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首页> 外文期刊>Chinese journal of physiological sciences >EXPRESSION OF INTERMEDIATE FILAMENT NESTIN DURING MOUSE BRAIN DEVELOPMENT
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EXPRESSION OF INTERMEDIATE FILAMENT NESTIN DURING MOUSE BRAIN DEVELOPMENT

机译：小鼠脑发育过程中中间丝巢蛋白的表达

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摘要

Nestin represents a newly recognized class of intermediate filaments found in neurocpithelial stem cells of the developing rat brain. In order to search for a nestin ho-molog in mouse neural precursor cells, one peptide from the central region of ratnestin was synthesized, and a polyclonal anti-peptide antiserum was produced in the rabbit. Class G immunoglobulin (lgG), designated as anti-Nes-1, was purified and characterized by enzyme-linked immunosorbent assay (ELISA), Western blot, and immunohistochemistry. No significant cross-reactions with other intermediate filaments were detected by ELISA and Western blots. Antibody specificities were confirmed by solid-phase absorption to its corresponding peptide. Western blot showed that anti-Nes-1 identified a polypeptide with a molecular weight of 240-kD, identical to that of rat nestin, in the extracts of fetal and postnatal mouse brain and in the primary cultures of neural precursor cells. In tissue sections of fetal mouse neural tube, anti-Nes-1 immunostained the proximal and distal processes of radially oriented neural precursor cells. Primary cultures of these cells labeled with bromodeoxyuridine contained actively proliferating, anti-Nes-1 positive bipolar cells. Anti-Nes-1 staining of tissue sections and short-term cultures of spina! cord cells from mid-gestatiorial embryos labeled with bromodeoxyuridine in vivo were also consistent with this basic pattern. No correlation was found between the immune-reactive patterns of anti-Nes-1 and anti-rnicrotubule associated protein 2 (MAP2). This anti-peptide antibody may provide a new tool for exploring functional roles of nestin during differentiation of mouse neural precursor cells.
机译：巢蛋白代表在发展中的大鼠大脑的神经上皮干细胞中发现的一类新的中间丝。为了在小鼠神经前体细胞中寻找巢蛋白同源物，合成了来自大鼠肾上腺素中心区域的一种肽，并在兔中产生了多克隆抗肽抗血清。纯化的G类免疫球蛋白（lgG），称为抗Nes-1，通过酶联免疫吸附测定（ELISA），Western印迹和免疫组化进行表征。通过ELISA和Western印迹未检测到与其他中间丝的显着交叉反应。通过固相吸收到其相应的肽来确认抗体的特异性。蛋白质印迹表明，抗Nes-1在胎儿和出生后小鼠脑的提取物中以及在神经前体细胞的原代培养物中鉴定出与大鼠Nestin相同的分子量为240 kD的多肽。在胎儿小鼠神经管的组织切片中，抗Nes-1免疫染色了径向定向神经前体细胞的近端和远端过程。这些用溴脱氧尿苷标记的细胞的原代培养物中含有活跃增殖的抗Nes-1阳性双极细胞。组织切片和脊柱短期培养物的抗Nes-1染色！体内由溴脱氧尿苷标记的妊娠中期胚胎的脐带细胞也与此基本模式一致。在Nes-1和抗小管相关蛋白2（MAP2）的免疫反应模式之间未发现相关性。该抗肽抗体可能为探索巢蛋白在小鼠神经前体细胞分化过程中的功能性作用提供新工具。

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