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

Xiangdi Mao; Sainan Min; Qihua He; Xin Cong

doi:10.3791/64167

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In Vivo Vascular Permeability Detection in Mouse Submandibular Gland

体内小鼠下颌下腺血管通透性检测

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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.

在本协议中，通过在双光子激光扫描显微镜下将不同分子加权荧光示踪剂注射到体内测试动物模型的角静脉中来评估下颌下腺（SMG）的内皮屏障功能。

本协议描述了一种体内细胞旁通透性检测措施，以评估小鼠下颌下腺中紧密内皮连接的功能。然而，双光子激光扫描显微镜不仅具有传统共聚焦显微镜的优势，而且还可用于更清晰地检测更深的组织和图像。本实验为评估不同组织特别是动物表面组织和器官的血管通透性提供了很好的方法测量方法。

首先选择合适的示踪剂，例如荧光素、异硫氰酸酯（标记的葡聚糖）和罗丹明B（标记的右旋糖酐）具有不同的激发发射光谱，以最大程度地减少渗透性测定中荧光信号之间的干扰。将痕量稀释到无菌磷酸盐缓冲盐水中，制成每毫升100毫克的储备液，并将等分试样储存在零下20摄氏度的避光处。麻醉8至10周龄雄性野生型小鼠后，轻轻握住小鼠的头部和颈部，使眼球的一侧略微突出。

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