Work

Mouse-derived Synaptosomes Trypsin Cleavage Assay to Characterize Synaptic Protein Sub-localization

Year: 2024

Type: article

Abstract: This protocol describes the isolation of synaptosomes from mouse brain tissue, followed by limited trypsin digestion to assess the compartmental localization of synaptic proteins. Synaptosomes, which are isolated via differential centrifugation, consist of resealed presynaptic terminals that are protected from proteolysis, while the exposed postsynaptic compartments are accessible to trypsin and undergo proteolytic cleavage. Following digestion, samples are analyzed using SDS-PAGE and western blotting. Proteins of interest are probed using specific antibodies to determine whether they are presynaptic or postsynaptic. Presynaptic proteins (e.g., synaptophysin, SNAP-25) remain intact, while postsynaptic proteins (e.g., GluA1, GluN2A) are cleaved.

Source: BIO-PROTOCOL

DOI: 10.21769/bioprotoc.5164

Authors: Jasmeet Shergill, Domenico Tehran

Institutions: Rheinland-Pfälzische Technische Universität Kaiserslautern-Landau, Leibniz-Forschungsinstitut für Molekulare Pharmakologie

Language: English

Topic: Neuroscience and Neuropharmacology Research

Subfield: Cellular and Molecular Neuroscience

Field: Neuroscience

Domain: Life Sciences

Open Access status: diamond

Funder: Klaus Tschira Stiftung

OpenAlex ID: W4405174480

FWCI:

Cites:

Cited by:

Related to: 10