XHTW 의료 연구에 WM 켁 재단 고유 영 학자이며, 또한 NIH / NHLBI 보조금 R01 - R01 - HL089598 및 R01HL091947, 그리고 근육질 영양 장애 협회 부여 # 69238에 의해 지원됩니다. RJvO는 생리 게놈의 2008-2010 미국 생리 학회 박사 과정 이수 해안에서받는 것입니다. 이 작품은 또한 CaMKII가 마음에 신호를위한 Fondation Leducq 동맹에 의해 부분적으로 지원됩니다.

1 부 : 운영 필드의 작성 <ol><li> 운영 필드 75% 이소 프로필 알코올로 소독합니다. </li><li> 가열 패드와 적당한 온도에 있는지 확인하십시오. 권장 시스템 ° C ± 1 ° C. 37 유지 치료 패드에 연결된 Gaymar 순환 펌프입니다 심장 박동의 급속한 감소를 피하기 위해 그것은 수술 동안 정상 체온을 유지하는 것이 중요합니다. </li><li> 수술 도구는 수술 전에 뜨거운 구슬 살균기로 소독을하고 있습니다. 무딘 가위, 물론 곡선 집게 X 2, 벌금 45 ° 직각 집게 X 2, 직각 봄 가위, 가슴 견인기, 그리고 바늘 홀더 :.이 절차 들어, 다음과 같은 수술 도구가 필요합니다 </li><li> 커튼 applicators은 출혈의 경우를 대비해해야합니다. </li></ol> 2 부 : 준비 및 마우스의 삽관법 <ol><li> 마우스로 유도 챔버에 anesthetized 아르 2퍼센트 isoflurane 0.5 -1.0 L / 분 백퍼센트 O 2 혼합. </li><li> 헤어 가위는 neckline 중순 가슴 수준으로 모피를 면도하는 데 사용됩니다. </li><li> 마우스는 체온을 유지하기 위해 가열 패드 위에 부정사 위치에 배치됩니다. </li><li> 고무 밴드는 목에을 연장하기 위해 동물의 앞니 위에 배치됩니다. 한 손으로 곡선 집게를 사용하여 혀가 부드럽게 측면으로 조작할 수 있습니다. 다른 한편으로, endotracheal 삽관법은 PE 90 튜브를 사용하여 수행됩니다. endotracheal 튜브 그런 다음 숨을 125-150 / 분에서 하버드 볼륨 순환될 쥐 환풍기 사이클링 0.1 - 0.3ml의 갯벌 볼륨에 연결되어 있습니다. 1.0 L / 분 백퍼센트 O 2 - 수술 도중 마취가 0.5와 isoflurane 1.5-2 %에서 관리하고 있습니다. 마취의 올바른 수준 마우스 손톱 침대 (발가락 - 핀치 반사)에 압력을 적용하여 확인됩니다. </li><li> 외과 분야는 70 % 알콜 다음 betadine 솔루션으로 소독합니다. 이 절차는 세 번 반복됩니다. </li><li> 작업하는 동안 수술 부위의 오염을 방지하기 위해 무균 드레이프는 작업 필드 노출 떠나는 마우스 배치됩니다. </li><li> 멸균 장갑 세트 각 마우스에 사용됩니다. </li></ol> 파트 3 : 트랜스 대동맥의 결합 <ol><li> 두 번째 늑골을 부분 thoracotomy은 수술 현미경을 수행하고 흉골은 가슴 견인기를 사용하여 철회 있습니다. </li><li> 좋은 팁 45 ° 직각 포셉가 부드럽게 대동맥 아치에서 thymus과 지방 조직을 분리하는 데 사용됩니다. </li><li> 가로 대동맥의 신분에 따라, 6.0 실크 봉합사의 작은 조각은 innominate과 왼쪽 경동맥 동맥 (그림 1) 사이에 배치됩니다. </li><li> 두 느슨한 노트는 가로 대동맥 주위에 연결되어 있으며, 27의 작은 조각은 ½ 게이지 뭉툭한 주사 바늘을 가로 대동맥에 평행하게 배치됩니다. 첫 번째 매듭은 빠르게, 바늘에 대해 연결된 두 번째로 다음과 바늘 즉시 직경 0.4mm의 수축을 항복하기 위해 제거됩니다. 