이 프로젝트는 부분적으로 윌리엄 L.Young, 윌리엄 L.Young과 후아 수, 후아 SU에 R21 NS070153에 P01의 NS44155에 에스 펜 워커 NIH T32 GM008440, R01 NS27713에 의해 후아 수에 미국 심장 협회 (American Heart Association) AHA 10GRNT3130004에 의해 지원됩니다. 박사 아나 로드리게스 - 에르난데스는 &quot;Obra는 사회 La Caixa는&quot;의 연구비 지원

1. 마우스를 준비 <ol><li> 이소 플루 란 가스와 마우스에서 전신 마취를 유도한다. 통증 관리 복강 brupenorphine 0.15 ml를 주입한다. 마취 레벨이 마우스의 발 샤프트로 만족스러운 경우 계속하기 전에 확인합니다. </li><li> 접착 테이프로 고정 사지와 지느러미 드러 누움에 마우스를 넣습니다. 목의 머리와 가위로 가슴 상부를 제거합니다. 피하 주사를 통해 수술 절차 동안 수화 마우스를 유지 0.9 % 식염수를 0.2 mL의 관리. </li><li> 엄격한 멸균 방법에 따라 수술 분야를 준비합니다. 피부 절개의 면적은 90 % 알코올로 청소해야합니다. </li></ol> 2. 경동맥 및 외부 경정맥을 해부 <ol><li> 마우스의 낮은 목 영역에서 수평 정중선 자궁 절개를합니다. 상처를 심화 한 후,에 사용하여 침샘과 자궁 경부 연부 조직을 상승raction 봉합 (그림 1). 흉쇄 근육 (SCM)에 횡 방향 오른쪽 외부 경정맥 (EJV)을 노출. 정맥 위에 과도한 견인력이 손상 및 혈전증을 유발 수 있으므로이 단계는, 현미경 하에서 수행되어야한다. </li><li> 조심스럽게 두개골베이스에 쇄골에서 그것의 과정을 따라 오른쪽 EJV을 해부하다. 일반적으로 전기 바이폴라 집게, 응고 이상 임시 클립 배치 및 문합을 위해 적절한 길이를 준비하는 모든 분기를 나눕니다. </li><li> SCM에 기관과 내측에 측면, 경동맥 (CCA)을 탐구한다. 그것은 신중하게 단지 외부 및 내부 경동맥 동맥으로의 분기점을 넘어 쇄골 노출되어야한다. 이 단계에서는,주의는 나중에 문합의 개통을 손상시킬 수 EJV에 과도한 마찰을 피하기 위해 다시 부여해야합니다. </li></ol> 3. 문합 준비 <ol><li> 10-0 뉴욕으로 CCA를 결찰그 분기에 경도 단지 인접. 다음으로, 가능한 한 쇄골에 가까운 근위부 CCA를 통해 임시 클립을 적용합니다. </li><li> 흐름이 중단되면, 그냥 분기 합자 아래에있는 동맥을 가로로 쪼개다과 루멘 내부에 남아있는 피를 씻어 식염수로 세척하십시오. 동맥 벽에 열 부상 위험에 미래 문합을 넣을 수 있기 때문에,이 단계에서 양극 응고을 피하십시오. </li><li> EJV의 에지의 시인성을 향상시키기 위하여, 중간 벽은 계획된 venotomy의 과정을 따라 청색 마킹 펜으로 표시된다. EJV가 표시되면, 가능한 한 꼬리 등 말단부 결찰 가능한 뇌와 근위 단부에 임시 혈관 클립을 적용 10-0 봉합사를 사용한다. </li><li> 벌금 30 G 바늘과 0.5 ml를 주사기로, EJV의 표시된 영역에 초기 구경을하고 바로 혈전 형성을 방지하기 위해 식염수로 루멘 관개. 다음으로, microscissors와 venotomy를 확장CCA 길이의 약 2-3 배의 직경까지이다. 스트레칭 폭력 피와 선명하고 깔끔한 최첨단을 유지하기 위해주의를 기울입니다. </li><li> EJV에 CCA의 끝을 대략적인. CCA의 기증자 결국 사이드 컷 절개 venotomy 크기로 길이 (그림 2)에 직경을 조정합니다. </li></ol> 4. 엔드 - 투 - 측 문합 <ol><li> CCA EJV 투 엔드 - 투 - 측 문합위한 11-0 나일론 모노 필라멘트 봉합사를 사용한다. craneo - 꼬리 방향으로 문합의 중간 벽을 봉합하는 초기 단계입니다. 모든 스티치는 정맥 벽에 외부-에서 첫 번째 (그림 3) 배치 및 다음 내부 아웃 동맥 벽 (그림 4)에서해야한다. 이것은 항상 (그림 5)에서 문합 혈관의 외부 표면에 매듭을 유지합니다. 중단되거나 연속 봉합사를 사용할 수 있지만, 모든 연속 봉합 마지막 단계로 조여 주어야 하나. </li><li> O중간 벽이 봉합 된 NCE, 측면 벽과 두개골에 꼬리부터 절차를 반복합니다. 이제 모든 스티치 외부-의 동맥 벽 처음부터 옆 정맥 벽 안쪽에서 바깥쪽으로 배치해야합니다. 식염수 관개 절차를 수행하는 동안 항상 볼 수 문합의 루멘을 유지하는 데 도움이됩니다. </li><li> 문합의 모든 단계를 완료 한 후, 제 옆에있는 동맥에서 정맥에서 임시 클립을 제거합니다. 동맥혈은 문합에서 전혀 또는 거의 돌아 다니며 함께 EJV 유입 (동영상 VH 모델과 VH 모델 2 참조)합니다. 최소한의 출혈은 문합을 통해면 압축을 사용하지 않고 중지해야합니다. 이는 민감한 정맥 혈전증을 방지하기 위해서 피해야한다. </li><li> 타악기는 문 합부를 통과하면 확인과 명백한 출혈이 관찰되지, 식염수 수술 필드를 관개하고 6-0 나일론 봉합사로 자궁 절개를 닫습니다. 마지막으로, intrap 0.15 ml를 더 관리수술 후 통증 관리 및 피하 0.9 % 식염수의 0.2 ~ 0.4 ml를위한 eritoneal brupenorphine는 수술 중에 혈액 손실을 보충합니다. </li></ol>

