We are grateful to Dr. Clowes for first developing and describing this method. We are also thankful to Dr. Tulis for his detailed protocol which has been fundamentally helpful to our previous, current and future work. This work was supported by grants R01HL097111 and R01HL123364 from the NIH to M.T., and by American Heart Association grant 14GRNT18880008 to M.T.
We would like to thank Rachel Newton for her expert technical support and for her valuable help during the filming process.

참고 : 다음과 같은 실험 동물의 사용을 검토하고 기관 동물 관리 및 사용위원회 (IACUC)에 의해 승인되었다. 1. 수술 전 절차 <ol style="margin-left: 40px;"><li> 사용하기 전에 수술 도구를 소독. <ol><li> 모든 수술기구를 수술 전 24 시간 이하 압력솥. 여러 수술 당일이 수행되는 경우, 수술 사이에서 건조 비드 멸균기로 멸균 악기. </li></ol></li><li> 필터 사용 전에 멸균 식염수. </li><li> 쥐를 달아 마취제 (케타민 80 ㎎ / kg과 자일 라진 (xylazine) 7 ㎎ / ㎏)의 용량을 계산한다. </li><li> 내 복강 (IP) 마취제를 관리. <ol><li> 5 ~ 10 분에 발가락 핀치에 의한 진정 작용의 적합성을 확인합니다. 진정 작용이 완료되지 않은 경우 약물 (케타민 7 ㎎ / kg과 자일 라진 (xylazine) 0.6 ㎎ / ㎏)의 추가 작은 복용량을 관리합니다. </li><li> 확인 바늘이 배달 약물 내 당에 충분한 깊이itoneally 복강에 전체 약물 솔루션을 제공하는 실패는 부적절한 진정 발생할 수 있기 때문이다. 주 : 주사 부위 피부 궤양과 탈모 인한 부주의 피하 (SC) 약액 주입 수술 후 수일 가시적 일 것이다. </li></ol></li><li> 3 ml의 멸균 생리 식염수를 피하 (SC)를 주입한다. 멸균 면봉을 사용하여 건조되는 것을 방지하기 위해 각막을 양쪽 눈에 안연고 소량의 적용. </li><li> 난방 기기를 준비합니다. 전자 레인지 또는 물을 욕조에 의해 사전 온열 패드. </li><li> 수술 플랫폼 드러 동물을 넣습니다. <ol><li> 복부 목 지역에서 머리를 제거합니다. , 면봉으로 머리 제거제를 적용 30 초 정도 기다린 후 거즈로 완전히 닦아. </li><li> 포비돈 요오드 스크럽 70 % 에탄올 목 면봉. </li></ol></li><li> 가운, 헤어 커버, 수술 마스크, 안경을 포함한 개인 보호 장비를 착용 할 것. 아이돌 착용무균 수술 장비 및 소모품을 취급하기 전에 끝에 수술 장갑을 ILE. </li><li> 노출 만 목 영역과 멸균 수술 시트와 쥐 드레이프. </li></ol> 2. 외과 적 치료 <ol style="margin-left: 40px;"><li> 수술하는 동안 15 분마다 발가락 핀치에 의해 동물의 진정 깊이를 확인합니다. 동물 발가락 핀치에 응답하는 경우, 케타민과 자일 라진의 추가 작은 용량 (초기 용량의 10 %)를 추가합니다. </li><li> 왼쪽 경동맥을 해부 (CCA) <ol><li> 목의 중앙에 직선 세로 절개를 만들기 위해 메스를 사용합니다. 피부 절개의 길이는 대략 동맥 1.5-2 cm의 부분을 분리 할 목적으로, 1.5 cm이다. 다를 수 있습니다 길이는 연구의 목적에 따라 달라집니다. </li><li> 무디게 피부 결합 조직의 해부. 집게 팁을 유지하고 피부 또는 하부 조직에 구멍을하지 않도록하십시오. </li><li> 길이 방향 아론 근육 층을 해부하다g 기관의 왼쪽. </li><li> 근육 층을 열 때, 밀착 미주 신경과 왼쪽 CCA를 시각화. 극단주의와 왼쪽 경동맥이 최소한의 스트레칭으로 미주 신경을 분리와 함께 퉁명스럽게 해부. </li><li> 분기까지 원심 CCA를 해부하다. 