This work was supported by NIH grants HL111121 (JDM) and TR000954 (JDM).

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The authors have nothing to disclose.

노화는 심장 혈관 석회화 1 점진적 증가와 연관되어있다. 혈역학 적으로 유의 한 대동맥 판막 협착 (65) 2 세 이상 인구의 3 %에 영향을 미치는, 심지어 중간 대동맥 판막 협착 (3~4m / s의 피크 속도)을 가진 환자는 40 % 미만의 5 년 무 사건 생존율이 3. 현재,이 대동맥 판막의 석회화의 진행을 느리게 할 효과적인 치료법은 없으며, 수술 대동맥 판막 치환술 고급 대동맥 판막 협착증 4 사용할 수있는 유일한 치료입니다. 개시 및 대동맥 판막의 석회화의 진행에 기여하는 메커니즘의 깊은 이해를 확보하기위한 연구는 대동맥 판막 협착 (5), (6)을 관리하는 약물과 비 수술 방법으로 이동에서 중요한 첫 번째 단계입니다. 유전학적인LY-변경 마우스는 다양한 질병에 기여 이제 대동맥 판막 협착 6, 7, 8의 생물학을 이해하기위한 역학적 연구의 최전선에 오는 메커니즘에 대한 우리의 이해를 개발에 중요한 역할을했다. 같은 다른 심혈 관계 질환과는 달리 죽상 동맥 경화증과 심장 장애 혈관과 심실 기능을 평가하기위한 표준 프로토콜이 잘 확립-이 생쥐의 심장 판막 기능의 생체 표현형과 관련된 고유 한 문제가 대부분입니다. 최근 리뷰 많은 이미징 및 설치류 9, 10, 11 밸브 기능을 평가하는 데 사용 침습적 양식의 장점과 단점에 대한 철저한 토론을 제공하고 있지만, 지금까지, 우리는 COMPRE을 제공하는 출판물을 인식하지 못합니다를 면밀히는 단계별 생쥐의 표현형의 심장 판막 기능을위한 프로토콜을. 이 논문의 목적은 쥐 심장 판막 기능 표현형 방법 및 프로토콜을 설명한다. 모든 방법과 절차는 메이요 클리닉 기관 동물 관리 및 사용위원회에 의해 승인되었습니다. 이 프로토콜의 핵심 요소는 마취 깊이 심장 기능의 평가 및 심장 판막 기능의 평가를 포함한다. 우리는이 보고서는 심장 판막 질환의 분야에서 연구를 추구에 관심 수사관을 안내하는 역할을하지 않지만이 빠르게 성장하는 분야에서 데이터의 재현성 및 유효성을 보장하기 위해 프로토콜 표준화에 관련된 국내 및 국제 대화를 시작하기를 바랍니다. 중요한 것은, 고해상도 초음파 시스템을 이용하여 성공적으로 이미징 (일반적으로 초음파 검사에 사용되는 용어) 초음파의 원리 작동 지식 프린 기본적인 이해를 필요심장 생리학 및 초음파 검사와 유의 한 경험 레는 설치류에서 심장 기능의 정확하고 시간을 효율적으로 평가를 허용합니다.

