Vannini Laura, Willis Judith H
Department of Cellular Biology, University of Georgia, Athens, GA 30602, USA.
Department of Cellular Biology, University of Georgia, Athens, GA 30602, USA.
Arthropod Struct Dev. 2016 Nov;45(6):519-535. doi: 10.1016/j.asd.2016.10.006. Epub 2016 Nov 4.
Previous work with EM immunolocalization examined the intracuticular placement of several antibodies directed against cuticular proteins (CPs) in various structures of Anopheles gambiae. Those structures had long stretches of fairly uniform cuticle. We have now used 19 antibodies directed against members of five CP families on two adult structures with considerable complexity, Johnston's organ and the corneal lens of the compound eye. We also localized chitin with colloidal-gold labeled wheat germ agglutinin. Twelve of these antibodies recognized structures in Johnston's organ. Only 6 were detected in the outer pedicel wall, but the internal structures were more complex with distinct distributions of members of the five CP families in six different structures. The corneal lens had four distinct regions of laminar cuticle. Thirteen of the 15 members of the CPR family were detected, none from the other CP families. Specific antibodies were localized to different regions and in different laminae within a region. The specificity of deployment of cuticular proteins revealed in this study is helping to explain why An. gambiae allocates about 2% of its protein coding genes to structural CPs.
之前关于免疫电镜定位的研究,检测了针对冈比亚按蚊不同结构中几种表皮蛋白(CPs)的抗体在表皮内的定位情况。这些结构具有很长一段相当均匀的表皮。我们现在使用了针对五个CP家族成员的19种抗体,作用于两个具有相当复杂性的成虫结构,即江氏器和复眼的角膜晶状体。我们还用胶体金标记的麦胚凝集素对几丁质进行了定位。其中12种抗体识别出了江氏器中的结构。在外柄壁中仅检测到6种,但内部结构更为复杂,五个CP家族的成员在六种不同结构中有不同的分布。角膜晶状体有四个不同的层状表皮区域。检测到了CPR家族15个成员中的13个,其他CP家族未检测到任何成员。特定抗体定位于不同区域以及区域内的不同层中。本研究中揭示的表皮蛋白分布特异性,有助于解释为什么冈比亚按蚊将其约2%的蛋白质编码基因分配给结构CPs。