내 경동맥 (ICA) 및 외부 경동맥 (ECA) - 조심스럽게 분기 두 가지를 해부하다. </li><li> 동맥 1.5-2 cm 정도의 부분이 주변 조직으로부터 분리 될 때까지, CCA 주위 해부 유지. </li></ol></li><li> 풍선 부상 <ol><li> 영구적으로 약 5mm 떨어진 분기점에서 ECA에 합자을합니다. 영구적으로 ECA와 ICA의 분기점에 가까운 ECA의 후두 분기를 결찰. 또한 영구적으로 분기 및 ECA 합자 사이에 위치, 다른 지점을 결찰있는 경우 - 예를 들어, 우수한 갑상선 분기를. 모든 합자에 사용되는 봉합사는 4-0 검은 색실크. CCA의 기단부 및 ICA의 선단부에 클립. 참고 : 자, 혈액의 흐름이 정지 된 영구적으로 결찰을 통해 (ECA)에 또는 일시적으로 (CCA와 ICA에) 클리핑을 통해. 분기 지역에서 내강 내용은 전신 순환에서 고립되었다. </li><li> 작은 마이크로 가위로 ECA에서 arteriotomy의 절개를합니다. 절개 말단 봉합 매듭 가까이 있는지 확인합니다. 식염수 면봉으로 닦아 혈액. </li><li> ECA 루멘으로 불지 2 층 풍선 카테터를 삽입합니다. CCA 루멘에 근위 풍선 카테터를 전진. 클립이 유지 곳의 끝이 도달 할 때까지 근위 카테터를 전진하십시오. </li><li> 3 방향 꼭지에 여성 루어 잠금으로 풍선 팽창 장치를 연결하고 풍선 카테터에 3 방향 스톱 콕의 남성 루어 잠금 장치를 연결합니다. </li><li> 천천히 직경의 1.5 배에 경동맥 넓히다하기 위해, 약 1.5 기압의 압력으로 풍선을 팽창. </lI&gt; <li> 부드럽게 회전 다시 분기에 풍선을 잡아 당깁니다. </li><li> 풍선을 수축하고 기단부에 다시​​ 진행합니다. 다시 팽창하고 당김 절차를 두 번 더 반복한다. </li><li> 동맥 내강에서 풍선 카테터를 철회. </li></ol></li><li> 시약의 관내 관리 (예를 들어, siRNA를, 마약) 주 : 여기에서, 사용 된 시약은 하나의 shRNA 기질 상호 작용은 분자 1 (STIM1) 또는 비 - 타겟팅 shRNA를 제어 대상으로 인코딩 렌티 바이러스 입자를 함유하는 용액이었다. STIM1이 세포막 칼슘 이온 채널의 활성화를 제어 ER 칼슘 센서이며 이동성 증식 표현형 12,30-33으로 혈관 평활근 탈분화 동안 조절 달려 단일 경막 소포체 (ER) 단백질이다. <ol><li> 주사기 (24 G, 1.6 cm)에 혈관 이상 - 더 - 니들 카테터를 연결합니다. 테스트 reag 기음 30 μL의 용액을엔트. ECA의 같은 절개에 카테터를 삽입합니다. </li><li> CCA에 카테터 팁을 발전 카테터를 수정하고 일시적으로 절개를 닫습니다 ECA에 하나의 매듭 봉합의 조각을 묶어. </li><li> CCA의 루멘 내로 테스트 시약 용액을 주입한다. 30 분 동안 혈관의 루멘 유지 용액. 식염수로 습기 노출 된 조직을 유지하고 젖은 거즈로 덮어. </li><li> 배양 후, 남은 용액을 대기음. 그 하나의 매듭을 풀고 카테터를 철회. </li><li> arteriotomy 절개에 근접하게 봉합의 조각으로 ECA 넥타이. 분기에 가능한 한 가깝게 매듭을 확인합니다. </li></ol></li><li> 상처를 닫습니다. <ol><li> 풍선 부상과 카테터 도입 혈관의 누출 또는 구멍을 일으킬 수 있기 때문에, ICA의 클립을 제거하고 누출이 있는지 확인합니다. 출혈이 관찰되는 경우, 거즈를 적용하고 출혈을 멈추게하기 위해 부드러운 압력을 적용합니다. </li><li> 제거CCA의 다른 클립. </li><li> 출혈의 흔적이 없는지 확인하고 모든 클램프 및 기타 수술 도구를 제거합니다. 초과 봉합을 잘라. </li><li> 닫기 상처 사용하여 피부 봉합 (4-0 블랙 실크). Povidon - 요오드 또는 다른 항 패혈증 / 살균제 / virucide 에이전트 폐쇄 상처의 모든 측면에 면봉. 멸균 식염수 (SC)의 3 ㎖를 주입 </li></ol></li></ol> 3. 수술 후 절차 <ol style="margin-left: 40px;"><li> 수술 후 열 패드에 쥐를 유지합니다. 쥐에 근육 주사 (IM)를 통해 0.05 ㎎ / ㎏ 부 프레 노르 핀 중 하나 복용량을 관리합니다. 복구 절차 동안 촉촉한 동물의 눈과 입을 유지합니다. 이 깨어 보행이 될 때까지 동물을 모니터링합니다. </li><li> 완전히 복구 될 때까지 모든 침구없이 깨끗한 케이지에서 동물을 놓습니다. 복구 후, 동물은 동물의 방으로 돌아 개별적으로 보관됩니다. </li></ol>

