This work was supported by the National Institutes of Health (HL114669). CM is a recipient of a postdoctoral research fellowship grant awarded by the Myocarditis Foundation, NJ.

윤리 선언문 : 모든 동물의 절차는 실험 동물의 관리 및 사용에 대한 지침에 따라 실시 및 네브래스카 링컨, 링컨, 네브래스카의 대학에 의해 승인되었습니다. 실험자가 면역 심근염 1. 유도 <ol><li> 이 mg/1.5 ml의 최종 농도 1X 인산 완충 식염수에 Myhc-α 334-352 용해시켜 펩티드 용액을 준비한다. </li><li> 50 NG / μL의 스톡 농도를 구하는 동결 건조 PT 50 μg을 함유하는 바이알에 무균 1X PBS 1 ㎖를 첨가하여, 백일해 독소 (PT)을 준비한다. 멸균 1.5 ML 튜브에 주식의 20 μl를 타고 1 NG / μL의 작업 농도를 얻기 위해 멸균 1X PBS 980 μl를 추가합니다. </li><li> 결핵균 H37Rv 40 밀리그램을 추가하여 완전한 프로 인트 보조제 (CFA)를 준비 (MTB)의 최종 농도를 얻기 위해 CFA 10 ㎖로 추출5 ㎎ / ㎖. </li><li> 펩티드-CFA 에멀젼을 준비합니다. 참고 : EAM 유도를 들어, 펩티드 CFA 에멀젼 Myhc-α 동물 당 334-352의 100 μg을 함유하는 150 ㎕를 투여했다. 예를 들어, 열 마우스 Myhc-α 334-352 1 ㎎을 함유하는 펩티드-CFA 에멀젼의 1.5 ML를 사용하여 면역합니다. <ol><li> 에멀젼의 1.5 ML를 준비하려면, 나누어지는 750 개의 1.5 ML 튜브에 334-352 펩타이드 솔루션을 Myhc가-α의 μL 및 CFA는 또 다른 1.5 ML 튜브에 MTB 보충. 3 ㎖의 루어-LOK 주사기를 사용하여, CFA / MTB 엑기스이어서 제 펩티드 용액을 그린다. </li><li> 3 방향 꼭지에 주사기를 연결하고 빈 3 ML의 주사기에 꼭지의 다른 콘센트에 연결합니다. 그래서 펩타이드 CFA 혼합물을 합리적으로 좋은 저항과 다른 하나의 주사기에서 흘러 크레인의 개통을 조정합니다. </li><li> ~ 1 분 동안 반복하여 다른 하나의 주사기에서 내용을 추진 한 다음 2 ~ 3 분 동안 얼음에 전체 어셈블리를 배치하여 혼합. 이 과정을 반복합니다10 배의 절차에 최소. </li><li> 주의 깊게 100 ㎖ 유리 비커에 아직도 물에 작은 방울을 배치하여 에멀젼의 일관성을 확인합니다. 방울은 물에 분산 할 것으로 예상되지 않습니다. 만약 그렇다면, 원하는 농도가 달성 될 때까지 계속 혼합한다. </li><li> 1 mL의 주사기와 마개에 부착 된 두 개의 3-ML 주사기 중 하나를 교체하여 1 ㎖ 루어 코리아 주사기로 3 ㎖ 주사기에서 에멀젼의 내용을 전송하고 1 ML의 주사기에 27 ½ G 바늘을 연결합니다. </li></ol></li><li> 6 년에서 8 주 된 여성 A / J 마우스 (~ 75 μL 각)의 두 사타구니 지역에 분할 복용 피하에있는 펩티드-CFA 에멀젼 150 μl를 주입한다. </li><li> 0 일 및 2 일 postimmunization에 복강 내 각 동물에 PT 서스펜션 (100 NG)의 100 μl를 관리 할 수​​ 있습니다. </li><li> 피하 t의 양쪽에 분할 복용 펩타이드 CFA 에멀젼 150 μl를 투여하여 7 일째에 예방 접종 절차를 반복그는 흉골 (~ 75 μL 각). 신선한 등의 이상이 에멀젼을 준비합니다. 그 후, 21 일에, MRM 영상에 동물을 대상 3 단계를 참조하십시오. </li></ol> 2. 동물 취급 <ol><li> 따뜻함을 유지하기 위해 아래에 배치 된 가열 패드 2 % 이소 플루 란 - 공기 혼합물을 포함하는 마취 유도 챔버 각 마우스를 배치하고 특별히 디자인 된 동물 홀더 (그림 1)에 동물을 전송합니다. </li><li> 동물 홀더에 발생하기 쉬운 위치에 동물을 무력화 그래서 주둥이는 마취 (그림 1)를 유지하기 위해 코 콘에 맞는. 마우스의 앞니에 부착 된 물린 바로 마우스의 머리를 고정합니다. </li><li> 실험 기간 동안 동물의 체온을 유지하기 위해 스캐너의 수직 구멍에 삽입의 출구 호스에 공기를 불어 히터의 전원을 켭니다. </li><li> 촬상 전체 세션 동안 2 ㎖ / 분의 유속으로 0.5 내지 2 %의 이소 플루 란으로 마취를 유지한다. 확인발가락 핀치 방법을 사용하여 마취, 아무 움직임이 예상. </li><li> 압축 공기를 넣은 베개 센서, 마우스 꼬리 / 발목 광섬유 맥박 산소 센서, 각각 호흡, 심장 박동과 체온, (그림 1)를 모니터링하는 직장 온도 프로브를 설정합니다. 참고 : 심장 게이팅이 동맥혈 산소 포화도의 비 침습적 모니터링을 허용 맥박 산소 측정기를 통해 수행된다. 산소 포화도 센서는 좌측 발목에 부착되어야하고, 다리 실 루프로 고정하고 센서의 발목을 유지하기 위해 녹화한다. MRM 촬상 어떤 조영제를 사용하지 않고 호흡과 심장 신호를 게이팅함으로써 달성된다. </li></ol> 3. 이미지 획득 <ol><li> 동물 (도 1)를 제조 한 후, 자기장의 균질성이 최대가 시야 (FOV)의 중앙에 위치 마음 MRM 스캐너의 중심에 마우스를 배치. 