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In ovo Electroporation in Chick Midbrain for Studying Gene Function in Dopaminergic Neuron Development
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Deep Proteome of the Developing Chick Midbrain.

Kaitlyn E Stepler1, Seth C Hannah1,2, Lisa A Taneyhill2

  • 1Department of Chemistry & Biochemistry, University of Maryland, College Park, Maryland 20742, United States.

Journal of Proteome Research
|August 24, 2023
PubMed
Summary
This summary is machine-generated.

This study provides the deepest proteome profile of the chick embryo midbrain, identifying over 5900 proteins. This deep proteomics analysis offers new insights into molecular mechanisms of neural crest cell development and craniofacial patterning.

Keywords:
chick embryocranial neural crestepithelial-to-mesenchymal transition (EMT)mass spectrometryproteomics

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Area of Science:

  • Developmental Biology
  • Proteomics
  • Molecular Biology

Background:

  • Cranial neural crest cell epithelial-to-mesenchymal transition (EMT) and migration are vital for craniofacial development.
  • Disruptions in these processes can lead to developmental defects and diseases.
  • The chick embryo is a key model for studying human embryonic development, but protein-level data for midbrain neural crest cell processes are lacking.

Purpose of the Study:

  • To establish a comprehensive protein-level profile of the chick embryo midbrain during early development.
  • To investigate the molecular underpinnings of cranial neural crest cell EMT and migration using proteomics.
  • To provide a deep proteome dataset for understanding craniofacial patterning.

Main Methods:

  • Utilized mass spectrometry (MS)-based proteomics for deep profiling of the chick midbrain.
  • Employed advanced techniques including optimal lysis, offline fractionation, μPAC separation, and high-resolution tandem MS.
  • Identified over 5900 proteins and 450 phosphoproteins.

Main Results:

  • Achieved the deepest coverage of the chick midbrain proteome to date.
  • Identified proteins involved in critical pathways for neural crest cell EMT and migration, including signaling, proteolysis, extracellular matrix remodeling, and transcriptional regulation.
  • Generated a valuable dataset for studying embryonic development at the protein level.

Conclusions:

  • This study presents a significant advancement in understanding the chick midbrain proteome.
  • The findings provide crucial molecular insights into embryonic craniofacial development and neural crest cell biology.
  • Demonstrates the power of deep proteomics for characterizing developmental tissue microenvironments.