Project Details
Description
Project Summary
Biomolecular condensates that arise from liquid-liquid phase separation have emerged as a central player in
numerous cellular processes. The material properties of these condensates are associated with various
biological roles. For example, the surface tension of liquid condensates governs the interaction between the
condensate and other cellular structures, regulating processes such as nucleoli organization, autophagy,
microtubule branching, P granule growth, and cell surface signaling. Under abnormal conditions, several types
of neuronal protein condensates change from liquid states to solid fibrils that resemble the hallmarks of
neurodegeneration. However, current understanding of biomolecular condensates is limited, mainly due to the
lack of accurate tools that can perturb and monitor the material properties of these microscale condensates.
Established techniques focus on individual aspects of condensate properties and are often susceptible to
instrumentation challenges or measurement artifacts. Moreover, quantifications of condensates in live cells are
still elusive. Recently, we demonstrated a micropipette-based technique that directly measures both the surface
tension and viscosity of purified protein condensates, free from common sources of artifacts. Importantly, our
technique shares a large part of its core hardware with patch-clamp, a well-established tool used by
neuroscientists to record electrical signals in live cells and animals.
In ongoing experiments, we have applied the technique to condensates of several neuronal proteins. This
includes not only proteins associated with neurodegeneration, but also synapsin, a highly abundant neuronal
protein that regulates synaptic vesicle clustering and transmission. Furthermore, we have tested the compatibility
between our micropipette-based technique and patch-clamp recording in live cells. Based on these preliminary
data, we hypothesize that micropipette is broadly applicable to measure condensates made of neuronal proteins,
allowing mechanistic understanding of the material properties of these condensates in live cells. Our Specific
Aims are: (1) Mechanistic understanding of the surface tension and viscoelasticity of neuronal protein
condensates through in vitro reconstitutions. (2) Quantification of neuronal protein condensates in live cells.
We anticipate the proposed technology can be easily adapted by the broader scientific community to study
biomolecule condensates in cells. Data from this project will give direct insights into the roles of condensate
material properties in mediating neurological processes and neurodegenerative diseases. The quantification of
condensates in cultured cells will also lie the basis for exploring condensate material properties in complex
biological systems.
Status | Finished |
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Effective start/end date | 8/15/22 → 7/31/24 |
Funding
- National Institute on Drug Abuse: $207,247.00
- National Institute on Drug Abuse: $185,019.00
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