MOSQUITE OOCYTES--MECHANISM OF PROTEIN ACCUMULATION

蚊子卵母细胞——蛋白质积累机制

• 批准号: 3322925
• 负责人: ALEXANDER SIMEON RAIKHEL
• 金额: $ 8.21万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-07-01 至 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3322925
• 关键词: Culicidae; antibody formation; arthropod borne communicable disease; binding proteins; clathrin; egg /ovum; egg proteins; egg yolk; genetic library; hormone regulation /control mechanism; ligands; monoclonal antibody; nucleic acid probes; oogenesis; phagocytosis; pinocytosis; radiotracer; vitellin

The specific accumulation of yolk proteins by the oocyte is a cornerstone of egg maturation and, therefore, a key element of reproductive physiology. The research proposed here concerns the elucidation of the molecular mechanism governing this process. We have generated a library of monoclonal antibodies against yolk protein at different processing stages in the mosquito oocyte. This gives us a powerful tool to analyse the mechanism of yolk accumulation and develop specific molecular probes. Purified vitellogenin (VG) will be labeled with radio-, fluorescent, and EM markers, and then used for binding and structural studies of its receptor. I intend to investigate the role of hormones and other factors in yolk accumulation. Radioligand binding assay will be used to determine the affinity and number of VG receptors on the oocyte membrane as well as the influence of various factors on these characteristics. I will study the nature of VG epitopes responsible for its recognition by the oocyte. Also, I propose to develop a probe specific for mosquito clathrin, a major component of coated vesicles, and then to study the developmental aspects of their biogenesis. I wish to determine: (1) when both VG receptors and clathrin are synthesized, and (2) what factors govern their production. An understanding of the VG-receptor system will set the stage for our next goal, purification and study of the molecular properties of the VG receptor. Information on the nature of VG recognition and its accumulation by the oocyte, as well as factors affecting this process, will provide an essential background in the search for new approaches in the control of this important arthropod vector.

卵母细胞对卵黄蛋白的特异性积累是一个重要的过程。 卵子成熟的基石，因此， 生殖生理学 这里提出的研究涉及 阐明了控制这一过程的分子机制。 我们已经建立了一个抗卵黄单克隆抗体库 蛋白质在蚊子卵母细胞的不同加工阶段。 这为我们分析蛋黄的作用机理提供了有力的工具 积累并开发特异性分子探针。 纯化 卵黄蛋白原（VG）将用放射性、荧光和EM标记 标记，然后用于其结合和结构研究， 受体的 我打算研究激素和其他因素在 卵黄积累 放射性配体结合试验将用于 确定卵母细胞上VG受体的亲和力和数量 膜以及各种因素对这些的影响 特色 我将研究VG表位的性质， 被卵母细胞识别。 此外，我建议制定一项 蚊子网格蛋白的特异性探针， 囊泡，然后研究它们的发育方面， 生物起源 我想确定：（1）当VG受体和 网格蛋白是合成的，（2）什么因素控制着它们 生产 对VG受体系统的理解将为以下研究奠定基础： 我们的下一个目标，纯化和分子性质的研究 VG受体。 关于确认脆弱商品性质的信息 及其在卵母细胞中的积累，以及影响 这一过程将为寻找 控制这种重要节肢动物媒介的新方法。