참고 : 와이드 보어(89mm) 트리플 100 G / cm의 축 그라디언트 및 4cm의 무선 주파수 (RF) 촬상 코일 탑재 9.4 T 수직 보어 자석 고해상도 3 차원 (3D) 이미지를 얻기 위해 사용한다. 주 : MRI 스캐너를 사용하는 경우 비자로 제공해야합니다. </li><li> 영상 인터페이스를 실행하고 &quot;연구 옵션&quot;메뉴에서 &quot;새로운 연구&quot;를 선택합니다. 명령 줄에서 &quot;mtune&quot;를 입력하고 &quot;조정 GUI&quot;를 끌어 실행합니다. &quot;시작 프로브 조정&quot;을 선택하고 &quot;자동 스케일&quot;버튼을 클릭합니다. 조정 GUI는 RFsignal를 표시하도록 변경됩니다. 튜닝 프로톤 공명 주파수 RF 코​​일 (400 MHz 이상)에서 코일의 단부에서 원격 튜닝 / 매치 노브를 사용. &quot;시작&quot;탭에서 관련 버튼을 클릭하여 주파수 및 전력 보정을 실행하려면 &quot;설치 전&quot;페이지로 이동합니다. shimming 페이지를 끌어 탭 &#39;연구 심 &quot;탭&quot;&#39;에 XYZ (빠른) 버튼을 누르십시오., 심 페이지로 이동 모든 반복을 선택하고 자동 shimming을 수행하기 위해 심 버튼을 누르십시오. </li><li> 마우스 마음을 집중하는 영상 인터페이스의 &quot;연구&quot;탭에서 스카우트 순서를 선택합니다. &quot;획득&quot;탭에서 35mm 2 FOV를 변경하고 컴퓨터의 기본 설정을 유지합니다. 순서를 실행하려면 시작을 클릭하고 심장은 FOV의 중심에 있지 않은 경우, 동물 홀더의 위치를​​ 조정할 RF 코​​일 리튠 다시 정찰 이미지를 얻는다. </li><li> &quot;연구&quot;탭에서 &quot;GEMS&quot;순서 버튼을 클릭 한 후 단축과 ​​마음 (20, 21)의 긴 축을 따라 두 개의 직교 평면을 얻기 위해 해당 &quot;획득&quot;탭을 획득 매개 변수를 입력합니다. 참고 : 그라데이션 에코 검사에 대한 일반적인 인수 매개 변수는 다음과 같습니다 슬라이스 두께 1 ㎜, 반복 시간 (TR), 200 밀리 초; 에코 시간 (TE), 2.67 밀리 초 (최소); 플립 각도 25 °; 면내 매트릭스 크기 128 x 128의; FOV, 22mm2; 인수의 수, 4; 1 분 30 초 대략 획득 시간. </li><li> &quot;전진&quot;탭에서 LV 해부학 및 기능 매개 변수를 측정하기 위해 맥박 산소 측정기 - 게이트 짧은 축 시네 MR 이미지를 수집하기 위해 &quot;CINE&quot;순서를 선택합니다. 마우스를 가져 가면 및 드래그하여 심장의 긴 축보기를 기반으로 영상 조각의 위치와 각도를 조정합니다. 그라데이션 에코 시네 순서 얻기 위해 &quot;획득&quot;탭에서 다음과 같은 이미지 매개 변수를 입력 : TR, 500 밀리 초; TE, 5 밀리 초; FOV, 22mm 2; 인수 매트릭스, 256 X 256; 슬라이스 두께 1mm; 인수의 수, 8; 프레임 수, 10; 17 분 ~의 획득 시간. 획득을 시작하려면 시작을 클릭합니다. </li><li> 이미징 소프트웨어의 &quot;I / O&quot;탭을 사용 DICOM 형식으로 획득 된 영상을 변환하고 처리를 위해 데이터 센터에 대응하는 파일을 전송. </li><li> 촬상 과정의 끝에서, 모든오우 마우스는 필터 탑 케이지 내에서 마취에서 복구 할 수 있습니다. 그들은 흉골 드러 누움을 유지하고 필요에 따라 추가 연구를 위해 그들을 유지하기 위해 충분한 의식을 되 찾을 때까지 무인 쥐를 두지 마십시오. </li></ol> 심장 자기 공명 현미경 이미징의 4. 데이터 분석 <ol><li> LV (19)의 해부학 적 및 기능적 매개 변수를 분석하는 세그먼트의 소프트웨어를 사용합니다. &quot;열기 이미지 파일 (들)&quot; &quot;파일&quot;메뉴의 하위 메뉴를 사용하여 소프트웨어에 DICOM 형식 시네 이미지를로드하여. GUI의 이미지 타입과 이미지 뷰 평면으로 &#39;짧은 축 중반 심실&#39;와 영상 기술, &#39;씨네&#39;로 &#39;MRGE&#39;를 선택합니다. </li><li> 를 통해 최종 심장 확장 및 최종 수축기에 사용되는 시간 프레임을 지정하는 &quot;다이 애스 털리을 끝으로 현재 기간을 설정&quot;을 각각 &quot;편집&quot;메뉴의 하위 메뉴 &quot;수축기을 끝으로 현재 기간을 설정합니다.&quot; </li><li>먼저 &quot;LV&quot;명령 단추를 클릭 한 다음 &quot;엔도&quot;과 오른쪽 하단 패널에 &quot;EPI&quot;명령 버튼을 수동으로 각각 좌심실 내막과 외막을 그립니다. 계산의 정확도를 증가시키기 위해 대응하는 커맨드 버튼을 눌렀을하여 유두근을 제거한다. </li><li> 이러한 이완기 볼륨, 수축기 볼륨, 뇌졸중 볼륨, 그리고 오른쪽 상단 패널의 방출 분율로 정량화 된 LV 매개 변수를 참조하십시오. 같은 두께, 및 심실 직경 22로 LV 매개 변수를 측정하는 &quot;기타&quot;명령 버튼을 누른 다음, &quot;측정 캘리퍼스&quot;명령 단추를 클릭하십시오. </li><li> 필요에 따라 이미지를 재 처리 &#39;. 매트&#39;형식의 세분화를 포함하여 이미지를 저장하기 위해 &quot;파일&quot;메뉴의 &quot;이미지 스택 및 분할 모두 저장&quot;메뉴를 클릭합니다. </li></ol>

