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A subpopulation of cardiomyocytes expressing alpha-skeletal actin is identified by a specific polyclonal antibody.
通过特异性多克隆抗体鉴定表达 α-骨骼肌动蛋白的心肌细胞亚群。

摘要

The NH(2)-terminal decapeptide of alpha-skeletal actin that contains a primary sequence specific for this isoform was used to raise a polyclonal antibody in rabbits. Using sequential affinity chromatography, we recovered from serum antibodies reacting exclusively with alpha-skeletal actin when tested by immunoblotting and immunofluorescence. Epitope mapping by means of competition assays with synthetic peptides indicated that the acetyl group and the first 9 amino acids are essential for specificity. The monospecific antibody was then used to investigate the distribution of alpha-skeletal actin in the myocardium of newborn and normal or hypertensive (with or without fibrotic areas) adult rats. Immunostaining of normal heart revealed that alpha-skeletal actin is diffusely distributed within practically all myocardial fibers of the newborn rat, whereas it is restricted to a small proportion of adult rat cardiomyocytes, which appear intensely stained. A correlation, albeit not complete, was found between the distribution of alpha-skeletal actin and beta-myosin heavy chain. During cardiac hypertrophy induced by aortic ligature between the renal arteries, the expressions of alpha-skeletal actin mRNA and protein were increased. The distribution of immunostaining had a focal pattern similar to that of normal adult rats, reactive fibers being more numerous and more intensely stained compared with normal myocardium. Positive fibers were particularly abundant at the periphery of fibrotic areas. Using this antibody, we have demonstrated for the first time the differential distribution of alpha-skeletal actin in heart tissues. Changes in the distribution of this isoform in hypertrophic heart provide new insight into the mechanisms by which the heart adapts to work overload. This antibody will prove useful in exploring the mechanisms of expression of alpha-skeletal actin and in defining its role in physiological and pathological situations.

译文

Α-骨骼肌动蛋白的 NH (2)-末端十肽含有对该同种型特异的一级序列,用于在兔体内培养多克隆抗体。使用顺序亲和层析法,当通过免疫印迹和免疫荧光检测时,我们从血清抗体中回收了仅与 α-骨骼肌动蛋白反应的抗体。通过与合成肽的竞争试验进行抗原定位表明,乙酰基和前 9 个氨基酸对特异性至关重要。然后用单一特异性抗体研究 α-骨骼肌动蛋白在新生大鼠和正常或高血压 (有或无纤维化区域) 成年大鼠心肌中的分布。正常心脏的免疫染色显示,α-骨骼肌动蛋白几乎广泛分布在新生大鼠的所有心肌纤维中,而它仅限于一小部分成年大鼠心肌细胞, 看起来非常脏。发现 α-骨骼肌动蛋白和 β-肌球蛋白重链的分布之间存在相关性,尽管不完整。在主动脉结扎肾动脉引起的心肌肥厚中,α-骨骼肌动蛋白 mRNA 和蛋白的表达增加。免疫染色的分布与正常成年大鼠的病灶模式相似,与正常心肌相比,反应性纤维数量更多,染色更强烈。阳性纤维在纤维化区域的外围特别丰富。使用这种抗体,我们首次证明了 α-骨骼肌动蛋白在心脏组织中的差异分布。这种同种型在肥厚性心脏中分布的变化为心脏适应超负荷工作的机制提供了新的见解。该抗体将被证明在探索 α-骨骼肌动蛋白的表达机制和确定其在生理和病理情况中的作用方面是有用的。

polyclonal antibody

免疫 抗体 临床研究术语
概述  :  

多克隆抗体是指用一种包含多种抗原决定簇的抗原免疫动物,刺激机体所产生的具有不同抗原表位、不同抗体特异性的抗体,所产生的免疫血清实际上是含有多种抗体的混合物。   抗体制备 多克隆抗体主要从动物免疫血清、恢复期患者血清或免疫接种人群的血清中获得。   优点 多克隆抗体由于刺激抗原表位多,其特异性较弱,刺激抗原多为自然源性抗原,故其抗体稳定性较好,其制备过程中对免疫原纯度要求较高,免疫原纯度越高越好

Polyclonal [,pɑli'klonəl]

释义    adj. 多克隆的;多元性繁殖系的;多细胞系的

例句    Polyclonal antibody preparation against the peptides of NDRG family proteins.抗人NDRG家族寡肽的多克隆抗体制备。

 

Antibody 英 [’æntibɒdi] 美 [’æntibɑːdi]

释义    n. [免疫] 抗体

例句    Antibodies are very specific in that each antibody will recognize and bind to one protein region only.抗体具有非常特异性,每种抗体只识别和结合一种蛋白质区域。

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