RUI: Cytoskeletal Systems and Calcium in Ooplasmic Movementsand Cytokinesis in the Egg of a Teleost Fish, the Medaka (Oryzias latipes)

本研究的目标是：1）确定 微丝和微管在卵质运动中的作用 青鳉卵，2）确定细胞质钙的作用 脉冲，梯度，和波动的运动的卵质的 青鳉卵; 3）确定 胞质分裂和微丝的机械事件，钙 脉冲，梯度和波在胚盘在前两个 青鳉卵的裂缝 卵质运动和胞质分裂 将使用延时视频显微镜进行研究，使用相位 对比度，Nomarski DIC和落射荧光光学，其中 适当 细胞骨架系统的作用将通过 1)应用或显微注射药物破坏这些系统， 2)用荧光定位微丝和微管 鬼笔环肽和间接免疫荧光法。 的 钙的作用将通过1）寻找钙脉冲， 梯度和波与钙敏感的光蛋白， 水母发光蛋白，和2）将钙缓冲液注射到细胞质中， 鸡蛋 利用鱼卵作为模型，这些研究应该提供新的 对两种基本活动的生理基础的深入了解 受精卵的细胞质：其结构 重组并分裂成子细胞。 本研究 还将为本科生提供参与的机会 在一个积极的研究计划中，使用最先进的技术， 光镜

The goals of the proposed research are: 1) to determine the roles of microfilaments and microtubules in movements of the ooplasm in the medaka egg, 2) to determine the role of cytoplasmic calcium pulses, gradients, and waves in movements of the ooplasm of the medaka egg, and 3) to determine the relationship between the mechanical events of cytokinesis and microfilaments, calcium pulses, gradients and waves in the blastodisc during the first two cleavages of the medaka egg. Ooplasmic movements and cytokinesis will be studied with time-lapse video microscopy, using phase contrast, Nomarski DIC, and epifluorescence optics where appropriate. The role of cytoskeletal systems will be studied by 1) applying or microinjecting drugs that disrupt these systems, and 2) localizing microfilaments and microtubules with fluorescent phalloidins and by indirect immunofluorescence, respectively. The role of calcium will be studied by 1) looking for calcium pulses, gradients, and waves with the calcium-sensitive photoprotein, Aequorin, and 2) injecting calcium buffers into the cytoplasm of the egg. Using the fish egg as a model, these studies should provide new insights into the physiological bases of two fundamental activities of the cytoplasm of the fertilized egg: its structural rearrangement and its division into daughter cells. This research will also provide the opportunity for undergraduates to participate in an active research program using state of the art techniques of light microscopy.