가짜 제어 마우스에서 전체 과정은 대동맥의 결합을 제외하고 동일합니다. </li><li> 가슴 견인기을 제거하고 다시 부풀려 폐 2 초 동안 환풍기 모근에서의 유출이다. </li><li> 갈빗대는 중지 봉합 패턴 6.0 prolene의 봉합사를 사용하여 닫힙니다. </li><li> 피부는 지속적인 치료 패턴 6.0 prolene의 봉합사를 사용하여 닫힙니다. </li></ol> 4 부 : 포스트 수술 복구 <ol><li> 이후 수술 진통제 들어, 마우스가 buprenorphine (0.1 밀리그램 / kg) intraperitoneally로 주입됩니다. 수술 후 탈수의 징후가있다면, 멸균 생리가 intraperitoneally 주어집니다. </li><li> 마취는 점차 해제 위치로 낮춘 및 자발적인 호흡의 징후가 발생할 때 endotracheal 튜브가 제거됩니다. </li><li> 마우스가 발생하기 쉬운 위치로 이동하고 손난로에서 복구하는 허용됩니다. </li></ol> 5 부 : 트랜스 대동맥의 성공적인 결합의 확인 <ol><li> TAC 후 일주일은 마우스가 가로 대동맥의 결합에 의해 유도된 압력 과부하에 노출된 정도를 확인하기 위해 다시 anesthetized입니다. 37 유지 1.0 L / 분 백퍼센트 O 2, 체온 ° C ± 1 ° C. - 위에서 설명한 바와 같이, 마취는 0.5과 isoflurane 1.5-2 %로 유지 </li><li> 20 MHz의 도플러 프로브 흐름 판매율을 탐지하기 위해 45 ° 각도로 목의 왼쪽과 오른쪽 측면에 배치됩니다. 컴퓨터 기반의 도플러 신호 프로세서 (인더스 인 스트 루먼트, 휴스턴, TX) 도플러 신호를 표시하고 저장하는 데 사용되었다. 5 </li><li> 실험 프로토콜에 필요한 압력 과부하의 정도에 따라 오른쪽 경동맥 (RC) / 좌측 경동맥 (LC) 특정 범위 내에서 유동 비율만을 생쥐는 추가 분석을 위해 포함됩니다. 심각한 압력 과부하의 결과 어깨를 수축이 T를 리드 반면에 예를 들어, 압력 과부하의 중간 정도, 5-8의 비율로 연결80-10의 OA 비율 (그림 5.3A). 가짜 동물은 (운영하지만, 출혈도 잡았하지 않음) 그러나, ~ 1 (그림 5.3B)의 비율을 것으로 예상된다. </li></ol> 6 : 대표 결과 야생 형 생쥐의 전술에 따라 일반 외과 생존은 80-90 % 정도입니다. 가로 대동맥의 성공적인 수술 내고 5-10의 오른쪽과 왼쪽 경동맥 사이의 도플러 유속 비율 (RC / LC) (그림 1)으로 이어질 것입니다. 가짜 운항 마우스 (그림 2A)에 비해 압력 과부하와 생쥐는 수축 (그림 2B)의 죄어져 있음에 따라, 6-8 주에 1~2주 및 심장 팽창 이내에 심장 비대를 개발 예정이다. 유전자 - 수정된 생쥐가 다른 생존 속도를 전시 수도 있지만 TAC 후 첫 달 동안 사망률은 야생 형 생쥐에 (&lt;20 %) 일반적으로 낮습니다. 6,7 <img src="/files/ftp_upload/1729/1729fig1tmb.jpg" alt="그림 1" border="0" /> 그림 1. 도플러 속도 측정을 사용하여 트랜스 대동맥의 결합의 확인. 오른쪽 대표 도플러 속도 신호 (RC)과 왼쪽 경동맥 (LC) 동맥 육일 포스트 TAC. 이 음반은 ~ 6.8의 RC / LC 유동 비율과 함께 성공적인 결합을 보여줍니다. 하시기 바랍니다 <a href="https://www.jove.com/files/ftp_upload/1729/1729fig1.jpg">여기를 클릭</a> 그림 1의 더 큰 버전을 볼 수 있습니다. <img src="/files/ftp_upload/1729/1729fig2.jpg" alt="그림 2" /> 그림 2. TAC 다음 대표 전심 이미지. . 섐 - 운영 야생 타입 마우스 심장. B. 와일드 타입 마우스 심장 16주 포스트 TAC. 각 라인 = 1mm.