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No conflicts of interests were declared.

이 연구는자가 면역 심근염으로 영향을받는 생쥐의 심장 이상을 확인할 수있는 비 침습적 인 도구로 MRM 절차 및 유틸리티를 설명합니다. EAM의 조직 학적 특징은 인간의 후 사구체 심근염과 유사하기 때문에, 마우스 모델은 일반적으로 심근 부상 23-25의 면역 메커니즘을 묘사하기 위해 사용된다. 그러나, 심근염 영향 동물은 임상 적으로 정상 나타나고, 진단은 실험 7의 종단에서 ?...

심장 마비가 사망의 주요 원인이며, 심근염 젊은 청소년 1 심장 마비의 하나의 주된 원인이다. 심근염으로 영향을받는 대부분의 환자는 증상이 남아 있고 질병은 저절로 2 해결됩니다. 그러나, 영향을받는 사람들의 10 ~ 20 %는 확장 성 심근증 (DCM) 3로 이어지는, 만성 질환을 개발할 수 있습니다. 다양한 동물 모델은 심근염의 면역 기전을 연구하기 위해 개발되었다. 질병은 심장 마이 오신 중쇄 (Myhc)-α 또는 immunodominant 펩티드 조각 또는 콕 사키 바이러스의 B3 4-9와 같은 병원체 감염에 의해 동물을 면역에 의해 심근염에 민감한 A / J와의 Balb / C 마우스에서 유도 할 수있다. 이 연구는 A / J 생쥐 334-352 유도 심근염을 Myhc가-α 포함한다. 심근 침입을 보여주는에도 불구하고, 심근염 영향을받는 동물은 임상 적으로 정상 표시; 진단은 염증 7에 대한 마음의 조직 학적 평가를 기반으로ND 심 초음파 10. 자기 공명 현미경 (MRM)을 분 혈관의 레벨 (최대 10 μm의 직경)에 기능적 세부의 평가를 허용하는, 고해상도 입체 평면 심혈관 이미징을 얻기 위해 일반적으로 사용되는 방법이지만, 해상력이 수준은 해상도는 일반적으로 1mm 11-14까지 수득되는 루틴 자기 공명 영상 (MRI) 스캔 절차로 달성하지. 이 질병 과정 (14)의 초기 시점에 성능 파라미터를 도출하는 것도 고해상도 이미지의 획득을 허용하는 상태로 MRM는 이점을 제공한다. 임상 적으로, MRM 영상은 광범위하게 병에 걸린 심장, 폐, 뇌 15-17의 기능 매개 변수를 연구에 적용하고있다. 본 연구에서는 면역 심근염으로 영향을받는 A / J 생쥐의 심장 이상을 확인할 수있는 비 침습적 인 도구로 MRM 기술의 사용이 표시됩니다. 특히, T그는 MRM 영상은 허용 등 합리적인 정확도 (18)와 좌심실 이완 기말 용적 및 박출계수 (EF) 등의 기능 매개 변수의 정량화. 각 파라미터의 정의는 : LV 단부 이완기 볼륨 이완기 사이클의 끝에서 좌심실에서 혈액의 양 및 박출계수, 행정 용적 / 이완 기말 볼륨. 데이터 분석은 자기 공명 스캐너 (19)에 의해 취득 처리 DICOM 호환 심장 혈관 이미지를 위해 개발 된 자유롭게 사용할 세그먼트 소프트웨어를 사용하여 수행됩니다. 데이터는 건강한 쥐에서 이러한 기능 매개 변수와 비교하여, 좌심실 이완 기말 용적, 뇌졸중 볼륨과 구혈률의 감소에 해당하는 myocarditic 동물의 좌심실 벽의 두께 증가를 공개했다.