<table><tbody><tr><td>High resolution ultrasound machine</td><td>VisualSonics, Fujifilm</td><td>Vevo 2100&amp;nbsp;</td><td></td></tr><tr><td>Isoflurane diffuser (capable of delivering 1 % to 1.5 % isoflurane mixed with 1 L/min 100% O&lt;sub&gt;2&lt;/sub&gt;</td><td>VisualSonics, Fujifilm</td><td>N/A</td><td></td></tr><tr><td>Transducers for small mice (550D) or larger mice (400)</td><td>MicroScan, VisualSonics, Fujifilm</td><td>MS 550D, MS 400</td><td></td></tr><tr><td>Animal platform</td><td>VisualSonics, Fujifilm</td><td>11503</td><td></td></tr><tr><td>Advanced physiological monitoring unit</td><td>VisualSonics, Fujifilm</td><td>N/A</td><td></td></tr><tr><td>Isoflurane</td><td>Terrell</td><td>NDC 66794-019-10</td><td></td></tr><tr><td>Nose cone and tubing connected to isoflurane diffuser and 100% O&lt;sub&gt;2&lt;/sub&gt;</td><td>Custom Engineered in-house</td><td>--</td><td></td></tr><tr><td>Hair razor</td><td>Andis Super AGR+ vet pack clipper</td><td>AD65340</td><td></td></tr><tr><td>Ultrasound gel</td><td>Parker Laboratories</td><td>REF 01-08</td><td></td></tr><tr><td>Electrode gel&amp;nbsp;</td><td>Parker Laboratories</td><td>REF 15-25</td><td></td></tr><tr><td>Adhesive tapes</td><td>Fisher Laboratories</td><td>1590120B</td><td></td></tr><tr><td>Paper towels</td><td></td><td></td><td></td></tr></tbody></table>

echocardiographic approaches and protocols for comprehensive phenotypic characterization of valvular heart disease in mice

The aim of this manuscript and accompanying video is to provide an overview of the methods and approaches used for imaging heart valve function in rodents, with detailed descriptions of the appropriate methods for anesthesia, the echocardiographic windows used, the imaging planes and probe orientations for image acquisition, the methods for data analysis, and the limitations of emerging technologies for the evaluation of cardiac and valvular function. Importantly, we also highlight several future areas of research in cardiac and heart valve imaging that may be leveraged to gain insights into the pathogenesis of valve disease in preclinical animal models. We propose that using a systematic approach to evaluating cardiac and heart valve function in mice can result in more robust and reproducible data, as well as facilitate the discovery of previously underappreciated phenotypes in genetically-altered and/or physiologically-stressed mice.

