이 작품은 과학, 기술 및 S. 한국의 교육부에서 NIH 기금 HL87012 (HJ), HL75209 (HJ), UO1HL80711 (DH, DG, HJ) 및 세계 클래스 대학 프로젝트 (HJ)의 기금에 의해 지원되었다.

1. 왼쪽 경동맥 부분 내고 <ol><li> 생쥐는 세 ~ 8 주가되면 사용됩니다. 생쥐의 무게는 최소한 18g으로 권장합니다. </li><li> 비드 악기 살균기 30 초 악기를 소독하고 진정 수 </li><li> 케타민을 (80mg/kg)과 Xylazine (10mg/kg) IACUC 승인으로 intraperitoneal (IP) 주입 또는 대체 마취와 마취를 유발. IP 주사, 복부의 왼쪽 낮은 사분면에 마우스를 주입. </li><li> 완전히 anesthetized까지 온난 챔버에 넣어 마우스. 발가락 핀치에 대한 응답과 마취의 깊이를 확인합니다. </li><li> 부정사 (배꼽 사이드까지) 위치에 수술 무대에서 플레이스 마우스. 테이프 포어 - 포스 야자수와 무대 아래로 뒷다리 - 싶단다 단독. </li><li> mandible과 흉골 사이에 목에 depilatory 에이전트 교양 금액 (예 : 나이르)을 적용합니다. 부드럽게 마사지는 모든 머리가 제거됩니다 때까지. </li><li> depilatory 에이전트를 청소하고 depilated 지역 Betadine의 자유 금액을 적용합니다. </li><li> ligations에 대한 6-0 실크 봉합사의 1 인치 길이의 섹션을 준비합니다. 각 마우스 치료의 두 ​​가지가 필요합니다. </li><li> 날카로운 작은 가위를 사용하여 목 가운데 4-6밀리미터 수직 절개를합니다. 피부와 기본 근막 모두 절단해야합니다. </li><li> 정중선에서 왼쪽으로 왼쪽 귀 부근의 선 (腺)을 이동하고 퉁명스럽게 바로 기관의 해부 시작합니다. 당신은 쉽게 pulsating 왼쪽 일반 경동맥을 식별할 수 있어야합니다. </li><li> 당신이 분기점을 찾을 때까지 caudally 일반적인 경동맥을 따르십시오. </li><li> 외부 경동맥의 동맥, 내부 경동맥, 후두 동맥과 우수한 갑상선 동맥 : 퉁명스럽게 왼쪽 일반적인 경동맥에서 다음 네 가지 말초 지점을 노출하는 분기점을 해부하다. 이러한 혈관, 특히 우수한 갑상선은 깨지기 수 있습니다. 절개 매우주의하십시오. </li><li> 6-0 실크 봉합사를 사용하여 뛰어난 갑상선 위에 외부 경동맥을 묶어. 이렇게하려면 동맥 아래 한 포셉을 통과하고 다른 집게의 도움으로 precut 치료의 한 조각을 들고. 부드럽게 동맥 아래 통과 치료를 가져옵니다. 단단히 매듭에서 잡은되는 모든 주변 조직을 피하고 타이. 당신이 효과적으로 크게 다른 모델 완전한 결합을 수행합니다 프로 시저의 끝 부분에 의해, 뛰어난 갑상선 동맥의 이륙 아래 당신이 찾아낸다면. </li><li> 같은 기술을 사용하는 한 매듭으로 내부 경동맥과 후두 동맥을 묶어. </li><li> 봉합과 뛰어난 갑상선 동맥의 명백의 위치를​​ 확인합니다. </li><li> 조직 수선의 소량을 사용하여 대략적인 피부와 가까운 피부. </li><li> 복구까지 온난 챔버에 넣어 마우스. 복구는 일반적으로 30-60분 걸립니다. </li><li> 동물 첫 번째 24 시간 이내에 곤경에 처한 경우 사용되는 마취가 연장 통증 (예 흡입 isoflurane)를 제공하거나하지 않는 경우 Buprenorphine (0.1mg/kg)의 하나 복용량은 게시 즉시 수술을 시행해야합니다. 우리의 경험에서, 마취에 대한 관리 케타민을 복용은 수술 경험이 풍부한 외과 의사에 의해 수행되고 더 이상의 진통제가 필요하지 않습니다 충분한 진통제를 제공합니다. </li></ol> 2. 