<table><tbody><tr><td>Myhc-a 334-352 (DSAFDVLSFTAEEKAGVYK)</td><td>Neopeptide, Cambridge, MA</td><td></td><td>Store at 4 &amp;#176;C</td></tr><tr><td>CFA</td><td>Sigma Aldrich, St Louis, MO</td><td>5881</td><td>Store at 4 &amp;#176;C</td></tr><tr><td>MTB H37Rv extract</td><td>Difco Laboratories, Detroit, MI</td><td>231141</td><td>Store at 4 &amp;#176;C</td></tr><tr><td>PT</td><td>List Biologicals Laboratories, Campbell, CA</td><td>181</td><td>Store at 4 &amp;#176;C</td></tr><tr><td>1x PBS</td><td>Corning, Manassas, VA</td><td>21-040-CV</td><td>Store at 4 &amp;#176;C</td></tr><tr><td>Isoflurane</td><td>Piramal Healthcare, Mumbai, India</td><td>NDC66794-013-25</td><td></td></tr><tr><td>Female A/J mice</td><td>Jackson Laboratories, Bar Harbor, ME</td><td>646</td><td></td></tr><tr><td>Luer-lok sterile 1 ml syrringe</td><td>BD, Franklin Lakes, NJ</td><td>309628</td><td></td></tr><tr><td>Luer-lok sterile 3 ml syrringe</td><td>BD, Franklin Lakes, NJ</td><td>309657</td><td></td></tr><tr><td>Sterile needle, 18 G</td><td>BD, Franklin Lakes, NJ</td><td>305195</td><td></td></tr><tr><td>Sterile needle, 27 1/2 G</td><td>BD, Franklin Lakes, NJ</td><td>305109</td><td></td></tr><tr><td>3-way stopcock</td><td>Smiths Medical ASD, Inc. Dublin, OH</td><td>MX5311L</td><td></td></tr><tr><td>Kerlix gauze bandage rolls</td><td>Covidien, Mansfield, MA</td><td>6720</td><td></td></tr><tr><td>Kimwipes</td><td>Kimberly-Clark Professional, Roswell, GA</td><td>34155</td><td></td></tr><tr><td>Protouch Stockinette</td><td>Medline Industries, Mundelein, IL</td><td>30-1001</td><td></td></tr><tr><td>Sterile surgical scissors and forceps</td><td>INOX tool Corporation</td><td></td><td></td></tr><tr><td>Micro oven</td><td>GE Healthcare,&amp;nbsp;</td><td></td><td></td></tr><tr><td>ThermiPAQ hot and cold therapy system&amp;nbsp;</td><td>Theramics Corporation, Springfield, IL</td><td></td><td></td></tr><tr><td>Reptile heating lamp</td><td>Energy Savers Unlimited, Inc. Carson, CA</td><td></td><td></td></tr><tr><td>3M Transpore tapes</td><td>Target Corporation, MN</td><td></td><td></td></tr><tr><td>Up and Up Polymyxin B sulfate/Bacitracin/Neomycin sulfate antibiotic ointment</td><td>Target Corporation, MN</td><td></td><td></td></tr><tr><td>North Safety DeciDamp-2PVC foam ear plugs</td><td>North Safety Products, Smithfield, RI</td><td></td><td></td></tr><tr><td>Cotton tipped applicator, 6&amp;rsquo;&amp;rsquo; wooden stem&amp;nbsp;</td><td>Jorgensen Laboratories, Inc. Loveland, CO</td><td></td><td></td></tr><tr><td>Anesthesia induction chamber&amp;nbsp;</td><td>Summit Anesthesia Solutions, Ann Harbor, MI</td><td></td><td></td></tr><tr><td>Summit Anesthesia Support system for regulating flow of anesthesia&amp;nbsp;</td><td>Summit Anesthesia Solutions, Ann Harbor, MI</td><td></td><td></td></tr><tr><td>Specially designed animal holder</td><td>Agilent Technologies, Santa Clara, CA</td><td></td><td></td></tr><tr><td>Bickford&amp;nbsp;Omnicon F/Air&amp;nbsp;anesthesia gas filter unit&amp;nbsp;</td><td>A.M. Bickford, Inc. Wales Center, NY</td><td></td><td></td></tr><tr><td>Pulse-oximeter module, MR compatible small animal monitoring and gating system&amp;nbsp;</td><td>Small Animal Instruments, Inc. Stony Brook, NY</td><td></td><td></td></tr><tr><td>Oxygen cylinder&amp;nbsp;</td><td>Matheson-Tri Gas, North-Central Zone, Lincoln, NE</td><td></td><td></td></tr><tr><td>Gas regulator&amp;nbsp;</td><td>Western Medica, West Lake, OH</td><td></td><td></td></tr><tr><td>Signal breaking module, MR compatible small animal monitoring and gating system</td><td>Small Animal Instruments, Inc. Stony Brook, NY</td><td></td><td></td></tr><tr><td>9.4 T (400 MHz) 89 mm vertical core bore MR scanner</td><td>Agilent Technologies, Santa Clara, CA</td><td></td><td></td></tr><tr><td>4 cm millipede micro-imaging RF coil</td><td>Agilent Technologies, Santa Clara, CA</td><td></td><td></td></tr><tr><td>SAM PC monitor</td><td>Small Animal Instruments, Inc. Stony Brook, NY</td><td></td><td></td></tr><tr><td>Quantitative Medical Image analysis software</td><td>http://segment.heiberg.se;&amp;nbsp; Segment v1.8 R1430,&amp;nbsp; Medviso, Oresunds region, Sweden</td><td></td><td></td></tr><tr><td>Matlab software</td><td>The Mathworks, Inc.&amp;nbsp; Natick, MA</td><td></td><td></td></tr><tr><td>Computer-Unix operating system</td><td></td><td></td><td></td></tr></tbody></table>

noninvasive assessment of cardiac abnormalities in experimental autoimmune myocarditis by magnetic resonance microscopy imaging in the mouse

