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Protocol for isolating and processing mouse sciatic nerve fibers for confocal immunohistochemistry.

Lacey B Sell1, Qian Shi1, Manzoor A Bhat1

  • 1Department of Cellular and Integrative Physiology, Joe R. and Teresa Lozano Long School of Medicine, University of Texas Health Science Center San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229, USA; IBMS Neuroscience Graduate Program, Joe R. and Teresa Lozano Long School of Medicine, University of Texas Health Science Center San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229, USA.

STAR Protocols
|January 26, 2024
PubMed
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This study details a protocol for preparing mouse sciatic nerves for confocal immunohistochemistry. This method aids in studying peripheral nervous system diseases affecting myelinated axons.

Area of Science:

  • Neuroscience
  • Pathology
  • Histology

Background:

  • Motor and neurodegenerative diseases frequently impact the peripheral nervous system (PNS).
  • Myelinated axons within the sciatic nerve are crucial for understanding PNS disease pathology.
  • Current methods require standardized protocols for detailed examination.

Purpose of the Study:

  • To present a detailed protocol for isolating and processing mouse sciatic nerves.
  • To enable researchers to perform confocal immunohistochemistry on these nerve samples.
  • To facilitate the characterization of pathologies in myelinated fibers relevant to PNS disorders.

Main Methods:

  • The protocol covers mouse perfusion, sciatic nerve isolation, and fixation.
  • It includes steps for nerve mounting on slides, immunohistochemical staining, and imaging.
Keywords:
Cell BiologyMicroscopyNeuroscience

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  • Confocal microscopy is utilized for high-resolution visualization.

    • Main Results:

    • A reproducible protocol for preparing mouse sciatic nerves for immunohistochemistry was established.
    • The method allows for detailed examination of myelinated axon pathology.
    • Successful staining and imaging of nerve structures were demonstrated.

    Conclusions:

    • This protocol provides a valuable tool for researchers studying peripheral nervous system diseases.
    • It enhances the ability to characterize pathologies affecting myelinated axons.
    • The method supports advancements in understanding and diagnosing neurodegenerative and motor diseases affecting the PNS.