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The authors declare that they have no competing financial interests.

쥐의 경동맥 풍선 손상은 잘. 2007 34 Tulis에 의해 기술 된 포괄적으로 박사 Tulis이 절차의 모든 세부 사항을 논의하고있다. 이 절차를 수행하기에 관심이있는 독자는 매우 Tulis &#39;프로토콜을 읽을 것을 권장합니다. 그러나, 우리는 박사 Tulis으로 동의하지 않는 한 가지가 대신 식염수 또는 액체의 종류와 풍선을 팽창, 우리는 공기와 함께 팽창 제안했다. 우리의 개인적인 경험에 따르면, 액체로 팽창하는 것...

풍선 카테터는 혈관에서의 혈전이나 죽종 막혀 사이트 (들)를 확대 할 목적으로, 혈관 성형술 절차에서 사용하는 의료 장치이다. 좁아진 혈관 내강이 팽창 된 풍선에 의해 강제로 개방되고, 혈액 공급은 협심증, 심근 경색, 다리 통증 등의 다운 스트림 허혈 증상을 완화하기 위해 순차적으로 복원 할 것입니다. 그럼에도 불구하고, 혈관 성형술의 큰 성공은 혈관 내강 (재 협착) 1의 재 협착 혈관 압력 손상 (풍선 부상), 즉 혈관 벽 리모델링을 일으키는 힘의 결과로 많은 경우에 수술 후 합병증으로 감소하고있다. 동물 모델의 수는 연구자가 풍선 부상 관련 혈관 벽 리모델링 (2) 기본 메커니즘을 이해하는 데 도움 혈관 성형술 절차를 흉내 개발되었다. 모델링에 이용 모든 동물 종 중에서, 래트는 가장 자주 사용되는 하나이다. C토끼, 개, 돼지에 ompared, 쥐의 장점은 낮은 비용, 사용의 상대적 용이성과 쥐의 생리의 현재 지식입니다. 마우스는 유전자 조작 된 균주의 넓은 범위에서 추가적인 장점을 가지고 있지만, 쥐 용기를 풍선 카테터를 삽입하기에는 너무 작다. 지난 3 년 동안 실험 쥐 연구자들은 3-6 리모델링 신생 내막 형성과 혈관의 토대 분자 및 세포 메커니즘을보다 잘 이해할 수있게되었다. 풍선 부상을 넘어, 혈관 재는 또한 죽상 동맥 경화증 7, 8, 고혈압 9 및 동맥류 (10) 등 대부분의 주요 혈관 질환에 참여하고 있습니다. 따라서, 지식 풍선 손상 모델을 통해 얻은 일반적으로 전체 혈관 벽 질환 연구에 도움이됩니다. 쥐 벌룬 손상 모델의 전체적인 목적은 단지 혈관 질환을 더욱 이해하기 위해뿐만 아니라, 신규 한 제제의 효능을 테스트하는 것이다질병 관리 (11, 12). 재 협착에 현재 임상 약물 치료 바로 혈관 성형술 후 혈관 내강을 통해 배치 약물 방출 스텐트에 의해 적용됩니다. 동물 모델에서 새 에이전트 시험을위한 효율적인 아직 더 경제적 인 방법은 잘 발달 된 지역 관​​내 관류 방법이다. 이 방법을 통해 테스트 한 후보 에이전트는 작은 분자 약물 복용 13, 14, 사이토 카인을 포함하거나 성장, 15, 16 인자 유전자 조작 에이전트 (의 cDNA 클론의 siRNA 등) 17 ~ 20, 및 새로운 의약품 제제 (21, 22). 지금까지 쥐 벌룬 손상 모델 혈관 질환 / 질병을 연구하는데 가장 유용한 모델의 하나로 남아있다. 이 첫 번째 단계는 생체 내에서 시험 관내에서 이동하는 일반적으로, 베드 사이드에서 벤치에 기초 단계이지만 마지막이어야한다. 쥐 실험의 결과는 더욱 심의 인간 번역 전에 특성화 될 필요때문에 혈관 침대와 혈관 해부학뿐만 아니라 인간과 쥐의 23 ~ 26 사이의 고유 종의 차이의 차이에 임상 적으로 사용. 그럼에도 불구하고, 여전히 병진 의학 연구에 필수적인 도구이다. 이러한 연구는 유전자 변형 된 쥐의 부족에 의해 제한하기 위해 사용하는 한편 징크 핑거로서 신규 게놈 접근법 29 녹아웃 생쥐는 쉽게 액세스 가능하게 한 27 클레아 TALENs 28 CRISPR-캐스 때문에 더 이상 문제가 없었다.

