Functional complexity of the axonal growth cone: a proteomic analysis

• Published in: PloS one Vol. 7; no. 2; p. e31858
• Main Authors: Estrada-Bernal, Adriana, Sanford, Staci D, Sosa, Lucas J, Simon, Glenn C, Hansen, Kirk C, Pfenninger, Karl H
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 27-02-2012; Public Library of Science (PLoS)
• Subjects: Analysis; Animal behavior; Animals; Annotations; Axon guidance; Axons - metabolism; Axons - physiology; Biology; Brain; Brain - embryology; Brain - metabolism; Brain research; Carbohydrate metabolism; Carbohydrates; Carbohydrates - chemistry; Catabolism; Cell culture; Cluster Analysis; Complexity; Computational Biology - methods; Cones; Confidence intervals; Cytoskeleton; Developmental disabilities; Endoplasmic reticulum; Fetuses; Folding; Golgi apparatus; Growth; Growth cones; Growth Cones - metabolism; Hippocampus; Hippocampus - metabolism; Immunofluorescence; Informatics; Localization; Medicine; Metabolism; Microscopy, Fluorescence - methods; Nervous system; Neurons; Pediatrics; Physics; Physiological aspects; Proteasomes; Protein biosynthesis; Protein Folding; Protein Processing, Post-Translational; Protein synthesis; Proteins; Proteome; Proteomics - methods; Rats; Rats, Sprague-Dawley; Trends; Ubiquitination; Wiring; United States > US; Colorado; Ohio
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0031858

The growth cone, the tip of the emerging neurite, plays a crucial role in establishing the wiring of the developing nervous system. We performed an extensive proteomic analysis of axonal growth cones isolated from the brains of fetal Sprague-Dawley rats. Approximately 2000 proteins were identified at ≥ 99% confidence level. Using informatics, including functional annotation cluster and KEGG pathway analysis, we found great diversity of proteins involved in axonal pathfinding, cytoskeletal remodeling, vesicular traffic and carbohydrate metabolism, as expected. We also found a large and complex array of proteins involved in translation, protein folding, posttranslational processing, and proteasome/ubiquitination-dependent degradation. Immunofluorescence studies performed on hippocampal neurons in culture confirmed the presence in the axonal growth cone of proteins representative of these processes. These analyses also provide evidence for rough endoplasmic reticulum and reveal a reticular structure equipped with Golgi-like functions in the axonal growth cone. Furthermore, Western blot revealed the growth cone enrichment, relative to fetal brain homogenate, of some of the proteins involved in protein synthesis, folding and catabolism. Our study provides a resource for further research and amplifies the relatively recently developed concept that the axonal growth cone is equipped with proteins capable of performing a highly diverse range of functions.