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Neuroscience
Akut Ictal faaliyet kemirgen ve insan dokusu üretimi ve isteğe bağlı başlatma
Akut Ictal faaliyet kemirgen ve insan dokusu üretimi ve isteğe bağlı başlatma
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Generation and On-Demand Initiation of Acute Ictal Activity in Rodent and Human Tissue

Akut Ictal faaliyet kemirgen ve insan dokusu üretimi ve isteğe bağlı başlatma

Full Text
9,431 Views
06:45 min
January 19, 2019

DOI: 10.3791/57952-v

Michael Chang1,2, Suzie Dufour1,3, Peter L. Carlen1,2,3,5,6, Taufik A. Valiante1,2,3,4

1Division of Fundamental Neurobiology,Krembil Research Institute, 2Institute of Medical Science, Faculty of Medicine,University of Toronto, 3Institute of Biomaterials and Biomedical Engineering,University of Toronto, 4Division of Neurosurgery, Department of Surgery,University of Toronto, 5Division of Neurology, Department of Medicine,University of Toronto, 6Department of Physiology,University of Toronto

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Overview

This study focuses on the use of acute seizure models, specifically utilizing 4-aminopyridine, to investigate the mechanisms of epileptiform activities in both mouse and human cortical tissues. The ability to induce electrographic seizure events on demand allows for detailed examination of neuronal behavior associated with seizure onset and termination.

Key Study Components

Area of Science

  • Neuroscience
  • Electrophysiology
  • Epilepsy research

Background

  • Understanding the underlying mechanisms of seizures is crucial for developing effective treatments.
  • Acute seizure models enable real-time observation of seizure dynamics.
  • Electrophysiological recordings reveal insights into seizure initiation and propagation.
  • The technique has potential applications in testing anti-seizure drugs.

Purpose of Study

  • To explore neural subpopulations responsible for the onset and termination of seizures.
  • To assess the efficacy of potential anti-seizure therapies in a controlled manner.
  • To investigate the characterization of ictal-like events in cortical brain slices.

Methods Used

  • The study utilized acute cortical brain slices prepared from mice and human tissues.
  • Electrophysiological recordings were conducted to measure seizure-like activities induced by 4-aminopyridine.
  • Detailed slicing and incubation protocols were employed to ensure tissue viability.
  • Optogenetic strategies were incorporated to trigger ictal events through precise light stimulation.
  • A MATLAB-based software was developed for detecting and classifying epileptiform events.

Main Results

  • The protocol reliably induced ictal-like events in high-quality brain slices within 15 minutes of 4-AP application.
  • It was found that 40% of slices successfully generated these events, indicating effective tissue preparation.
  • Responses included a characteristic sentinel spike and various phases of firing indicative of seizure activity.
  • Similar results were observed across different mouse models, highlighting the robustness of the technique.

Conclusions

  • This study demonstrates a reliable method for inducing and studying electrographic seizures, providing insights into neuronal mechanisms involved in epilepsy.
  • The findings support the exploration of seizure dynamics in vivo and in vitro.
  • Understanding these mechanisms could lead to better treatment strategies for epilepsy and other related disorders.

Frequently Asked Questions

What are the advantages of using acute seizure models?
Acute seizure models allow for on-demand generation of seizures, facilitating real-time studies of neuronal activity and the underlying mechanisms of seizure onset and termination.
How are the cortical slices prepared for experimentation?
Cortical slices are prepared by carefully slicing preclinical brain tissue into 450 μm sections and incubating them in carbogenated ACSF to maintain viability.
What types of data are obtained using this method?
Electrophysiological data, including the characterization of ictal-like events and measures of neuronal firing patterns, are obtained through recordings during induced seizures.
How can the method be adapted for testing anti-seizure drugs?
This method can be adapted by applying various anti-seizure drug candidates during the induced seizures and observing their effects on seizure characteristics.
What are some limitations of this approach?
Tissue quality is critical; damaged tissues are less likely to generate reliable seizure-like events, necessitating careful handling during slice preparation.

Akut kriz modelleri epileptiform olaylar altında yatan mekanizmaları eğitim için önemlidir. Ayrıca, epileptiform olaylar üstünde-istek oluşturma yeteneği kendi başlatma altında yatan olay tam sırası çalışmaya son derece verimli bir yöntem sağlar. Burada, fare ve insan dokusu içinde kurulan akut 4-aminopyridine kortikal nöbet modelleri açıklar.

Bu yöntem, elektrografik nöbet başlangıcı ve sonlandırmadan hangi sinirsel alt popülasyonların sorumlu olduğu gibi nöbet mekanizmalarının incelenmesinde anahtar soruların yanıtlandırılmasına yardımcı olabilir. Bu tekniğin en büyük avantajı, elektrografik nöbet olaylarını, isteğe bağlı olarak ve klinik olarak gözlenenleri anımsatan in vivo ve in vitro modellerini yeniden üretebiliyor olmasıdır. Bu teknik aynı zamanda potansiyel nöbet önleyici tedavileri aramak için de kullanılabilir, farklı anti-nöbet ilaç adaylarının uygulanması sırasında elektrografik nöbet olayları tetiklemeye çalışarak.

Kortikal dilimleri hazırlamak için, preklinik beyin dokusunu anında yapışkan tutkal kullanarak Vibratome aşamasına yapıştırın. Specimin tutucuyu yavaşça tampon tepsiye yerleştirin. Beynin sırt kısmının Vibratom'un bıçağına baktığından emin ol.

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