경동맥의 초음파 검사 <ol><li> 부분 내고이 흐름을 방해 유도 여부를 확인하기 위해 경동맥 흐름은 하루 후 내고 검사를해야합니다 </li><li> isoflurane과 마취를 유도 - 필요한 경우 마취 유도 상자가 사용할 수 </li><li> 최대 이미징 단계, 배꼽쪽에 anesthetized 쥐를 플레이스 </li><li> 이미징 무대에서 ECG 센서에 테이프 팔, 다리 </li><li> 나이르의 교양 프로그램 (이미 내고 절차 완료하지 않을 경우이 단계에만 필요)의 목을 잎이 진 </li><li> 온도를 기록하는 직장​​ 온도계를 삽입 </li><li> 목에 젤 에코 신청 </li><li> B - 모드에서 시작합니다. </li><li> 이미징 비행기는 90도 (프로브에 손잡이가 마우스의 코를 향해 가리 켜야합니다)해야합니다 </li><li> 이미지가 얻을 때까지 마우스 목에에 프로브를 낮춥니다. </li><li> 무대 왼쪽과 오른쪽기도까지 (중간 라인 구조) 식별 조작 </li><li> 왼쪽 일반적인 경동맥을 식별 </li><li> 틸트 영상 무대 너무 각도는 이미징 비행기와 경동맥 사이에 만들어집니다. 큰 이런 각도에서 강한 도플러 신호가 될 것입니다. </li><li> 왼쪽 일반적인 경동맥의 중간에 자리 펄스 웨이브 도플러. </li><li> 필요에 따라 각도 보정을 사용합니다. </li><li> 오른쪽 공통 경동맥에서 측정을 반복합니다. </li><li> 성공적인 부분 경동맥 내고는 심장 확장하는 동안 대동맥 입구쪽으로 흐름의 반전과 함께 오른쪽에 비해 왼쪽 일반적인 경동맥의 흐름 전반적인 감소 (~ 80-90%)가 발생합니다. </li><li> 만족스러운 이미지를 취득했습니다 일단 마취를 해제, 테이프 레스에서 마우스와 무료 생쥐에서 젤 에코 닦아traints. 예열 복구 챔버의 생쥐를 배치합니다. </li></ol> 3. 경동맥 내막에서 RNA 격리 <ol><li> 기관 IACUC 프로토콜에 따라 CO 2 흡입하여 마우스를 희생 </li><li> 종이 타월에 테이프 발. </li><li> 복부에서 흉부의 상단에 마우스의 피부를 잘라. </li><li> 가위의 날카로운 쌍의로 ribcage 아래의 복부 벽을 엽니다. </li><li> 다이어프램을 집게로 흉골을 들어 컷 후 심장을 노출하는 ribcage 버려야. </li><li> 가위로 베나 카바 컷. </li><li> 폐, 간 창백한 될 때까지 좌심실을 통해 10 단위 / ML 헤파린을 포함한 정상적인 생리와 2-3 분 압력 perfuse (120 mmHg). </li><li> 목 피부를 잘라 경동맥 동맥이 노출되기 전까지는 모든 지방, 근육 및 결합 조직을 제거하십시오. </li><li> 해부 현미경으로 마우스를 놓습니다. </li><li> 찔린 오른쪽 두번째 재관류를 위해 왼쪽 경동맥에 내고 사이트 아래에 구멍. </li><li> 좌측 경동맥이 잘되어 있는지 확인한하고, 좌심실을 통해 10 단위 / ML 헤파린을 포함한 정상적인 생리와 함께 ~ 1 분 다시 압력 perfuse. </li><li> 좋은 팁 포셉 작은 봄 가위를 사용하여, 신중하게 carotids를 둘러싼 페리 - adventitial 조직을 제거합니다. 이 세척 단계에서 carotids를 당기거나 스트레칭하지 않도록주의하십시오. </li><li> 대동맥 아치와 경동맥 분기점 위에 내고 지점 사이의 좌측 경동맥을 잘라. </li><li> 오른쪽 쇄골 하 동맥 분기 지점과 경동맥 분기점 사이의 오른쪽 경동맥을 잘라. </li><li> 얼음처럼 차가운 HBSS를 포함, 35mm 문화 요리에 carotids를 전송합니다. 