MICROSCOPIC LASER SPECTROSCOPY OF CHICK EMBRYO LENSES

细胞分化和发育的关键因素之一是 分化细胞特异性大分子的细胞内合成 细胞。鸡胚晶状体被广泛用作研究中的模型系统。 对细胞分化的影响。它的形态和细胞质蛋白 构图很简单，在开发的所有阶段都可以访问。 相当大的努力集中在晶状体的蛋白质化学上。 使用与白内障相关的分子遗传学技术。 我们最近开发了一种显微激光技术 散射光谱学，可以观察到布朗运动。 单个活细胞胞质内的大分子。从 散射光涨落的幅度和驰豫时间 这些大分子，有可能确定它们的细胞内 浓度和粒度分布。有了这种非侵入性技术，我们 已经成功地确定了细胞内蓄积的时间进程 在发育过程中鸡胚胎晶状体中Delta-晶体蛋白的含量。这个 对于状态的研究，技术是敏感和定量的 在一项关于血红蛋白的研究中发现了细胞质蛋白 聚集在人类正常红细胞和镰刀状红细胞内。 根据我们的初步数据，我们建议研究在体内的发展 鸡胚晶状体体外分化视网膜色素沉着 在克隆细胞培养过程中将上皮细胞转化为晶状体细胞。使用这项技术， 晶状体蛋白的细胞内组成将被确定。 在分化过程中。比较研究使用 还将进行生化技术。这项技术还将 可用于研究晶体蛋白的聚集过程 晶状体细胞的渗透性肿胀。建议的研究将会导致 从数量上更好地了解发展和 眼睛的转分化，并在物理基础上照亮 细胞质异常。

One of the key factors in cell differentiation and development is the intracellular synthesis of macromolecules specific to the differentiated cells. The chick embryo lens is widely used as a model system in the study of cell differentiation. Its morphology and cytoplasmic protein composition are simple and it is accessible at all stages of development. Considerable effort has been focused on the protein chemistry of the lens using molecular genetic techniques in relation to cataract. We have recently developed a technique of microscopic laser light scattering spectroscopy that allows observation of the Brownian motion of macromolecules within the cytoplasm of a single, live cell. From the amplitude and relaxation time of the fluctuations of light scattered from the macromolecules, it is possible to determine their intracellular concentration and size distribution. With this non-invasive technique, we have succeeded in determining the time course of intracellular accumulation of Delta-crystallin within the chick embryo lens during development. The technique is sensitive and quantitative for the study of the state of cytoplasmic proteins as was demonstrated in a study of hemoglobin aggregation inside human normal and sickle red blood cells. Based on our preliminary data we propose t study the in vivo development of the chick embryo lens, in vitro differentiation of retinal pigmented epithelium into lens cells upon clonal cell culture. Using the technique, the intracellular composition of crystallin proteins will be determined during the differentiation processes. Comparative studies using biochemical techniques will also be carried out. The technique will also be used to study the aggregation process of crystallins associated with osmotic swelling of the lens cells. The studies as proposed will lead to quantitatively better understanding of the development and transdifferentiation of the eye, and shed light on the physical basis for cytoplasmic abnormalities.