Name: assessing hepatic metabolic changes during progressive colonization of germ-free mouse by 1h nmr spectroscopy
Uploaded: 2011-12-15T13:00:00
Description: Watch this Scientific Journal Video about Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy at JoVE.com

설명의 예제로 사용된 모든 NMR 스펙트럼은 재정적으로 비벼 대다가 지원되었습니다 이전에 출판 연구 7에서 파생됩니다.

1. 세균없는 동물과 샘플 컬렉션의 식민지 <ol><li> 컨트롤 (그림 1)로 될 것입니다 종래의 동물 앞에 필터를 갖춘 연습장에서 종래의 축산 룸에 isolators하고 집에서 그들로부터 세균없는 동물을 제거합니다. </li><li> 쓰레기의 혼합 절반 (옛 3 일) 세균없는 동물의 쓰레기와 컨트롤 기존의 케이지에서 찍은. 항상 더러운 일반 쓰레기의 1 / 3이 박테리아의 수준 (적어도 3 일 동안 그것을 보관) 유지하기 ?...

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이 프로토콜에서는, 우리는 더 이상 그대로 생검의 1 H 인사 MAS NMR의 프로 파일링에 의해 평가 간장 대사에 내장 microbiota의 영향을 조사하기 위해 열린 환경에서 진보 식민지 절차를 설명했다. 식민의 다양한 방법은 문헌에 설명되어있다. 정의된 microbiota과 동물을 식민지하는 가장 일반적인 방법은 구두 gavage이나 오염된 식수 19,20입니다. 이전 21 설명된대로 방패 접종도 사...

특정 시약 및 장비의 테이블 : <table border="1"><tbody><tr><th> 시약의 이름 </th><th> 회사 </th><th> 카탈로그 번호 </th><th> 댓글 </th></tr><tr><td> 2.5 mm의 microtube </td><td> 새로운 시대 </td><td> NE-H5/2.5-V-Br </td><td></td></tr><tr><td> 1.7 mm 모세관 </td><td> 시그마 - 올드 리치 </td><td> NORS175001 </td><td></td></tr><tr><td> 모세관 어댑터 </td><td> 새로운 시대 </td><td> NE-325-5/1.7 </td><td></td></tr><tr><td> 추출로드 </td><td> 새로운 시대 </td><td> NE - 341-5 </td><td></td></tr><tr><td> 50 μL와 HR - MAS 로터 BL4 구면 테플론 스페이서 키트 </td><td> Bruker </td><td> HZ07213 </td><td></td></tr><tr><td> 50 μL 삽입을위한 도구 키트 </td><td> Bruker </td><td> B2950 </td><td></td></tr><tr><td> 사전 III 600MHz의 NMR </td><td> Bruker </td><td></td><td></td></tr><tr><td> 1 H 인사 MAS NMR 견고한 프로브 </td><td> Bruker </td><td></td><td></td></tr><tr><td> 중수소 산화 99.9 % </td><td> 시그마 - 올드 리치 </td><td> 530867 - 1L </td><td></td></tr><tr><td> 3 - (trimethylsilyl) 프로피 산성 - D 4 (TSP) </td><td> 시그마 - 올드 리치 </td><td> 269​​,913 </td><td></td></tr></tbody></table>

assessing hepatic metabolic changes during progressive colonization of germ-free mouse by 1h nmr spectroscopy

그것은 내장 박테리아가 이러한 면역 보호와 비타민 합성과 같은 혜택의 범위를 제공하는 호스트 항상성에 크게 기여 잘 알려져있다. 그들은 또한 생태계 필수적인 대사 오르간 제작, 영양소의 상당한로 호스트를 제공합니다. 호스트와 내장 microbiota 사이의 신진 대사 상호 작용을 이해하는 창자의 식물과 대사 증후군 사이의 링크의 증가 증거의 맥락에서 현대 생물학의 중요한 과제가되고있다. 1-4 식민지는 (또한 정상화 과정으로 함) 전 세균없는 동물의 미생물의 설립을 지정합니다. 그것이 출생시 발생하는 자연적인 과정이지만, 그것은 또한 창자 꽃 생태계를 관리하고 추가로 호스트 신진 대사에 미치는 영향을 결정하기 위해 성인 세균없는 동물에 사용됩니다. 식민 프로세스를 제어하는​​ 일반적인 절차는 singl로 gavage 방법을 사용하는 것입니다E 또는 미생물의 혼합물. 이 방법은 매우 빠른 식민지의 결과와 5 매우 스트레스가되는 단점을 선물한다. 그것은 스트레스를 최소화하고 점차 호스트 신진 대사에 세균 설립의 영향을 관찰하기 위해 느린 식민지 과정을 취득하는 것이 유용합니다. 이 원고에서는, 우리는 비파괴 신진 대사 프로 파일링 기법을 사용하여 점차 식민지 과정에서 간장 대사의 수정을 평가하는 절차를 설명합니다. 우리는 1 H NMR 기반의 신진 대사 프로 파일링 공동 metabolites 미생물의 비뇨기 배설에 의해 반영 내장 미생물의 신진 대사 활동을 평가하여 내장 미생물의 식민지를 모니터링 제안합니다. 이것은 일반적으로 DGGE (기울기 겔 전기 영동을 denaturing)에 의해 배설물 박테리아를 모니터링하여 평가 직감의 미생물 생태계의 안정 설립 넘어 직감의 미생물 활동의 안정성을 감사하실 수 있습니다. 6식민지는 종래의 열린 환경에서 열립니다하고 더러운 쓰레기 컨트롤로 될 것입니다 기존의 동물에 의해 더러워진에 의해 시작됩니다. coprophagous 동물 설치류되고, 이쪽은 이전에 설명한 것처럼 동질적인 식민지를 보장합니다. 7 간장 대사 프로 파일링은 NMR 분광법을 치는것 1 H 고해상도 매직 각도를 사용하여 손상 간 생검에서 직접 측정됩니다. 이 세미 양적 기술은 세포 구조를 손상시키지 않고, 평가하는 빠른 방법을 제공, 그러한 트리 글리세 리드, 포도당과 글리코겐으로 주요 metabolites 추가하기 위해서는 식민지 과정과 70-10 간장 신진 대사 사이의 복잡한 상호 작용을 예상하고있다. 이 방법은 모든 조직 생검 11,12에도 적용할 수 있습니다.

Watch this Scientific Journal Video about Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy at JoVE.com

Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy

진보 식민 절차는 더욱 호스트 간장 대사에 미치는 영향을 평가하기 위해 설명되어 있습니다. 간장 신진 대사가 고해상도 매직 앵글 스피닝 (HR의 MAS) 그대로 생검의 NMR 프로 파일링에 의해 평가하는 동안 식민지는 NMR 기반의 신진 대사 프로 파일링 공동 metabolites 미생물의 비뇨기 배설을 평가하여 invasively 아닌 모니터입니다.

하여 세균없는 마우스 프로 그레시브 콜로니 동안 간장 신진 대사 변경 사항 평가 1 H NMR 분광학

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Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by 1H NMR Spectroscopy

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