심근염 심근의 염증이지만, 질병의 그러한 영향 쇼 임상 증상 만 ~ 10 %. 심근 손상의 면역 이벤트를 연구하기 위해, 심근염의 다양한 마우스 모델이 널리 사용되어왔다. 이 연구는 심장 마이 오신 중쇄 (Myhc)-α A / J 생쥐에서 334-352로 유발 된 면역 심근염 (EAM)를 포함; 영향을받는 동물은 림프 구성 심근염을 개발하지만 뚜렷한 임상 증상과 함께. 이 모델에서, 비 침습 양상으로 자기 공명 현미경 (MRM)의 유틸리티가 표시됩니다 Myhc-α 334-352으로 예방 접종을 동물의 심장 구조 및 기능 변화를 확인합니다. EAM 건강한 마우스는 9.4 T (400 메가 헤르츠) 4cm 노래기 무선 주파수 이미징 프로브와 100 G / cm 트리플 축 그라데이션을 갖춘 89mm 수직 코어 구멍 스캐너를 사용하여 몇 군데 있었다. 심장 이미지는 애니보기 구배 에코 기반 시네 펄스 시퀀스를 사용하여 마취 된 동물로부터 취득하고 있었다LS는 호흡과 맥박 산소 측정기에 의해 감시했다. 분석은 건강한 쥐에 비해 심실의 내부 직경에 상응하는 감소로, EAM 마우스의 심실 벽의 두께 증가를 공개했다. 데이터는 염증이 마음에 형태 학적 및 기능적 변화는 살아있는 동물에서 비 침습적 모니터링 MRM으로 할 수있는 것이 좋습니다. 결론적으로, MRM은 진행과 회귀 병원체에 의한 질병에 심근 부상뿐만 아니라 치료에 반응을 평가하는 이점을 제공한다.

이 보고서에서, EAM으로 영향을받는 동물의 마음의 구조와 기능의 변화를 결정하는 비 침습 양상으로 MRM 기술의 유틸리티가 표시됩니다. 심근염은 CFA 7 Myhc-α 334-352으로 동물을 면역에 의해 A / J 생쥐에서 유도하고, 동물은 21 일 후 예방 접종에 MRM 실험을 실시 하였다. MRM 영상은 트리플 축 기울기 (최대 강도 100 G / cm)가 장착 된 89mm 수직 구멍 자석을 사용하여 9.4 T (양성자 400 메가 헤르츠)에...

Watch this Scientific Journal Video about Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse at JoVE.com

Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse

This study demonstrates the successful establishment of magnetic resonance microscopy imaging as a non-invasive tool to assess the cardiac abnormalities in mice affected with autoimmune myocarditis. The data indicate that the technique can be used to monitor the disease-progression in live animals.

마우스에있는 자기 공명 현미경 영상으로 실험자가 면역 심근염에있는 심장의 이상 유무 비 침습적 평가

Myocarditis is an inflammation of the myocardium, but only ~10% of those affected show clinical manifestations of the disease. To study the immune events ...

noninvasive-assessment-cardiac-abnormalities-experimental-autoimmune

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9.9K 조회수.  University of Nebraska-Lincoln. This study demonstrates the successful establishment of magnetic resonance microscopy imaging as a non-invasive tool to assess the cardiac abnormalities in mice affected with autoimmune myocarditis. The data indicate that the technique can be used to monitor the disease-progression in live animals.

비디오: Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse

High-frequency High-resolution Echocardiography: First Evidence on Non-invasive Repeated Measure of Myocardial Strain, Contractility, and Mitral Regurgitation in the Ischemia-reperfused Murine Heart

Ischemia-reperfusion (IR) was surgically performed in murine hearts which were then subjected to repeated imaging to monitor temporal changes in functional parameters of key clinical significance. Two-dimensional movies were acquired at high frame rate (8 kHz) and were utilized to estimate high-quality myocardial strain. Two-dimensional elastograms (strain images), as well as strain profiles, were visualized. Results were powerful in quantitatively assessing IR-induced changes in cardiac events including left-ventricular (LV) contraction, LV relaxation and isovolumetric phases of both pre-IR and post-IR beating hearts in intact mice. In addition, compromised sector-wise wall motion and anatomical deformation in the infarcted myocardium were visualized.  The elastograms were uniquely able to provide information on the following parameters in addition to standard physiological indices that are known to be affected by myocardial infarction in the mouse: internal diameters of mitral valve orifice and aorta, effective regurgitant orifice, myocardial strain (circumferential as well as radial), turbulence in blood flow pattern as revealed by the color Doppler movies and velocity profiles, asynchrony in LV sector, and changes in the length and direction of vectors demonstrating slower and asymmetrical wall movement. This work emphasizes on the visual demonstration of how such analyses are performed. 