<ol>
	<li>Rockman, H. A., Ross, R. S., Harris, A. N., Knowlton, K. U., Steinhelper, M. E., Field, L. J., Ross, J., Chein, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Segregation+of+atrial-specific+and+inducible+expression+of+an+atrial+natriuretic+factor+transgene+in+an+in+vivo+murine+model+of+cardiac+hypertrophy.">Segregation of atrial-specific and inducible expression of an atrial natriuretic factor transgene in an in vivo murine model of cardiac hypertrophy.</a> Proc. Natl. Acad. Sci. USA. 88, 8277-8281 (1991).</li><li>Heineke, J., Molkentin, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+cardiac+hypertrophy+by+intracellular+signaling+pathways.">Regulation of cardiac hypertrophy by intracellular signaling pathways.</a> Nat. Rev. Mol. Cell. Biol. 7, 589-600 (2006).</li><li>Hartley, C. J., Reddy, A. K., Madala, S., Michael, L. H., Entman, M. L., Taffett, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doppler+estimation+of+reduced+coronary+flow+reserve+in+mice+with+pressure+overload+cardiac+hypertrophy.">Doppler estimation of reduced coronary flow reserve in mice with pressure overload cardiac hypertrophy.</a> Ultrasound Med. Biol. 34, 892-901 (2008).</li><li>Reddy, A. K., Taffett, G. E., Li, Y. -. H., Lim, S. -. W., Pham, T. T., Pocius, J. S., Entman, M. L., Michael, L. H., Hartley, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doppler+signal+processing+for+use+in+mice:+applications.">Doppler signal processing for use in mice: applications.</a> IEEE Trans. Biomed. Eng. 52, 1771-1783 (2005).</li><li>Li, Y. -. H., Reddy, A. K., Ochoa, L. N., Pham, T. T., Hartley, C. J., Micheal, L. H., Entman, M. L., Taffett, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+age+on+peripheral+vascular+response+to+transverse+aortic+banding+in+mice.">Effect of age on peripheral vascular response to transverse aortic banding in mice.</a> J. Gerontol. 58A, 895-899 (2003).</li><li>Bourajjaj, M., Armand, A. S., Martins, d. a. C. o. s. t. a., Weijts, P. A., Nagel, B. V. a. n. d. e. r., Heeneman, R., Wehrens, S., Windt, X. H. D. e., J, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NFATc2+is+a+necessary+mediator+of+calcineurin-dependent+cardiac+hypertrophy+and+heart+failure.">NFATc2 is a necessary mediator of calcineurin-dependent cardiac hypertrophy and heart failure.</a> J. Biol. Chem. 28, 22295-22303 (2008).</li><li>Oort, R. J. v. a. n., Respress, J. L., Li, N., Reynolds, C., Almeida, A. C. D. e., Skapura, D. G., Windt, L. J. D. e., Wehrens, X. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+development+of+pressure+overload+induced+cardiac+hypertrophy+and+dysfunction+in+an+RyR2-R176Q+knockin+mouse+model.">Accelerated development of pressure overload induced cardiac hypertrophy and dysfunction in an RyR2-R176Q knockin mouse model.</a> Hypertension. 55, (2010).</li><li>Tarnavski, O., McMullen, J. R., Schinke, M., Nie, Q., Kong, S., Izumo, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+cardiac+surgery:+comprehensive+techniques+for+the+generation+of+mouse+models+of+human+diseases+and+their+application+for+genomic+studies.">Mouse cardiac surgery: comprehensive techniques for the generation of mouse models of human diseases and their application for genomic studies.</a> Physiol Genomics. 16, 349-360 (2004).</li></ol>

인간의 대동맥 협착증을 모방한 것이었 전술은 생쥐에서 심장 비대 및 심장 마비를 유발하는 일반적인 방법입니다. 대동맥 수축에 대한 대체 사이트는 오름차순 및 복부 대동맥을 포함합니다. 대동맥 수축 오름차순하면 좌심실 (LV)에 극도 더 빠른 과부하를 제공합니다. 반면, 복부 대동맥 수축 가능한 보상하는 수단으로 순환 그대로 큰 부분을 떠납니다.이 조사를 더받을 수있는 시간 종속 방식?...

