# Understanding higher-order glycine receptor assemblies in neurotransmission

> **NIH NIH R35** · UT SOUTHWESTERN MEDICAL CENTER · 2024 · $410,000

## Abstract

Overview. Electrical signal transmission between cells underlies almost all physiological processes in human,
from heartbeat, to learning and memory. Neurotransmitter receptors are organized in clusters at micron-sized
inter-cellular machineries, namely synapses, throughout our nervous system to enable signal transmission.
Decades of intense research have characterized the working mechanism of most individual receptors. However,
due to technical difficulties, the clusters of receptors remain enigmatic – neither the structure, nor the functional
significance is known. In this proposal, my lab will use state-of-the-art methods and a reconstitution system to
systematically characterize the architecture of synaptic glycine receptors, which is the last major neurotransmitter
receptors whose architecture remained elusive before our work. We will also develop novel technologies for
quantitative characterization of higher-order assemblies of glycine receptors, and other proteins in the 2D setting
of lipid membranes. Following is a brief description of the proposed work.
Glycine receptors and its clustering with gephyrin scaffold. Glycine receptor (GlyR) belongs to the Cys-loop
family of pentameric ligand-gated ion channels. GlyRs in adult tissue are heteromeric receptors composed of
both the α and β subunits, which form clusters with scaffold protein gephyrin at synapses through a specific
interaction between the β subunit and gephyrin. Disfunction of GlyR signaling is the major cause of the rare
congenital disease hyperekplexia and related to chronical neurological pain and autism spectrum disorders. The
architecture of heteromeric GlyRs and how they form clusters with gephyrin is very limited – even the α:β subunit
stoichiometry has been under debate for decades. We have discovered an unexpected subunit composition of
GlyR, explaining the unique function of heteromeric GlyRs and clearing long-lasting confusion. We will use this
established platform to characterize all major types of GlyRs and how they interact with gephyrin.
Develop novel technologies for characterizing protein clusters in lipid membrane. My lab will develop a novel
correlated raised-total internal refection fluorescence microscopy (TIRF) and electrophysiology reconstitution
system to characterize ion channel/receptor clusters. This system has the high signal/noise ratio and single-
molecule sensitivity as traditional TIRF, and allow complex electrophysiological experiments to be performed
simultaneously with imaging. We will learn how clusters form, how they are regulated, and whether clustering
gives rise to functional effects and regulate physiological activities. The knowledge gained here will guide the
reconstitution of functional clusters for structural characterization using cryo-EM single particle and/or
tomography methods. These new methods will allow quantitative characterization of the spatial organization and
functional significance of clustering, as well as the mechanisms ...

## Key facts

- **NIH application ID:** 10841674
- **Project number:** 5R35GM146860-03
- **Recipient organization:** UT SOUTHWESTERN MEDICAL CENTER
- **Principal Investigator:** Weiwei Wang
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $410,000
- **Award type:** 5
- **Project period:** 2022-08-01 → 2027-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10841674

## Citation

> US National Institutes of Health, RePORTER application 10841674, Understanding higher-order glycine receptor assemblies in neurotransmission (5R35GM146860-03). Retrieved via AI Analytics 2026-06-11 from https://api.ai-analytics.org/grant/nih/10841674. Licensed CC0.

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