High-frequency High-resolution Echocardiography: First Evidence on Non-invasive Repeated Measure of Myocardial Strain, Contractility, and Mitral Regurgitation in the Ischemia-reperfused Murine Heart 

High frequency Doppler ultrasound is a novel technology for assessing regional myocardial function. This work presents first evidence demonstrating applicability of this versatile imaging platform for the repeated measure of myocardial strain, dp/dt, and mitral regurgitation in the ischemia-reperfused (IR) murine heart.

고주파 고해상도 Echocardiography : 국소 빈혈 - reperfused Murine 중심부에 심근 스트레인, 수축성 및 승모판 병리 현상의 비침습 반복 측정에 대한 최초의 증거

Ischemia-reperfusion (IR) was surgically performed in murine hearts which were then subjected to repeated imaging to monitor temporal changes in ...

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의학

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery (SALLI) MRI in Rats

Small animal magnetic resonance imaging is an important tool to study cardiac function and changes in myocardial tissue. The high heart rates of small animals (200 to 600 beats/min) have previously limited the role of CMR imaging. Small animal Look-Locker inversion recovery (SALLI) is a T1 mapping sequence for small animals to overcome this problem 1. T1 maps provide quantitative information about tissue alterations and contrast agent kinetics. It is also possible to detect diffuse myocardial processes such as interstitial fibrosis or edema 1-6. Furthermore, from a single set of image data, it is possible to examine heart function and myocardial scarring by generating cine and inversion recovery-prepared late gadolinium enhancement-type MR images 1.
	The presented video shows step-by-step the procedures to perform small animal CMR imaging. Here it is presented with a healthy Sprague-Dawley rat, however naturally it can be extended to different cardiac small animal models.

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery SALLI MRI in Rats

Contrast enhanced cardiac magnetic resonance (CMR) imaging allows comprehensive in vivo assessment of the heart in small animal models of cardiovascular disease. Here we detail the procedures to perform CMR imaging, reconstruction and analysis using Small Animal Lock-Locker Inversion Recovery (SALLI) in rats.

작은 동물 룩 로커 반전 복구 (SALLI) 쥐의 MRI를 사용하여 심장 기능 및 심근 형태학의 평가

Small animal magnetic resonance  imaging is an important tool to study cardiac function and changes in myocardial  tissue. The high heart rates of small ...

MRI and PET in Mouse Models of Myocardial Infarction

Myocardial infarction is one of the leading causes of death in the Western world. The similarity of the mouse heart to the human heart has made it an ideal model for testing novel therapeutic strategies.
In vivo magnetic resonance imaging (MRI) gives excellent views of the heart noninvasively with clear anatomical detail, which can be used for accurate functional assessment. Contrast agents can provide basic measures of tissue viability but these are nonspecific. Positron emission tomography (PET) is a complementary technique that is highly specific for molecular imaging, but lacks the anatomical detail of MRI. Used together, these techniques offer a sensitive, specific and quantitative tool for the assessment of the heart in disease and recovery following treatment.
In this paper we explain how these methods are carried out in mouse models of acute myocardial infarction. The procedures described here were designed for the assessment of putative protective drug treatments. We used MRI to measure systolic function and infarct size with late gadolinium enhancement, and PET with fluorodeoxyglucose (FDG) to assess metabolic function in the infarcted region. The paper focuses on practical aspects such as slice planning, accurate gating, drug delivery, segmentation of images, and multimodal coregistration. The methods presented here achieve good repeatability and accuracy maintaining a high throughput.

We describe how to perform MRI and PET imaging of the mouse heart. The protocol is tailored to assess treatment efficacy in models of myocardial infarction and heart failure.

Myocardial infarction is one of the leading causes of death in the Western world. The similarity of the mouse heart to the human heart has made it an ...

In Vivo Quantitative Assessment of Myocardial Structure, Function, Perfusion and Viability Using Cardiac Micro-computed Tomography

The use of Micro-Computed Tomography (MicroCT) for in vivo studies of small animals as models of human disease has risen tremendously due to the fact that MicroCT provides quantitative high-resolution three-dimensional (3D) anatomical data non-destructively and longitudinally. Most importantly, with the development of a novel preclinical iodinated contrast agent called eXIA160, functional and metabolic assessment of the heart became possible. However, prior to the advent of commercial MicroCT scanners equipped with X-ray flat-panel detector technology and easy-to-use cardio-respiratory gating, preclinical studies of cardiovascular disease (CVD) in small animals required a MicroCT technologist with advanced skills, and thus were impractical for widespread implementation. The goal of this work is to provide a practical guide to the use of the high-speed Quantum FX MicroCT system for comprehensive determination of myocardial global and regional function along with assessment of myocardial perfusion, metabolism and viability in healthy mice and in a cardiac ischemia mouse model induced by permanent occlusion of the left anterior descending coronary artery (LAD).

In Vivo Quantitative Assessment of Myocardial Structure, Function, Perfusion and Viability Using Cardiac Micro-computed Tomography

This method outlines the use of Quantum Micro-Computed Tomography (MicroCT) to assess cardiac morphology, function, perfusion, metabolism and viability with iodinated contrast agent in mice with experimentally-induced myocardial ischemia. The technique can be applied for non-destructive high-throughput longitudinal in vivo imaging of various animal models of human heart disease.

심근 구조, 기능의 생체 양적 평가에서 관류 및 생존 능력은 심장 마이크로 컴퓨터 단층 촬영을 사용하여

The use of Micro-Computed Tomography (MicroCT) for in vivo studies of small animals as models of human disease has risen tremendously due to the fact that ...