필요한 경우, 신중하게 남아있는 페리 - adventitial 조직을 제거합니다. </li><li> 각 경동맥 당 150 μl QIAsol의 용해 버퍼 (Qiagen)를 작성하여 29g 바늘로 인슐린 주사기 (10분의 3 ML 주사기)를 준비합니다. </li><li> 조심스럽게 경동맥의 한쪽에 바늘의 끝부분을 삽입합니다. </li><li> 포셉과 경동맥과 바늘의 끝을 잡고 있지만, 신속하게 (~ 1 초) (intima의 eluate) 1.5ml 튜브에 QIAsol 용해 완충액 (150 μl)를 플러시. </li><li> 경동맥 남은 (미디어 + adventitia) 한 HBSS에서 1.5 ML 튜브에 넣어 액체 질소 스냅인 동결을 씻어. </li><li> Intima의 eluate 및 냉동 남은 그 다음 제조 업체의 지침에 따라 miRNeasy 미니 키트 (QIAGEN)를 사용 내막 또는 미디어 + adventitia RNA 분리에 사용됩니다. </li></ol> 4. 대표 결과 : 우리는 부분적으로 마우스 경동맥 동맥과 초음파 검사는 다음과 같은 절차 언젠가 실시되었다 왼쪽 출혈도 잡았. 그림 1에 나타난 성공적인 부분 내고 혈액 흐름 속도를 감소시키고 심장 확장하는 동안 왼쪽 일반적인 경동맥의 흐름을 (방해 흐름) 반대. 높은 지방 음식과 함께 ApoE 녹아웃 (KO) 생쥐의 부분 내고는 왼쪽 일반 경동맥 (LCA)에 강력한 죽상 경화증을 유도하지만 contralateral 오른쪽 경동맥에서 2 주 (그림 2). 경동맥 intima에서 수집한 RNA는 PECAM - 1과 같은 내피 마커 유전자를 포함, 예 : A - SMA (그림 3)와 같은 중간 평활근 세포 마커 유전자 무료 동안. <img src="/files/ftp_upload/1861/1861fig1.jpg" alt="그림 1" /> 그림 1 : 경동맥 해부학 및 ligations의 도식 : 우수한 갑상선 동맥 (STA)가 열려 떠나면서 왼쪽 일반적인 경동맥의 세 지점 (LCA) [외부 경동맥의 동맥 (ECA), 내부 경동맥 (ICA), 그리고 후두 동맥 (OA)가], 출혈도 잡았했다. <img src="/files/ftp_upload/1861/1861fig2.jpg" alt="그림 2" /> 그림 2 : 초음파 이미지 유속 프로필을 표시하고 부분 내고는 심장 확장하는 동안 LCA의 흐름 반전을 (화살표로 표시) 유도 것을 공개. RCA의 흐름을 내고 이후에 변경되지 남아 있습니다. <img src="/files/ftp_upload/1861/1861fig3.jpg" alt="그림 3" /> 그림 3 : 부분 내고 높은 지방 다이어트는 빠르게 ApoE KO 마우스의 LCA에 죽상 경화증을 유도 ApoE KO 마우스 부분적으로 출혈도 잡았 1로 3 주간 높은 지방 음식을 공급했다.. LCA의 냉동 섹션은 오일 - 레드 - O 물들일되었습니다. <img src="/files/ftp_upload/1861/1861fig4.jpg" alt="그림 4" /> 그림 4. 내막 RNA의 준비 방법. 내막 RNA와 중간 + adventitial (M +) RNA가 가짜 운항 RCA 및 LCA C57BL에 / 6 생쥐에서 얻은되었습니다. RNAS는 PECAM - 1과 α - SMA는 내부 통제로 18 기의를 사용하는 세미 양적 RT - PCR (A) 및 qPCR (B, C)에 의해 분석되었다. 막대 그래프는 ± SEM, n은 = 3 건 있습니다.

