Regulation of kainate receptor expression in cone bipolar cells

视锥双极细胞中红藻氨酸受体表达的调节

基本信息

批准号：
10367733
负责人：
Anis Contractor
金额：
$ 43.44万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-30 至 2026-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10367733
关键词：
AMPA Receptors Acids Acute Address Adult Affinity Anatomy Area Autoimmune Diseases Binding Brain CRISPR/Cas technology Cell Culture Techniques Cells Chief Cell Communication Cone Contractor Data Databases Development Epilepsy Excision Excitatory Synapse Frequencies Genetic Diseases Glutamate Receptor Glutamates Health Human Individual Interneurons Kainic Acid Receptors Kinetics Knock-out Light Measures Mediating Metabotropic Glutamate Receptors Microscopy Modality Modification Molecular Molecular Genetics Mood Disorders Mus Pathway interactions Perfusion Pharmaceutical Preparations Photoreceptors Physiologic pulse Physiological Play Primates Process Property Proteins Proteomics RNA Splicing Receptor Cell Recovery Regulation Resolution Retina Retinal Cone Role Signal Transduction Spermophilus Structure Synapses System Vision Work authority base cell type desensitization experimental study genetic manipulation in vivo kainate mouse model outer plexiform layer pain signal preservation receptor receptor expression receptor function response tool transcriptomics virus genetics visual information

项目摘要

The long term objective of this work is to understand the molecular genetic basis for how the retina transmits and processes the signals produced when lights are turned off. Cone photoreceptors mediate vision during daylight and use glutamate as their transmitter. The cone signals are carried across the retina by two functional classes of bipolar cells, On and Off, which have different glutamate receptors that cause them to depolarize (ie, signal) either at light-on or -off, respectively. Each functional class of bipolar cell can be subdivided into 5-6 anatomical types. This study focuses on the Off bipolar cells that chiefly express sign-preserving kainate-type glutamate receptors (subunits GluK1-5). Surprisingly, while AMPA receptors predominate at most CNS synapses, kainate receptors predominate at the cone synapses of most of the Off bipolar cell types. They are important to study for at least four reasons. First, the temporal signaling properties of kainate receptors differ from those of AMPA receptors, and these differences are likely to change the size and timing of the Off signals traversing the retina in as yet poorly understood ways. Second, the kainate receptors at most well-studied synapses almost always contain the GluK2 subunit, whereas the Off bipolar cells choose the less well-studied GluK1 subunit as their main subunit instead. Third, Off bipolar cells express Neto1, an auxiliary receptor subunit that profoundly alters the in vivo responses of GluK2-containing receptors. The effect of Neto1 on endogenous GluK1-containing receptors has not yet been studied. And fourth, while the functional properties of different kainate receptors on the Off bipolar cell have been described, the molecular basis for their diverse responses has not been established. To determine the subunit basis for signaling at the Off bipolar cell synapse, we propose to study synaptic responses in mice in which receptor subunits (GluK1-GluK5) are individually knocked out either permanently or acutely. Similar studies will be performed to define the role of the auxiliary subunit Neto1 in communication. Three specific aims will 1) determine the subunits that mediate synaptic responses in Off bipolar cell types 1a, 2, and 3a; 2) determine the function of Neto proteins in signaling at the Off bipolar cell synapse; and, 3) determine the physiological function of the GluK1-1d subunit, which is unique to the retina, the molecular factors that localize the GluK1-1d subunit to the synapse.

这项工作的长期目标是了解视网膜的分子遗传基础传输并处理关闭灯时产生的信号。锥光受体介导白天的视力，并使用谷氨酸作为发射器。圆锥信号载有通过两个功能性类别的双极单元，开关的两种功能性类别，它们具有不同的谷氨酸受体分别在轻度或-OFF处将其去极化（即信号）。每个功能性的双极细胞都可以细分为5-6个解剖类型。这项研究专注于主要表达符号保护海藻酸盐型谷氨酸的双极细胞受体（亚基Gluk1-5）。令人惊讶的是，大多数CNS占主导地位的AMPA受体突触，海藻酸盐受体在大多数OFF双极细胞的锥突触上占主导地位类型。由于至少四个原因，研究很重要。首先，时间信号属性海藻酸盐受体的不同与AMPA受体不同，这些差异很可能改变关闭信号的大小和时机，遍历视网膜方式。其次，大多数研究的突触下的海藻酸盐受体几乎总是包含 gluk2亚基，而离多极性细胞选择较少良好的gluk1亚基作为其主要亚基。第三，双极细胞表达neto1，这是一个辅助受体亚基，深刻地改变了含Gluk2受体的体内反应。 Neto1对尚未研究含内源性Gluk1的受体。第四，而功能已经描述了双极细胞上不同海豚受体的特性，分子尚未建立各种反应的基础。确定亚基的基础在离多双极细胞突触的信号传导，我们建议研究小鼠的突触反应受体亚基（Gluk1-Gluk5）被单独击倒或敏锐地击倒。将进行类似的研究，以定义辅助亚基Neto1在通信中的作用。三个特定目标将1）确定介导双极性突触反应的亚基细胞类型1a，2和3a； 2）确定neto蛋白在双极性信号传导中的功能细胞突触； 3）确定Gluk1-1d亚基的生理功能，这是独特的对于视网膜，将Gluk1-1d亚基定位到突触的分子因子。