생체공학

Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed in mice is high. Phenotypic characterization of these mouse models also requires the examination of cardiac function and morphology. Echocardiography and magnetic resonance imaging (MRI) are commonly used approaches to characterize cardiac function and morphology in mice. Echocardiographic and MRI equipment specialized for use in small rodents is&#160;expensive and requires a dedicated space. This protocol describes cardiac measurements in mice using a clinical echocardiographic system with a 15 MHz human vascular probe. Measurements are performed on anesthetized adult mice. At least three image sequences are recorded and analyzed for each animal in M-mode in the parasternal short-axis view. Afterwards, cardiac histological examination is performed, and cardiomyocyte diameters are determined on hematoxylin-eosin- or wheat germ agglutinin (WGA)-stained paraffin sections. Vessel density is determined morphometrically after Pecam-1 immunostaining. The protocol has been applied successfully to pharmacological studies and different genetic animal models under baseline conditions, as well as after experimental myocardial infarction by the permanent ligation of the left anterior descending coronary artery (LAD). In our experience, echocardiographic investigation is limited to anesthetized animals and is feasible in adult mice weighing at least 25 g.

Echocardiographic examination is frequently used in mice. Expensive high-resolution ultrasound devices have been developed for this purpose. This protocol describes an affordable echocardiographic procedure combined with histological morphometric analyses to determine cardiac morphology.

마우스의 심장 형태학의 echocardiographic 및 조직학 검사

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed ...

Quantification of Mouse Heart Left Ventricular Function, Myocardial Strain, and Hemodynamic Forces by Cardiovascular Magnetic Resonance Imaging

Mouse models have contributed significantly to understanding genetic and physiological factors involved in healthy cardiac function, how perturbations result in pathology, and how myocardial diseases may be treated. Cardiovascular magnetic resonance imaging (CMR) has become an indispensable tool for a comprehensive in vivo assessment of cardiac anatomy and function. This protocol shows detailed measurements of mouse heart left ventricular function, myocardial strain, and hemodynamic forces using 7-Tesla CMR. First, animal preparation and positioning in the scanner are demonstrated. Survey scans are performed for planning imaging slices in various short- and long-axis views. A series of prospective ECG-triggered short-axis (SA) movies (or CINE images) are acquired covering the heart from apex to base, capturing end-systolic and end-diastolic phases. Subsequently, single-slice, retrospectively gated CINE images are acquired in a midventricular SA view, and in 2-, 3-, and 4-chamber views, to be reconstructed into high-temporal resolution CINE images using custom-built and open-source software. CINE images are subsequently analyzed using dedicated CMR image analysis software.
Delineating endomyocardial and epicardial borders in SA end-systolic and end-diastolic CINE images allows for the calculation of end-systolic and end-diastolic volumes, ejection fraction, and cardiac output. The midventricular SA CINE images are delineated for all cardiac time frames to extract a detailed volume-time curve. Its time derivative allows for the calculation of the diastolic function as the ratio of the early filling and atrial contraction waves. Finally, left ventricular endocardial walls in the 2-, 3-, and 4-chamber views are delineated using feature-tracking, from which longitudinal myocardial strain parameters and left ventricular hemodynamic forces are calculated. In conclusion, this protocol provides detailed in vivo quantification of the mouse cardiac parameters, which can be used to study temporal alterations in cardiac function in various mouse models of heart disease.

This study describes a comprehensive cardiovascular magnetic resonance imaging (CMR) protocol to quantify the left ventricular functional parameters of the mouse heart. The protocol describes the acquisition, post-processing, and analysis of the CMR images as well as assessment of different cardiac functional parameters.

심혈관 자기 공명 영상에 의한 마우스 심장 좌심실 기능, 심근 균주 및 혈역학력의 정량화

Mouse models have contributed significantly to understanding genetic and physiological factors involved in healthy cardiac function, how perturbations ...

Quantification of Biventricular Function and Morphology by Cardiac Magnetic Resonance Imaging in Mice with Pulmonary Artery Banding

Right ventricular (RV) function and failure are major determinants of outcome in acquired and congenital heart diseases, including pulmonary hypertension. Assessment of RV function and morphology is complex, partly due to the complex shape of the RV. Currently, cardiac magnetic resonance (CMR) imaging is the golden standard for noninvasive assessment of RV function and morphology. The current protocol describes CMR imaging in a mouse model of RV pressure load induced by pulmonary artery banding (PAB). PAB is performed by placing a 6-0 suture around the pulmonary artery over a 23 G needle. The PAB gradient is determined using echocardiography at 2 and 6 weeks. At 6 weeks, the right and left ventricular morphology and function is assessed by measuring both end-systolic and end-diastolic volumes and mass by ten to eleven cine slices 1 mm thick using a 9.4 T magnetic resonance imaging scanner equipped with a 1,500 mT/m gradient. Representative results show that PAB induces a significant increase in RV pressure load, with significant effects on biventricular morphology and RV function. It is also shown that at 6 weeks of RV pressure load, cardiac output is maintained. Presented here is a reproducible protocol for the quantification of biventricular morphology and function in a mouse model of RV pressure load and may serve as a method for experiments exploring determinants of RV remodeling and dysfunction.

To understand the pathophysiology of right ventricular (RV) adaptation to abnormal loading, experimental models are crucial. However, assessment of RV dimensions and function is complex and challenging. This protocol provides a method to perform cardiac magnetic resonance imaging (CMR) as a noninvasive benchmark procedure in mice subjected to RV pressure load.

