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Biology
酸化重水素を用いた褐色脂肪組織における de novo 脂肪酸合成の定量的測定(英語)
酸化重水素を用いた褐色脂肪組織における de novo 脂肪酸合成の定量的測定(英語)
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JoVE Journal Biology
Quantitative Determination of De Novo Fatty Acid Synthesis in Brown Adipose Tissue Using Deuterium Oxide

酸化重水素を用いた褐色脂肪組織における de novo 脂肪酸合成の定量的測定(英語)

Full Text
1,542 Views
07:34 min
May 12, 2023

DOI: 10.3791/64219-v

Rory Turner*1, Rajib Mukherjee*2, Martina Wallace1, Joan Sanchez-Gurmaches2,3,4

1UCD Conway Institute and UCD Institute of Food and Health, School of Agriculture and Food Science,University College Dublin, 2Division of Endocrinology,Cincinnati Children’s Hospital Medical Center, Cincinnati, 3Division of Developmental Biology,Cincinnati Children’s Hospital Medical Center, Cincinnati, 4Department of Pediatrics,University of Cincinnati College of Medicine

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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 presents a novel quantitative method using deuterium oxide and gas chromatography mass spectrometry (GCMS) to analyze fatty acid de novo lipogenesis in brown adipose tissue in vivo. The method offers a non-invasive approach to accurately assess fatty acid synthesis flux, providing insights into metabolic status.

Key Study Components

Research Area

  • Metabolic pathways
  • Fatty acid synthesis
  • Brown adipose tissue analysis

Background

  • Fatty acid synthesis is under-studied compared to glucose disposal.
  • Existing methods may not accurately reflect synthesis flux due to impacts from upstream glucose handling.
  • This method is applicable across different mouse models and various tissues.

Methods Used

  • Sample preparation and extraction techniques using deuterium oxide.
  • In vivo analysis of brown adipose tissue in mouse models.
  • Gas chromatography mass spectrometry (GCMS) for measurement of fatty acid methyl esters (FAMEs).

Main Results

  • Documented deuterium enrichment levels in body water and fatty acids.
  • Observed differences in fatty acid enrichment based on temperature conditions.
  • Findings validate the efficiency of the method in assessing total fatty acidity synthesis in brown adipose tissue.

Conclusions

  • The study successfully demonstrates an effective method for evaluating fatty acid synthesis in vivo.
  • It contributes significantly to the understanding of metabolic processes and has potential applications in further biological research.

Frequently Asked Questions

What is the significance of using deuterium oxide in this method?
Deuterium oxide allows for the precise measurement of fatty acid synthesis by incorporating deuterium into fatty acids during metabolic processes.
How does this method improve upon traditional techniques?
This method provides a non-invasive approach that reflects fatty acid flux more accurately than techniques relying on isotope-labeled glucose.
Can this method be used in other tissues apart from brown adipose tissue?
Yes, the method is potentially adaptable for studying fatty acid synthesis in various tissues in different mouse models.
What was observed regarding temperature's effect on fatty acid enrichment?
The study found higher fatty acid enrichment at thermoneutral conditions compared to lower temperatures.
What are the potential implications of this research?
Understanding fatty acid synthesis's role in metabolism could inform research on obesity and metabolic diseases.
Is the method suitable for analyzing all fatty acids?
Yes, the method analyzes total fatty acids across multiple lipid classes, although additional separation techniques may be needed for specific studies.
What precautions should be taken during GCMS analysis?
It is essential to avoid overloading samples and to check for background peaks to ensure accurate results.

ここでは、in vivoで褐色脂肪組織における全脂肪酸de novo脂肪生成を分析するために、酸化重水素とガスクロマトグラフィー質量分析(GCMS)を利用した安価な定量法を紹介します。

グルコース処理などの他の代謝経路とは対照的に、脂肪酸合成は日常的に評価されていないため、代謝状態の解釈が不完全です。この方法の主な利点は、比較的非侵襲的であり、上流のグルコース処理の変化が結果に影響を与える可能性のある同位体標識グルコースを使用するよりも、脂肪酸合成フラックスをよりよく反映する可能性が高いことです。ここでは褐色脂肪組織に焦点を当てていますが、この方法は、あらゆるマウスモデルのあらゆる組織を研究するのに役立つ可能性があります。

まず、サンプルをドライアイスの上に置きます。事前にラベル付けされた微量遠心チューブを用意し、分析天びんの上に置き、天びんを風袋引きします。次に、ピンセットとメスまたはスチールカミソリの刃をドライアイスの上に10〜20秒間置き、冷まします。

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生物学 第195号 褐色脂肪組織 酸化重水素 代謝経路 機能的役割 全身代謝ホメオスタシス 生理学的プロセス 病理学的プロセス グルコース処理 機能評価 詳細なプロトコル 定量的方法 ガスクロマトグラフィー質量分析(GCMS) 全脂肪酸de novo合成 炭素源 サンプル調製 ダウンストリーム計算 褐色脂肪 代謝ホメオスタシス

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