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Investigating the Molecular Mechanisms that Drive Electrical Synapse Development

研究驱动电突触发育的分子机制

基本信息

批准号：
10679980
负责人：
Lila E Kaye
金额：
$ 4.68万
依托单位：
UNIVERSITY OF OREGON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10679980
关键词：
Acceleration Adult Animals Antibodies Architecture Automobile Driving Binding Binding Proteins Biochemical Biochemistry Biological Biological Assay Biological Models Brain C-terminal Cell Culture Techniques Cells Chemical Synapse Chemicals Chemistry Client Communication Complex Connexins Cytoplasm DLG4 gene Data Development Disease Electrical Synapse Electrons Embryo Epithelium Foundations Gap Junctions Genetic Genetic Techniques Health Human Image Imaging Techniques Impairment Integral Membrane Protein Investigation Knowledge Link Liquid substance Mediating Membrane Metabolic Modeling Molecular N-terminal Neurons Optics Pathway interactions Phase Physical condensation Postsynaptic Membrane Property Protein Binding Domain Protein Biochemistry Protein C Protein Dynamics Proteins Publishing Role Scaffolding Protein Series Stains Stereotyping Structure Synapses Synaptic Membranes Tail Testing Tight Junctions Training Transgenic Animals Translating Visualization Zebrafish analog density experimental study fascinate fluorescence imaging genomic locus in vivo in vivo Model innovation loss of function molecular assembly/self assembly molecular domain mutant neural circuit neurodevelopment neurotransmission postsynaptic presynaptic recruit scaffold synaptic function synaptogenesis

项目摘要

PROJECT SUMMARY Electrical synapses are complex cellular and biochemical structures with important roles in health and disease. They are composed of neuronal gap junctions that link the cytoplasm of synapsing neurons through channels composed of transmembrane Connexin proteins (Cx). However, there are striking gaps in knowledge surrounding the identification of non-Connexin electrical synapse proteins and the characterization of molecular mechanisms driving electrical synapse formation. Electron micrograph images first revealed the now well-characterized molecular assemblies of the chemical synapse, showing large electron dense regions beneath pre- and postsynaptic membranes now known as the Active Zone (AZ) and the Postsynaptic Density (PSD). Similar cytoplasmic electron dense regions have been observed at neuronal gap junctions, suggesting the presence of additional machinery regulating electrical synapses. The Miller lab recently identified ZO1b as being necessary and sufficient for Cx localization and electrical synapse function in the zebrafish Mauthner neural circuit. ZO1b is a multidomain molecular scaffold known for organizing cytosolic and transmembrane proteins at epithelial tight junctions. Interestingly, ZO proteins at tight junctions display the fascinating biochemical property known as liquid-liquid phase separation (LLPS) allowing them to create a non- membrane-bound compartments within the cell and concentrate binding partners to build fluid molecular assemblies. Indeed, ZO1b’s chemical synapse analog PSD95, as well as other synaptic scaffolds of the AZ and PSD, have also been suggested to organize chemical synapse architecture through LLPS. However, functional characterization for LLPS in vivo has been difficult due to the absence of an assessable model system. This proposal uses a combination of protein dynamics, binding assays, and structure/function mutants in cell culture to first determine the functional domains involved in Cx-scaffolding and LLPS of ZO1b and then translates those findings in vivo to the optically transparent, genetically tractable Mauthner cell circuit. Together the results will provide a foundational model for the molecules required for electrical synapse development and the biochemical interactions that drive it, as well as accelerated training for the applicant in neurodevelopment, protein biochemistry, and zebrafish genetics.

项目总结