Name: Stable Isotope In-Vivo Labeling for Mass-Spectrometry Identification of Paternal Metabol Ites Transport to Spermation During fertilization (수정 중 정자에서 난모세포로 전달된 부계 대사산물의 질량 분광법 식별을 위한 안정 동위원소 생체 내 라벨링) (동영상) | JoVE Journal
Uploaded: 2025-06-17T14:00:00.000+00:00
Duration: 5 min 55 s
Description: 배우세요 안정 동위원소 생체 내 라벨링을 활용한 질량 분광법으로 수정 중 정자에서 난모세포로 전달된 부계 대사산물을 식별하는 방법을. 이 연구는 초기 배아 발달과 후성 유전에서 부계 기여의 역할을 밝힙니다.

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5 minutes

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<li>Wilkinson, D. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+validation+of+the+application+of+D2O+stable+isotope+tracer+techniques+for+monitoring+day-to-day+changes+in+muscle+protein+subfraction+synthesis+in+humans%5BTitle%5D)+AND+%22Am+J+Physiol+Endocrinol+Metab%22%5BJournal%5D)">A validation of the application of D2O stable isotope tracer techniques for monitoring day-to-day changes in muscle protein subfraction synthesis in humans.</a> Am J Physiol Endocrinol Metab. 306 (5), E571-E579 (2014).</li>
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<table><tbody><tr><td>2H2O 여과, 99.8 원자 % 2H</td><td>시그마 알드리치</td><td>756822</td><td></td></tr><tr><td>아세톤</td><td>시그마 알드리치</td><td>179124</td><td></td></tr><tr><td>애질런트&nbsp; 실-8</td><td>애질런트(미국 캘리포니아주 산타클라라)</td><td></td><td></td></tr><tr><td>C16-세라마이드-13C3-15N</td><td>LGC 표준</td><td>TRC-C342770-25MG</td><td></td></tr><tr><td>디클로로메탄</td><td>시그마 알드리치</td><td>34856</td><td></td></tr><tr><td>디에틸 에테르&nbsp;</td><td>시그마 알드리치</td><td>673811</td><td></td></tr><tr><td>에탄올</td><td>시그마 알드리치</td><td>1,11,727</td><td></td></tr><tr><td>HCG(인간 융모막 성선 자극 호르몬,</td><td>시그마 알드리치</td><td>CG5-1VL</td><td></td></tr><tr><td>L-라이신-&엡실론;-15N 염산염</td><td>시그마 알드리치</td><td>608971</td><td></td></tr><tr><td>남성과 여성&nbsp; C57BL/6JOlaHsd 마우스</td><td>이노티브&nbsp;</td><td>C57BL/6JOlaHsd</td><td></td></tr><tr><td>메틸 tert-부틸 에테르(MTBE)</td><td>시그마 알드리치</td><td>306975</td><td></td></tr><tr><td>기상OH</td><td>시그마 알드리치</td><td>1,06,035</td><td></td></tr><tr><td>N,O-비스(트리메틸실릴(TMS) 트리플루오로아세트아미드(BSTFA)</td><td>수펠코</td><td>15238</td><td></td></tr><tr><td>PBS</td><td>응용</td><td>A0965-9010</td><td></td></tr><tr><td>PMSG(임신한 암말' S 혈청)</td><td>리 바이오솔루션즈</td><td>493-10</td><td></td></tr><tr><td>피리딘</td><td>시그마 알드리치</td><td>360570</td><td></td></tr><tr><td>시마즈 GCMS 2010plus&nbsp;</td><td>시마즈</td><td></td><td></td></tr><tr><td>시토스타놀-5,6,22,23-d4</td><td>시그마 알드리치</td><td>680028</td><td></td></tr><tr><td>트리플루오로아세테이트 나트륨</td><td>시그마 알드리치</td><td>1615138</td><td></td></tr><tr><td>솔라리엑스 XR 7T&nbsp;</td><td>브루커 달토닉스</td><td></td><td></td></tr><tr><td>TimsTOF 프로 Q-TOF&nbsp;</td><td>브루커 달토닉스</td><td></td><td></td></tr><tr><td>U13C 포도당&nbsp;</td><td>시그마 알드리치</td><td>389374</td><td></td></tr><tr><td>궁극의 RS3000 UHPLC&nbsp;</td><td>써모 피셔 사이언티픽</td><td></td><td></td></tr><tr><td>VanGuard CSHTM 프리컬럼(5.0 &times; 2.1 mm; 1.7 &mu; m, 130 &Aring;)</td><td>바다</td><td></td><td></td></tr></tbody></table>

stable isotope in-vivo labeling for mass-spectrometry identification of paternal metabol ites transport to spermation during fertilization (수정 중 정자에서 난모세포로 전달된 부계 대사산물의 질량 분광법 식별을 위한 안정 동위원소 생체 내 라벨링)

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

정자에서 난모세포로의 대사산물 이동을 모니터링하기 위해 LABELED 및 UNLABELED 그룹의 씨수말과 짝짓기한 C57 BL/6J 암컷에서 분리된 모룰라를 질량 분광법 기반 대사체학에 의해 수집하고 분석합니다. 대사 산물은 6-7 단계에서 설명한 대로 추출하고 8 단계에서 설명한 대로 TMS로 유도체화했습니다. LABELED morulae, UNLABELED morulae, UNLABELED 난모세포, LABELED 및 UNLABELED 정자 세포의 대사체 및 지질체 프로파일링을 수행했습니다. 결과 프로파일은 LABELED morulae 및 LABELED 정자 세포 모두에 구체적으로 존재하고 UN morulae 및 UNLABLLED 난모세포에는 없는 화학 종을 식별하기 위해 통계적으로 분석, 정렬 및 비교되었습니다. 후보 화학 종은 더 무거운 분자 질량(주로 2-4 Da의 Δm/z)을 부여받아야 하며 자연적으로 발생하는 화학 종과 함께 용리되어야 합니다. 이러한 더 무거운 종의 존재는 수정 시 특정 부계 대사 산물이 난모세포...

