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Neuroscience
Vivo kan - beyin bariyerini geçirgenliği tahlil Fluorescently kullanarak farelerde izley...
Vivo kan - beyin bariyerini geçirgenliği tahlil Fluorescently kullanarak farelerde izley...
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

Vivo kan - beyin bariyerini geçirgenliği tahlil Fluorescently kullanarak farelerde izleyiciler etiketli

Full Text
26,340 Views
09:35 min
February 26, 2018

DOI: 10.3791/57038-v

Kavi Devraj1,2, Sylvaine Guérit1, Jakranka Macas1, Yvonne Reiss1

1Institute of Neurology (Edinger Institute),Goethe University Hospital, 2Pharmazentrum Frankfurt, Institute for General Pharmacology and Toxicology,Goethe University Hospital

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Overview

This study presents a method for assessing mouse brain vascular permeability using intraperitoneally injected fluorescent tracers. It aims to investigate blood-brain barrier dysfunction, relevant in conditions such as stroke and Alzheimer's disease, by providing quantitative and visual data on neurovascular permeability.

Key Study Components

Area of Science

  • Neuroscience
  • Vascular Biology
  • Biomedical Research

Background

  • Assessing blood-brain barrier dysfunction is critical for understanding various neurological disorders.
  • The use of fluorescent tracers allows for both quantitative and qualitative analysis of permeability.
  • This method can provide insights on recovery from vascular dysfunction after therapy.
  • Implications extend towards the diagnosis and treatment of brain edema occurring in multiple diseases.

Purpose of Study

  • To develop a reliable assay for measuring vascular permeability in mouse brain models.
  • To evaluate therapeutic interventions aimed at restoring blood-brain barrier functionality.
  • To facilitate deeper investigations into the underlying mechanisms of neurovascular permeability changes.

Methods Used

  • This study employs an intraperitoneal injection method using fluorescent tracers.
  • A mouse model is utilized to examine the integrity of the blood-brain barrier.
  • Concurrent quantitative assessment of permeability is performed via fluorometry, alongside fluorescence microscopy for regional visualization.
  • The complete procedure, from injection to analysis, takes approximately 5–6 hours for 10 mice.
  • Detailed steps include anesthesia, cardiac perfusion, tissue harvesting, and subsequent fluorescence measurements.

Main Results

  • The method allows for effective quantification of blood-brain barrier permeability.
  • Fluorescent microscopy provides visual confirmation of tracer localization and permeability assessments.
  • The procedure has shown potential applicability in studying recovery processes following various neurological injuries.

Conclusions

  • This study demonstrates a specific, efficient assay for examining vascular permeability in mouse models.
  • It enables researchers to further explore therapeutic impacts on blood-brain barrier integrity.
  • The method contributes to understanding the mechanisms of vascular dysfunction in neurological disorders.

Frequently Asked Questions

What are the advantages of using this permeability assay?
This assay provides a quantitative approach to measure vascular permeability and allows for both fluorometric and microscopic methods for comprehensive analysis.
How is the mouse model prepared for the assay?
Mice are injected with a fluorescent tracer, deeply anesthetized, and undergo cardiac perfusion before tissue harvesting for analysis.
What types of data can be obtained from this method?
The method yields quantitative permeability data from fluorometric readings and detailed visualizations from fluorescence microscopy.
How can this technique be applied in research?
It can be used to study the blood-brain barrier's response to treatments and its role in various neurological diseases, like strokes or brain tumors.
Are there any limitations to this method?
The assay's effectiveness relies on proper perfusion technique and timely processing, which requires careful attention to procedural details.
What are the key safety considerations for handling mice during the procedure?
Ensure adherence to institutional animal care protocols, including proper anesthesia and humane treatment throughout the procedure.

Burada bir fare beyin damar geçirgenliği tahlil mayi enjeksiyon kan - beyin bariyerini disfonksiyon hayvan modelleri için geçerlidir perfüzyon ardından floresan tarayıcıları kullanarak mevcut. Bir hemi-beyin geçirgenliği kantitatif değerlendirilmesi ve diğer izleme görselleştirme/immunostaining için kullanılır. Yordamı 5-6 h 10 fareler için alır.

Bu prosedürün genel amacı, hayvan modellerinde kan-beyin bariyeri disfonksiyonunu değerlendirmek için floresan etiketli izleyiciler kullanarak farelerde beyin damar sisteminin geçirgenliğini değerlendirmektir. Bu yöntem, hastalıklarda nöro-vasküler geçirgenlik ve tedavi üzerine iyileşmesi ile ilgili kan-beyin bariyeri alanındaki temel soruların yanıtlanmasına yardımcı olabilir. Bu yöntemin temel avantajı basit ve nicel olmasıdır.

Kantitatif olan florometri ve ayrıca bölgesel savunmalar için floresan mikroskobu ile geçirgenliğin eşzamanlı olarak değerlendirilmesi imkanı verir. Bu tekniğin etkileri, beyin tümörleri, felç ve Alzheimer hastalığı dahil olmak üzere çeşitli nörolojik bozuklukların tanı ve tedavisine kadar uzanır, çünkü bu hastalıklar yaşamı tehdit eden beyin ödemi ve önemli bir terapötik hedef olarak BBD fonksiyonunun iyileştirilmesi ile ilişkilidir. Prosedürün bir kısmını göstermek, enstitüde bilimsel teknoloji görevlisi olan Bayan Jadranka Macas olacak.

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