통상적 동물 심장 초음파 영상으로부터 얻어진 이미지의 예는이 원고에 포함된다. 동물의 가슴에 트랜스 듀서의 배치를 보여 독자에게 변환기 바와 같은 이미지를 얻기 위해 배치되는 위치에 대한 명확한 이해를 제공하기 위해 제공된다. 초음파 실험실 셋업의 사진은 적당한 장치, 특히 사용되는 초음파 변환기 마취의 방법의 중요성을 강조하기 위해 포함된다. 2 차원 / B 모드, M 모드, 그리고 정상 및 비정상 밸브, 좌우 심실과 대동맥 근부의 색상과 도플러 표시가 제대로 표시되어 있습니다. 스트레인 속도 이미징 일상적으로 실행되지 않지만, 예를 들어도 포함된다. 승모판 막 폐쇄 부전증은 S 동안 밸브를 가로 질러 높은 보통 비 층류 혈류 속도 (모자이크 착색)을 특징으로ystole (그림 5). 심전도 추적의 QRS 컴플렉스 후에 발생 MV 걸쳐 좌심방에 좌심실로부터 이러한 모자이크 컬러 도플러 유동 패턴의 존재는, MR의 명확한 진단을 허용한다. 이 대동맥판 부전 및 / 또는 좌심실 기능 장애의 부재에서 발생하는 경우, 이것은 절연 승모판 탈출증 특징으로 할 수있다. 좌심실의 현저한 팽창이 (인한 심부전 또는 마취 깊이 과도한 실험적으로 유도하기 위해)이 있다면,이 허혈성 승모판 역류 (또는 부전, 심장 기능 장애 보조)로 특성화 될 수있다. 펄스 파 스펙트럼 도플러 디스플레이 혈류의 역류 제트의 존재 및 타이밍을 확인하기 위해 사용될 수있다. 정상적인 대동맥 밸브를 열고 각 심장주기 동안 적절하게 닫습니다 세 얇고 유연한 교두가 있습니다. 대동맥 밸브 끝의 분리는 2D 유도로 측정됩니다장축보기 대동맥 판막의 M 모드. 전자 캘리퍼스 왼쪽 대동맥 첨단의 첨단에 오른쪽 대동맥 끝 (그림 3)의 선단에서 측정하는 데 사용됩니다. 정상 마우스에서 대동맥 판막 첨단 분리 거리가 0.9 ~ 1.3 mm이다. 컬러 도플러 수축기 대동맥 밸브 내로 걸쳐 층류를 나타낸다. 난류는 대동맥 판막 폐쇄 부전의 증가 흐름의 조건에서 평가, 또는 대동맥 판막 협착증에서와 같이, 압력을 증가시킬 수있다. 이는 유출 기관에 모자이크 색상으로 설명된다. 대동맥 판막 폐쇄 부전의 아주 작은 양으로 인해 hyperdynamic 심장 기능에 피크 transvalvular 속도의 상당한 증가가 발생할 수 있고, 심실 프리로드를 왼쪽으로 증가. 1.50 m / s의 0.90 m / s의 정상 마우스의 범위에서 피크 대동맥 속도. &gt; 5m의 피크 대동맥 판막 속도는 / s의 심한 대동맥 판막 협착 마우스에 기록되었습니다. (12) (도 8)의 인덱스를 제공하기 위해 사용될 수있다. 폐동맥 가속 시간은 최대 유속 수축기 폐동맥 유동 개시까지의 시간 간격이다. 우심실 토출 시간은 수축기 폐동맥 수축기 흐름이 중단 된 지점을 우심실 토출의 개시 사이의 간격이다. 우심실 토출 시간 폐동맥 가속 시간의 비율의 저하와 함께 짧아 폐동맥 가속 시간의 조합 (폐동맥 또는 우심실 압력 침습 또는 직접 측정을 이용하여 확인 될 수있다) 폐동맥 고혈압의 존재를 시사 . <img alt="그림 1" src="/files/ftp_upload/54110/54110fig1.jpg" /> 그림 1 : 동물 Cardiac 초음파 연구소. 실험실은 고주파수 (30 MHz와 40 MHz의) 센서 (MS (400) 및 MS 550D), 이소 플루 확산 동물 플랫폼, 온도, 심박수 모니터, 1 % 내지 1.5 %의 이소 플루 란 소 동물 전용 초음파 장비에 장착되어 1 L / 분으로 100 % O 2, 노우즈 콘 및 이소 플루 확산 및 100 % O 2, 헤어 면도기 초음파 겔 전극 겔, 접착 테이프, 종이 타월에 연결된 튜브와 혼합. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig1large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 2" src="/files/ftp_upload/54110/54110fig2.jpg" /> 그림 2 : 기본 변환기 위치. (A) 흉골 창. 변환기 헤드 미부 향하는 변환기의 화상 인덱스 마커 왼쪽 흉골 테두리에 위치된다. FR이 위치 OM, 좌심실, 대동맥 판막 및 대동맥 및 폐동맥 밸브의 짧은 축 볼의 장축 뷰가 얻어 질 수있다. 창에서 흉골 (B)는 트랜스 듀서 헤드 후방 방향 홈과 반 시계 방향으로 회전된다. 이 위치에서 좌심실과 대동맥 판막과 폐동맥 밸브의 장축 뷰의 짧은 축 뷰가 얻어 질 수있다. (C) 혀끝 창. 변환기 헤드는 심장의 정점에 위치한다. 이 위치에서 좌우 심실과 승모판 막과 삼첨판 밸브의 장축 뷰가 얻어 질 수있다. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig2large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 3" src="/files/ftp_upload/54110/54110fig3.jpg" /> 그림 3 : 규격에 대동맥 밸브 기능의 평가석회화 대동맥 밸브 질환을 가진 마우스에서 대동맥 밸브 기능 대 등 마우스. 장축 보아 정상 대동맥 밸브 (A) 2 차원 이미지. 대동맥 판막은 수축기 동안 잘 열리고 있습니다. (B) M 모드 영상에서는 정상 대동맥 판막 기능 (상자 형 모양)을 묘사. 첨단 분리 거리가 1.12 mm로 측정합니다. 정상 대동맥 판막에서 최고 속도 (C) 스펙트럼 도플러 디스플레이는 1.3 m / s로 meaured했다. 저밀도 지단백 수용체의 긴 축보기에서 석회화 된 대동맥 밸브 (D) 2D 이미지 결핍 (LDLR - / -)와 서부 다이어트가 공급 아포 지단백 B100 전용 (형 ApoB 100/100) 마우스입니다. 교두가 두꺼워하고 수축기 동안 제한된 개방 결과 에코를 증가하고있다. 동일한 협착 대동맥판 묘사 (E)는 M 모드 영상 0.7 mm의 교두 분리 거리 측정을 나타낸다. (F &lt;)&gt; / 강한 협착 대동맥 판막에서 최고 속도의 스펙트럼 도플러 디스플레이는 4.6 m / s로 meaured했다. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig3large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 4" src="/files/ftp_upload/54110/54110fig4.jpg" /> 그림 4 : 정상 승모판의 M 모드. 꼭대기 창에서 승모판의 장축 뷰가 얻어진다. 심문의 M 모드 라인은 승모판 막 전단지에 걸쳐 적용된다. 승모판 전단 두께 이론적 전자 캘리퍼스를 사용하여 측정 될 수 있지만, 이는 정상적인 승모판의 전단을 빠르게 이동하는 얇은 저조한 에코 주어진 매우 어려울하고있다. 