Neurohypophyseal and fluid homeostasis in transgenic rats expressing a tagged rat vasopressin prepropeptide in hypothalamic neurons.
Neurohypophyseal and fluid homeostasis in transgenic rats expressing a tagged rat vasopressin prepropeptide in hypothalamic neurons.
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在下丘脑神经元中表达标记的大鼠加压素前原肽的转基因大鼠的神经垂体和液体稳态。
DOI：
10.1210/endo.137.11.8895381
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发表时间：
1996
期刊：
影响因子：
4.8
通讯作者：
D. Murphy
中科院分区：
文献类型：
--
作者：
S. Waller;K. Fairhall;J. Xu;I. Robinson;D. Murphy
我们已经开发出一种转基因系统，第一次，有利于监测的监管动态的中央肽能系统从神经肽基因的转录到存储和释放的成熟分泌产物。大鼠血管加压素（VP）转基因（5-VCAT-3）的表达仅限于大鼠下丘脑血管加压素能大细胞神经元，含有一个序列，如果翻译，将在VP前体的C-末端放置一个独特的十六肽（DRSAGYYGLFKDRKEK，缩写为DR-12-EK）。我们已经提出了一种针对这个“标签”的抗体，并使用免疫组织化学，电子显微镜，RIA和HPLC，已首次表明，VP转基因RNA被翻译成蛋白质产物中发现，在一个处理的形式，在分泌颗粒在后垂体的转基因大鼠。肽标签对VP前体C-末端的破坏耐受性良好，不会破坏VP产生或干扰盐和水平衡。渗透压刺激增加下丘脑DR-12-EK水平，但垂体后叶DR-12-EK水平的变化更为复杂。在盐加载5天后，DR-12-EK水平下降，如如果其释放与VP的释放协调所预期的。然而，10天的盐负荷后，垂体后叶DR-12-EK水平增加，尽管较低水平的VP。这可能反映了转基因在RNA水平上对渗透压挑战的更大响应，相对于来自内源性基因的那些，增加了转运到垂体后叶的含DR-12-EK的翻译产物的比例。在RNA和蛋白质水平的标记的转基因对渗透压挑战的夸张的反应提供了一个新的机会，在分子水平上研究VP系统的调控动力学，但在生理上有利的情况下的完整的动物。
We have developed a transgenic system that, for the first time, facilitates monitoring of the regulatory dynamics of a central peptidergic system from transcription of a neuropeptide gene to the storage and release of the mature secretory product. A rat vasopressin (VP) transgene (5-VCAT-3), the expression of which is restricted to hypothalamic vasopressinergic magnocellular neurons in rats, contains a sequence that, if translated, would place a unique hexadecapeptide (DRSAGYYGLFKDRKEK, abbreviated to DR-12-EK) at the C-terminus of the VP precursor. We have raised an antibody against this "tag" and, using immunohistochemistry, electron microscopy, RIA, and HPLC, have shown for the first time that a VP transgene RNA is translated into a protein product found, in a processed form, in secretory granules in the posterior pituitaries of transgenic rats. Disruption of the C-terminus of the VP precursor by the peptide tag is well tolerated and does not disrupt VP production or disturb salt and water balance. An osmotic stimulus increased hypothalamic DR-12-EK levels, but changes in posterior pituitary DR-12-EK levels were more complex. After 5 days of salt-loading, DR-12-EK levels fell, as would be expected if its release was coordinate with that of VP. However, after 10 days of salt-loading, posterior pituitary DR-12-EK levels increased, despite the lower level of VP. This probably reflects the greater response of the transgene to osmotic challenge at the RNA level, increasing the proportion of DR-12-EK-containing translation products transported to the posterior pituitary relative to those derived from the endogenous gene. The exaggerated response of the tagged transgene to osmotic challenge at both RNA and protein levels affords a new opportunity to study the regulatory dynamics of the VP system at the molecular level, but within the physiologically advantageous context of the intact animal.
转基因小鼠中大鼠精氨酸加压素基因的表达。
DOI：
10.1210/mend.7.5.8100353
发表时间：
1993
期刊：
Molecular endocrinology (Baltimore, Md.)
影响因子：
--
作者：
Grant,FD;Reventos,J;Gordon,JW;Kawabata,S;Miller,M;Majzoub,JA
通讯作者：
Majzoub,JA