작품은 Translational 의학 및 치료학에 대한 연구소의 화목과에서 펜실베니아 대학과 마를린 L. 코헨과 제롬 H. Fleisch 학술 그랜트 의학 및 공학 (T32HL007954)의 연구소에서 교육 교부금에 의해 투자되었다 코네티컷 대학 보건 센터 (LHS)와 NIH NS - 029331 (FAW).

1. 장비 준비 <ol><li> 박테리아 오염을 최소화하기 위해 75 % 에탄올로 stereotaxic 프레임과 펌프를 아래로 닦아주십시오. </li><li> 해밀턴 주사기와 융합된 실리카 바늘을 소독. 참고 : 화학 살균을 사용하면, 사용하기 전에 멸균 물에 여러 번 씻어해야합니다. </li><li> 75 % 에탄올로 파라핀 왁스 종이의 표면을 닦아 건조 수 있습니다. </li></ol> 2. 분사에 대한 마우스의 준비 참고 : 그들이 새로운 환경에 적응하고 스트레스를 줄일 수 있도록 귀하의 동물 시설에 대한 수술 전 최소 7 일 전달 생쥐 되세요. <ol><li> 사전 수술 기본에 대한 마우스의 무게를. </li><li> 꼬리 핀치에 응답하기 전까지 Isoflurane 30 % 산소 70 % 질소 산화물, 4 %를 마취를 유발 </li><li> 이후 수술 진통제에 대한 buprenorphine 0.1mg/kg intraperitoneally와 마우스를 주사 </li><li> 두피를 쉐이브 </li><li> 무균 석유 젤리로 코트의 눈 </li><li> betadine × 3 잎사귀와 두피를 준비 후, 두피가 건조 수 </li><li> 멸균 수술 메스를 사용하여 두피의 1cm 절개 정중선 화살 만들기 참고 : 관대한 절개는 두개골의 랜드마크의 완전한 노출을 수 있습니다. </li><li> 베이스에서 1cm를 시작 꼬리 복부 표면의 1cm를 면도하고 betadine와 × 3 잎사귀 준비 </li><li> stereotaxic 프레임에 마우스 플레이스 참고 : 마우스 bregma의 오른쪽에있는 우수 bregma의 노출과 최소 3mm와 프레임의 기본과 두개골 병렬의 표면과 프레임에 안전하게하는 것이 중요합니다. </li></ol> 3. 했습니다만 출혈 수술 참고 : 전체 수술하는 동안 마우스를 지속적으로 37 유지 Isoflurane 30 % 산소 70 % 질소 산화물, 그리고 1-3%와 anesthetized있다 ± 0.5 ° C는 서미 스터 제어 가열 패드를 사용하고 직장 온도계에 의해 감시. <ol><li> 프레임 1 CC의 주사기에 살균 27g 바늘을 연결합니다. </li><li> 바늘 정확히 bregma 끝날 때까지 stereotaxic 팔을 조정합니다. </li><li> 그 바늘은 두개골의 표면에 bregma에 수평 낮은 2.5 mm에서되도록 팔을 조정합니다. </li><li> 수동으로 완전히 두개골을 구멍하지 프레임 복용 치료의 완만한 하향 운동을 적용하는 동안 머리의 표면에 버 구멍을 만들기 위해 주사기를 회전. </li><li> 주사기 / 바늘로 수동으로 바늘하고 완전한 버 구멍을 제거 참고 : 두개골과 뇌 실질 실수로 바늘을 밀어의 최소화 위험의 내부 테이블을 천공이있을 때 손으로 버 구멍을 완료하면 즉시 인식을 허용합니다. </li><li> 멸균 수술 블레이드를 사용하여 꼬리의 복부 표면에 가로 절개를하고 동맥혈로부터 2-3 큰 방울은 파라핀 왁스 종이에 떨어질 수 있습니다. 