<ol>
	<li>Korshunov, V. A., Berk, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow-induced+vascular+remodeling+in+the+mouse:+a+model+for+carotid+intima-media+thickening.">Flow-induced vascular remodeling in the mouse: a model for carotid intima-media thickening.</a> Arterioscler Thromb Vasc Biol. 23 (12), 2185-2191 (2003).</li><li>Sullivan, C. J., Hoying, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow-dependent+remodeling+in+the+carotid+artery+of+fibroblast+growth+factor-2+knockout+mice.">Flow-dependent remodeling in the carotid artery of fibroblast growth factor-2 knockout mice.</a> Arterioscler Thromb Vasc Biol. 22 (7), 1100-1105 (2002).</li><li>Nam, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+carotid+ligation+is+a+model+of+acutely+induced+disturbed+flow,+leading+to+rapid+endothelial+dysfunction+and+atherosclerosis.">Partial carotid ligation is a model of acutely induced disturbed flow, leading to rapid endothelial dysfunction and atherosclerosis.</a> Am J Physiol Heart Circ Physiol. 297 (4), H1535-H1543 (2009).</li></ol>

이 절차에서는 외부 경동맥, 내부 경동맥, 후두 동맥과 우수한 갑상선 동맥을 포함한 일반적인 경동맥의 모든 지점의 적절한 신분증이 필요합니다. 시연으로 초음파를 사용하여 유동 패턴을 확인하는 것도 중요합니다. 우리의 경험 손에, 500 개 이상의 생쥐의 초음파 연구에 따라 부분 내고 수술의 성공 비율은 90 %보다 낫습니다. 경동맥 내막 RNA 준비 들어, 첫 번째 단계 재관류 후 경동맥의 모양에...

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		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Normal Saline (0.9% sodium chloride)</td>
<td>&nbsp;</td>
<td>Baxter</td>
<td>2B1323</td>
<td>&nbsp;</td>
<tr>
<td>Insulin syringe (3/10 ml 29g syringe)</td>
<td>&nbsp;</td>
<td>Tyco Health care</td>
<td>8881600145</td>
<td>&nbsp;</td>
<tr>
<td>HBSS</td>
<td>&nbsp;</td>
<td>Cellgro, Mediatech, Inc.</td>
<td>21-023</td>
<td>&nbsp;</td>
<tr>
<td>miRNeasy mini kit (including, QIAzol)</td>
<td>&nbsp;</td>
<td>QIAGEN</td>
<td>217004</td>
<td>&nbsp;</td>
<tr>
<td>Vevo 770 High-Resolution Micro-Imaging System</td>
<td>&nbsp;</td>
<td>Visualsonics</td>
<td>a.Visualsonics 770 system b.Imaging stage c.Vevo integrated rail system d.RMV scan head for mice</td>
<td>or similar system</td>
</tr>
<tr>
<td>6-0 Silk Suture, sterile</td>
<td>&nbsp;</td>
<td>Covidien</td>
<td>s-1172 c2</td>
<td>&nbsp;</td>
<tr>
<td>Small scissors</td>
<td>&nbsp;</td>
<td>Roboz</td>
<td>RS-5675</td>
<td>or similar</td>
</tr>
<tr>
<td>Dissecting Forceps</td>
<td>&nbsp;</td>
<td>Roboz</td>
<td>RS-5005</td>
<td>or similar</td>
</tr>
<tr>
<td>Tissue Mend II</td>
<td>&nbsp;</td>
<td>Webster Vet Supply</td>
<td>07-856-7946</td>
<td>&nbsp;</td>		</tbody>
		</table>
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a model of disturbed flow-induced atherosclerosis in mouse carotid artery by partial ligation and a simple method of rna isolation from carotid endothelium

