Name: In-vivo-Markierung stabiler Isotope zur massenspektrometrischen Identifizierung von väterlichen Metaboliten, die während der Befruchtung von Spermien auf die Eizelle übertragen werden (Video) | JoVE Journal
Uploaded: 2025-06-17T14:00:00.000+00:00
Duration: 5 min 55 s
Description: Erfahren Sie die massenspektrometrische Identifizierung väterlicher Metaboliten bei der Befruchtung mittels stabiler Isotop-Markierung in Mäusen. Diese Methode eröffnet neue Einblicke in epigenetische Vererbungsmechanismen.

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5 minutes

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<table><tbody><tr><td>2H2O gefiltert, 99,8 Atom % 2H</td><td>Sigma Aldrich</td><td>756822</td><td></td></tr><tr><td>Aceton</td><td>Sigma Aldrich</td><td>179124</td><td></td></tr><tr><td>Agilent&nbsp; SIL-8</td><td>Agilent (Santa Clara, CA, USA)</td><td></td><td></td></tr><tr><td>C16-Ceramid-13C3-15N</td><td>LGC-Norm</td><td>TRC-C342770-25MG</td><td></td></tr><tr><td>Dichlormethan</td><td>Sigma Aldrich</td><td>34856</td><td></td></tr><tr><td>Diethylether</td><td>Sigma Aldrich</td><td>673811</td><td></td></tr><tr><td>Ethanol</td><td>Sigma Aldrich</td><td>1,11,727</td><td></td></tr><tr><td>HCG (Humanes Choriongonadotropin,</td><td>Sigma Aldrich</td><td>CG5-1VL</td><td></td></tr><tr><td>L-Lysin-&epsilon;-15N-Hydrochlorid</td><td>Sigma Aldrich</td><td>608971</td><td></td></tr><tr><td>Männlich und weiblich&nbsp; C57BL/6JOlaHsd-Mäuse</td><td>Inotiv&nbsp;</td><td>C57BL/6JOlaHsd</td><td></td></tr><tr><td>Methyl-tert-butylether (MTBE)</td><td>Sigma Aldrich</td><td>306975</td><td></td></tr><tr><td>MetOH</td><td>Sigma Aldrich</td><td>1,06,035</td><td></td></tr><tr><td>N,O-Bis(trimethylsilyl(TMS) Trifluoracetamid (BSTFA)</td><td>Supelco</td><td>15238</td><td></td></tr><tr><td>PBS</td><td>Applichem</td><td>Nr. A0965-9010</td><td></td></tr><tr><td>PMSG (Trächtige Stute" s Serum)</td><td>Lee Biosolutions</td><td>493-10</td><td></td></tr><tr><td>Pyridin</td><td>Sigma Aldrich</td><td>360570</td><td></td></tr><tr><td>Shimadzu GCMS 2010plus&nbsp;</td><td>Shimadzu</td><td></td><td></td></tr><tr><td>Sitostanol-5,6,22,23-d4</td><td>Sigma Aldrich</td><td>680028</td><td></td></tr><tr><td>Natriumtrifluoracetat</td><td>Sigma Aldrich</td><td>1615138</td><td></td></tr><tr><td>SolariX XR 7T&nbsp;</td><td>Bruker Daltonics</td><td></td><td></td></tr><tr><td>TimsTOF Pro Q-TOF&nbsp;</td><td>Bruker Daltonics</td><td></td><td></td></tr><tr><td>U13C Glukose&nbsp;</td><td>Sigma Aldrich</td><td>389374</td><td></td></tr><tr><td>Ultimative RS3000 UHPLC&nbsp;</td><td>Thermo Fisher Scientific</td><td></td><td></td></tr><tr><td>VanGuard CSHTM Vorsäule (5,0 x 2,1 mm; 1,7 μm; m, 130 &Aring;)</td><td>Gewässer</td><td></td><td></td></tr></tbody></table>

in-vivo-markierung stabiler isotope zur massenspektrometrischen identifizierung von väterlichen metaboliten, die während der befruchtung von spermien auf die eizelle übertragen werden

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

Um den Metabolitentransfer von den Spermien auf die Eizelle zu überwachen, werden Morulae, die von C57 BL/6J-Weibchen isoliert wurden, die mit Bullen aus MARKIERTEN und UNMARKIERTEN Gruppen gepaart wurden, gesammelt und mittels Massenspektrometrie-basierter Metabolomik analysiert. Die Metaboliten wurden wie in Schritt 6-7 beschrieben extrahiert und wie in Schritt 8 beschrieben mit TMS derivatisiert. Es wurde ein metabolomisches und lipidomisches Profiling von MARKIERTEN Morulae, UNMARKIERTEN Morulae, UNMARKIERTEN Eizelle...