화살표는 수축기의 승모판 막 전단지의 M 모드를 가리 킵니다. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig4large.jpg" target="_blank">cli를하시기 바랍니다</a>이 그림의 더 큰 버전을 보려면 여기를 (CK). <img alt="그림 5" src="/files/ftp_upload/54110/54110fig5.jpg" /> 그림 5 : 색 도플러 영상을 이용한 승모판 막 역류 제트의 증거. 흉골 창에서 승모판의 변형 장축 뷰가 얻어진다. 컬러 도플러 심문은 (화살표로 강조) 수축기 동안 승모판에서 모자이크 컬러 잉크젯를 보여줍니다. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig5large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 6" src="/files/ftp_upload/54110/54110fig6.jpg" /> 그림 6 : 긴 축 주 폐 동맥의 전망과 주요 지점. 주요 폐동맥의 장축 뷰 (MPA) 및 우측 (RPA)과 왼쪽 (LPA) 분기는 paraste 얻을 수있다<html> rnal 창. 오른쪽 심실 유출 관 (우심실 유출로), 폐동맥 밸브 (PV) 및 대동맥 (AO)는 부분적으로 볼 수 있습니다. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig6large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 7" src="/files/ftp_upload/54110/54110fig7.jpg" /> 그림 7 : 일반 폐동맥 밸브를 묘사 M 모드의 이미지. 흉골 창에서 폐동맥 밸브 모두 단기 및 장축 뷰가 얻어 질 수있다. 심문 M 모드 라인은 폐동맥 밸브를 가로 질러인가된다. 폐동맥 밸브 첨단 분리 (화살표) 거리가이 뷰에서 측정 할 수있다. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig7large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> 10fig8.jpg &quot;/&gt; 그림 8 : 폐동맥 밸브의 맞은 편에 흐름의 펄스 파 도플러 심문. 폐동맥 가속 시간 (PAAT)의 최대 유속 수축기 폐동맥 유동 개시까지의 시간 간격이다. 우심실 토출 시간 (RVET)의 흐름이 중단 된 지점을 우심실 토출의 개시 사이의 간격이다. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig8large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 9" src="/files/ftp_upload/54110/54110fig9.jpg" /> 그림 9 : 좌심실의 짧은 축보기를 묘사 M 모드의 이미지. 흉골 창에서 좌심실의 짧은 축보기 반 변환기 헤드를 회전시킴으로써 얻어 지도록 뒤쪽 또는 등쪽 화상 인덱스 마커 포인트. 제 m 개조심문 E 라인은 유두근 수준에서 좌심실 사이에인가된다. 좌심실 이완 기말 치수 (LVEDD), 좌심실 수축 기말 치수 (LVESD), 및 전방 벽 (AW)과 후벽 (PW) 두께를 용이하게 측정 할 수있다. 모든 측정에서 유두근 (*)를 포함하지 않도록주의하십시오. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig9large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 10" src="/files/ftp_upload/54110/54110fig10.jpg" /> 그림 10 : 색 도플러 평가 및 승모판 유입의 펄스 파 도플러 스펙트럼 디스플레이. (A) 꼭대기 장축보기 승모판 유입 컬러 도플러 평가를 나타내는 이미지. 2 차원 컬러 도플러 이미지가 g위한 중요한 도구가 될 수 있습니다(패널 B에 도시) 펄스 파 도플러 트레이싱의 취득에 해당하는 샘플 볼륨 위치를 uiding. (B) 펄스 파 도플러를 이용하여 승모판 유입의 스펙트럼 표시됩니다. (혀끝 장축 뷰) 승모판 걸쳐 혈류 파 펄스 도플러 평가는 좌심실 확장기 기능을 평가하기 위해 수행된다. 샘플 볼륨은 승모판 막 전단지의 조언에 배치됩니다. isovolumic 완화 시간 (IVRT) isovolumic 수축 시간 (IVCT) 좌심실 시간 (LVET) 피크 승모판 유입 속도 (E)는 모든 승모판 걸쳐 펄스 파 도플러 속도의 스펙트럼 표시에서 유도 될 수있다. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig10large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 11" src="/files/ftp_upload/54110/54110fig11.jpg" /> 인터넷gure 11 : 중격 승모판 같이 고리의 조직 ​​도플러 영상. 꼭대기 창에서 승모판의 장축 뷰가 얻어진다. 조직 도플러 샘플 량은 승모판 막륜의 중격 부위에 위치한다. 피크 승모판 유입 속도 (도 10b에서 변수 E) 및 (E 피크 승모판 막륜의 조직 속도의 비율 &#39;(흰색 화살표로 표시는) 좌심실 확장기 함수를 평가하기 위해 사용되는 공통 E / E라고&#39;). <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig11large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a> <img alt="그림 12" src="/files/ftp_upload/54110/54110fig12.jpg" /> 그림 12 : 좌심실 심근의 변형과 변형 속도의 평가. 존재 가능한 전문 분석 소프트웨어 패키지는 시판하며, 변형률과 변형률 속도 변수 일 수있다고유 심근 수축 특성의 초기 또는 하위 임상 변경 조치로 얻을. 위의 예는 마우스에서 일반적으로 획득 된 영상 평면에서 반경 방향 변형 및 변형 속도를 묘사. 이 영상면 (및 변형 트레이싱의 후속 형상이) 자주 혀끝의 긴 축 또는 4 챔버보기에서 취득 인간의 이미지와 다를 수 있습니다. <a href="https://ecsource-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54110/54110fig12large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭하십시오.</a>