잘 알려진 가까운 협회에도 불구하고, atherogenesis에 방해 흐름을 연결하는 직접적인 증거가 부족했습니다. 우리는 최근 경동맥 부분 결합 방식의 수정된 버전을 사용해야 [1,2]는 심하게 마우스 일반적으로 경동맥의 낮은 oscillatory 흐름 조건 방해 흐름의 두 주요 특성을 유도 것을 보여주기 위해. 이 모델을 사용, 우리는 방해 흐름이 실제로 Apolipoprotein E의 녹아웃 마우스에서 신속하고 강력한 죽상 경화증의 개발 [3] 단서가 확실한 증거를 제공하고 있습니다. 우리는 또한 마우스 경동맥 intima [3]에서 고순도로 내피 RNA 준비하는 방법을 개발했습니다. 이 마우스 모델과 방법을 사용하여, 우리는 그 부분 내고는 사주에 의해 그러한 intraplaque neovascularization 같은 복잡한 병변 형성의 기능과 함께 hyperlipidemic 마우스 모델에서 2 주내에 강력하고 빠른 atheroma의 형성에 의해 다음, 일주일에 내피 기능 장애의 원인이 발견했습니다. 이 급속한<em&gt; 생체내에</em&gt; 모델과 내피 RNA 준비 방법은 혈관 생물학과 질병의 흐름에 의존 규정을 기본 분자 메커니즘을 결정하는 데 사용할 수 있습니다. 또한, 그것은 상당히 감소 학습 기간의 내피 기능 장애와 죽상 경화증을 대상으로 다양한 치료 개입을 테스트하는 데 사용할 수 있습니다.

Watch this Scientific Journal Video about A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium at

A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium

이것은 일반적인 마우스 경동맥의 intraplaque 네오 - vascularization (4 주)와 방해 흐름 조건 및 후속 죽상 경화증 개발 (2 주) 원인 부분 경동맥 내고 수술을 설명합니다. 우리는 또한 고순도 내피 RNA를 제공하고, 경동맥 intima에서 RNA의 분리의 소설 방법을 설명합니다.

부분 결합하여 마우스 경동맥 동맥에있는 방해 흐름 유발 죽상 경화증과 경동맥 내피로부터 RNA 분리의 간단한 방법의 모델

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified ...

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28.1K 조회수.  Emory University. 이것은 일반적인 마우스 경동맥의 intraplaque 네오 - vascularization (4 주)와 방해 흐름 조건 및 후속 죽상 경화증 개발 (2 주) 원인 부분 경동맥 내고 수술을 설명합니다. 우리는 또한 고순도 내피 RNA를 제공하고, 경동맥 intima에서 RNA의 분리의 소설 방법을 설명합니다.

비디오: A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce atherosclerosis (reviewed in 1 and 2). The role of shear stress has been extensively studied in vitro investigating the influence of flow dynamics on cultured endothelial cells 1,3,4 and in vivo in larger animals and humans 1,5,6,7,8. However, highly reproducible small animal models allowing systematic investigation of the influence of shear stress on plaque development are rare. Recently, Nam et al. 9 introduced a mouse model in which the ligation of branches of the carotid artery creates a region of low and oscillatory flow. Although this model causes endothelial dysfunction and rapid formation of atherosclerotic lesions in hyperlipidemic mice, it cannot be excluded that the observed inflammatory response is, at least in part, a consequence of endothelial and/or vessel damage due to ligation.
In order to avoid such limitations, a shear stress modifying cuff has been developed based upon calculated fluid dynamics, whose cone shaped inner lumen was selected to create defined regions of low, high and oscillatory shear stress within the common carotid artery 10. By applying this model in Apolipoprotein E (ApoE) knockout mice fed a high cholesterol western type diet, vascular lesions develop upstream and downstream from the cuff. Their phenotype is correlated with the regional flow dynamics 11 as confirmed by in vivo Magnetic Resonance Imaging (MRI) 12: Low and laminar shear stress upstream of the cuff causes the formation of extensive plaques of a more vulnerable phenotype, whereas oscillatory shear stress downstream of the cuff induces stable atherosclerotic lesions 11. In those regions of high shear stress and high laminar flow within the cuff, typically no atherosclerotic plaques are observed.
 