다음 신속하게 멸균 거즈를 사용하여 압력과 출혈 중지합니다. </li><li> 펌프에 해밀턴 주사기 및 장소 주사기에 17 μL 혈액을 철회. </li><li> 포인트 5 stereotaxic 팔을 조정 ° 수직 축에 상대 medially. </li><li> 바늘의 끝은 두개골에 버 구멍을 통해 그리고 낮은 바늘 3.5 mm가되도록 조심스럽게 stereotaxic 팔을 조정합니다. </li><li> 2 분 후 바늘 0.5 mm를 철회 잠깐 만요, (그래서 끝을 깊이 3mm입니다) </li><li> 두뇌가 바늘 주위에 다시 확장하고 주입하는 동안 바늘 삽입 트랙을 refluxing 혈액의 위험을 최소화할 수 있도록 5 분 동안 기다리십시오. </li><li> 7.5 μL를위한 혈액 1 μL / 분 주사. </li><li> 초기 혈액 응고 허용하고 조직 교대에 대한 intracranial 압력 고도를 최소화하기 위해 발생하는 5 분 동안 기다리십시오. </li><li> 1 μL / 분에 남아있는 7.5 μL를 주입 </li><li> 바늘이 응고 혈액 수 있도록 25 분 장소에서 유지하도록 허용 참고 : 응고 혈액은 바늘을 삽입 사이트를 refluxing 혈액집니다 위해 바늘을 철수 때 기다려야 실패 </li><li> 천천히 주사기를 철회하고 응고에서 바늘에 잔류 혈액을 방지하고 바늘의 재사용을 보장하기 위해 즉시 온수 헹굼. </li><li> 프레임에서 마우스를 제거하고 수의학 수술 접착제와 꼬리와 두피 incisions를 닫습니다. </li><li> 마취의 전원을 끕니다. </li><li> 지속적으로 음식을 moistened을 무료로 이용 예열되는 동안 학살 마우스 수 있습니다. </li><li> littermates 때 완전히 깨어와 케이지에 마우스를 반환합니다. 음식에 대한 액세스 동물 도움 새장 바닥에 서부 유럽 표준시 음식 펠릿를 놓습니다. </li></ol> 4. 대표 결과 : <img src="/files/ftp_upload/2618/2618fig1.jpg" alt="그림 1"/> 그림 1. 십오분 ICH 수술 후 마우스 뇌 코로나 섹션을 참조하십시오. 즉시 두뇌가 바늘을 삽입 사이트에서 코로나 섹션의 총 점검에 따라 ICH의 성공을위한 검사되었습니다 희생. 코퍼스의 callosum 과거, 또는 심실에 바늘 트랙까지, 두뇌의 기본까지 추적 출혈이 실패한 것으로 간주되었고 그 생쥐는 모든 분석에서 제거되었습니다. 전체 ICH succeSS 요금은 0 % 사망률과 함께 50 생쥐에 75~85%되었습니다. <img src="/files/ftp_upload/2618/2618fig2.jpg" alt="그림 2"/> 그림 2. 실린더 시험 바로 기저 신경의 ICH 후 왼쪽 hemiparesis을 보여줍니다. ICH 수술 후 (A) 샘플 마우스 후면. 왼쪽 기초 신경의 ICH 후 실린더의 벽에에서만 오른쪽 forelimb의 위치를​​ 적어 둡니다. (B) 실린더 테스트 그래프 후 마우스 일대에서 1 결과 ICH 수술 (N = 5) 가짜에 비해 (n은 = 4). 위장 마우스는 혈액 주입 (바늘이 두뇌에 삽입되었다)를 제외한 모든 절차를했다. 각 마우스는 12 cm 직경 투명 유리 실린더에 배치 20 투우사에 대한 관찰되었다. 실린더의 벽면에있는 앞발의 초기 배치는 후면마다 득점했다. 마우스 바닥에 반환하고 다음 뒤쪽가 득점하기 전까지 후속 움직임은 (예 : 수평 탐사 등) 득점되지 않았습니다. 0 더 forelimb 선호 1은 오른쪽 forelimb가 사용되었습니다 표시하지 표시된 곳에, (# 오른쪽 + # 왼쪽 + # 모두) / - laterality 지수는 계산 (# forelimb 게재 위치 왼쪽 실린더의 측면에 # 오른쪽 forelimb 위치)가 .