<table border="0" cellpadding="0" cellspacing="0" width="1059">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:384px;">Name of Material/ Equipment</td>
			<td style="width:389px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
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			<td height="21" style="height:21px;">Fogarty balloon embolectomy catheters, 2 French&#160;</td>
			<td>Edwards Lifesciences, Germany</td>
			<td>&#160;120602F</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Deltaphase Operating Board - Includes 2 Pads &#38; 2 Insulators</td>
			<td>Braintree Scientific, Inc.</td>
			<td>39OP</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;LED light source</td>
			<td>Fisher Scientific</td>
			<td>12-563-501&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hartmann Mosquito Forceps 4&#8221; curved</td>
			<td>Apiary Medical, Inc. San Diego, CA</td>
			<td>gS 22.1670</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Crile Retractor 4&#8221; double ended</td>
			<td>Apiary Medical, Inc.</td>
			<td>gS 34.1934</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Other surgical instruments</td>
			<td>Roboz Surgical Instrument Company, Inc., Gaithersburg, MD</td>
			<td></td>
			<td></td>
		</tr>
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			<td height="21" style="height:21px;">Peripheral Intravenous (I.V.) Cannula, 24G</td>
			<td>BD</td>
			<td>381312</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">Ketamine HCl, 100mg/mL, 10mL</td>
			<td>Ketaset- Patterson Vet</td>
			<td>07-803-6637&#160;</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Xylazine (AnaSed),20mg/mL,20mL</td>
			<td>Ketaset- Patterson Vet</td>
			<td>07-808-1947</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Buprenex, 0.3mg/1ml (5 Ampules/Box)</td>
			<td>Ketaset- Patterson Vet</td>
			<td>07-850-2280</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nair Baby Oil Hair Removal Lotion-9 oz</td>
			<td>Amazon/Walmart/CVS</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Inflation Device</td>
			<td>Demax Medical</td>
			<td>DID30</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">D300 3-way Stopcock</td>
			<td>B.Braun Medical Inc.</td>
			<td>4599543</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Artificial Tears Ointment&#160;</td>
			<td>Rugby Laboratories, Duluth, GA</td>
			<td>N/A</td>
			<td></td>
		</tr>
	</tbody>
</table>

