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JoVE Journal
Neuroscience
Isolement axoplasme du nerf sciatique du rat
Isolement axoplasme du nerf sciatique du rat
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Axoplasm Isolation from Rat Sciatic Nerve

Isolement axoplasme du nerf sciatique du rat

Full Text
14,132 Views
05:29 min
September 24, 2010

DOI: 10.3791/2087-v

Ida Rishal1, Meir Rozenbaum1, Mike Fainzilber1

1Department of Biological Chemistry,Weizmann Institute of Science

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article presents a detailed protocol for isolating axoplasm from the adult rat sciatic nerve. The method involves dissection of nerve fascicles and incubation in a hypotonic medium to effectively release myelin and lyse non-axonal structures, leading to the extraction of axon-enriched material.

Key Study Components

Area of Science

  • Neuroscience
  • Cell Biology
  • Axonal Research

Background

  • Isolation of axonal cytoplasm is crucial for studying neuronal function.
  • The sciatic nerve serves as a model for peripheral nerve studies.
  • Understanding axonal components can aid in neurological research.
  • Previous methods may not yield pure axonal material.

Purpose of Study

  • To develop a reliable protocol for axoplasm isolation.
  • To enhance the purity of axonal cytoplasm for further analysis.
  • To facilitate the study of axonal components using biochemical techniques.

Methods Used

  • Dissection of the sciatic nerve to access nerve fascicles.
  • Removal of connective tissue surrounding the nerve.
  • Incubation of nerve segments in a hypotonic medium.
  • Subsequent centrifugation and incubation in isotonic solution.

Main Results

  • The protocol successfully isolates axonal cytoplasm.
  • Western blot analysis confirms high enrichment of axonal components.
  • Results demonstrate the effectiveness of the method.
  • Potential applications in studying axonal biology are highlighted.

Conclusions

  • This method provides a robust approach for axoplasm isolation.
  • It can be utilized in various studies related to neuronal function.
  • The findings contribute to the understanding of axonal structure and function.

Frequently Asked Questions

What is the main goal of this protocol?
The main goal is to isolate pure axonal cytoplasm from peripheral nerve.
Why is the sciatic nerve used in this study?
The sciatic nerve is a well-established model for studying peripheral nerves.
What techniques are used to analyze the axonal components?
Western blot analysis is used to confirm the enrichment of axonal components.
How does the hypotonic medium contribute to the isolation process?
The hypotonic medium helps to lyse non-axonal structures and release myelin.
What are the potential applications of this protocol?
It can be used in various studies related to neuronal function and axonal biology.

Nous démontrons un protocole pour l'isolement du nerf axoplasme sciatique de rat adulte repose sur la dissection des fascicules nerveux et l'incubation dans le milieu hypotonique pour libérer de la myéline et Lyse non axonale structures, suivie de l'extraction de l'axone reste-matériau enrichi.

L’objectif global de cette procédure est d’isoler le cytoplasme axonal pur ou l’ectoplasme du nerf périphérique. Ceci est accompli en disséquant d’abord le nerf sciatique. La deuxième étape de la procédure consiste à retirer le tissu conjonctif du nerf et à séparer les plaques.

La troisième étape de la procédure est l’incubation de courts segments de cellules nerveuses dans un milieu hypotonique. La dernière étape de la procédure est l’incubation en solution isotonique, la centrifugation et l’opsm milian. En fin de compte, on peut obtenir des résultats qui montrent un degré élevé d’enrichissement des composants axonaux par l’analyse par transfert Western.

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