Stable Isotope In-Vivo Labeling for Mass-Spectrometry Identification of Paternal Metabol Ites Transport to Spermation During fertilization (수정 중 정자에서 난모세포로 전달된 부계 대사산물의 질량 분광법 식별을 위한 안정 동위원소 생체 내 라벨링)

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

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JoVE Journal

Biology

생물학

이미징 CA의 기술 2 + 인간 정자의 시그널링

중간 신진 대사 유량의 가스 크로마 토그래피 질량 spectrometric 분석하여 안정 원소 프로파일은 선충류에 설명되어...

개발하고 성인 단계에서 중개 신진 대사 플럭스 안정 원소 프로파일 Caenorhabditis 엘레간스</em

이 프로토콜은 세포에서 유리 및 에스테르화된 생체 활성 지방산의 추출 및 분석을 보여줍니다. 지방산은 안정 동위원소 희석, 키랄 액체 크로마토그래피, 전자 포획, 대기 화학 이온화, 다중 반응 모니터링 질량 분석법(SID-LC-ECAPCI-MRM/MS)을 사용하여 정확하게 정량화됩니다.

타겟 안정 동위 원소 희석 액체 크로마 토그래피 - 질량 분광법 분석을위한 세포 지질 추출

Bisulfite의 돌연변이 유발이의 DNA methylation 분석을위한 황금 표준입니다. 우리 수정된 프로토콜은 단일 세포 수준에서 DNA의 methylation 분석을 허용하고 특별히 개별 oocytes를 위해 설계되었습니다. 또한 절단 단 배아 사용할 수 있습니다.

단일 Oocyte의 Bisulfite의 돌연변이 유발

모든 세포 유형에 대한 단백질체 분석은 액체 크로마토그래피 질량 분석법(MS) 전에 시료의 복잡성을 줄이기 위해 출발 물질의 순도와 사전 분획에 크게 의존합니다. 역 세척 기술을 사용하면 설치류에서 순수한 정자를 얻을 수 있습니다. 분해 후 인산펩티드는 TiO2를 사용하여 농축할 수 있습니다.

설치류 부고환 정자의 포스 포 분석

우리는 빠른 시내 현장 샘플링의 높은 정밀도와 최소한의 침공에 신진 대사 활동의 스냅샷의 화학 특성을 촉진 하기 위하여 모 세관 기반 마이크로-샘플링를 사용 하 여 개별 셀의 작은 부분을 사용할 수 있는 단계를 설명 배아를 사용자 단일 셀 모 세관 전기 이동 법 질량 분석 플랫폼을 사용 하 여 라이브.

단일 셀 대사체학 라이브 개구리 (Xenopus laevis) 배아에 대 한 현미경 모 세관 전기 이동 법 질량 분석

지방산 영양 마커 접근, 즉, 전체 분자와 소비자 조직으로 전송으로 지방산의 동화 없이 또는 사소한 수정, 작은 토양 무척 추 동물의 지방 산 성 대사에 지식 격차에 의해 방해 된다. Isotopologue 프로 파일링 풀 영양 상호 작용 하는 귀중 한 도구로 제공 됩니다.

영양 탄소 전송 및 무척 추 동물 소비자의 지질 대사에 대 한 통찰력을 제공 합니다 지방산 13C Isotopologue 프로 파일링

여기에서 우리는 척추동물 Xenopus laevis 배아에서 알려진 조직 운명을 가진 세포 계통의 질량 분석 기반 단백질체 특성 분석을 설명합니다.

발달 중인 (개구리) 배아의 세포 계통 유도 질량 분석법 단백질체학

여기에는 머무름 시간, 이동성 및 단편화를 사용하여 안정 동위원소 라벨링을 통해 지질 구조를 스크리닝하는 방법이 제시되어 있습니다.

안정 동위원소 라벨링 LC-TIMS-TOF MS/MS를 사용한 새로운 지질 추적

이 비디오는 매트릭스를 이용한 레이저 탈착 이온화 질량 분석법 (든 - MS)에 의해 펩티드와 단백질 분석을위한 울트라 얇은 매트릭스 / analyte 레이어의 준비를 보여줍니다.

든 샘플 준비 : 울트라 씬 레이어 방법

배우세요 안정 동위원소 생체 내 라벨링을 활용한 질량 분광법으로 수정 중 정자에서 난모세포로 전달된 부계 대사산물을 식별하는 방법을. 이 연구는 초기 배아 발달과 후성 유전에서 부계 기여의 역할을 밝힙니다.

Stable Isotope In-Vivo Labeling for Mass-Spectrometry Identification of Paternal Metabol Ites Transport to Spermation During fertilization (수정 중 정자에서 난모세포로 전달된 부계 대사산물의 질량 분광법 식별을 위한 안정 동위원소 생체 내 라벨링) (동영상) | JoVE Journal

Stable Isotope In-Vivo Labeling for Mass-Spectrometry Identification of Paternal Metabol Ites Transport to Spermation During fertilization (수정 중 정자에서 난모세포로 전달된 부계 대사산물의 질량 분광법 식별을 위한 안정 동위원소 생체 내 라벨링)

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