vascular balloon injury and intraluminal administration in rat carotid artery

쥐의 경동맥 풍선 손상 모델이 아니라 20 년 이상 설립되었습니다. 이는 혈관 평활근 탈분화, 신생 혈관 내막 형성 및 혈관 재 형성에 관여하는 분자 세포 메커니즘을 연구하는 중요한 방법 남아있다. 수컷 스프 라그 - 돌리 래트 모델에 대한 가장 자주 사용되는 동물이다. 여성 호르몬은 혈관 질환에 대한 보호하고, 따라서이 절차에 변화를 소개로 여성 쥐 바람직하지 않다. 좌측 경동맥은 전형적 우측 경동맥은 음성 대조군으로서의으로 부상된다. 왼쪽 경동맥 손상은 내피 세포를 denudes과의에게 혈관 벽을 넓히다 팽창 된 풍선에 의해 발생합니다. 손상 후, 이러한 화합물 및 약학 적 유전자 또는 shRNA를 전송 중의 사용과 같은 잠재적 인 치료 전략을 평가할 수있다. 일반적으로 유전자 또는 shRNA를 전송, 혈관 내강의 손상된 부분은 로컬 V를 30 분 동안 형질 도입한다손상된 혈관 벽에 전달 및 발현 단백질 또는 shRNA를 하나를 인코딩 iral 입자. 증식 성 혈관 평활근 세포를 나타내는 신생 내막 비후 보통 손상 후 이주에서 봉우리. 용기는 주로 세포 및 분자 세포 신호 전달 경로의 분석뿐만 아니라 유전자 및 단백질 발현이 시점에서 수확. 용기는 또한 의도 된 실험 목적에 따라, 특정 단백질 또는 경로의 발현 및 / 또는 활성의 개시를 결정하기 위해 이전의 시점에서 수확 될 수있다. 혈관 조직 학적 염색, 면역 단백질 / mRNA의 분석, 및 활성 분석을 사용하는 것을 특징으로하고 평가할 수있다. 같은 동물에서 그대로 오른쪽 경동맥 이상적인 내부 통제이다. 분자 세포 매개 부상 유발 변화는 내부 우측 제어 동맥 동맥 부상을 비교함으로써 평가 될 수있다. 마찬가지로, 치료 양식은이 다칠를 비교함으로써 평가할 수있다컨트롤에 D 및 치료 동맥은 동맥 부상.

두 주 부상 후, 경동맥은 수확 단면 및 형태 학적 분석을 실시한다. 동맥이 교차 절단 및 H &amp; E 염색있다 (그림 1, 2B, C 3). 쥐 경동맥 벽 분홍색 선으로 표시 탄성 얇은 판의 4 개의 레이어가 포함되어 있습니다. 라미 최 외부 탄성 박판 (EEL)과 최 박판 사이의 영역은, 내부 탄성 박판 (IEL)이 미디어 평활근 층 (도 1)이다. IEL 내부 영역 내막 그대로, 혈관 내피 세포의 단층이고;...

Watch this Scientific Journal Video about Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery at JoVE.com

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

이 프로토콜은 쥐의 경동맥에 관내 부상의 원인이 이제부터는 신생 내막의 증식을 유도하는 풍선 카테터를 사용합니다. 이 부상에 대한 응답으로 혈관 재 형성의 메커니즘을 연구하는 잘 확립 된 모델입니다. 또한 널리 잠재적 인 치료 방법의 유효성을 결정하는데 사용된다.

쥐 경동맥의 혈관 풍선 부상과 관내 관리

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and ...

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Research

JoVE Journal

Medicine

16.2K 조회수.  State University of New York College of Nanoscale Science and Engineering (SUNY CNSE).  이 프로토콜은 쥐의 경동맥에 관내 부상의 원인이 이제부터는 신생 내막의 증식을 유도하는 풍선 카테터를 사용합니다. 이 부상에 대한 응답으로 혈관 재 형성의 메커니즘을 연구하는 잘 확립 된 모델입니다. 또한 널리 잠재적 인 치료 방법의 유효성을 결정하는데 사용된다. 

비디오: Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

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

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Intraluminal 중간 대뇌 동맥 폐색의 쥐 뇌졸중 모델에서 뇌 관류 모니터링에 최적화 된 시스템

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

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 ...