In-vivo-Markierung stabiler Isotope zur massenspektrometrischen Identifizierung von väterlichen Metaboliten, die während der Befruchtung von Spermien auf die Eizelle übertragen werden

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

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Techniques for Imaging Ca 2 + Signaling in menschlichen Spermien

Stabile Isotope Profilierung mittels Gaschromatographie-Massenspektrometrie des Vermittlers Stoffwechselflusses ist in dem Fadenwurm...

Stabilen Isotopen Profiling von Intermediary Metabolic Flux in Entwicklungs-und Erwachsenen-Stadium Caenorhabditis elegans</em

Dieses Protokoll demonstriert die Extraktion und Analyse von freien und veresterten bioaktiven Fettsäuren aus Zellen. Fettsäuren werden mittels stabiler Isotopenverdünnung, chiraler Flüssigkeitschromatographie und Elektroneneinfang der atmosphärischen chemischen Ionisation, Mehrfachreaktionsüberwachung, Massenspektrometrie (SID-LC-ECAPCI-MRM/MS) genau quantifiziert.

Cellular Lipidextraktion für Targeted Stable Isotope Dilution Flüssigchromatographie-Massenspektrometrie-Analyse

Bisulfit-Mutagenese ist der Goldstandard für die Analyse von DNA-Methylierung. Unsere modifizierte Protokoll ermöglicht DNA-Methylierungs-Analyse auf der Ebene einzelner Zellen und wurde speziell für die einzelnen Eizellen entwickelt. Es kann auch für die Spaltung-Embryonen verwendet werden.

Einzel Oocyte Bisulfit-Mutagenese

Die proteomische Analyse eines jeden Zelltyps hängt stark von der Reinheit und der Vorfraktionierung des Ausgangsmaterials ab, um die Probe vor der Flüssigchromatographie-Massenspektrometrie (MS) zu entkomplexifizieren. Durch den Einsatz von Rückspültechniken können reine Spermien aus Nagetieren gewonnen werden. Nach der Verdauung können Phosphopeptide mit TiO2 angereichert werden.

Phosphopeptid Analyse der Nebenhoden Nagetier Spermien

Wir beschreiben Schritte, mit denen schnell in Situ Probenahme von einen kleinen Teil einer einzelnen Zelle mit hoher Präzision und minimalen Invasion mit Kapillar-basierten Mikro-Probenahme, chemische Charakterisierung einer Momentaufnahme der metabolischen Aktivität in zu erleichtern Embryonen mit einer speziell angefertigten Einzelzelle Kapillarelektrophorese und Massenspektrometrie-Plattform zu leben.

Mikrosonde Capillary Electrophoresis Mass Spectrometry für einzellige Metabolomik in Live-Frosch (Xenopus Laevis) Embryonen

Die Fettsäure trophische Marker Ansatz, d. h. die Aufnahme von Fettsäuren als gesamte Molekül und Transfer in das Gewebe der Verbraucher ohne oder geringfügige Änderung, durch Wissenslücken im Fettsäure-Stoffwechsel der kleinen Boden Wirbellosen behindert wird. Isotopologue Profilierung dient als ein wertvolles Instrument zur trophische Interaktionen zu entwirren.

Fettsäure- 13C Isotopologue Profilierung gibt Einblick in trophische Carbon Transfer und Fettstoffwechsel der Wirbellosen Verbraucher

In dieser Arbeit beschreiben wir eine massenspektrometriebasierte proteomische Charakterisierung von Zelllinien mit bekannten Gewebeschicksalen im Embryo von Xenopus laevis im Wirbeltier.

Zellliniengesteuerte Massenspektrometrie-Proteomik im sich entwickelnden (Frosch-)Embryo

Hier wird eine Methode zum Screening von Lipidstrukturen durch Markierung stabiler Isotope unter Verwendung ihrer Retentionszeit, Mobilität und Fragmentierung vorgestellt.

De-novo-Verfolgung von Lipiden mittels stabiler Isotopenmarkierung LC-TIMS-TOF MS/MS

Dieses Video demonstriert die Herstellung von ultra-dünnen Matrix / Analyt-Schicht für die Analyse von Peptiden und Proteinen durch Matrix-Assisted Laser Desorption Ionisation-Massenspektrometrie (MALDI-MS).

MALDI-Probenvorbereitung: Die Ultra Thin-Layer-Methode

Erfahren Sie die massenspektrometrische Identifizierung väterlicher Metaboliten bei der Befruchtung mittels stabiler Isotop-Markierung in Mäusen. Diese Methode eröffnet neue Einblicke in epigenetische Vererbungsmechanismen.

In-vivo-Markierung stabiler Isotope zur massenspektrometrischen Identifizierung von väterlichen Metaboliten, die während der Befruchtung von Spermien auf die Eizelle übertragen werden (Video) | JoVE Journal

In-vivo-Markierung stabiler Isotope zur massenspektrometrischen Identifizierung von väterlichen Metaboliten, die während der Befruchtung von Spermien auf die Eizelle übertragen werden

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