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Biology
マウス鎖骨上褐色脂肪組織の解剖、組織学的処理、遺伝子発現解析
マウス鎖骨上褐色脂肪組織の解剖、組織学的処理、遺伝子発現解析
JoVE Journal
Biology
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JoVE Journal Biology
Dissection, Histological Processing, and Gene Expression Analysis of Murine Supraclavicular Brown Adipose Tissue

マウス鎖骨上褐色脂肪組織の解剖、組織学的処理、遺伝子発現解析

Full Text
3,255 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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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.

ここでは、マウスの鎖骨上褐色脂肪組織の組織学的および遺伝子発現解析を解剖および実行するための実用的な手順を提供します。

1 私たちの研究室は、その解剖学的類似性から、褐色脂肪分析のための貴重なデポ 4 として、鎖骨上 2 褐色脂肪組織 3 または SCBAT を確立することに焦点を当てています。5 ヒトの褐色脂肪組織との類似性 6 このプロトコルは、SCBAT 研究のアクセシビリティ 7 を向上させることを目的としています。8 このデポは、サイズが小さいため 9 首の大きな静脈に近いため、威圧的かもしれません。 10したがって、SCBATを抽出し処理する簡単な方法を11に提供することが不可欠です。

12 我々の方法論は、まず、SCBATを抽出する際に解剖顕微鏡14を使用することによって際立っている。15 これにより、精度16と抽出効率が向上する。17 これは、このような小さな組織貯蔵庫で作業するときに重要である17。19 次に、遺伝子発現分析を行う前に、凍結SCBATサンプル20を均質化して、RNA収量を増やし、利用可能な組織の量が少ないことを補うために22。

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