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首页> 外文期刊>Journal of neuroendocrinology >Processing of frameshifted vasopressin precursors.
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Processing of frameshifted vasopressin precursors.

机译：框架加工型血管加压素前体。

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Biosynthesis of the vasopressin (VP) prohormone in magnocellular neurones of the hypothalamo-neurohypophysial system comprises endoplasmic reticulum (ER) transit, sorting into the regulated secretory pathway and subsequent processing in the individual proteins VP, neurophysin and a glycoprotein. These processes are severely disrupted in the homozygous diabetes insipidus (di/di) Brattleboro rat, which expresses a mutant VP precursor due to a single nucleotide deletion in the neurophysin region of the VP gene resulting in VP deficiency. Previous studies have shown the presence of additional frameshift mutations in VP transcripts, in solitary magnocellular neurones of the di/di rat due to a GA dinucleotide deletion resulting in two different mutant VP precursors with partly restored reading frame. Frameshifted VP precursors are also expressed in several magnocellular neurones in wild-type rats. In this study, we determined if the +1 frameshifted precursors from di/di and wild-type rats can lead to biosynthesis of the hormone VP. Therefore, eukaryotic expression plasmids containing the frameshifted VP cDNAs were transiently expressed in peptidergic tumour cell lines, and cells were analysed by reversed phase high-performance liquid chromatography and specific radioimmunoassays, and by immunofluoresence. Neuro2A neuroblastoma cells expressing the +1 frameshifted precursors of di/di rats retained products in the cell body. Only precursor or insignificant quantities of neurophysin-immunoreactive products were detected. In contrast, in AtT20 cells, frameshifted VP precursors were at least partly processed to yield the VP peptide, indicating that they have access to the regulated secretory pathway. Comparison between the two cell lines showed a very slow ER transit of the wild-type prohormone combined with inefficient processing in Neuro2A cells. The results show that mutant precursors can reach the regulated secretory pathway if ER transport is sufficiently rapid as in the case of AtT20 cells. This suggests that the di/di rat may regain the capacity to biosynthesize authentic VP through these +1 frameshifted precursors in magnocellular neurones.
机译：上下甲酰胺 - 神经内体系的甲型内甲素（VP）前型的生物合成，包括内质网（ER）转动，分类为调节分泌途径，随后在单个蛋白质VP，神经蛋白和糖蛋白中加工。这些方法在纯合糖尿病（DI / DI）Brorthboro大鼠中严重破坏，其由于VP基因的神经蛋白区域中的单个核苷酸缺失而表达突变VP前体，导致VP缺乏症。以前的研究表明，由于GA二核苷酸缺失，在DI / DI大鼠的孤立甲型甲状腺细胞神经元中存在额外的帧突变突变，​​其由于GA二核苷酸缺失导致具有部分恢复的读取框架的两种不同的突变VP前体。框架vp前体也以野生型大鼠的几种甲型粒状神经元表达。在这项研究中，我们确定了来自DI / DI和野生型大鼠的+1帧的前体是否可以导致激素VP的生物合成。因此，含有框架的VP CDNA的真核表达质粒在PeTimergic肿瘤细胞系中瞬时表达，并通过反相高效液相色谱和特异性放射线测量和通过免疫脉冲来分析细胞。 Neuro2A神经母细胞瘤细胞表达+1帧框架前体的DI / DI大鼠保留在细胞体中的产物。仅检测到唯一的神经素 - 免疫反应产物的前体或微不足道的量。相反，在ATT20细胞中，至少部分地将帧阈值的VP前体至少部分加工以产生VP肽，表明它们可以进入受调节的分泌途径。两种细胞系之间的比较显示了野生型前夸醌的易动转移，与神经2A细胞中的低效加工结合。结果表明，如果在ATT20细胞的情况下，如果ER传输足够快地，突变前体可以到达调节的分泌途径。这表明DI / DI RAT可以通过磁细胞神经元中的这些+1架构前体来重新获得生物合成的能力。

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