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Protocol for integrated phosphoproteomic and proteomic analysis of the mouse ventral tegmental area with functional and kinase motif enrichment

Year: 2026

Type: article

Abstract: We present a protocol for label-free proteomic and phosphoproteomic analysis of the mouse ventral tegmental area (VTA). We describe steps for combining region-specific biopsy-punch collection, parallel proteomic and phosphoproteomic processing, liquid chromatography-tandem mass spectrometry (LC-MS/MS), and downstream enrichment analysis. We detail procedures for SynGO-based synaptic localization, g:Profiler/DAVID pathway enrichment, and iceLogo/pLogo motif analysis. This protocol enables quantitative assessment of protein abundance, phosphorylation dynamics, and functional interpretation from limited brain region samples. For complete details on the use and execution of this protocol, please refer to Diezel et al. 1 1. Diezel, C.C. ∙ Majuta, L.A. ∙ Bahramnejad, E. ... Temporal multi-omic exploration of the ventral tegmental area in chronic pain and passive coping behaviors iScience. 2026; 29 , 116085 Full Text Full Text (PDF) Scopus (0) PubMed Google Scholar

Source: STAR Protocols

DOI: 10.1016/j.xpro.2026.104911

Authors: Cody C. Diezel, Paul R. Langlais, Todd W. Vanderah, Arthur C. Riegel

Institution: University of Arizona

Language: English

Topic: Advanced Proteomics Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: gold

Estimated APC paid: $1,550

Funders: University of Arizona, Health Sciences, University of Arizona, National Institute on Drug Abuse