MOLECULAR MECHANISMS OF SIGNAL TRANSDUCTION IN RETINA
MOLECULAR MECHANISMS OF SIGNAL TRANSDUCTION IN RETINA
批准号:
2163127
负责人:
ROBERT M DUVOISIN
金额:
$20.09万
依托单位国家:
美国
项目类别:
财政年份:
1992
资助国家:
美国
项目状态:
已结题
起止时间:
1992-04-01 至 1995-03-31
关键词:
biological signal transduction cell differentiation cell type complementary DNA developmental neurobiology electrophysiology genetic library genetically modified animals glutamate receptor immunocytochemistry in situ hybridization laboratory mouse molecular biology molecular cloning neural information processing neural transmission nucleic acid probes nucleic acid sequence polymerase chain reaction receptor expression retina retina degeneration retinal bipolar neuron
中文摘要
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。
英文摘要
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.
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海外基金