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Related Concept Videos

Reporter Genes02:11

Reporter Genes

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Related Experiment Video

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Functional Imaging of Brown Fat in Mice with 18F-FDG micro-PET/CT
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Fluorescent Genetic Tools for Studying Brown Fat Development and Function in Mice.

Rajib Mukherjee1, Joan Sanchez-Gurmaches2,3,4

  • 1Division of Endocrinology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA.

Methods in Molecular Biology (Clifton, N.J.)
|February 15, 2022
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Researchers developed new methods to track brown fat cells and their precursors using reporter mice. These techniques aid in understanding brown adipose tissue development and function in various conditions.

Keywords:
Adipocyte precursor cellsClearingConfocal microscopyDevelopmentFlow cytometryImmunofluorescenceLineage tracingmTmG

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Isolating Brown Adipocytes from Murine Interscapular Brown Adipose Tissue for Gene and Protein Expression Analysis
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Isolating Brown Adipocytes from Murine Interscapular Brown Adipose Tissue for Gene and Protein Expression Analysis
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Area of Science:

  • Cell Biology
  • Developmental Biology
  • Metabolic Research

Background:

  • Understanding brown adipose tissue (BAT) development and function is crucial for metabolic health.
  • Identifying and isolating brown adipocyte precursors and mature adipocytes is key to studying BAT.
  • Current techniques may have limitations in fully characterizing these cell populations during development and disease.

Purpose of the Study:

  • To establish and validate novel protocols for tracing and isolating brown adipocyte precursors and adipocytes.
  • To utilize the R26R-mTmG reporter mouse model for enhanced visualization and quantification of brown adipose cells.
  • To provide tools for investigating the impact of genetic and environmental factors on brown adipocyte biology.

Main Methods:

  • Utilized R26R-mTmG reporter mice for lineage tracing and cell identification.
  • Employed confocal microscopy for high-resolution imaging of brown adipose tissue.
  • Applied fluorescence-based flow cytometry for quantitative analysis and cell sorting.

Main Results:

  • Successfully developed and optimized protocols for isolating and analyzing brown adipocyte precursors and adipocytes.
  • Demonstrated the efficacy of R26R-mTmG reporter system in visualizing distinct cell populations within thermogenic tissues.
  • Established a workflow combining microscopy and flow cytometry for comprehensive characterization.

Conclusions:

  • The reported protocols offer robust methods for studying brown adipocyte development and function.
  • These techniques are valuable for investigating alterations in brown adipose tissue biology.
  • The findings provide essential tools for future research in metabolic diseases and thermogenesis.