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Gaymar T/pump</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>Model TP-500</td>
<td>Circulating water pump</td>
</tr>
<tr>
<td>Temp. therapy pads</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>60-3414</td>
<td>&nbsp;</td>
<tr>
<td>Hot bead sterilizer</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>18000-45</td>
<td>&nbsp;</td>
<tr>
<td>Blunt scissors</td>
<td>&nbsp;</td>
<td>Roboz</td>
<td>RS-5980</td>
<td>&nbsp;</td>
<tr>
<td>Angled spring scissors</td>
<td>&nbsp;</td>
<td>Roboz</td>
<td>RS-5668</td>
<td>&nbsp;</td>
<tr>
<td>Chest retractor</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>17002-02</td>
<td>&nbsp;</td>
<tr>
<td>Course curved forceps</td>
<td>&nbsp;</td>
<td>Roboz</td>
<td>RS-5138</td>
<td>&nbsp;</td>
<tr>
<td>Dumont fine 45&deg; angled forceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11253-25</td>
<td>&nbsp;</td>
<tr>
<td>Needle holder</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>12565-14</td>
<td>&nbsp;</td>
<tr>
<td>Electric shaver</td>
<td>&nbsp;</td>
<td>General Electric</td>
<td>&nbsp;</td>
<td>For shaving mouse fur</td>
</tr>
<tr>
<td>PE 90 tubing</td>
<td>&nbsp;</td>
<td>BD</td>
<td>427420</td>
<td>For intubation</td>
</tr>
<tr>
<td>MiniVent (ventilator)</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>Type 845</td>
<td>&nbsp;</td>
<tr>
<td>Betadine</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>19-027132</td>
<td>May be purchased at medical supply store</td>
</tr>
<tr>
<td>Sterile gloves</td>
<td>&nbsp;</td>
<td>Mckesson</td>
<td>20-1565</td>
<td>&nbsp;</td>
<tr>
<td>Stereo microscope</td>
<td>&nbsp;</td>
<td>Unico</td>
<td>ZM186</td>
<td>&nbsp;</td>
<tr>
<td>6-0 silk suture</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>18020-60</td>
<td>&nbsp;</td>
<tr>
<td>27 &frac12; gauge needle</td>
<td>&nbsp;</td>
<td>BD</td>
<td>305109</td>
<td>&nbsp;</td>
<tr>
<td>6-0 prolene suture</td>
<td>&nbsp;</td>
<td>Mckesson</td>
<td>3286</td>
<td>&nbsp;</td>
<tr>
<td>Buprenorphine</td>
<td>&nbsp;</td>
<td>Institutional Animal Facility</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

transverse aortic constriction in mice

마우스의 가로 대동맥 수축 (TAC)은 압력 과부하 유발 심장 비대 및 심장 마비에 대해 일반적으로 사용되는 실험적인 모델입니다.<sup&gt; 1</sup&gt; TAC 처음 종종 심장 수축성의 임시 향상과 관련된 심장의 보상 비대로 연결됩니다. 시간이지나면서, 그러나, 만성 hemodynamic 과부하에 대한 반응은 심장 팽창과 심장 마비의 결과, maladaptive됩니다.<sup&gt; 2</sup&gt; murine TAC 모델이 처음으로 확인되었다 락맨<em&gt; 외</em&gt;.<sup&gt; 1</sup&gt; 이후 광범위하게 인간의 심장 혈관 질병을 모방하고 심장 비대 반응 및 심장 마비로 개발에 참여 근본적인 신호 프로세스를 명료하게하다하는 중요한 도구로 사용되고 있습니다. 등 (내사) 관상 동맥을 내림차순 왼쪽 앞쪽에의 완전한 폐색과 같은 심장 마비의 다른 실험 모델에 비해, 전술은 심장 비대 및 심장 마비의 개발에보다 점진적 시간 코스보다 재현성 모델을 제공합니다. 여기, 우리는 생쥐에서 외과 전술을 수행하는 단계별 절차를 설명합니다. 대동맥 결합에 의해 만들어진 압력 과부하의 수준을 확인하려면, 고주파 도플러 프로브가 오른쪽과 왼쪽 경동맥 동맥의 혈액 흐름 판매율 간의 비율을 측정하는 데 사용됩니다.<sup&gt; 3, 4</sup&gt; 80-90%의 수술 생존 속도, banding 가로 대동맥은 생쥐에서 왼쪽 심실 비대 및 심장 마비를 유도하는 효과적인 기술입니다.

