Abstract
Mammalian axonal development begins in embryonic stages and continues postnatally. After birth, axonal proteomic landscape changes rapidly, coordinated by transcription, protein turnover, and post-translational modifications. Comprehensive profiling of axonal proteomes across neurodevelopment is limited, with most studies lacking cell-type and neural circuit specificity, resulting in substantial information loss. We create a Cre-dependent APEX2 reporter mouse line and map cell-type-specific proteome of corticostriatal projections across postnatal development. We synthesize analysis frameworks to define temporal patterns of axonal proteome and phosphoproteome, identifying co-regulated proteins and phosphorylations associated with genetic risk for human brain disorders. We discover proline-directed kinases as major developmental regulators. APEX2 transgenic reporter proximity labeling offers flexible strategies for subcellular proteomics with cell type specificity in early neurodevelopment, a critical period for neuropsychiatric disease.
| Original language | English (US) |
|---|---|
| Article number | e78847 |
| Journal | eLife |
| Volume | 11 |
| DOIs | |
| State | Published - Oct 2022 |
Funding
This work was supported by the NSF CAREER Award 1846234, NIMH R56MH113923, NINDS R01NS107539, NIMH R01MH117111, the Beckman Young Investigator Award, Searle Scholar Award, Rita Allen Foundation Scholar Award, and Sloan Research Fellowship (all YK), and NIMH R01MH118497 (MLM). VD is a predoctoral fellow of the American Heart Association (19PRE34380056) and an affiliate fellow of the NIH 2T32GM15538. The authors are grateful to Lindsey Butler for mouse colony management, Dr. Thomas Bozza for advice during mouse line generation, Dr. Thom Saundersm, Wanda Filipiak, and Galina Gavrilina for preparation of gene-edited mice in the Transgenic Animal Model Core of the University of Michigan\u2019s Biomedical Research Core Facilities, Northwestern Biological Imaging Facility (RRID:SCR_017767) and Dr. Tiffany Schmidt for confocal microscope access, Northwestern High Throughput Analysis laboratory for the microplate reader, and Dr. Anastasia Yocum and her team at A2IDEA, LLC as data analysis consultants. Some schematics were created with https://biorender.com/. This work was supported by the NSF CAREER Award 1846234, NIMH R56MH113923, NINDS R01NS107539, NIMH R01MH117111, the Beckman Young Investigator Award, Searle Scholar Award, Rita Allen Foundation Scholar Award, and Sloan Research Fellowship (all YK), and NIMH R01MH118497 (MLM). VD is a predoctoral fellow of the American Heart Association (19PRE34380056) and an affiliate fellow of the NIH 2T32GM15538.
ASJC Scopus subject areas
- General Neuroscience
- General Biochemistry, Genetics and Molecular Biology
- General Immunology and Microbiology
