Targeting the Corticospinal Tract in Neonatal Rats with a Double-Viral Vector using Combined Brain and Spine Surgery

Rupert D. Smit; Thomas J. Campion III; Rachel L. Stingel; Pushti H. Shah; Jie Chen; George M. Smith

doi:10.3791/62698

뇌 및 척추 결합 수술을 사용하여 이중 바이러스 벡터로 신생아 쥐의 피질척수관 표적화

JoVE Journal
Neuroscience

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Neuroscience

Targeting the Corticospinal Tract in Neonatal Rats with a Double-Viral Vector using Combined Brain and Spine Surgery

뇌 및 척추 결합 수술을 사용하여 이중 바이러스 벡터로 신생아 쥐의 피질척수관 표적화

Full Text

2,601 Views

07:27 min

June 30, 2021

DOI: 10.3791/62698-v

Rupert D. Smit¹, Thomas J. Campion III¹, Rachel L. Stingel¹, Pushti H. Shah¹, Jie Chen¹, George M. Smith¹

¹Shriners Hospitals Pediatric Research Center, Lewis Katz School of Medicine,Temple University

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

Overview

This protocol demonstrates a novel method for applying gene therapies to specific subpopulations of neurons in neonatal rats by utilizing chemogenetic modifiers. The study investigates the contributions of these neurons to the recovery from spinal cord injury, providing insights into the effectiveness of various techniques in promoting recovery in the developing nervous system.

Key Study Components

Area of Science

Neuroscience
Neurobiology
Gene Therapy

Background

Investigates neuronal subpopulations' roles in recovery post-spinal cord injury.
Focuses on neonatal rat models to understand developmental neuroscience.
Details a double viral vector technique for targeted transduction of neurons.

Purpose of Study

To develop a method for gene therapy targeting specific neuronal populations.
To enhance understanding of mechanisms driving recovery from spinal cord injuries.
To address challenges in accurately locating spinal anatomy during the procedure.

Methods Used

This study employs surgical procedures involving viral vector injections into the cortex and spinal cord.
Neonatal Sprague Dawley rats serve as the biological model for testing gene therapies.
Step-by-step instructions for viral vector injection into the corticospinal tract are provided.
Critical steps include identification of anatomical landmarks and precise surgical techniques.

Main Results

The method successfully targets and transduces neuronal subpopulations for investigative purposes.
Findings support the use of excitatory DREADDs to aid recovery after spinal cord injury.
Highlights the importance of accurate anatomical manipulation and injection methodology for effective results.

Conclusions

This innovative technique enables precise targeting of neuronal populations, crucial for studying recovery mechanisms.
Significant implications for therapeutic applications in neurological recovery and underlying injury treatments.
Supports advancing methodologies in developmental neuroscience research.

Frequently Asked Questions

What are the advantages of using neonatal rats?

Neonatal rats offer a model for studying developmental processes and recovery mechanisms in the central nervous system due to their neuroplasticity.

How is the gene therapy administered in this study?

Gene therapy is administered via surgical injections of viral vectors into specific regions of the brain and spinal cord, targeting key neuronal subpopulations.

What types of outcomes can this method provide?

This method can provide insights into neuronal recovery processes, effectiveness of gene therapies, and potential mechanisms of action for spinal cord injury treatment.

How can this method be adapted to other studies?

The surgical and injection techniques can be adapted for different viral vectors or targets, allowing for various experimental designs in neuroscience research.

What are key considerations in performing this protocol?

Careful anatomical identification and precision in surgical techniques are crucial to avoid damaging surrounding tissues and to ensure successful transduction.

이 프로토콜은 전방 화학 유전 변형제를 체성 운동 피질에 주입하고 역행 수송 가능한 Cre 재조합 효소를 자궁 경부 척수에 주입하여 출생 후 5-10 일에 신생아 쥐의 세포 하위 집단에 유전자 요법을 적용하는 새로운 방법을 보여줍니다.

이것은 신생아 쥐에서 피질 척수관의 성공적인 공동 감염을 입증하는 최초의 프로토콜입니다. 어린 포유류에서 세포의 개별 하위 집단이 척수 손상으로부터의 회복에 어떻게 기여하는지 조사하기위한 정확한 청사진을 제공합니다. 다른 이중 바이러스 벡터 기술과 마찬가지로이 방법을 사용하면 사용자가 뉴런의 특정 하위 집단을 정확하게 형질 도입 할 수 있으므로 연구원은 회복을 촉진하는 다양한 화학 발생 기술의 효과를 확인할 수 있습니다.

우리는이 기술에 익숙하지 않은 연구자들이 적절한 척추 해부학을 찾는 데 어려움을 겪을 것으로 기대합니다. 따라서 척추 부분을 지속적으로 자주 계산하고 항상 랜드 마크를 다시 참조하는 것이 좋습니다. 신생아 Sprague Dawley 강아지의 마취 깊이를 확인한 후 두피 상단의 정중선에있는 집게를 눌러 절개 부위를 확인하고 시상 봉합사를 느낍니다.

View the full transcript and gain access to thousands of scientific videos