Sun B, Salvaterra P M
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, CA 91010, USA.
J Neurochem. 1995 Jul;65(1):434-43. doi: 10.1046/j.1471-4159.1995.65010434.x.
Antibodies to the plant glycoprotein horseradish peroxidase (HRP) are used extensively to identify neurons in Drosophila and other insects. We are interested in characterizing the gene product(s) recognized by anti-HRP antibodies because it may be important for nervous system function and/or development. Here we identify and purify from adult Drosophila heads an anti-HRP-reactive Mr 42K glycoprotein that is likely to be the major contributor to neuronal specific anti-HRP staining. Several different monoclonal antibodies to the purified 42K glycoprotein recognize up to three proteins with distinct mobilities between Mr 38K and 42K that vary as a function of developmental age. We have collectively named these components Nervana (nerve antigen), because the monoclonal antibodies also specifically stain cultured neurons and embryonic nervous system with a pattern indistinguishable from anti-HRP staining. Western blots indicate the presence of immunologically similar proteins in a wide variety of insect species and in nac (neurally altered carbohydrate) mutant Drosophila flies that lack anti-HRP staining in adult nervous system. It should now be possible to undertake a full biochemical and functional characterization of Nervana in Drosophila.
针对植物糖蛋白辣根过氧化物酶(HRP)的抗体被广泛用于鉴定果蝇和其他昆虫中的神经元。我们对表征抗HRP抗体识别的基因产物感兴趣,因为它可能对神经系统功能和/或发育很重要。在这里,我们从成年果蝇头部鉴定并纯化了一种抗HRP反应性的42K糖蛋白,它可能是神经元特异性抗HRP染色的主要贡献者。针对纯化的42K糖蛋白的几种不同单克隆抗体识别出多达三种迁移率不同的蛋白质,其分子量在38K到42K之间,随发育年龄而变化。我们将这些成分统称为Nervana(神经抗原),因为单克隆抗体也能特异性地对培养的神经元和胚胎神经系统进行染色,其染色模式与抗HRP染色无法区分。蛋白质印迹法表明,在多种昆虫物种以及在成年神经系统中缺乏抗HRP染色的nac(神经改变碳水化合物)突变果蝇中存在免疫相似的蛋白质。现在应该可以对果蝇中的Nervana进行全面的生化和功能表征。