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Real-Time Fluorescent Measurement of Synaptic Functions in Models of Amyotrophic Lateral Sclerosis

Real-Time Fluorescent Measurement of Synaptic Functions in Models of Amyotrophic Lateral Sclerosis

Published on: July 16, 2021

Using FASS-LTP in postmortem mice brain tissues to assess pathological synaptic function.

Joshua Currie¹, Shaneilahi Budhwani², Klarissa H Garza³

¹John Sealy School of Medicine, University of Texas Medical Branch at Galveston, USA; Mitchell Center for Neurodegenerative Diseases, Department of Neurology, University of Texas Medical Branch, USA.

Journal of Neuroscience Methods

|July 16, 2024

Summary

This study enhances the Fluorescence Assisted Single-Synapse Long-Term Potentiation (FASS-LTP) method for assessing synaptic integrity in frozen brain tissue. The improved technique offers robust, reproducible analysis for neurodegeneration research and potential clinical applications.

Keywords:

FASS-LTP Flow cytometry Frozen brain tissues Glutamatergic synapse High throughput Synaptic neurotransmission

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Published on: March 23, 2011

Area of Science:

Neuroscience
Pharmacology
Biochemistry

Background:

Synaptic integrity is crucial for assessing neurodegeneration, with electrophysiology as a gold standard.
Fluorescence Assisted Single-Synapse Long-Term Potentiation (FASS-LTP) offers high-throughput synaptic assessment for evaluating drug efficacy in translational models.

Purpose of the Study:

To adapt and validate the FASS-LTP technique for analyzing synaptic integrity in frozen brain tissue.
To demonstrate the method's utility in a 3xTg-AD mouse model treated with a small molecule inhibitor.

Main Methods:

Adapted FASS-LTP by incorporating a 30-minute rocker-based thawing step for uniform synaptosome separation.
Applied the enhanced FASS-LTP to four brain regions of 3xTg-AD mice at 6 and 12 months, treated with VU0155069 or vehicle.
Utilized frozen brain tissue for longitudinal analysis.

Main Results:

The FASS-LTP technique robustly characterized functional synaptic integrity in multiple brain regions.
Demonstrated the ability to assess the impact of disease-treatment therapies longitudinally.
Confirmed the method's effectiveness in frozen brain tissue samples.

Conclusions:

The enhanced FASS-LTP protocol provides a user-friendly and reproducible method for analyzing synaptic function in frozen tissues.
The technique shows potential for translational use in human clinical samples to understand therapeutic impacts on synaptic performance.
FASS-LTP offers a robust analysis of synaptosomes, improving reproducibility in rodent and human synapses across physiological and pathological states.