MICROSCOPIC LASER SPECTROSCOPY OF CHICK EMBRYO LENSES

细胞分化和发育中的关键因素之一是 分化细胞特异性大分子的细胞内合成 细胞 鸡胚透镜被广泛用作研究的模型系统 细胞分化。 其形态和胞浆蛋白 组成是简单的，它是在发展的各个阶段。 相当大的努力集中在透镜的蛋白质化学上 利用分子遗传学技术研究白内障。 我们最近开发了一种显微激光技术 散射光谱，允许观察的布朗运动， 单个活细胞的细胞质内的大分子。 从 光散射的波动的振幅和弛豫时间 大分子，可以确定它们的细胞内 浓度和尺寸分布。 通过这种非侵入性技术，我们 已经成功地确定了细胞内积累的时间过程， 鸡胚透镜发育过程中δ-晶状体蛋白的表达。 的 技术是敏感和定量的研究状态， 细胞质蛋白质，如血红蛋白研究中所证实的 人正常和镰状红细胞内的聚集。 根据我们的初步数据，我们建议研究体内发育的 鸡胚透镜，视网膜色素的体外分化 在克隆细胞培养时，上皮细胞转化为透镜细胞。 使用该技术， 将测定晶状体蛋白的细胞内组成 在分化过程中。 比较研究使用 还将进行生物化学技术。 该技术还将 可用于研究晶体蛋白的聚集过程， 透镜细胞的渗透性肿胀。 拟议的研究将导致 从数量上更好地了解发展情况， 转分化的眼睛，并阐明了物理基础， 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.