<ol>
	<li>Hua, Y., Schallert, T., Keep, R. F., Wu, J., Hoff, J. T., Xi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+Tests+After+Intracerebral+Hemorrhage+in+the+Rat.">Behavioral Tests After Intracerebral Hemorrhage in the Rat.</a> Stroke. 33, 2478-2484 (2002).</li><li>James, M., Warner, D., Laskowitz, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+Models+of+Intracerebral+Hemorrhage:+A+Translational+Perspective.">Preclinical Models of Intracerebral Hemorrhage: A Translational Perspective.</a> Neurocritical Care. 9, 139-152 (2008).</li><li>Foerch, C., Arai, K., Jin, G., Park, K. -. P., Pallast, S., van Leyen, K., Lo, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+Model+of+Warfarin-Associated+Intracerebral+Hemorrhage.">Experimental Model of Warfarin-Associated Intracerebral Hemorrhage.</a> Stroke. 39, 3397-3404 (2008).</li><li>Nakamura, T., Xi, G., Hua, Y., Schallert, T., Hoff, J. T., Keep, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracerebral+hemorrhage+in+mice:+model+characterization+and+application+for+genetically+modified+mice.">Intracerebral hemorrhage in mice: model characterization and application for genetically modified mice.</a> Journal of Cerebral Blood Flow and Metabolism. 24, 487-494 (2004).</li><li>Rynkowski, M. A., Kim, G. H., Komotar, R. J., Otten, M. L., Ducruet, A. F., Zacharia, B. E., Kellner, C. P., Hahn, D. K., Merkow, M. B., Garrett, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mouse+model+of+intracerebral+hemorrhage+using+autologous+blood+infusion.">A mouse model of intracerebral hemorrhage using autologous blood infusion.</a> Nat. Protocols. 3, 122-128 (2008).</li><li>Wang, J., Fields, J., Dor&#233;, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+development+of+an+improved+preclinical+mouse+model+of+intracerebral+hemorrhage+using+double+infusion+of+autologous+whole+blood.">The development of an improved preclinical mouse model of intracerebral hemorrhage using double infusion of autologous whole blood.</a> Brain Research. 1222, 214-221 (2008).</li><li>Zhao, X., Sun, G., Zhang, J., Strong, R., Song, W., Gonzales, N., Grotta, J. C., Aronowski, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hematoma+resolution+as+a+target+for+intracerebral+hemorrhage+treatment:+Role+for+peroxisome+proliferator-activated+receptor+&#947;+in+microglia/macrophages.">Hematoma resolution as a target for intracerebral hemorrhage treatment: Role for peroxisome proliferator-activated receptor &#947; in microglia/macrophages.</a> Annals of Neurology. 61, 352-362 (2007).</li></ol>

관심 없음 충돌 선언하지 않습니다.

자연 기초 신경의 출혈을 재현할 모델 autologous 꼬리 동맥 혈액 검사 결과를 사용하여 생쥐에서 했습니다만 출혈이 수술 모델입니다. 생쥐의 ICH 모델은 pathophysiology을 조사하기 위해 유전자 변형 동물의 이용의 장점을 제공하지만, 그들의 작은 크기가 큰 동물에 비해 neurosurgical 절차가 더 기술적으로 어려울 수 있습니다. collagenase 모델과 autologous 혈액 주입 모델은 실험 ICH 두 ?...