Watch this Scientific Journal Video about Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice at JoVE.com

Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice

This protocol provides a detailed description of the echocardiographic approach for comprehensive phenotyping of heart and heart valve function in mice.

마우스의 심장 판막 질환의 종합 표현형 특성에 대한 심 초음파 접근 및 프로토콜

The aim of this manuscript and accompanying video is to provide an overview of the methods and approaches used for imaging heart valve function in ...

echocardiographic-approaches-protocols-for-comprehensive-phenotypic

Universidad San Sebastian

Research

JoVE Journal

Medicine

15.9K 조회수.  Mayo Clinic. This protocol provides a detailed description of the echocardiographic approach for comprehensive phenotyping of heart and heart valve function in mice. 

비디오: Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice

Transthoracic Echocardiography in Mice

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene targeting methods can be used to generate mice with altered cardiac size and function,1-3 and as a result, in vivo techniques are needed to evaluate their cardiac phenotype. Transthoracic echocardiography, pulse wave Doppler (PWD), and tissue Doppler imaging (TDI) can be used to provide dimensional measurements of the mouse heart and to quantify the degree of cardiac systolic and diastolic performance. Two-dimensional imaging is used to detect abnormal anatomy or movements of the left ventricle, whereas M-mode echo is used for quantification of cardiac dimensions and contractility.4,5 In addition, PWD is used to quantify localized velocity of turbulent flow,6 whereas TDI is used to measure the velocity of myocardial motion.7 Thus, transthoracic echocardiography offers a comprehensive method for the noninvasive evaluation of cardiac function in mice.

