Vasopressin mRNA in situ hybridization: localization and regulation studied with oligonucleotide cDNA probes in normal and Brattleboro rat hypothalamus.
Vasopressin mRNA in situ hybridization: localization and regulation studied with oligonucleotide cDNA probes in normal and Brattleboro rat hypothalamus.
复制标题
加压素 mRNA 原位杂交：使用寡核苷酸 cDNA 探针在正常和布拉特尔伯勒大鼠下丘脑中研究定位和调节。
DOI：
10.1073/pnas.82.16.5555
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发表时间：
1985
影响因子：
11.1
通讯作者：
Habener,JF
中科院分区：
文献类型：
--
作者：
Uhl,GR;Zingg,HH;Habener,JF
可杂交的加压素mRNA可定量定位于大鼠下丘脑切片的原位。放射性标记的寡核苷酸cDNA探针，通过化学和酶的手段合成，提供强杂交区域已知含有加压素细胞体。多个单链32 P-、35 S-或3 H-标记的寡核苷酸显示随着探针从8个碱基延长至75个碱基而增加的局部杂交。竞争研究、RNase实验、解剖定位和使用多种探针都支持杂交特异性。在视上核和室旁核中的可杂交mRNA的近似加倍可以检测到动物脱水。可杂交的mRNA密度几乎是正常的Brattleboro大鼠下丘脑核团自由进入水。这些方法可以提供洞察区域mRNA的动态，并可能反映肽能神经元的功能活性。
Hybridizable vasopressin mRNA may be quantitatively localized in situ in sections from rat hypothalamus. Radiolabeled oligonucleotide cDNA probes, synthesized by chemical and enzymatic means, provide strong hybridization in zones known to contain vasopressin cell bodies. Multiple single-stranded 32P-, 35S-, or 3H-labeled oligonucleotides demonstrate localized hybridization that increases as probes are lengthened from 8 to 75 bases. Competition studies, RNase experiments, anatomic localization, and use of multiple probes all support hybridization specificity. An approximate doubling of hybridizable mRNA in both supraoptic and paraventricular nuclei can be detected with dehydration of the animals. Hybridizable mRNA densities are virtually normal in hypothalamic nuclei of Brattleboro rats given free access to water. These methods can provide insight into regional mRNA dynamics and may reflect functional activity of peptidergic neurons.