Right ventricular (RV) function and failure are major determinants of outcome in acquired and congenital heart diseases, including pulmonary hypertension. ...

Induction of Myocardial Infarction and Myocardial Ischemia-Reperfusion Injury in Mice

Acute myocardial infarction is a common cardiovascular disease with high mortality. Myocardial reperfusion injury can counteract the beneficial effects of heart reflow and induce secondary myocardial injury. A simple and reproducible model of myocardial infarction and myocardial ischemia-reperfusion injury is a good tool for researchers. Here, a customizable method to create a myocardial infarction (MI) model and MIRI by precision ligation of the left anterior descending coronary artery (LAD) through micromanipulation is described. Accurate and reproducible ligature positioning of the LAD helps obtain consistent results for heart injury. ST-segment changes can help to identify model accuracy. The serum level of cardiac troponin T (cTnT) is used to assess the myocardial injury, cardiac ultrasound is employed to evaluate the myocardial systolic function, and Evans-Blue/triphenyl tetrazolium chloride staining is used to measure infarct size. In general, this protocol reduces procedure duration, ensures controllable infarct size, and improves mouse survival.

Here we describe a simple and reproducible method that can induce myocardial infarction or myocardial ischemia-reperfusion injury in mice by precision ligation of the left anterior descending coronary artery through micromanipulation.

마우스의 심근경색 및 심근 허혈-재관류 손상 유도

Acute myocardial infarction is a common cardiovascular disease with high mortality. Myocardial reperfusion injury can counteract the beneficial effects of ...

An Experimental Model of Myocardial Infarction for Studying Cardiac Repair and Remodeling in Knockout Mice

Cardiovascular disease is the most prevalent cause of death in Western countries, with acute myocardial infarction (MI) being the most prevalent form. This paper describes a protocol for studying the role of galectin 3 (Gal-3) in the temporal evolution of cardiac healing and remodeling in an experimental animal model of MI. 
The procedures described include an experimental model of MI with a permanent coronary ligature in male C57BL/6J (control) and Gal-3 knockout (KO) mice, an echocardiography procedure to study cardiac remodeling and systolic function in vivo, a histological evaluation of interstitial myocardial fibrosis with picrosirius red-stained and rhodamine-conjugated lectin-stained sections for studying myocyte hypertrophy by the cross-sectional area (MCSA), and the quantification of infarct size and cardiac remodeling (scar thinning, septum thickness, and expansion index) by planimetry in slices stained with Masson's trichrome and triphenyl tetrazolium chloride. Gal-3 KO mice with MI showed disrupted cardiac remodeling and an increase in the scar thinning ratio and the expansion index. At the onset of MI, myocardial function and cardiac remodeling were also severely affected. At 4 weeks post MI, the natural evolution of fibrosis in infarcted Gal-3 KO mice was also affected. 
In summary, the experimental model of MI is a suitable model for studying the temporal evolution of cardiac repair and remodeling in mice with the genetic deletion of Gal-3 and other animal models. The lack of Gal-3 affects the dynamics of cardiac repair and disrupts the evolution of cardiac remodeling and function after MI.

Here, we describe an experimental model of myocardial infarction, an echocardiography procedure to study cardiac remodeling and function, and procedures for quantifying fibrosis and hypertrophy in picrosirius red-stained and rhodamine-stained sections, as well as the infarct size and expansion index in slices stained with Masson's trichrome.

Cardiovascular disease is the most prevalent cause of death in Western countries, with acute myocardial infarction (MI) being the most prevalent form. ...

Mesoscopic Optical Imaging of Whole Mouse Heart

Both genetic and non-genetic cardiac diseases can cause severe remodeling processes in the heart. Structural remodeling, such as collagen deposition (fibrosis) and cellular misalignment, can affect electrical conduction, introduce electromechanical dysfunctions and, eventually, lead to arrhythmia. Current predictive models of these functional alterations are based on non-integrated and low-resolution structural information. Placing this framework on a different order of magnitude is challenging due to the inefficacy of standard imaging methods in performing high-resolution imaging in massive tissue. In this work, we describe a methodological framework that allows imaging of whole mouse hearts with micrometric resolution. The achievement of this goal has required a technological effort where advances in tissue transformation and imaging methods have been combined. First, we describe an optimized CLARITY protocol capable of transforming an intact heart into a nanoporous, hydrogel-hybridized, lipid-free form that allows high transparency and deep staining. Then, a fluorescence light-sheet microscope able to rapidly acquire images of a mesoscopic field of view (mm-scale) with the micron-scale resolution is described. Following the mesoSPIM project, the conceived microscope allows the reconstruction of the whole mouse heart with micrometric resolution in a single tomographic scan. We believe that this methodological framework will allow clarifying the involvement of the cytoarchitecture disarray in the electrical dysfunctions and pave the way for a comprehensive model that considers both the functional and structural data, thus enabling a unified investigation of the structural causes that lead to electrical and mechanical alterations after the tissue remodeling.

We report a method for mesoscopic reconstruction of the whole mouse heart by combining new advancements in tissue transformation and staining with the development of an axially scanned light-sheet microscope.

전체 마우스 심장의 메조스코픽 광학 이미징

Both genetic and non-genetic cardiac diseases can cause severe remodeling processes in the heart. Structural remodeling, such as collagen deposition ...