylated oligonucleotide probes are useful for the analysis of pituitary endocrine function because they are applicable to the three hybridization methods, namely, Northern blot hybridization and ISH at the light and electron microscope levels.
嗜铬蛋白 A 和 B 是垂体和其他正常和肿瘤性人类内分泌组织中的信使核糖核酸。
DOI: --
发表时间: 1989
期刊: Laboratory investigation; a journal of technical methods and pathology
影响因子: --
作者:
Lloyd,RV;Iacangelo,A;Eiden,LE;Cano,M;Jin,L;Grimes,M
通讯作者: Grimes,M
通过原位杂交分析人垂体肿瘤。
DOI: 10.1016/s0344-0338(88)80009-8
发表时间: 1988
期刊: Pathology, research and practice
影响因子: --
作者:
Lloyd,RV
通讯作者: Lloyd,RV
使用生物素化寡核苷酸探针检测常规处理的组织切片中的信使 RNA。
DOI: 10.1093/ajcp/92.2.166
发表时间: 1989
影响因子: 3.5
作者:
Hankin,RC;Lloyd,RV
通讯作者: Lloyd,RV
通过原位杂交分析无效细胞腺瘤、嗜酸细胞瘤和促性腺激素腺瘤中的垂体激素和嗜铬粒蛋白 A mRNA。
DOI: --
发表时间: 1991
期刊: The American journal of pathology
影响因子: --
作者:
Lloyd,RV;Jin,L;Fields,K;Chandler,WF;Horvath,E;Stefaneanu,L;Kovacs,K
通讯作者: Kovacs,K
通过原位杂交分析内分泌活跃和临床沉默的促肾上腺皮质腺瘤。
DOI: --
发表时间: 1990
期刊: The American journal of pathology
影响因子: --
作者:
Lloyd,RV;Fields,K;Jin,L;Horvath,E;Kovacs,K
通讯作者: Kovacs,K