MOLECULAR MECHANISMS OF SIGNAL TRANSDUCTION IN RETINA

描述：(申请人摘要)该实验室的目标是 更好地理解神经加工的分子机制 视网膜。众所周知，光刺激分为开和关 通路是在双极细胞水平上完成的。因为人们认为 光感受器细胞释放谷氨酸作为神经递质， On和Off双极电池之间的功能差异建议为 它们表达不同的谷氨酸受体的结果。特指 这项建议旨在探讨表达与 不同谷氨酸受体与大鼠双极细胞的多样性 小鼠视网膜。 1.非双极细胞的谷氨酸受体。多样性和分布性 的谷氨酸受体亚基的数量将通过In 水平线和水平线的原位杂交和免疫组织化学分析 视网膜垂直切片和分离的视网膜细胞。谷氨酸受体 电生理识别个体Off亚基的表达 两极细胞将使用聚合酶链式反应技术来确定。 2.双极细胞上的谷氨酸受体。APB型谷氨酸 受体，它被认为是介导双极的符号倒置 细胞，将被克隆。抗体将被用来对抗病毒的片段 APB受体在细菌中表达。的多样性和分布性 谷氨酸受体将在双极细胞上进行原位检测 水平和垂直的杂交和免疫组织化学分析 视网膜切片和分离的视网膜细胞。谷氨酸受体 电生理鉴定个体在双极细胞上的表达 将使用聚合酶链式反应技术进行分析。 3.发育中的谷氨酸受体。研究表明，早期的 发育中的大脑中神经递质受体的表达， 暗示在神经回路的建立中起到了一定的作用。这个 谷氨酸受体表达调控与血管紧张素转换酶 双极细胞的分化将使用原位分析 杂交和免疫组织化学。 这里提出的研究将为分析 谷氨酸受体在视网膜退行性变小鼠视网膜组织中的表达 评估受体在发育过程中和胚胎发育过程中的变化 使用转基因小鼠的成年视网膜。

DESCRIPTION: (Applicant's Abstract) The goal of the laboratory is to better understand the molecular mechanisms of neural processing in the retina. It is known that the segregation of light stimuli into ON and OFF pathways is done at the level of the bipolar cells. Since it is thought that photoreceptor cells release glutamate as a neurotransmitter, the functional difference between ON and OFF bipolar cells is proposed to be the result of their expressing distinct glutamate receptors. Specifically this proposal aims to explore the relationship between the expression of different glutamate receptors and the diversity of bipolar cells in the mouse retina. 1.Glutmate receptors in OFF bipolar cells. The diversity and distribution of glutamate receptor subunits will be examined in OFF bipolar cells by in situ hybridization and immunonohistochemical analyses of horizontal and vertical retinal sections and dissociated retinal cells. Glutamate receptor subunit expression in electrophysiologically identified individual OFF bipolar cells will be determined using the PCR technique. 2.Glutamate receptors in ON bipolar cells. The APB-type glutamate receptor, which is thought to mediate the sign inversion in ON bipolar cells, will be cloned. Antibodies will be raised against fragments of the APB receptor expressed in bacteria. The diversity and distribution of glutamate receptors will be examined in ON bipolar cells by in situ hybridization and immunohistochemical analyses of horizontal and vertical retinal sections and dissociated retinal cells. Glutamate receptor expression in electrophysiologically identified individual ON bipolar cells will be analyzed using the PCR technique. 3.Glutamate receptors during development. Studies have shown an early exrpession of neurotransmitter receptors in the developing brain, suggesting some role in the establishment of neural circuitry. The regulation of glutamate receptor expression in relation to the differentiation of bipolar cells will be analyzed using in situ hybridization and immunohistochemistry. The research proposed here will provide the basis with which to analyze the expression of glutamte receptors in mice with retinal degenerations and to assess the effects of receptor alterations during development and in the adult retina using transgenic mice.