마우스 신경 외과에 대한 Stereotaxic 프레임 (스토엘팅, 51925) Microinfusion 펌프 및 프로세서 (지금 뭐 - 3 Micro4, 세계 정밀 계측기, 사라소타, 플로리다) 마우스 따뜻한 (스토엘팅, 50300) Inhalational 마우스 마취 (브레인 트리 과학, EZ - AF9000) 25 μL gastight borosilicate 해밀턴 코팅 플런저와 주사기없이 바늘 (해밀턴 회사, 리노, NV, 1702RN 주사기 : 765401, 물미 : 30,949, 스페이서 : 30,946) 융합 실리카 바늘은 2cm 길이 (해밀턴, 17739)에 컷 전에 각각의 수술을 소독하는 경우 * 참고 해밀턴 주사기와 융합된 실리카 바늘은 여러 수술을 위해 재사 용할 수 있습니다. 이 자료는 혈액이 응고되지 않도록하는 데있어 매우 중요합니다. 무균 수술 장갑 수술 가운, 모자 및 마스크 Betadine 75 % 에탄올 무균 27g 바늘 (싱글 사용) 무균 한 CC 주사기 (싱글 사용) 무균 수술 블레이드 Cidex 멸균 물 buprenorphine 및 isoflurane 멸균 거즈 파라핀 왁스 용지 사각형 베티 수술 접착제 (Vetbond, 3M, 세인트 폴, MN)

autologous blood injection to model spontaneous intracerebral hemorrhage in mice

Investigation of the pathophysiology of injury after intracerebral
hemorrhage (ICH) requires a reproducible animal model. While ICH
accounts for 10-15% of all strokes, there remains no specific
effective therapy. The autologous blood injection model in mice
involves the stereotaxic injection of arterial blood into the basal
ganglia mimicking a spontaneous hypertensive hemorrhage in man. The
response to hemorrhage can then be studied in vivo and the
neurobehavioral deficits quantified, allowing for description of the
ensuing pathology and the testing of potential therapeutic agents. The
procedure described in this protocol uses the double injection
technique to minimize risk of blood reflux up the needle track, no
anticoagulants in the pumping system, and eliminates all dead space
and expandable tubing in the system.

Watch this Scientific Journal Video about Autologous Blood Injection to Model Spontaneous Intracerebral Hemorrhage in Mice at JoVE.com

Autologous Blood Injection to Model Spontaneous Intracerebral Hemorrhage in Mice

이 프로토콜에서 설명하는 생쥐에서 했습니다만 출혈의 autologous 혈액 주입 모델은 바늘 트랙까지 혈액 역류, 펌핑 시스템에서 anticoagulants의 위험을 최소화하기 위해 이중 사출 기술을 사용하고, 시스템의 모든 죽은 공간 및 확장 튜브를 제거합니다.

마우스에서 자연 했습니다만 출혈을 모델로 Autologous 혈액 주입

Investigation of the pathophysiology of injury after intracerebral
hemorrhage (ICH) requires a reproducible animal model.  While ICH
accounts for 10-15% ...

autologous-blood-injection-to-model-spontaneous-intracerebral

Research

JoVE Journal

Neuroscience

14.7K 조회수.  University of Connecticut Health Center. 이 프로토콜에서 설명하는 생쥐에서 했습니다만 출혈의 autologous 혈액 주입 모델은 바늘 트랙까지 혈액 역류, 펌핑 시스템에서 anticoagulants의 위험을 최소화하기 위해 이중 사출 기술을 사용하고, 시스템의 모든 죽은 공간 및 확장 튜브를 제거합니다.

비디오: Autologous Blood Injection to Model Spontaneous Intracerebral Hemorrhage in Mice

Methods to Characterize Spontaneous and Startle-induced Locomotion in a Rotenone-induced Parkinson's Disease Model of Drosophila

Parkinson&#8217;s disease is a neurodegenerative disorder that results from the degeneration of dopaminergic neurons in the central nervous system, primarily in the substantia nigra. The disease causes motor deficiencies, which present as rigidity, tremors and dementia in humans. Rotenone is an insecticide that causes oxidative damage by inhibiting the function of the electron transport chain in mitochondria. It is also used to model Parkinson&#8217;s disease in the Drosophila. Flies have an inherent negative geotactic response, which compels them to climb upwards upon being startled. It has been established that rotenone causes early mortality and locomotion defects that disrupt the flies&#8217; ability to climb after they have been tapped downwards. However, the effect of rotenone on spontaneous movement is not well documented. This study outlines two sensitive, reproducible, and high throughput assays to characterize rotenone-induced deficiencies in short-term startle-induced locomotion and long-term spontaneous locomotion in Drosophila. These assays can be conveniently adapted to characterize other Drosophila models of locomotion defects and efficacy of therapeutic agents.

