Mes-1 and Asymmetric Germline Divisions in C. elegans

描述（由申请人提供）：不对称划分是基本的 产生具有不同发育命运的子细胞的过程。 C.胚胎发生过程中生殖系的形成。elegans提供了一个理想的 系统来研究这个过程。生殖系是通过一系列 需要MES-1蛋白的不对称分裂。MES-1，受体 酪氨酸激酶样蛋白，作为这些事件的方向线索 并且其本身是不对称局部化的。它被本地化到一个区域， 生殖系细胞，边界的肠细胞和下一个生殖系细胞将 form.目的1研究细胞间接触在MES-1定位中的作用 以及在生殖细胞发育过程中发生的两极分化事件。个人 胚胎的细胞将被分离并重新结合，以测试这些细胞是否 需要互动。MES-1和其他不对称定位的细胞 将通过免疫荧光观察组分，以查看它们的模式是否 在单独发育的细胞中发生改变。目标2将决定哪个区域 MES-1是本地化所必需的。 MES-1部分的融合蛋白 产生绿色荧光蛋白（GFP）并转化为 蠕虫，其中这些融合的位置将被评估。的假设 预测MES-1的胞外区与细胞膜上的配体相互作用， 邻近的肠细胞，这反过来又导致它在边界处积聚 细胞之间。目标3将鉴定参与MES-1的基因， 在这些部门中发挥作用。基因将被筛选的技术， RNA干扰导致缺陷的基因，表明其在生殖系分裂中的作用 将被追捕。 本申请可能发现，细胞-细胞接触是生殖细胞发育所必需的。 发展，这将显着改变早期C的公认观点。 线虫胚胎发生人们一直认为生殖细胞的分裂 细胞自主因此，这些实验可能会发现一种新的信号传导机制， 通路鉴定在这一过程中起作用的其他基因将不会 不仅为未来的实验奠定了基础，而且还将推动我们的 理解不对称的划分。

DESCRIPTION (Provided by Applicant): Asymmetric division is a fundamental process that generates daughter cells with different developmental fates. Formation of the germline during embryogenesis in C. elegans offers an ideal system in which to study this process. The germline is generated via a series of asymmetric divisions that require the MES-1 protein. MES-1, a receptor tyrosine kinase-like protein, functions as an orientation cue for these events and is itself asymmetrically localized. It is localized to a region of the germline cell that borders the gut cell and where the next germline cell will form. Aim 1 will investigate the role of cell-cell contact in localizing MES-1 and polarizing the events that occur during germline development. Individual cells of the embryo will be separated and reassociated to test if these interactions are required. MES-1 and other asymmetrically positioned cell components will be visualized by immunofluorescence to see if their pattern is altered in cells that develop in isolation. Aim 2 will determine which region of MES-1 is required for localization. Fusion proteins of portions of MES-1 and the green fluorescent protein (GFP) will be created and transformed into worms, where the position of these fusions will be assessed. The hypothesis predicts the extracellular region of MES-1 interacts with a ligand on the adjacent gut cell, which in turn causes it to accumulate at the boundary between the cells. Aim 3 will identify genes that participate with MES-1 and that function in these divisions. Genes will be screened by the technique of RNAi. Genes that cause defects indicative of their role in germline divisions will be pursued. This application may find that cell-cell contact is required for germline development, which would significantly alter the accepted view of early C. elegans embryogenesis. It has been thought that the germline divisions occur cell-autonomously. Thus these experiments may identify a new signaling pathway. Identification of other genes that function in this process will not only lay the foundation for future experiments but will also advance our understanding of asymmetric divisions.