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:(申请人的摘要)实验室的目标是
更好地理解神经处理的分子机制,
视网膜。 已知的是,将光刺激分离为开和关
是在双极细胞的水平上完成的。 因为人们认为
感光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被认为是
这是它们表达不同谷氨酸受体的结果。 具体
这一建议旨在探讨表达
不同的谷氨酸受体和双极细胞的多样性,
小鼠视网膜。
1. OFF双极细胞中的谷氨酸受体。 之多样性和分布情况
谷氨酸受体亚单位的研究将在OFF双极细胞中进行,
原位杂交和免疫组织化学分析水平和
垂直视网膜切片和分离的视网膜细胞。谷氨酸受体
电生理学鉴定个体OFF中亚基表达
将使用PCR技术确定双极细胞。
2. ON双极细胞中的谷氨酸受体。 APB型谷氨酸盐
受体,其被认为介导ON双极中的符号反转
细胞,将被克隆。 抗体将针对
APB受体在细菌中表达。 之多样性和分布情况
谷氨酸受体将在ON双极细胞中通过原位杂交检测。
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
在电生理学鉴定的个体ON双极细胞中的表达
将使用PCR技术进行分析。
3.发育过程中的谷氨酸受体。 研究表明,早期
发育中的大脑中神经递质受体的表达,
这表明它在神经回路的建立中起了一定的作用。 的
谷氨酸受体表达的调节与
双极细胞的分化将使用原位杂交分析。
杂交和免疫组织化学。
本文提出的研究将为分析
谷氨酸受体在视网膜变性小鼠中的表达,
评估发育过程中受体改变的影响,
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