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실험실 동물 실험 절차는 캘리포니아 대학의 기관 동물 관리 및 사용위원회, 샌프란시스코 (UCSF)에 의해 승인되었다. 저자는이 절차에 사용되는 약물과 재료에 관한 관심의 잠재적 인 충돌이 없습니다.

지속적인 뇌 정맥 고혈압이 밀접하게 더 심한 임상 증상 및 경막에서 avfs 뇌 동정맥 기형 (3) 환자의 불량한 예후와 관련이있다. CVH의 이러한 효과는 널리 쥐 모델 1,2,8에서 연구되고있다. 마우스에 상당 모델 궁극적 정맥 고혈압의 병인 및 경막 AVF 뇌 AVM와의 관계에 관련된 분자 경로의 분석을 허용하는 유전자 변형 동물의 사용을 허용한다. 여기서, 우리는 엔...

대뇌 정맥 고혈압 동물 모델은 뇌 동정맥 기형 동정맥 루 1-7의 병태 생리에 대한 이해의 핵심 도구로 증명했다. 가장 널리 사용은 총 경동맥 (CCA) 및 래트에서 일관된 1,8-10 대뇌 정맥 고혈압 (CVH)을 유발 외부 경정맥 (EJV) 사이 인공 누공 통해 생성 쥐 모델이다. 등가 생쥐 모델은 서로 다른 트랜스 제닉 마우스 균주를 사용할 수있는 가능성을 열어,뿐만 아니라 이러한 병태 생리 아니라 뇌 혈관 질환에 대한 유전자 치료에 잠재적 인 추가 연구를 허용한다. 또한, 실험은 상기 구조물이 마우스 모델을 구축 H 편두통과 같은 정맥 고혈압, 일시적인 글로벌 건망증, 과도 단안 실명 등 (11) 그러나, 이전의 시도와 관련된 기타 뇌 혈관 질환의 연구에 적용 할 수있다아베 인해 소형의 해부학 5,12-에 누공의 개통의 어려움을 보여 주었다. 여기서 우리는 장기적인 특허 루와 마우스의 내구성 정맥 고혈압으로 변환 쥐 CCA와 EJV의 성공적인 문합을위한 단계별 프로토콜을 설명합니다.

