Induction of Complete Transection-Type Spinal Cord Injury in Mice

Ronak Reshamwala; Tanja Eindorf; Megha Shah; Graham Smyth; Todd Shelper; James St. John; Jenny Ekberg

doi:10.3791/61131

Induzione della lesione completa del midollo spinale di tipo transezione nei topi

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Neuroscience

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JoVE Journal Neuroscience

Induction of Complete Transection-Type Spinal Cord Injury in Mice

Induzione della lesione completa del midollo spinale di tipo transezione nei topi

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06:51 min

May 6, 2020

DOI: 10.3791/61131-v

Ronak Reshamwala^1,2,3, Tanja Eindorf^2,3, Megha Shah^2,3, Graham Smyth^2,3, Todd Shelper^2,3, James St. John*^1,2,3, Jenny Ekberg*^2,3

¹Griffith Institute for Drug Discovery,Griffith University, ²Menzies Health Institute Queensland,Griffith University, ³Clem Jones Centre for Neurobiology and Stem Cell Research,Griffith University

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Overview

This study outlines a protocol for conducting a precise laminectomy to induce a stable transection-type spinal cord injury in a mouse model. The approach aims to minimize collateral damage while maximizing reproducibility and survival rates for subsequent spinal cord injury research.

Key Study Components

Area of Science

Neuroscience
Spinal cord injury models
Surgical protocols

Background

Understanding spinal cord injuries is critical for developing potential therapies.
Accurate induction methods allow consistent assessments of injury and recovery.
This protocol offers a structured approach to performing a laminectomy without excessive damage.

Purpose of Study

To establish a reliable method for inducing spinal cord transection in mice.
To ensure high reproducibility and survival rates in the model.
To create a framework for subsequent studies on treatment responses after spinal cord injury.

Methods Used

The protocol employs a surgical technique for full spinal cord transection using a fine drill.
The key biological model consists of eight to ten-week-old female C57BL/6 mice.
Important steps include confirming anesthesia, skin incision, muscle dissection, and lamina removal.
Detailed descriptions of surgical precision, irrigation, and suturing are provided to ensure clarity.

Main Results

The protocol leads to a consistent size defect at the transection site with reliable behavioral outcomes.
Immunohistochemistry reveals clear demarcation of the spinal cord boundaries post-injury.
Behavioral testing indicated complete loss of hind limb functionality, crucial for assessing treatment efficacy.
The protocol demonstrates reliable reproducibility for subsequent research applications.

Conclusions

The standardized approach allows for accurate modeling of spinal cord injuries and facilitates understanding of recovery strategies.
This method can underpin future studies aimed at improving spinal cord injury outcomes.
Implications of the findings extend to potential therapeutic applications and further investigations into neuronal repair mechanisms.

Frequently Asked Questions

What are the advantages of using this surgical model?

This surgical model provides a controlled environment for studying spinal cord injuries with minimal collateral damage, enhancing the reliability of research outcomes.

How is the spinal cord injury induced in mice?

The injury is induced via a precise transection at the center of the laminectomy window using a narrow cutting blade after thorough surgical exposure.

What types of data are obtained from this protocol?

Data obtained includes histological analyses through immunohistochemistry and behavioral assessments to evaluate recovery and functionality post-injury.

How does this method facilitate studies on treatment responses?

By establishing a reliable injury model, researchers can assess various treatment interventions and their efficacy in promoting recovery following spinal cord injury.

What are key considerations during the surgery?

Maintaining a stable hand during drilling and ensuring complete hemostasis before suturing are critical for surgical success and animal welfare.

Are there limitations to this protocol?

While the method is precise, variability in individual mouse responses to surgery and anesthesia could affect outcomes, highlighting the need for standardized procedures across studies.

Questo protocollo descrive come creare una laminectomia precisa per l'induzione di lesioni stabili del midollo spinale di tipo transezione nel modello del topo, con danni collaterali minimi per la ricerca sulle lesioni del midollo spinale.

Questo protocollo fornisce una lesione precisa e controllata chirurgicamente per indurre una traspirazione completa del midollo spinale nei topi con un'elevata riproducibilità e tassi di sopravvivenza molto elevati. L'uso di un trapano fine per la lesione ossea riduce al minimo il danno al tessuto circostante, consentendo di modellare le risposte alle lesioni e al trattamento con maggiore precisione. Per eseguire una laminectomia, dopo aver confermato una mancanza di risposta al riflesso del pedale in un topo femmina C57 nero 6 di otto-10 settimane anestetizzato, radere la pelliccia posteriore per esporre l'area chirurgica sulla colonna vertebrale dorsale e sterilizzare l'area rasata con tamponi di cotone sterili imbevuti di liquido antisettico di povidone iodio e spirito chirurgico.

Utilizzando un bisturi, effettuare un'incisione verticale della linea mediana a livello vertebrale T10 e utilizzare forcep dritte per sollevare la pelle dalla fascia sottostante per facilitare il posizionamento del riavvolgitore. Per esporre le spine delle vertebre T9-T11, utilizzare il bordo smussato del bisturi per fare una piccola incisione della linea mediana nel tessuto sottocutaneo e nella fascia sottostante e utilizzare forcelle sottili non affilate per smussare la dissezione e riflettere la fascia. Per esporre le lamina, utilizzare la punta smussata del bisturi per dividere il tronco dorsale e i muscoli paraspinosi lungo le spine delle vertebre T9-T11 e utilizzare le forcep smussate della punta fine per smussare i muscoli a strati per esporre le laminae delle vertebre.

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