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Biology
Dissezione, elaborazione istologica e analisi dell'espressione genica del tessuto adiposo bruno s...
Dissezione, elaborazione istologica e analisi dell'espressione genica del tessuto adiposo bruno s...
JoVE Journal
Biology
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JoVE Journal Biology
Dissection, Histological Processing, and Gene Expression Analysis of Murine Supraclavicular Brown Adipose Tissue

Dissezione, elaborazione istologica e analisi dell'espressione genica del tessuto adiposo bruno sopraclavicolare murino

Full Text
3,266 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.

In questo articolo, forniamo una procedura pratica per la dissezione e l'esecuzione di analisi istologiche e di espressione genica del tessuto adiposo bruno sopraclavicolare murino.

1 Il nostro laboratorio si concentra sulla definizionedel tessuto adiposo bruno sopraclavicolare 3 o SCBAT come un prezioso deposito 4 per l'analisi del grasso bruno a causa delle sue somiglianze anatomiche 5 con il tessuto adiposo bruno umano. 6 Questo protocollo ha lo scopo di aumentare l'accessibilità7 della ricerca SCBAT. 8 Questo deposito può essere intimidatorio a causa delle sue piccole dimensioni 9 e della vicinanza a grandi vene nel collo, 10 quindi è imperativo fornire 11 un metodo semplice di estrazione e lavorazione SCBAT.

12 La nostra metodologia si distingue13 in primo luogo per l'uso di un microscopio da dissezione 14 durante l'estrazione dell'SCBAT. 15 Ciò aumenta la precisione 16 e l'efficienza di estrazione, 17 che sono entrambi critici quando si lavora 18 con un deposito di tessuti così piccolo. 19 In secondo luogo, abbiamo omogeneizzato i campioni di SCBAT congelati 20 prima di eseguire l'analisi dell'espressione genica 21 per aumentare la resa dell'RNA per compensare 22 una quantità così bassa di tessuto disponibile.

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