Watch this Scientific Journal Video about Transverse Aortic Constriction in Mice at JoVE.com

Transverse Aortic Constriction in Mice

마우스의 가로 대동맥 수축 (TAC)은 심장 비대 및 심장 마비 개발을 기본 메커니즘을 연구하기 위해 일반적으로 사용되는 실험적인 모델입니다. 여기, 우리는 절차가 생쥐에서 심장 비대의 재현성 학위를 만들 수있는 대동맥을 수축에 대해 설명합니다.

마우스의 가로 대동맥 수축

Transverse aortic constriction (TAC) in the mouse is a commonly used experimental model for pressure overload-induced cardiac hypertrophy and heart ...

Research

JoVE Journal

Biology

66.7K 조회수.  Baylor College of Medicine (BCM). 마우스의 가로 대동맥 수축 (TAC)은 심장 비대 및 심장 마비 개발을 기본 메커니즘을 연구하기 위해 일반적으로 사용되는 실험적인 모델입니다. 여기, 우리는 절차가 생쥐에서 심장 비대의 재현성 학위를 만들 수있는 대동맥을 수축에 대해 설명합니다.

비디오: Transverse Aortic Constriction in Mice

Small Bowel Transplantation In Mice

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive medications, infection prophylaxis, and suitable patient selection helped improve actuarial graft and patient survival rates for all types of intestine transplantation. Patients with irreversible intestinal failure and complications of parenteral nutrition should now be routinely considered for small intestine transplantation. However, Survival rates for small intestinal transplantation have been slow to improve compares increasingly favorably with renal, liver, heart and lung. The small bowel transplantation is still unsatisfactory compared with other organs. Further progress may depend on better understanding of immunology and physiology of the graft and can be greatly facilitated by animal models. A wider use of mouse small bowel transplantation model is needed in the study of immunology and physiology of the transplantation gut as well as efficient methods in diagnosing early rejection. However, this model is limited to use because the techniques involved is an extremely technically challenging. We have developed a modified technique. When making anastomosis of portal vein and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s portal vein. The left wall of the inferior vena cava and donor s portal vein is closed with continuing sutures in the inside of the inferior vena cava after, after one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s portal vein are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis.  In this article, we provide details of the technique to supplement the video.

The mouse small bowel transplantation model has been recognized as an important tool to study mechanismes of immune rejection and screen new immunosuppressive drugs. However, this model is limited to use because the techniques involved is an extremely technically challenge. Now we introduce the modified technique.

마우스에 작은 창자 이식

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive ...

연구

생물학

Aortic Ring Assay

Angiogenesis, the sprouting of blood vessels from preexisting vasculature is associated with both natural and pathological processes. Various angiogenesis assays involve the study of individual endothelial cells in culture conditions (1). The aortic ring assay is an angiogenesis model that is based on organ culture. In this assay, angiogenic vessels grow from a segment of the aorta (modified from (2)). Briefly, mouse thoracic aorta is excised, the fat layer and adventitia are removed, and rings approximately 1 mm in length are prepared. Individual rings are then embedded in a small solid dome of basement matrix extract (BME), cast inside individual wells of a 48-well plate. Angiogenic factors and inhibitors of angiogenesis can be directly added to the rings, and a mixed co-culture of aortic rings and other cell types can be employed for the study of paracrine angiogenic effects. Sprouting is observed by inspection under a stereomicroscope over a period of 6-12 days. Due to the large variation caused by the irregularities in the aortic segments, experimentation in 6-plicates is strongly advised. Neovessel outgrowth is monitored throughout the experiment and imaged using phase microscopy, and supernatants are collected for measurement of relevant angiogenic and anti-angiogenic factors, cell death markers and nitrite.

