IMMUNOLOGIC STUDY OF GERM GRANULES IN C. ELEGANS EMBRY0S

早期胚胎发育过程中不对称性的建立和 不同发育信息在不同胚胎中的分布 细胞是每个多细胞生物发育的关键过程 有机体 然而，胚胎信息分子的性质和 支配其差别分布的机制尚不清楚。 这项建议描绘了一个结合免疫学，遗传学和生物化学 探讨了线虫C.优雅 一组 针对生殖系特异性颗粒的单克隆抗体（P 颗粒）已经生成。 为了应对不对称的 在胚胎中，这些颗粒被专门分离到生殖系或P 在早期胚胎发育过程中。 P颗粒作为一种优良的 用于研究对称分布的机制的标记物 细胞质成分到特定的胚胎细胞。 此外，他们可能 决定生殖细胞的命运。 抗P颗粒抗体 将与分子、细胞和遗传学相结合， 技术来解决以下问题：1）什么是P的性质 颗粒？ 抗体将用于纯化P颗粒，鉴定 它们的多肽成分，并确定它们是否含有核酸 酸 2)P颗粒是生殖细胞发育所必需的吗？如果是，是如何必需的 它们是否起作用？ 抗体将用于特异性恢复和 表征P-颗粒突变体。 抗体也将被显微注射到 活的蠕虫和胚胎来尝试功能性地包裹P颗粒。 第三章 P-granule基因表达是如何调控的？ 抗体将用于 克隆编码P-颗粒多肽的基因，克隆的序列 将用于研究基因的组织和转录。 四、 P颗粒如何分离到种系中？ 因为肌动蛋白丝 似乎参与P-颗粒分离，肌动蛋白的组织 纤维似乎参与P-颗粒偏析，组织 肌动蛋白丝在C.线虫胚胎及肌动蛋白的作用 将检查对颗粒分离的抑制剂。 长期目标是 提出的实验方法是为了更好地了解细胞的命运， 在胚胎发育的早期就被指定了。

The establishment of asymmetry during early embryogenesis and the distribution of different developmental information to different embryonic cells are crucial processes in the development of every multicellular organism. Yet the nature of embryonic informational molecules and the mechanisms governing their differential distribution are not understood. This proposal delineates a combined immunologic, genetic, and biochemical approach to studying these problems in the nematode C. elegans. A set of monoclonal antibodies directed against germ-line-specific granules (P granules) has already been generated. In response to the asymmetry of the embryo, these granules are segregated specifically to the germ line or P lineage during early embryogenesis. P granules serve as an excellent marker for studying the mechanism of a symmetric distribution of cytoplasmic components to specific embryonic cells. Furthermore, they may act to specify germ-cell fates. The antibodies directed against P granules will be used in combination with molecular, cellular, and genetic techniques to address the following questions: 1) What is the nature of P granules? The antibodies will be utilized to purify P granules, identify their polypeptide components, and determine whether they contain nucleic acid. 2) Are P granules required for germ-cell development and, if so, how do they function? The antibodies will be used to specifically recover and characterize P-granule mutants. Antibodies will also be microinjected into living worms and embryos to try to functionally inactivate P granules. 3) How is P-granule gene expression regulated? The antibodies will be used to clone the genes encoding P-granule polypeptides, and the cloned sequences will be used to study the organization and transcription of the genes. 4) How are P granules segregated to the germ lineage? Because actin filaments appear to be involved in P-granule segregation, the organization of actin filaments appear to be involved in P-granule segregation, the organization of actin filaments in C. elegans embryos and the effects of actin inhibitors on granule segregation will be examined. The long range goal of the proposed experimental approach is to better understand how cell fates are specified early in embryonic development.