Transthoracic echocardiography offers a noninvasive method for the evaluation of cardiac function in mice. A combination of ultrasound and Doppler imaging modalities can be used to obtain dimensional measurements of the heart and intracardiac blood flow, which together provide an assessment of cardiac systolic and diastolic performance.

마우스에 Transthoracic Echocardiography

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene ...

연구

의학

Murine Echocardiography and Ultrasound Imaging

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1 Echocardiography provides a powerful, non-invasive tool to serially assess cardiac morphometry and function in a living animal.2 However, using this technique on mice poses unique challenges owing to the small size and rapid heart rate of these animals.3 Until recently, few ultrasound systems were capable of performing quality echocardiography on mice, and those generally lacked the image resolution and frame rate necessary to obtain truly quantitative measurements. Newly released systems such as the VisualSonics Vevo2100 provide new tools for researchers to carefully and non-invasively investigate cardiac function in mice. This system generates high resolution images and provides analysis capabilities similar to those used with human patients. Although color Doppler has been available for over 30 years in humans, this valuable technology has only recently been possible in rodent ultrasound.4,5 Color Doppler has broad applications for echocardiography, including the ability to quickly assess flow directionality in vessels and through valves, and to rapidly identify valve regurgitation. Strain analysis is a critical advance that is utilized to quantitatively measure regional myocardial function.6 This technique has the potential to detect changes in pathology, or resolution of pathology, earlier than conventional techniques. Coupled with the addition of three-dimensional image reconstruction, volumetric assessment of whole-organs is possible, including visualization and assessment of cardiac and vascular structures. Murine-compatible contrast imaging can also allow for volumetric measurements and tissue perfusion assessment.

This video demonstrates use of a rail-mounted high-frequency ultrasound probe to perform echocardiography on an anesthetized mouse. The methods describe both conventional two-dimensional and M-mode measurements of cardiac function in addition to newer, more powerful tools such as color Doppler, strain analysis, as well as general and targeted contrast imaging.

Murine Echocardiography 및 초음파 이미징

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1  ...

Echocardiographic Assessment of the Right Heart in Mice

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential therapies. Given the expense and time involved in creating animal models of disease, it is critical that researchers have tools to accurately assess phenotypic expression of disease. Right ventricular dysfunction is the major manifestation of pulmonary hypertension. Echocardiography is the mainstay of the noninvasive assessment of right ventricular function in rodent models and has the advantage of clear translation to humans in whom the same tool is used. Published echocardiography protocols in murine models of PAH are lacking.
In this article, we describe a protocol for assessing RV and pulmonary vascular function in a mouse model of PAH with a dominant negative BMPRII mutation; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We provide a detailed description of animal preparation, image acquisition and hemodynamic calculation of stroke volume, cardiac output and an estimate of pulmonary artery pressure.

This article provides a protocol for the echocardiographic assessment of right ventricular size and pulmonary hypertension in mice. Applications include phenotype determination and serial assessment in transgenic and toxin-induced mouse models of cardiomyopathy and pulmonary vascular disease.

