Abstract
Characterizing neuronal heterogeneity at the single-cell level can improve our understanding of developmental and disease processes and lead to novel therapeutic strategies. While single-cell genomic and transcriptomic measurements have matured in recent years, mass spectrometry-based proteome profiling has lagged due to sensitivity limitations. However, recent progress in miniaturizing sample processing and separations have made it possible to profile >1000 proteins from single mammalian cells including neurons. Here, we present a detailed procedure that has provided the greatest depth of coverage for proteome profiling of single neurons to date. Single motor neurons and interneurons were isolated from human spinal tissue sections by laser capture microdissection, processed for proteomic analysis within nanoliter volumes and analyzed by ultralow-flow nanoLC-MS/MS. The protocol should be adaptable to a variety of applications and measurement platforms to further neuronal research.
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Acknowledgments
This work was supported by the National Institutes of Health under award numbers R33 CA225248 and R01 GM138931, and through a sponsored research agreement from Biogen, Inc.
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Misal, S.A., Kelly, R.T. (2022). Single-Cell Proteome Profiling of Neuronal Cells. In: Sweedler, J.V., Eberwine, J., Fraser, S.E. (eds) Single Cell ‘Omics of Neuronal Cells. Neuromethods, vol 184. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2525-5_3
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DOI: https://doi.org/10.1007/978-1-0716-2525-5_3
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