In conclusion, the shear stress-modifying cuff procedure is a reliable surgical approach to produce phenotypically different atherosclerotic lesions in ApoE-deficient mice.

The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.

마우스의 전단 응력 유발 죽상 경화증을 실행을위한 경동맥 혈압계의 주입

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

연구

의학

Training a Sophisticated Microsurgical Technique: Interposition of External Jugular Vein Graft in the Common Carotid Artery in Rats

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in peripheral arterial bypass surgery as well as in arteriovenous fistulas.1-5 The experimental procedure of vein graft interposition in the common carotid artery by using the cuff-technique has been applied in several research projects to examine the aetiology of neointimal hyperplasia and therapeutic options to address it. 6-8 The cuff prevents vessel anastomotic remodeling and induces turbulence within the graft and thereby the development of neointimal hyperplasia.
Using the superior caval vein graft is an established small-animal model for venous arterialization experiment.9-11 This current protocol refers to an established jugular vein graft interposition technique first described by Zou et al., 9 as well as others.12-14 Nevertheless, these cited small animal protocols are complicated.
To simplify the procedure and to minimize the number of experimental animals needed, a detailed operation protocol by video training is presented. This video should help the novice surgeon to learn both the cuff-technique and the vein graft interposition. Hereby, the right external jugular vein was grafted in cuff-technique in the common carotid artery of 21 female Sprague Dawley rats categorized in three equal groups that were sacrificed on day 21, 42 and 84, respectively. Notably, no donor animals were needed, because auto-transplantations were performed. The survival rate was 100 % at the time point of sacrifice. In addition, the graft patency rate was 60 % for the first 10 operated animals and 82 % for the remaining 11 animals. The blood flow at the time of sacrifice was 8&plusmn;3 ml/min. In conclusion, this surgical protocol considerably simplifies, optimizes and standardizes this complicated procedure. It gives novice surgeons easy, step-by-step instruction, explaining possible pitfalls, thereby helping them to gain expertise fast and avoid useless sacrifice of experimental animals.

Neointimal hyperplasia is the primary cause of stenosis in arterialized veins. We propose a new protocol whereby the right external jugular vein is grafted using the cuff technique in the common carotid artery of Sprague Dawley rats. The survival rate was 100 % at the time point of sacrifice.

쥐 온목 동맥에 개재 외부 경정맥 정맥 이식의 : 정교한 Microsurgical 기술을 교육

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

재 협착의 연구에 대한 경동맥 스텐트 시술의 쥐과 모형

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

마우스 폐 삽관 법의 간단한 방법

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

성인 마우스 정맥 ​​고혈압 모델 : 외부 경정맥 정맥 문합에 경동맥.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. Intra-tracheal inoculation of a known amount of bacteria has been successfully used to reproduce the pathogenesis of pneumonia which then develops into sepsis. Monitoring hemodynamic parameters and providing standard clinical treatment including infusion of antibiotics, fluids and drugs to maintain blood pressure is critical to simulate routine supportive care in this model but to do so requires both arterial and venous vascular access. The video details the surgical technique for implanting carotid artery and common jugular vein catheters in an anesthetized rat. Following a 72 hr recovery period, the animals will be re-anesthetized and connected to a tether and swivel setup attached to the rodent housing which connects the implanted catheters to the hemodynamic monitoring system. This setup allows free movement of the rat during the study while continuously monitoring pressures, infusing fluids and drugs (antibiotics, vasopressors) and performing blood sampling.

