MICROENVIRONMENT AND PRIMARY MESENCHYME GENE EXPRESSION

高等生物的发育和分化在一定程度上 单元格与来自 环境。这些信号可以是细胞内的，也可以是细胞外的。这个 这些信号及其作用机制的一致性仍然是 未知，并且仍然是发展中的一个基本问题 生物学。这项提案的主要目标是确定任何 细胞间相互作用、细胞外分子或因素 海胆胚胎原代间充质细胞的环境 它们对胚胎骨骼的分化很重要 并分析这些环境因素是如何调节该计划的 在分子水平上的分化。 具体目标 1.研究细胞间的相互作用在 海洋微粒和初级间充质细胞的分化 海胆胚胎。 2.检测细胞外膜修饰的效果 胚胎中基质(ECM)环境对胚胎分化的影响 纯净的骨髓间充质。这将涉及到使用抑制剂、药物 以及已知的干扰合成、分泌和 各种细胞外基质大分子的加工。 根据目标1和目标2的特定实验设计， 差异化将由以下一项或多项监视 参数。 A.针状体的形态特征。 B.特定针状体基质基因的转录 C.积累一种特定的针状体基质mRNA。 针状体基质蛋白的合成。 3.表征微球和微球合成的ECM组分 初级间充质细胞。

Development and differentiation in higher organisms are in part the result of cells interacting with and interpreting signals from the environment. These signals can be cellular and extracellular. The identity of these signals and their mechanism of action is still unknown and remains a fundamental question in developmental biology. The primary goal of this proposal is to identify any cellular interactions, molecules or factors in the extracellular environment of the primary mesenchyme cells of sea urchin embryos that are important for differentiation of the embryonic skeleton and analyze how these environmental factors regulate the program of differentiation at the molecular level. SPECIFIC AIMS 1. Examine the role of cell-cell interactions on the differentiation of micromeres nd primarymesenchyme cells of the sea urchin embryo. 2. Examine the effects of modification sof the extracellular matrix (ECM) environment in the embryo on differentiation of the primry mesenchyme. The will involve the use of inhibitors, drugs and analogs known to interfere with the synthesis, secretion and processing of various extracellular matrix macromolecules. Depending on the particular experimental design for Aims 1 and 2, differentation will be monitored by one or more of the following parameters. a. morphological appearance of spicules. b. transcription of a specific spicule matrix gene c. accumulation of a specific spicule matrix mRNA. d. synthesis of spicule matrix proteins. 3. Characterize the ECM components synthesized by micromeres and primary mesenchyme.