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

Xiangdi Mao; Sainan Min; Qihua He; Xin Cong

doi:10.3791/64167

In vivo Detecção de Permeabilidade Vascular em Glândula Submandibular de Camundongo

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In Vivo Vascular Permeability Detection in Mouse Submandibular Gland

In vivo Detecção de Permeabilidade Vascular em Glândula Submandibular de Camundongo

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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.

No presente protocolo, a função de barreira endotelial da glândula submandibular (SMG) foi avaliada por meio da injeção de diferentes marcadores fluorescentes de peso molecular nas veias angulares de modelos animais de teste in vivo sob um microscópio de varredura a laser de dois fótons.

O presente protocolo descreve uma medida de detecção de permeabilidade paracelular in vivo para avaliar a função de junções endoteliais apertadas em glândulas submandibulares de camundongos. A microscopia de varredura a laser de dois fótons, no entanto, não só tem a vantagem da microscopia confocal convencional, mas também pode ser usada para detectar tecidos e imagens mais profundos com mais clareza. Este experimento fornece uma boa medida de método para avaliar a permeabilidade vascular de diferentes tecidos, especialmente os tecidos superficiais e órgãos de animais.

Comece escolhendo marcadores apropriados, como fluoresceína, isotiocianato, dextrano marcado e rodamina B, dextran marcado com espectros distintos de emissões de excitação, para minimizar a interferência entre os sinais de fluorescência para o ensaio de permeabilidade. Diluir os vestígios em solução salina tamponada com fosfato estéril em estoques de 100 miligramas por mililitro e armazenar as alíquotas protegidas da luz a menos 20 graus Celsius. Depois de anestesiar o rato macho do tipo selvagem de 8 a 10 semanas de idade, segure suavemente a cabeça e o pescoço do rato para fazer com que um lado do globo ocular se projete ligeiramente.

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