마우스의 오른쪽 심장의 심 초음파 평가

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

A Pipeline to Characterize Structural Heart Defects in the Fetal Mouse

Congenital heart diseases (CHDs) are major causes of infant death in the United States. In the 1980s and earlier, most patients with moderate or severe CHD died before adulthood, with the maximum mortality during the first week of life. Remarkable advances in surgical techniques, diagnostic approaches, and medical management have led to marked improvements in outcomes. To address the critical research needs of understanding congenital heart defects, murine models have provided an ideal research platform, as they have very similar heart anatomy to humans and short gestation rates. The combination of genetic engineering with high-throughput phenotyping tools has allowed for the replication and diagnosis of structural heart defects to further elucidate the molecular pathways behind CHDs. The use of noninvasive fetal echocardiography to screen the cardiac phenotypes in mouse models coupled with the high fidelity of Episcopic fluorescence image capture (EFIC) using Episcopic confocal microscopy (ECM) histopathology with three-dimensional (3D) reconstructions enables a detailed view into the anatomy of various congenital heart defects. This protocol outlines a complete workflow of these methods to obtain an accurate diagnosis of murine congenital heart defects. Applying this phenotyping protocol to model organisms will allow for accurate CHD diagnosis, yielding insights into the mechanisms of CHD. Identifying the underlying mechanisms of CHD provide opportunities for potential therapies and interventions.

This article details murine congenital heart disease (CHD) diagnostic methods using fetal echocardiography, necropsy, and Episcopic fluorescence image capture (EFIC) using Episcopic confocal microscopy (ECM) followed by three-dimensional (3D) reconstruction.

태아 마우스의 구조적 심장 결함을 특성화하는 파이프라인

Congenital heart diseases (CHDs) are major causes of infant death in the United States. In the 1980s and earlier, most patients with moderate or severe ...

발생학

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable noninvasive method is essential to assess real-time cardiac morphological and functional changes in animal models of heart disease. Transthoracic echocardiography represents an important tool for noninvasive assessment of cardiac structure and function2. Here we used a high-resolution ultrasound imaging system to monitor myocardial remodeling and heart failure progression over time in a mouse model of TAC. B-mode, M-mode, and Doppler imaging were used to precisely assess cardiac hypertrophy, ventricular dilatation, and functional deterioration in mice following TAC. Color and pulse wave (PW) Doppler imaging was used to noninvasively measure pressure gradient across the aortic constriction created by TAC and to assess transmitral blood flow in mice. Thus transthoracic echocardiographic imaging provides comprehensive noninvasive measurements of cardiac dimensions and function in mouse models of heart disease.

The goal of this protocol is to noninvasively assess cardiac structural and functional changes in a mouse model of heart disease created by transverse aortic constriction, using B- and M-mode echocardiography and color/pulse wave Doppler imaging.

심 초음파 이미징에 의해 가로 대동맥 수축의 마우스 모델에서 심장 형태 학적 평가 및 기능 변경

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic pressure through echocardiography, as well as to obtain measurements of the Aorta and PA diameter in mice.
Mice older than 10 d of age can be analyzed using the present technique. The technique is composed of 3 main steps: set-up, image acquisition, and image analysis. The set-up step consists of getting the mouse anesthetized with 1% isoflurane, shaving it, and taping it in a supine position to a heated EKG board where the image acquisition will take place. The image acquisition step consists of learning to identify the cardiac structures and obtaining all the required images with its correspondent probe and axis in order to be able to calculate volumes and diameters. The image analysis step consists of measuring the previously acquired images with the aid of computer software.
Advantages of the proposed technique include a fast (15 min) procedure that would allow the researcher to evaluate interventions in a non-invasive, non-terminal approach and therefore follow the same mouse over time; each mouse can be used as its own control. This fact plus having the same operator perform all the acquisition and analysis for the entire experiment minimizes the limitation of operator-dependency. The present methodology is useful for mouse researchers in cardiovascular and pulmonary medicine.

The following protocol describes the methodology for the acquisition and analysis of echocardiographic images used to obtain the Left Atrial Volume (LAV), Aorta (Ao) diameter, and Pulmonary Artery (PA) diameter in mice. This technique is a non-invasive, non-terminal procedure that allows assessment of the cardiopulmonary function.

왼쪽 아트리움, 대동맥과 폐동맥의 쥐 심 초음파

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic ...

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 ...

