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Using Rotary Shadow Electron Microscopy to Characterize Semaphorin-Mediated Growth Cone Collapse.
1Department of Neuroscience, Washington University School of Medicine, Box 8108, 660 S Euclid, Saint Louis, MO, 63110, USA. bridgmap@wustl.edu.
Methods in Molecular Biology (Clifton, N.J.)
|October 28, 2016
Summary
Rotary shadow electron microscopy offers high-resolution insights into growth cone cytoskeleton changes. This method details neuronal responses to guidance factors like semaphorin 3A.
Area of Science:
- Neuroscience
- Cell Biology
- Biophysics
Background:
- Growth cone navigation is crucial for neural development.
- Cytoskeletal dynamics underlie growth cone responses to guidance cues.
- Understanding these dynamics requires high-resolution imaging techniques.
Purpose of the Study:
- To describe procedures for preparing cultured neurons for rotary-shadow electron microscopy (EM).
- To enable detailed comparisons of cytoskeletal structure in growth cones.
- To facilitate the study of neuronal responses to repulsive guidance factors.
Main Methods:
- Culturing neurons for microscopy.
- Preparing samples using rotary shadowing technique.
- High-resolution electron microscopy (EM) imaging of cytoskeletons.
Main Results:
- Established a protocol for rotary-shadow EM of growth cone cytoskeletons.
- Demonstrated the ability to detect global and macromolecular cytoskeletal changes.
- Provided detailed structural comparisons of cytoskeletons.
Conclusions:
- Rotary shadow EM is a powerful tool for visualizing growth cone cytoskeleton organization.
- This technique allows for precise analysis of cytoskeletal responses to semaphorin 3A.
- The described procedures enhance the study of neuronal guidance mechanisms.
