<em>In Vivo</em> Vascular Permeability Detection in Mouse Submandibular Gland

Xiangdi Mao; Sainan Min; Qihua He; Xin Cong

doi:10.3791/64167

İn Vivo Fare Submandibuler Bezinde Vasküler Geçirgenlik Tespiti

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In Vivo Vascular Permeability Detection in Mouse Submandibular Gland

İn Vivo Fare Submandibuler Bezinde Vasküler Geçirgenlik Tespiti

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07:10 min

August 4, 2022

DOI: 10.3791/64167-v

Xiangdi Mao¹, Sainan Min², Qihua He³, Xin Cong¹

¹Department of Physiology and Pathophysiology, Peking University School of Basic Medical Sciences, Key Laboratory of Molecular Cardiovascular Sciences,Ministry of Education, and Beijing Key Laboratory of Cardiovascular Receptors Research, ²Department of Oral and Maxillofacial Surgery,Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Reseah Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, ³State Key Laboratory of Natural and Biomimetic Drugs,Peking University

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Overview

This study evaluates the endothelial barrier function of the submandibular gland (SMG) using in vivo imaging techniques. Fluorescent tracers of varying molecular weights were injected into test animal models, demonstrating how molecular permeability can be assessed using two-photon laser scanning microscopy.

Key Study Components

Research Area

Endothelial barrier function
Vascular permeability studies
Fluorescent tracer application

Background

Importance of tight junctions in endothelial cells
Non-invasive imaging methods for tissue analysis
The role of submandibular glands in vascular studies

Methods Used

In vivo paracellular permeability detection
Mouse submandibular glands
Two-photon laser scanning microscopy

Main Results

Demonstrated varying leakage of fluorescent tracers based on molecular weight
Identified disruptions in endothelial barrier integrity due to duct ligation
Validated findings through fluorescence intensity quantification

Conclusions

The method provides insights into endothelial barrier function in tissues
Enhances understanding of vascular permeability related to physiological and pathological conditions

Frequently Asked Questions

What is the purpose of using different molecular weight tracers?

Using tracers of different molecular weights allows for assessing the permeability of tight junctions in endothelial cells.

How does two-photon laser scanning microscopy improve imaging?

This method enables deeper tissue imaging with less scattering compared to conventional techniques, providing clearer images.

What effects were observed when duct ligation was performed?

Duct ligation increased permeability, allowing larger molecular tracers to leak out, indicating a disruption in the endothelial barrier.

Why is proper SMG isolation critical in this protocol?

Careful isolation is essential to minimize artifacts and ensure accurate imaging and permeability assessments.

What are the applications of this imaging technique?

This technique can be applied to study vascular diseases, drug delivery systems, and tissue engineering.

What does the study reveal about vascular permeability in the SMG?

The findings provide a new understanding of how the SMG's endothelial barrier behaves under physiological challenges.

Bu protokolde, submandibuler bezin (SMG) endotel bariyer fonksiyonu, iki fotonlu lazer taramalı mikroskop altında test hayvanı modellerinin açısal damarlarına in vivo olarak farklı moleküler ağırlıklı floresan izleyiciler enjekte edilerek değerlendirilmiştir.

Bu protokol, fare submandibuler bezlerindeki sıkı endotel kavşaklarının işlevini değerlendirmek için in vivo parasellüler geçirgenlik tespit önlemini açıklamaktadır. Bununla birlikte, iki foton lazer tarama mikroskobu, sadece geleneksel konfokal mikroskopinin avantajına sahip olmakla kalmaz, aynı zamanda daha derin doku ve görüntüyü daha net bir şekilde tespit etmek için de kullanılabilir. Bu deney, farklı dokuların vasküler geçirgenliğini, özellikle de hayvanların yüzey dokularını ve organlarını değerlendirmek için iyi bir yöntem ölçüsü sağlar.

Geçirgenlik testi için floresan sinyalleri arasındaki paraziti en aza indirmek için floresein, izotiyosiyanat, etiketli dekstran ve farklı uyarma emisyon spektrumlarına sahip dekstran etiketli Rhodamine B gibi uygun izleyicileri seçerek başlayın. İzleri steril fosfat tamponlu salin halinde mililitre stoklar başına 100 miligram halinde seyreltin ve ışıktan korunan alikotları eksi 20 santigrat derecede saklayın. 8 ila 10 haftalık erkek vahşi tip fareyi anestezi altına aldıktan sonra, göz küresinin bir tarafının hafifçe çıkıntı yapmasını sağlamak için farenin başını ve boynunu yavaşça tutun.

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