Echocardiographic Measurement of Right Ventricular Diastolic Parameters in Mouse

Diastolic dysfunction is a prominent feature of right ventricular (RV) remodeling associated with conditions of pressure overload. However, the RV diastolic function is rarely quantified in experimental studies. This might be due to technical difficulties in the visualization of the RV in the apical four-chamber view in rodents. Here we describe two positions facilitating the visualization of the apical four-chamber view in mice to assess the RV diastolic function.
The apical four-chamber view is enabled by tilting the mouse fixation platform to the left and caudally (LeCa) or to the right and cranially (RiCr). Both positions provide images of comparable quality. The results of the RV diastolic function obtained from two positions are not significantly different. Both positions are comparably easy to perform. This protocol can be incorporated into published protocols and enables detailed investigations of the RV function.

Here we describe and compare two positions for obtaining the apical four-chamber view in mice. These positions enable the quantification of the right ventricular function, provide comparable results, and can be used interchangeably.

마우스의 우심실 확장기 매개변수의 심초음파 측정

Diastolic dysfunction is a prominent feature of right ventricular (RV) remodeling associated with conditions of pressure overload. However, the RV ...

Revealing Subtle Changes in Cardiac Function using Transthoracic Dobutamine Stress Echocardiography in Mice

Left ventricular (LV) dysfunction paves the final pathway for a multitude of cardiac disorders. With the non-invasive high-frequency transthoracic dobutamine stress echocardiography in humans, a reductionist investigation approach to unmask subtle changes in cardiac function has become possible. Here, we provide a protocol for using this technique in mice to facilitate expanded analysis of LV architecture and function in physiology and pathology enabling the observation of alterations in models of cardiac disease hidden in unstressed hearts. This investigation can be performed in one and the same animal and allows both, basal and pharmacologically stress-induced measurements. We outline detailed criteria for appropriate anesthesia, imaging-based LV analysis, consideration of intra- and interobserver variability, and obtaining positive inotrope response that can be attained in mice after intraperitoneal injection of dobutamine under near physiological conditions. To recapitulate the characteristics of human physiology and disease in small animal models, we highlight critical pitfalls in evaluation, e.g., a pronounced Bowditch effect in mice. To further meet translational objectives, we compare stress-induced effects in humans and mice. When used in translational studies, attention must be paid to physiological differences between mice and human. Experimental rigor dictates that some parameters assessed in patients can only be used with caution due to restrictions in spatial and temporal resolution in mouse models.

Left ventricular dysfunction constitutes the final common pathway for a host of cardiac disorders. We here provide a detailed protocol of transthoracic dobutamine stress echocardiography approach for comprehensive evaluation of the left ventricular function of mouse models of cardiac disease as well as cardiac phenotyping.

Left ventricular (LV) dysfunction paves the final pathway for a multitude of cardiac disorders. With the non-invasive high-frequency transthoracic ...

Echocardiographic Characterization of Left Ventricular Structure, Function, and Coronary Flow in Neonate Mice

Echocardiography is a non-invasive procedure that enables the evaluation of structural and functional parameters in animal models of cardiovascular disease and is used to assess the impact of potential treatments in preclinical studies. Echocardiographic studies are usually conducted in young adult mice (i.e., 4-6 weeks of age). The evaluation of early neonatal cardiovascular function is not usually performed because of the small size of the mouse pups and the associated technical difficulties. One of the most important challenges is that the short length of the pups&#39; limbs prevents them from reaching the electrodes in the echocardiography platform. Body temperature is the other challenge, as pups are very susceptible to changes in temperature. Therefore, it is important to establish a practical guide for performing echocardiographic studies in small mouse pups to help researchers detect early pathological changes and study the progression of cardiovascular disease over time. The current work describes a protocol for performing echocardiography in mouse pups at the early age of 7 days old. The echocardiographic characterization of cardiac morphology, function, and coronary flow in neonatal mice is also described.

The present protocol describes the echocardiographic assessment of left ventricular morphology, function, and coronary blood flow in 7-day old neonate mice.

신생아 마우스의 좌심실 구조, 기능 및 관상 동맥 흐름의 심 초음파 특성

Echocardiography is a non-invasive procedure that enables the evaluation of structural and functional parameters in animal models of cardiovascular ...