Angiogenesis, the sprouting of blood vessels from pre-existing vasculature, is associated with both natural and pathological processes. Here we demonstrate an aortic ring assay that allows angiogenic potentiators and inhibitors to be directly added to aortic rings in culture. Sprouting and neovessel outgrowth can be determined by inspecting the aortic rings over a period of 6-12 days.

대동맥 링 분석

Angiogenesis, the sprouting of blood vessels from preexisting vasculature is associated with both natural and pathological processes. Various angiogenesis ...

Ambulatory ECG Recording in Mice

Telemetric ECG recording in mice is essential to understanding the mechanisms behind arrhythmias, conduction disorders, and sudden cardiac death. Although the surface ECG is utilized for short-term measurements of waveform intervals, it is not practical for long-term studies of heart rate variability or the capture of rare episodes of arrhythmias. Implantable ECG telemeters offer the advantages of simple surgical implantation, long-term recording of electrograms in ambulatory mice, and scalability with simultaneous recordings of multiple animals. Here, we present a step-by-step guide to the implantation of telemeters for ambulatory ECG recording in mice. Careful adherence to aseptic technique is required for favorable survival results with the possibility of implantation and recording from weeks to months. Thus, implantable ECG telemetry is a valuable tool for detection of critical information on cardiac electrophysiology in ambulatory animal models such as the mouse. 

Telemetric ECG has emerged as an essential tool in evaluating animal models for cardiac arrhythmias and sudden cardiac death. Here, we present a stepwise guide to telemetric ECG recordings for application in long-term ambulatory ECG monitoring in mice.

마우스의 외래 ECG 기록

Telemetric ECG recording in mice is essential to understanding the mechanisms behind arrhythmias, conduction disorders, and sudden cardiac death. Although ...

Intravenous Injections in Neonatal Mice

Intravenous injection is a clinically applicable manner to deliver therapeutics. For adult rodents and larger animals, intravenous injections are technically feasible and routine. However, some mouse models can have early onset of disease with a rapid progression that makes administration of potential therapies difficult. The temporal (or facial) vein is just anterior to the ear bud in mice and is clearly visible for the first two days after birth on either side of the head using a dissecting microscope. During this window, the temporal vein can be injected with volumes up to 50 &#956;l. The injection is safe and well tolerated by both the pups and the dams. A typical injection procedure is completed within 1-2 min, after which the pup is returned to the home cage. By the third postnatal day the vein is difficult to visualize and the injection procedure becomes technically unreliable. This technique has been used for delivery of adeno-associated virus (AAV) vectors, which in turn can provide almost body-wide, stable transgene expression for the life of the animal depending on the viral serotype chosen.

Animal models of pediatric disease can experience early onset and aggressive disease progression. Clinically relevant therapy delivery to young mouse models can be technically difficult. This protocol describes a non-invasive intravenous injection method for newborn mice within the first two postnatal days of life.

신생아 마우스의 정맥 주사

Intravenous injection is a clinically applicable manner to deliver therapeutics. For adult rodents and larger animals, intravenous injections are ...

Isolation and Primary Culture of Mouse Aortic Endothelial Cells

The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. The mouse is an important model for cardiovascular disease research. This study demonstrates a simple method to isolate and culture endothelial cells from the mouse aorta without any special equipment. To isolate endothelial cells, the thoracic aorta is quickly removed from the mouse body, and the attached adipose tissue and connective tissue are removed from the aorta. The aorta is cut into 1 mm rings. Each aortic ring is opened and seeded onto a growth factor reduced matrix with the endothelium facing down. The segments are cultured in endothelial cell growth medium for about 4 days. The endothelial sprouting starts as early as day 2. The segments are then removed and the cells are cultured continually until they reach confluence. The endothelial cells are harvested using neutral proteinase and cultured in endothelial cell growth medium for another two passages before being used for experiments. Immunofluorescence staining indicated that after the second passage the majority of cells were double positive for Dil-ac-LDL uptake, Lectin binding, and CD31 staining, the typical characteristics of endothelial cells. It is suggested that cells at the second to third passages are suitable for in vitro and in vivo experiments to study the endothelial biology. Our protocol provides an effective means of identifying specific cellular and molecular mechanisms in endothelial cell physiopathology.

