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针对软体动物小的心脏活性肽SCPB的单克隆抗体:不同无脊椎动物中神经元的免疫标记

Monoclonal antibodies to the molluscan small cardioactive peptide SCPB: immunolabeling of neurons in diverse invertebrates.

作者信息

Masinovsky B, Kempf S C, Callaway J C, Willows A O

机构信息

Department of Zoology, University of Washington, Seattle 98195.

出版信息

J Comp Neurol. 1988 Jul 22;273(4):500-12. doi: 10.1002/cne.902730406.

Abstract

We reported a development of murine monoclonal antibodies to a molluscan small cardioactive peptide (SCPB) and their application to immunolabeling of neurons in several molluscan and arthropod species. In vitro stimulations of mouse lymphocytes with SCPB conjugated to a carrier protein yielded exclusively IgM class antibodies; in vivo stimulation resulted in generation of both IgM and IgG classes of antibodies. Monoclonal antibodies of the IgM class labeled identified SCP-containing neuron B11 in the frozen sections of the buccal ganglia of Tritonia diomedia. These antibodies failed to stain any neurons in whole mount preparations. A monoclonal antibody of IgG1 subclass selectively labeled neurons in both frozen sections and whole mount preparations of diverse invertebrate species. Thus, neurons B11, B12, and GE1 and several other neurons of the buccal and gastroesophageal ganglia of T. diomedia bound the antibody, and a similar pattern of immunolabeling was found in the closely related gastropod Tritonia festiva. We also observed SCPB-like immunoreactivity in the central neurons of other nudibranch and pulmonate molluscs and in examples of insect (Acheta domesticus and Tehrmobia domestica) and crustacean (Semibalanus cariosus) classes of the Arthropoda. Our results suggest a specific pattern of distribution of SCPB-like immunoreactivity in the gastropod nervous system and a broad occurrence of SCPB-like antigenicity in the diverse invertebrates.

摘要

我们报道了针对一种软体动物小活性心脏肽(SCPB)的鼠单克隆抗体的研制及其在几种软体动物和节肢动物物种神经元免疫标记中的应用。用与载体蛋白偶联的SCPB体外刺激小鼠淋巴细胞仅产生IgM类抗体;体内刺激则导致产生IgM和IgG两类抗体。IgM类单克隆抗体在多角海蛞蝓颊神经节的冰冻切片中标出了已确认的含SCP的神经元B11。这些抗体在整装标本中未能标记任何神经元。IgG1亚类的一种单克隆抗体在不同无脊椎动物物种的冰冻切片和整装标本中都能选择性地标记神经元。因此,多角海蛞蝓的神经元B11、B12和GE1以及颊神经节和胃食管神经节的其他几个神经元与该抗体结合,并且在亲缘关系相近的腹足纲动物festiva多角海蛞蝓中也发现了类似的免疫标记模式。我们还在其他裸鳃亚目和肺螺亚纲软体动物的中枢神经元以及节肢动物门昆虫纲(家蟋蟀和家白蚁)和甲壳纲(粗糙藤壶)的实例中观察到了SCPB样免疫反应性。我们的结果表明SCPB样免疫反应性在腹足纲动物神经系统中有特定的分布模式,并且SCPB样抗原性在多种无脊椎动物中广泛存在。

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