Cell type-specific biotin labeling in vivo resolves regional neuronal and astrocyte proteomic differences in mouse brain

Sruti Rayaprolu, Sara Bitarafan, Juliet V. Santiago, Ranjita Betarbet, Sydney Sunna, Lihong Cheng, Hailian Xiao, Ruth S. Nelson, Prateek Kumar, Pritha Bagchi, Duc M. Duong, Annie M. Goettemoeller, Viktor János Oláh, Matt Rowan, Allan I. Levey, Levi B. Wood, Nicholas T. Seyfried () and Srikant Rangaraju ()
Additional contact information
Sruti Rayaprolu: Emory University
Sara Bitarafan: Georgia W. Woodruff School of Mechanical Engineering, Parker H. Petit Institute for Bioengineering and Bioscience, and Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology
Juliet V. Santiago: Emory University
Ranjita Betarbet: Emory University
Sydney Sunna: Emory University
Lihong Cheng: Emory University
Hailian Xiao: Emory University
Ruth S. Nelson: Emory University
Prateek Kumar: Emory University
Pritha Bagchi: Emory University
Duc M. Duong: Emory University
Annie M. Goettemoeller: Emory University
Viktor János Oláh: Emory University
Matt Rowan: Emory University
Allan I. Levey: Emory University
Levi B. Wood: Georgia W. Woodruff School of Mechanical Engineering, Parker H. Petit Institute for Bioengineering and Bioscience, and Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology
Nicholas T. Seyfried: Emory University
Srikant Rangaraju: Emory University

Nature Communications, 2022, vol. 13, issue 1, 1-18

Abstract: Abstract Proteomic profiling of brain cell types using isolation-based strategies pose limitations in resolving cellular phenotypes representative of their native state. We describe a mouse line for cell type-specific expression of biotin ligase TurboID, for in vivo biotinylation of proteins. Using adenoviral and transgenic approaches to label neurons, we show robust protein biotinylation in neuronal soma and axons throughout the brain, allowing quantitation of over 2000 neuron-derived proteins spanning synaptic proteins, transporters, ion channels and disease-relevant druggable targets. Next, we contrast Camk2a-neuron and Aldh1l1-astrocyte proteomes and identify brain region-specific proteomic differences within both cell types, some of which might potentially underlie the selective vulnerability to neurological diseases. Leveraging the cellular specificity of proteomic labeling, we apply an antibody-based approach to uncover differences in neuron and astrocyte-derived signaling phospho-proteins and cytokines. This approach will facilitate the characterization of cell-type specific proteomes in a diverse number of tissues under both physiological and pathological states.

Date: 2022
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DOI: 10.1038/s41467-022-30623-x

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