<table border="1" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>10-0 Sterile Microsuture</td>
			<td>Arosurgical Ic.</td>
			<td>VT5A010Q10</td>
		</tr>
		<tr>
			<td>11-0 Sterile Microsuture</td>
			<td>Arosurgical Ic</td>
			<td>VT4A00N07</td>
		</tr>
		<tr>
			<td>DUROTIP Scissors</td>
			<td>Aesculap</td>
			<td>BC210R</td>
		</tr>
		<tr>
			<td>Micro-Adson Tissue Forceps</td>
			<td>Aesculap</td>
			<td>BD510R</td>
		</tr>
		<tr>
			<td>Microscissors</td>
			<td>Aesculap</td>
			<td>OC496R</td>
		</tr>
		<tr>
			<td>Micro Forceps #5 Jewelers</td>
			<td>Aesculap</td>
			<td>BD331R</td>
		</tr>
		<tr>
			<td>Angled Jewelers Forceps</td>
			<td>Aesculap</td>
			<td>BD329R</td>
		</tr>
		<tr>
			<td>Micro Suture Forceps</td>
			<td>Aesculap</td>
			<td>BD338R</td>
		</tr>
		<tr>
			<td>DUROGRIP Needle Holder</td>
			<td>Aesculap</td>
			<td>BM009R</td>
		</tr>
	</tbody>
</table>

adult mouse venous hypertension model: common carotid artery to external jugular vein anastomosis.

뇌 동정맥 기형 동정맥 루의 병태 생리의 이해는 동물 모델 덕분에 개선했다. 경동맥 (CCA)와 외부 경정맥 (EJV) 사이에 인공 루를 만드는 쥐 모델은 널리 설명하고 기술적으로 가능 입증되었다. 이 구조는 일관성있는 대뇌 정맥 고혈압 (CVH)을 유발, 따라서 형성, 임상 증상, 뇌 동정맥 기형 및 경막에서 avfs의 예후에 정맥 고혈압의 기여를 공부하고있었습니다. 등가 마우스 모델은 거의 설명하지 않았습니다 및 누공의 협착에 문제를 보여 주었다. 설립 쥐 모델은 이러한 뇌 혈관 질환에 대한뿐만 아니라 병태 생리의 연구뿐만 아니라 잠재적 인 유전자 치료를 허용합니다. 우리는 마우스의 내구성 두개 내 정맥 고혈압을 생산 동정맥루의 모델을 제시한다. 미세 수술 문합 오뮤린 및 f를 CCA EJV 인해 소형의 해부학하기 어려울 수 있고, 종종 비 특허 누관을 초래한다. 이 단계별 프로토콜에서 우리는이 과정 중에 발생하는 모든 중요한 문제를 해결. 대신 봉합 11-0 10-0을 이용 노광시의 과도한 혈관 수축을 방지하고 신중하게 계획 측 단문 합을 만드는 중요한 단계 중 일부이다. 이 방법은 미세 수술 기술 진보와 쥐 당량,이 지속적으로 개발 될 수있는 더 긴 학습 곡선이 필요하지만. 이 새로운 모델은 뇌 동정맥 기형 및 경막에서 avfs을 연구하는 데 유용 입증되었습니다 이전에 확립 된 실험 시스템과 유전자 변형 마우스 기술을 통합 할 수 있도록 설계되었습니다. 트랜스 제닉 마우스를 사용할 수있는 가능성을 열어, 유효한 모델의 광범위한 스펙트럼이 달성 될 수 있고, 유전자 치료는 또한 테스트 할 수있다. 실험 구조는 또한 상기 오티의 연구에 적용 할 수있다편두통과 같은 정맥 고혈압, 과도 글로벌 기억 상실, 과도 단안 실명 등 관련 그녀의 뇌 혈관 질환

모델의 성공적인 결과는 쥐의 뇌 정맥 고혈압을 유도하는 특허 동정맥루입니다. 모델을 검증하기 위해 우리는 처음 수술 후 2, 3, 4 주에 쥐의 sagital 동에있는 두개 내 정맥압을 측정 하였다. 6 다른 생쥐마다 투여군. 부비동 압력 군 8.8 ± 1.2 mmHg로는 수술 후 2 주 정도를 측정 하였다. 삼주 수술 후 측정 된 6 마우스에서, 부비동 압력은 4.7 ± 1.4 mmHg로했다. 수술은 3.9 ± 0.6 mmHg로 (도 6)의 동...

Watch this Scientific Journal Video about Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis. at JoVE.com

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

우리는 성인 마우스에서 대뇌 정맥 고혈압 안정적인 모델을 생성하는 방법을 설명한다. 이 모델은 광범위 래트에서 설명되고 시험되었다. 마우스에서이 새로운 상대 유전자 변형 동물을 사용할 수있는 가능성을 열어하여 모델의 애플리케이션을 넓게.

