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Biology
小鼠锁骨上棕色脂肪组织的解剖、组织学处理及基因表达分析
小鼠锁骨上棕色脂肪组织的解剖、组织学处理及基因表达分析
JoVE Journal
Biology
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JoVE Journal Biology
Dissection, Histological Processing, and Gene Expression Analysis of Murine Supraclavicular Brown Adipose Tissue

小鼠锁骨上棕色脂肪组织的解剖、组织学处理及基因表达分析

Full Text
3,253 Views
06:50 min
March 29, 2024

DOI: 10.3791/66475-v

Mark G Waterstraat1,2, ZiYi Wang1, Mari Kogiso1, Rommel Caballero-Juarez1,2, Miao-Hsueh Chen1

1Children's Nutrition Research Center, Department of Pediatrics,Baylor College of Medicine, 2Department of BioSciences,Rice University

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study provides a detailed protocol for the dissection of supraclavicular brown adipose tissue (scBAT) from mice, which is deemed a valuable model for analyzing brown fat due to its anatomical similarities to human brown adipose tissue. The focus is on improving accessibility to scBAT research through precise dissection and subsequent histological and gene expression analyses.

Key Study Components

Research Area

  • Supraclavicular brown adipose tissue (scBAT)
  • Gene expression analysis
  • Histological techniques

Background

  • scBAT is recognized for its potential importance in metabolic research.
  • The anatomical proximity to major veins poses challenges in dissection.
  • A need for accessible and accurate extraction methods has been identified.

Methods Used

  • Dissection using a microscope for precision.
  • Gene expression analysis following tissue homogenization.
  • Hematoxylin and eosin staining for histological assessment.

Main Results

  • The protocol described improves extraction efficiency and the quality of gene expression data.
  • scBAT displays characteristic tissue structures found in brown adipose tissue.
  • The study lays groundwork for understanding scBAT's genetic lineage and physiological roles.

Conclusions

  • This protocol enables more effective research into the roles of scBAT in metabolism.
  • The findings contribute to the broader field of adipose tissue biology.

Frequently Asked Questions

What is supraclavicular brown adipose tissue (scBAT)?
scBAT is a specific type of brown fat located near the clavicles, significant for its metabolic functions.
Why is scBAT important for research?
Due to its anatomical similarities to human brown adipose tissue, scBAT serves as an excellent model for studying metabolic processes.
What challenges exist when dissecting scBAT?
The small size and its location near major veins can complicate the dissection process.
How does the protocol increase extraction efficiency?
It employs a dissecting microscope for enhanced precision and utilizes tissue homogenization to improve RNA yield.
What are the key techniques used in the study?
The study uses dissection, histological staining, and gene expression analysis techniques.
What are the main findings regarding scBAT's cellular structure?
Hematoxylin and eosin staining confirmed the presence of typical brown adipose tissue characteristics in scBAT samples.
What future research directions does this study suggest?
The study suggests defining the genetic lineage and physiological functions of scBAT, as well as developing ex-vivo models for further research.

在这里,我们提供了一个实用的程序,用于解剖和执行小鼠锁骨上棕色脂肪组织的组织学和基因表达分析。

1 我们的实验室专注于建立2 锁骨上棕色脂肪组织 3 或 SCBAT 作为棕色脂肪分析的有价值的仓库 4,因为它的解剖学相似性 5 与人类棕色脂肪组织相似。6 该协议旨在提高SCBAT 研究的可及性 7。8 这个仓库可能令人生畏,因为它的尺寸小 9 并且靠近颈部的大静脉, 10 因此,必须提供 11 一种提取和处理 SCBAT 的简单方法。

12 我们的方法脱颖而出13 首先是在提取 SCBAT 时使用解剖显微镜 14。15 这提高了精度 16 和提取效率,17 这在使用如此小的组织仓库时都至关重要 18。19 其次,我们在进行基因表达分析 21 之前对冷冻的 SCBAT 样品 20 进行了匀浆,以提高 RNA 产量,以补偿 22 如此少量的可用组织。

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