Name: 受精中に精子から卵子に移された父方代謝産物の質量分析同定のための安定同位体 生体 内標識 (動画) | JoVE Journal
Uploaded: 2025-06-17T14:00:00.000+00:00
Duration: 5 min 55 s
Description: 学ぶ 受精時の父方代謝産物の卵子への移動をマウスを用いた質量分析と安定同位体標識で同定する方法。早期発生とエピジェネティック遺伝における父親の役割を解明します。

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5 minutes

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<table><tbody><tr><td>2H2O ろ過、99.8 原子 % 2H</td><td>シグマ・アルドリッチ</td><td>756822</td><td></td></tr><tr><td>アセトン</td><td>シグマ・アルドリッチ</td><td>179124</td><td></td></tr><tr><td>アジレント&nbsp;シル-8</td><td>Agilent (米国カリフォルニア州サンタクララ)</td><td></td><td></td></tr><tr><td>C16-セラミド-13C3-15N</td><td>LGC規格</td><td>TRC-C342770-25MG</td><td></td></tr><tr><td>ジクロロ メタン</td><td>シグマ・アルドリッチ</td><td>34856</td><td></td></tr><tr><td>ジエチルエーテル&nbsp;</td><td>シグマ・アルドリッチ</td><td>673811</td><td></td></tr><tr><td>エタノール</td><td>シグマ・アルドリッチ</td><td>1,11,727</td><td></td></tr><tr><td>HCG(ヒト絨毛性ゴナドトロピン、</td><td>シグマ・アルドリッチ</td><td>CG5-1VLシリーズ</td><td></td></tr><tr><td>L-リジン-&イプシロン;-15N塩酸塩</td><td>シグマ・アルドリッチ</td><td>608971</td><td></td></tr><tr><td>男性と女性&nbsp;C57BL/6JOlaHsdマウス</td><td>イノチフ&nbsp;</td><td>C57BL/6JOlaHsd</td><td></td></tr><tr><td>メチルtert-ブチルエーテル(MTBE)</td><td>シグマ・アルドリッチ</td><td>306975</td><td></td></tr><tr><td>気象庁</td><td>シグマ・アルドリッチ</td><td>1,06,035</td><td></td></tr><tr><td>N,O-ビス(トリメチルシリル(TMS) トリフルオロアセトアミド(BSTFA)</td><td>スペルコ</td><td>15238</td><td></td></tr><tr><td>PBSの</td><td>応用</td><td>A0965-9010</td><td></td></tr><tr><td>PMSG(妊娠中の牝馬S血清)</td><td>リー・バイオソリューションズ</td><td>493-10</td><td></td></tr><tr><td>ピリジン</td><td>シグマ・アルドリッチ</td><td>360570</td><td></td></tr><tr><td>島津製作所 GCMS 2010plus&nbsp;</td><td>島津製作所</td><td></td><td></td></tr><tr><td>シトスタノール-5,6,22,23-d4</td><td>シグマ・アルドリッチ</td><td>680028</td><td></td></tr><tr><td>トリフルオロ酢酸ナトリウム</td><td>シグマ・アルドリッチ</td><td>1615138</td><td></td></tr><tr><td>ソラリックスXR 7T&nbsp;</td><td>ブルカーダルトニックス</td><td></td><td></td></tr><tr><td>TimsTOF Pro Q-TOF&nbsp;</td><td>ブルカーダルトニックス</td><td></td><td></td></tr><tr><td>U13Cグルコース&nbsp;</td><td>シグマ・アルドリッチ</td><td>389374</td><td></td></tr><tr><td>究極のRS3000 UHPLC&nbsp;</td><td>サーモフィッシャーサイエンティフィック</td><td></td><td></td></tr><tr><td>VanGuard CSHTM プレカラム(5.0 & 2.1 mm; 1.7 & mu;m、130 &Aring;)</td><td>水</td><td></td><td></td></tr></tbody></table>

受精中に精子から卵子に移された父方代謝産物の質量分析同定のための安定同位体 生体 内標識

Stable Isotope In-Vivo Labeling for Mass-Spectrometry Identification of Paternal Metabolites Transferred from Sperm to Oocyte During Fertilization