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 ...

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Research

JoVE Journal

Medicine

14.0K 조회수.  University of California, San Francisco.  우리는 성인 마우스에서 대뇌 정맥 고혈압 안정적인 모델을 생성하는 방법을 설명한다. 이 모델은 광범위 래트에서 설명되고 시험되었다. 마우스에서이 새로운 상대 유전자 변형 동물을 사용할 수있는 가능성을 열어하여 모델의 애플리케이션을 넓게. 

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

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 ...

연구

의학

Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We have established bilateral common carotid artery occlusion as a preconditioning stimulus to induce early ischemic tolerance to transient focal cerebral ischemia in C57Bl6/J mice. In this video, we will demonstrate the methodology used for this study.

There is accumulating evidence, that ischemic preconditioning (PC) &#x2013; a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We established bilateral common carotid artery occlusion (BCCAO) as a preconditioning stimulus to induce early ischemic tolerance (IT) to transient focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) in C57Bl6/J mice.

초점 대뇌 국소 빈혈에 조기 허혈성 공차를 유도하기 위해 적절한 전처리 자극으로 양측 경동맥 동맥 폐색

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a ...

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. ...

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 ...

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, obesity and diabetes. Accordingly, when evaluating therapies for stroke in animals, it is important to select a model with excellent face validity. Ischemic stroke accounts for 80% of all strokes, and the majority of these occur in the territory of the middle cerebral artery (MCA), often inducing infarcts that affect the sensorimotor cortex, causing persistent plegia or paresis on the contralateral side of the body. We demonstrate in this video a method for producing ischemic stroke in elderly rats, which causes sustained sensorimotor disability and substantial cortical infarcts. Specifically, we induce permanent distal middle cerebral artery occlusion (MCAO) in elderly female rats by using diathermy forceps to occlude a short segment of this artery. The carotid artery on the ipsilateral side to the lesion was then permanently occluded and the contralateral carotid artery was transiently occluded for 60 min. We measure the infarct size using structural T2-weighted magnetic resonance imaging (MRI) at 24 hr and 8 weeks after stroke. In this study, the mean infarct volume was 4.5% &#177; 2.0% (standard deviation) of the ipsilateral hemisphere at 24 hr (corrected for brain swelling using Gerriet&#8217;s equation, n = 5). This model is feasible and clinically relevant as it permits the induction of sustained sensorimotor deficits, which is important for the elucidation of pathophysiological mechanisms and novel treatments.

Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke.

세 쥐의 경동맥 폐색으로 영구 말초 중간 대뇌을 수행하면 두피 허혈 지속적인 장애를 연구하는

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

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, ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

쥐 일반적인 경 동맥에 단계 대조 자기 공명 영상

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Myography 와이어를 사용 하 여 마우스 Mesenteric 동맥의 isometric 수축 측정

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Middle Cerebral Artery Occlusion Allowing Reperfusion via Common Carotid Artery Repair in Mice

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue plasminogen activator (tPA) as an approved drug treatment to target ischemic strokes. Current research in the field of ischemic stroke focuses on better understanding the pathophysiology of stroke, to develop and investigate novel pharmaceutical targets. Reliable experimental stroke models are crucial for the progression of potential treatments. The middle cerebral artery occlusion (MCAO) model is clinically relevant and the most frequently used surgical model of ischemic stroke in rodents. However, the outcomes of this model, such as lesion volume, are associated with high levels of variability, particularly in mice. The alternative MCAO model described here allows the reperfusion of the common carotid artery (CCA) and the increased perfusion of the middle cerebral artery (MCA) territory, using a tissue pad with fibrinogen-based sealant to repair the vessel, and the improved welfare of the mice by avoiding external carotid artery (ECA) ligation. This reduces the reliance on the Circle of Willis, which is known to be highly anatomically variable in mice. Representative data show that using this alternative surgical approach decreases the variability in lesion volumes between the traditional MCAO approach and the alternative approach described here.

Intraluminal filament occlusion of the middle cerebral artery is the most frequently used in vivo model of experimental stroke in rodents. An alternative surgical approach to allow common carotid artery repair is performed here, which allows the reperfusion of the common carotid artery and a full reperfusion to the middle cerebral artery territory.

쥐에 있는 일반적인 경 동맥 수리 통해 Reperfusion를 허용 하는 중간 대뇌 동맥 폐색

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue ...

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

성인 마우스 정맥 고혈압 모델 : 외부 경정맥 정맥 문합에 경동맥.