The vascular endothelial cells play a significant role in many important cardiovascular disorders. This article describes a simple method to isolate and expand endothelial cells from the mouse aorta without using any special equipment. Our protocol provides an effective means of identifying mechanisms in endothelial cell physiopathology.

분리 및 마우스 대동맥 내피 세포의 차 문화

The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. The mouse is an ...

Induction of Right Ventricular Failure by Pulmonary Artery Constriction and Evaluation of Right Ventricular Function in Mice

The mechanism of right ventricular failure (RVF) requires clarification due to the uniqueness, high morbidity, high mortality, and refractory nature of RVF. Previous rat models imitating RVF progression have been described. Compared with rats, mice are more accessible, economical, and widely used in animal experiments. We developed a pulmonary artery constriction (PAC) approach which is comprised of banding the pulmonary trunk in mice to induce right ventricular (RV) hypertrophy. A special surgical latch needle was designed that allows for easier separation of the aorta and the pulmonary trunk. In our experiments, the use of this fabricated latch needle reduced the risk of arteriorrhexis and improved the surgical success rate to 90%. We used different padding needle diameters to precisely create quantitative constriction, which can induce different degrees of RV hypertrophy. We quantified the degree of constriction by evaluating the blood flow velocity of the PA, which was measured by noninvasive transthoracic echocardiography. RV function was precisely evaluated by right heart catheterization at 8 weeks after surgery. The surgical instruments made inhouse were composed of common materials using a simple process that is easy to master. Therefore, the PAC approach described here is easy to imitate using instruments made in the lab and can be widely used in other labs. This study presents a modified PAC approach that has a higher success rate than other models and an 8-week postsurgery survival rate of 97.8%. This PAC approach provides a useful technique for studying the mechanism of RVF and will enable an increased understanding of RVF.

Here, we provide a useful approach for studying the mechanism of right ventricular failure. A more convenient and efficient approach to pulmonary artery constriction is established using surgical instruments made inhouse. In addition, methods to evaluate the quality of this approach by echocardiography and catheterization are provided.

폐 동맥 수축에 의한 우심실 장애 유도 및 마우스의 우심실 기능 평가

The mechanism of right ventricular failure (RVF) requires clarification due to the uniqueness, high morbidity, high mortality, and refractory nature of ...

Inducing Apical Periodontitis in Mice

The mechanisms involved in local induced inflammation can be studied using several available animal models. One of these is the induction of apical periodontitis (AP). Apical periodontitis is a common pathology of an inflammatory nature in the periodontal tissues surrounding the tooth root. In order to better understand the nature and mechanism of this pathology it is advantageous to perform the procedure in mice. The induction of this odontogenic inflammation is achieved by drilling into the mouse tooth until the dental pulp is exposed. Next, the tooth pulp remains exposed to be contaminated by the natural oral flora over time, causing apical periodontitis. After this time period, the animal is sacrificed, and the tooth and the jaw bone can be analyzed in various ways. Typical analyses include micro-CT imaging (to evaluate bone resorption), histological staining, immunohistochemistry, and RNA expression. This protocol is useful for research in the field of oral biology to better understand this inflammatory process in an in vivo experimental setting with uniform conditions. The procedure requires a careful handling of the mice and the isolated jaw, and a visual demonstration of the technique is useful. All technical aspects of the procedures leading to induced apical periodontitis and its characterization in a mouse model are demonstrated.

Here, we present a protocol to locally induce apical periodontitis in mice. We show how to drill a hole in the mouse&#39;s tooth and expose its pulp, in order to cause local inflammation. Analysis methods to investigate the nature of this inflammation, such as micro-CT and histology, are also demonstrated.

마우스에 있는 정점 치주염 유도

The mechanisms involved in local induced inflammation can be studied using several available animal models. One of these is the induction of apical ...