精子から卵子への代謝産物の移動をモニターするために、C57 BL/6J雌から単離された桑実胚をLABELEDおよびUNLABELEDグループの種牡馬と交配させ、質量分析に基づくメタボロミクスによって収集および分析します。代謝物は、ステップ6〜7に記載したように抽出し、ステップ8に記載したTMSで誘導体化した。LABELD桑実胚、UNLABEL桑実胣艇、UNLABELED卵子、LABELED、およびUNLABELED精子細胞のメタボロームおよびリピドミクスプロファイリングを実施しました。得られたプロファイルを統計的に分析、整列、および比較して、特に標識された桑実胚細胞と標識された精子細胞の両方に存在し、標識されていない桑実胚とUNLABLLED卵子には存在しない化学種を特定しました。候補となる化学種は、より重い分子量(主に2〜4 DaのΔm/z)を持ち、天然に存在する対応する分子種と共溶出する必要があります。これらのより重い種の存在は、受精時に特定の父性代謝産物が卵母細胞に移行していることを示している可能性があります。
図2A</strong...

Paternal contribution to embryo genetics has so far been limited to allelic sequences for decades. A decade of results suggests, instead, that epigenetic ...

stable-isotope-vivo-labeling-for-mass-spectrometry-identification

Research

JoVE Journal

Biology

生物学

イメージングカルシウムのためのテクニック 2 +人間の精子でシグナリング

中間代謝フラックスのガスクロマトグラフィー質量分析による安定同位体のプロファイリングは、線虫で説明されています...

開発と成人期における中間代謝フラックスの安定同位体プロファイル線虫（Caenorhabditis elegans）</em

このプロトコールは、細胞からの遊離およびエステル化された生理活性脂肪酸の抽出および分析を実証します。脂肪酸は、安定同位体希釈、キラル液体クロマトグラフィー、電子捕獲、大気化学イオン化、多重反応モニタリング質量分析(SID-LC-ECAPCI-MRM/MS)を用いて正確に定量されます。...

ターゲット安定同位体希釈液体クロマトグラフィー - 質量分析のための細胞の脂質の抽出

亜硫酸塩の突然変異誘発は、DNAメチル化を解析するためのゴールドスタンダードです。私たちの変更されたプロトコルは、単一細胞レベルでのDNAメチル化解析を可能にし、特に個々の卵のために設計されました。また、開裂段階の胚に使用することができます。...

単一の卵重亜硫酸塩の突然変異誘発

プロテオミクス分析は、液体クロマトグラフィー質量分析(MS)の前にサンプルを分解するために、出発物質の純度と事前分画の両方に大きく依存します。バックフラッシング技術を使用することにより、げっ歯類から純粋な精子を得ることができます。消化後、リン酸化ペプチドはTiO2を使用して濃縮できます。...

齧歯類精巣上体精子のリン酸化ペプチドの分析

高精度、キャピラリーを用いたマイクロ サンプリングを使用して代謝活動のスナップショットの化学的特性を容易に最小の侵入の個々...

生きているカエル (アフリカツメガエル) 胚の単一細胞メタボロミクスのマイクロ プローブ キャピラリー電気泳動質量分析法

すなわち、分子全体消費者組織への伝達と脂肪酸の同化脂肪酸の栄養マーカー アプローチ変更が小さい土壌無脊椎動物の脂肪酸代謝の知識のギャップによって妨げられて無またはマイナー。Isotopologue ・...

脂肪酸13C Isotopologue プロファイリング栄養炭素転送および無脊椎動物消費者の脂質代謝への洞察力を提供します。

ここでは、脊椎動物 アフリカツメガエル...

発生中の(カエル)胚における細胞系譜ガイド質量分析プロテオミクス

ここでは、安定同位体標識により、その保持時間、移動度、およびフラグメンテーションを利用した脂質構造をスクリーニングする方法を紹介します。...

安定同位体標識 LC-TIMS-TOF MS/MS を用いた de novo 脂質の追跡

このビデオでは、マトリックス支援レーザー脱離イオン化質量分析法（MALDI -...

MALDIサンプル調製：超薄層法

学ぶ 受精時の父方代謝産物の卵子への移動をマウスを用いた質量分析と安定同位体標識で同定する方法。早期発生とエピジェネティック遺伝における父親の役割を解明します。

受精中に精子から卵子に移された父方代謝産物の質量分析同定のための安定同位体生体内標識

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受精中に精子から卵子に移された父方代謝産物の質量分析同定のための安定同位体 生体 内標識

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受精中に精子から卵子に移された父方代謝産物の質量分析同定のための安定同位体生体内標識