A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is also a valuable model system to test, and to evaluate therapeutics and drugs that negate maladaptive remodeling in the vessel. The injury, or barotrauma, in the vessel lumen caused by an inflated balloon via an inserted catheter induces subsequent neointimal growth, often leading to hyperplasia or thickening of the vessel wall that narrows, or obstructs the lumen. The method described here is sufficiently sensitive, and the results can be obtained in relatively short time (2 weeks after the surgery). The efficacy of the drug or therapeutic against the induced-remodeling can be evaluated either by the post-mortem pathological and histomorphological analysis, or by ultrasound sonography in live animals. In addition, this model system has also been used to determine the therapeutic window or the time course of the administered drug. These studies can leadto the development of a better administrative strategy and a better therapeutic outcome. The procedure described here provides a tool for translational studies that bring drug and therapeutic candidates from bench research to clinical applications.

The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described.

생쥐 경동맥 풍선 손상 모델은 반대로 혈관 리모델링의 치료를 테스트하는

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is ...

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The simple murine AVF model was combined with intraluminal administration of drug solution to the venous endothelium at the same time as fistula creation. Technical aspects of this model are discussed. Under general anesthesia, an abdominal incision is made and the aorta and inferior vena cava (IVC) are exposed. The infra-renal aorta and IVC are dissected for clamping. After proximal and distal clamping, the puncture site is exposed and a 25 G needle is used to puncture both walls of the aorta and into the IVC. Immediately after the puncture, a reporter gene-expressing viral vector was infused in the IVC via the same needle, followed by 15 min of incubation. The intraluminal administration method enabled more robust viral gene delivery to the venous endothelium compared to administration by the external route. This novel method of delivery will facilitate studies that explore the role of the endothelium in AVF maturation and enable intraluminal drug delivery at the time of surgical operation.

After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

마우스 동정맥루 내피에 관내 약물 전달

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the industrialized world. New Zealand White (NZW) rabbits are widely used as an animal model for the study of atherosclerosis. They develop spontaneous lesions when fed with atherogenic diet; however, this requires long time of 4 - 8 months. To further enhance and accelerate atherogenesis, a combination of atherogenic diet and mechanical endothelial injury is often employed. The presented procedure for inducing atherosclerotic plaques in rabbits uses a balloon catheter to disrupt the endothelium in the left iliac artery of NZW rabbits fed with atherogenic diet. Such mechanical damage caused by the balloon catheter induces a chain of inflammatory reactions initiating neointimal lipid accumulation in a time dependent fashion. Atherosclerotic plaque following balloon injury show neointimal thickening with extensive lipid infiltration, high smooth muscle cell content and presence of macrophage derived foam cells. This technique is simple, reproducible and produces plaque of controlled length within the iliac artery. The whole procedure is completed within 20 - 30 min. The procedure is safe with low mortality and also offers high success in obtaining substantial intimal lesions. The procedure of balloon catheter induced arterial injury results in atherosclerosis within two weeks. This model can be used for investigating the disease pathology, diagnostic imaging and to evaluate new therapeutic strategies.

Animal models of atherosclerosis are essential to understand the mechanism and to investigate newer approaches to prevent plaque development or rupture, a leading cause of death in the industrialized world. This protocol uses a combination of balloon injury and cholesterol rich diet to induce atherosclerotic plaques in rabbit iliac artery.

가속된 동맥 경화의 토끼 모델: 장 골 동맥 풍선 상해의 방법 론 적 관점

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the ...

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 ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, we have translated a vascular injury model from a small animal into a large animal (canine), with slight modifications that can be used for pre-clinical screening of prophylactic and thrombolytic agents. In addition to the surgical methods, the modified protocol describes the step-by-step methods to assess carotid artery canalization by angiography, detailed instructions to process both the brain and carotid artery for histological analysis to verify carotid canalization and cerebral hemorrhage, and specific parameters to complete an assessment of downstream thromboembolic events by utilizing magnetic resonance imaging (MRI). In addition, specific procedural changes from the previously well-established small animal model necessary to translate into a large animal (canine) vascular injury are discussed.

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

제 2 철 염화 유도 개 경 동맥 혈전 증: 혈관 손상의 큰 동물 모델

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...