Transverse Fracture of the Mouse Femur with Stabilizing Pin

Fracture repair is an essential function of the skeleton that cannot be reliably modeled in vitro. A mouse injury model is an efficient approach to test whether a gene, gene product or drug influences bone repair because murine bones recapitulate the stages observed during human fracture healing. When a mouse or human breaks a bone, an inflammatory response is initiated, and the periosteum, a stem cell niche surrounding the bone itself, is activated and expands. Cells residing in the periosteum then differentiate to form a vascularized soft callus. The transition from the soft callus to a hard callus occurs as the recruited skeletal progenitor cells differentiate into mineralizing cells, and the bridging of the fractured ends results in the bone union. The mineralized callus then undergoes remodeling to restore the original shape and structure of the healed bone. Fracture healing has been studied in mice using various injury models. Still, the best way to recapitulate this entire biological process is to break through the cross-section of a long bone that encompasses both cortices. This protocol describes how a stabilized, transverse femur fracture can be safely performed to assess healing in adult mice. A surgical protocol including detailed harvesting and imaging techniques to characterize the different stages of fracture healing is also provided.

This protocol describes a method to perform fractures on adult mice and monitor the healing process.

안정화 핀을 가진 마우스 대퇴골의 횡단 골절

Fracture repair is an essential function of the skeleton that cannot be reliably modeled in vitro. A mouse injury model is an efficient approach to test ...

A Modified Technique for Transverse Aortic Constriction in Mice

Transverse aortic constriction (TAC) is a frequently used surgery in research regarding heart failure and cardiac hypertrophy based on the formation of pressure overload in mouse models. The main challenge of this procedure is to clearly visualize the transverse aortic arch and precisely band the target vessel. Classical approaches perform a partial thoracotomy to expose the transverse aortic arch. However, it is an open-chest model that causes a rather large surgical trauma and requires a ventilator during the surgery. To prevent unnecessary trauma and simplify the operating procedure, the aortic arch is approached via the proximal proportion of the sternum, reaching and binding the target vessel using a small self-made retractor that contains a snare. This procedure can be conducted without entering the pleura cavity and does not need a ventilator or microsurgical operation, which leaves the mice with physiological breathing patterns, simplifies the procedure, and significantly reduces operation time. Due to the less invasive approach and less operation time, mice can undergo fewer stress reactions and recover rapidly.

The present protocol describes a modified and simplified technique with a minimally invasive transverse aortic constriction (TAC) procedure using a self-made retractor. This procedure can be conducted without a ventilator or microscope and introduces pressure overload, eventually leading to cardiac hypertrophy or heart failure.

생쥐의 횡방향 대동맥 수축에 대한 수정된 기술

Transverse aortic constriction (TAC) is a frequently used surgery in research regarding heart failure and cardiac hypertrophy based on the formation of ...

A Calcium Phosphate-Induced Mouse Abdominal Aortic Aneurysm Model

An abdominal aortic aneurysm (AAA) is a life-threatening cardiovascular disease that occurs worldwide and is characterized by irreversible dilation of the abdominal aorta. Currently, several chemically induced murine AAA models are used, each simulating a different aspect of the pathogenesis of AAA. The calcium phosphate-induced AAA model is a rapid and cost-effective model compared to the angiotensin II- and elastase-induced AAA models. The application of CaPO4 crystals to the mouse aorta results in elastic fiber degradation, loss of smooth muscle cells, inflammation, and calcium deposition associated with aortic dilation. This article introduces a standard protocol for the CaPO4-induced AAA model. The protocol includes material preparation, the surgical application of the CaPO4&#160;to the adventitia of the infrarenal abdominal aorta, the harvesting of aortas to visualize aortic aneurysms, and histological analyses in mice.

This protocol describes a calcium phosphate-induced abdominal aortic aneurysm (AAA) mouse model to study the pathological features and molecular mechanisms of AAAs.

인산칼슘 유도 마우스 복부 대동맥류 모델

An abdominal aortic aneurysm (AAA) is a life-threatening cardiovascular disease that occurs worldwide and is characterized by irreversible dilation of the ...