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

경동맥과 경정맥의 도관은 의식 쥐 모델에서, 혼합 및 혈액 샘플링을 혈역학 적 조치를 수행하는

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

철 염화물에 의한 경동맥 및 장간막 용기에 혈전증 마우스 모델

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves focal endothelial-selective transgenesis, thereby allowing an investigator to determine the biological roles of endothelial-expressed transgenes and to quantify the in vivo transcriptional activity of DNA sequences in large artery endothelial cells. The method uses surgical isolation of rabbit common carotid arteries and an arteriotomy to deliver a transgene-expressing viral vector into the arterial lumen. A short incubation period of the vector in the lumen, with subsequent aspiration of the lumen contents, is sufficient to achieve efficient and durable expression of the transgene in the endothelium, with no detectable transduction or expression outside of the isolated arterial segment. The method allows assessment of the biological activities of transgene products both in normal arteries and in models of human vascular disease, while avoiding systemic effects that could be caused either by targeting gene delivery to other sites (e.g. the liver) or by the alternative approach of delivering genetic constructs to the endothelium by germ line transgenesis. Application of the method is limited by the need for a skilled surgeon and anesthetist, a well-equipped operating room, the costs of purchasing and housing rabbits, and the need for expertise in gene-transfer vector construction and use. Results obtained with this method include: transgene-related alterations in arterial structure, cellularity, extracellular matrix, or vasomotor function; increases or reductions in arterial inflammation; alterations in vascular cell apoptosis; and progression, retardation, or regression of diseases such as intimal hyperplasia or atherosclerosis. The method also allows measurement of the ability of native and synthetic DNA regulatory sequences to alter transgene expression in endothelial cells, providing results that include: levels of transgene mRNA, levels of transgene protein, and levels of transgene enzymatic activity.

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

This method is to introduce a transgene into the endothelium of rabbit carotid arteries. Introduction of the transgene allows the assessment of the biological role of the transgene product either in normal arteries or disease models. The method is also useful for measuring activity of DNA regulatory sequences.

Vivo에서 토끼 일반적인 경 동맥 내 피에 유전자 이동

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves ...

A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, intimal hyperplasia, and atherosclerosis. The goal of this article is to demonstrate a method for placing bilateral venous interposition grafts in a rabbit, then transducing the grafts with a gene transfer vector that achieves durable transgene expression. The method allows the investigation of the biological roles of genes and their protein products in normal vein graft homeostasis. It also allows the testing of transgenes for the activities that could prevent vein graft failure, e.g., whether the expression of a transgene prevents the neointimal growth, reduces the vascular inflammation, or reduces atherosclerosis in rabbits fed with a high-fat diet. During an initial survival surgery, the segments of right and left external jugular vein are excised and placed bilaterally as reversed end-to-side common carotid artery interposition grafts. During a second survival surgery, performed 28 days later, each of the grafts is isolated from the circulation with vascular clips and the lumens are filled (via an arteriotomy) with a solution containing a helper-dependent adenoviral (HDAd) vector. After a 20-min incubation, the vector solution is aspirated, the arteriotomy is repaired, and flow is restored. The veins are harvested at time points dictated by individual experimental protocols. The 28-day delay between the graft placement and the transduction is necessary to ensure the adaptation of the vein graft to the arterial circulation. This adaptation avoids rapid loss of transgene expression that occurs in vein grafts transduced before or immediately after grafting. The method is unique in its ability to achieve durable, stable transgene expression in grafted veins. Compared to other large animal vein graft models, rabbits have advantages of low cost and easy handling. Compared to rodent vein graft models, rabbits have larger and easier-to-manipulate blood vessels that provide abundant tissue for analysis.

This method describes the placement of interposition vein grafts in rabbits, the transduction of the grafts, and the achievement of durable transgene expression. This allows the investigation of physiological and pathological roles of transgenes and their protein products in grafted veins, and testing of gene therapies for vein graft disease.

일반적인 경 동맥 중재 이식 정 맥 경에서에서 튼튼한 Transgene 식의 토끼 모델

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, ...