Methods to Characterize Spontaneous and Startle-induced Locomotion in a Rotenone-induced Parkinson&#039;s Disease Model of Drosophila

Parkinson&#8217;s disease is a neurodegenerative disorder that results from the degeneration of dopaminergic neurons in the central nervous system, causing locomotion defects. Rotenone models Parkinson&#8217;s disease in Drosophila. This paper outlines two assays that characterize both spontaneous and startle-induced locomotion deficiencies caused by rotenone.

방법은 자발적인 특성화 및 놀라게 유도 로코을의 로테 논에 의한 파킨슨 병 모델에서합니다<em&gt; 초파리</em

Parkinson&#8217;s disease is a neurodegenerative disorder that results from the degeneration of dopaminergic neurons in the central nervous system, ...

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Establishing Mouse Models for Zika Virus-induced Neurological Disorders Using Intracerebral Injection Strategies: Embryonic, Neonatal, and Adult

The Zika virus (ZIKV) is a flavivirus currently endemic in North, Central, and South America. It is now established that the ZIKV can cause microcephaly and additional brain abnormalities. However, the mechanism underlying the pathogenesis of ZIKV in the developing brain remains unclear. Intracerebral surgical methods are frequently used in neuroscience research to address questions about both normal and abnormal brain development and brain function. This protocol utilizes classical surgical techniques and describes methods that allow one to model ZIKV-associated human neurological disease in the mouse nervous system. While direct brain inoculation does not model the normal mode of virus transmission, the method allows investigators to ask targeted questions concerning the consequence after ZIKV infection of the developing brain. This protocol describes embryonic, neonatal, and adult stages of intraventricular inoculation of ZIKV. Once mastered, this method can become a straightforward and reproducible technique that only takes a few hours to perform.

Here we describe a method for establishing a model of Zika virus-induced microcephaly in mouse. This protocol includes methods for embryonic, neonatal, and adult-stage intracerebral inoculation of the Zika virus.

Zika 바이러스 유도 신경 장애 Intracerebral 주입 전략을 사용 하 여 마우스 모델을 수립: 배아, 신생아, 및 성인

The Zika virus (ZIKV) is a flavivirus currently endemic in North, Central, and South America. It is now established that the ZIKV can cause microcephaly ...

Rat Model of Widespread Cerebral Cortical Demyelination Induced by an Intracerebral Injection of Pro-Inflammatory Cytokines

Multiple sclerosis (MS) is the most common immune-mediated disease of the central nervous system (CNS) and progressively leads to physical disability and death, caused by white matter lesions in the spinal cord and cerebellum, as well as by demyelination in grey matter. Whilst conventional models of experimental allergic encephalomyelitis are suitable for the investigation of the cell-mediated inflammation in the spinal and cerebellar white matter, they fail to address grey matter pathologies. Here, we present the experimental protocol for a novel rat model of cortical demyelination allowing the investigation of the pathological and molecular mechanisms leading to cortical lesions. The demyelination is induced by an immunization with low-dose myelin oligodendrocyte glycoprotein (MOG) in an incomplete Freund&#39;s adjuvant followed by a catheter-mediated intracerebral delivery of pro-inflammatory cytokines. The catheter, moreover, enables multiple rounds of demyelination without causing injection-induced trauma, as well as the intracerebral delivery of potential therapeutic drugs undergoing a preclinical investigation. The method is also ethically favorable as animal pain and distress or disability are controlled and relatively minimal. The expected timeframe for the implementation of the entire protocol is around 8 - 10 weeks.

The protocol presented here allows the reproduction of a widespread grey matter demyelination of both cortical hemispheres in adult male Dark Agouti rats. The method comprises of intracerebral implantation of a catheter, subclinical immunization against myelin oligodendrocyte glycoprotein, and intracerebral injection of a pro-inflammatory cytokine mixture through the implanted catheter.

프로 염증 성 사이토카인의 인트레이스렐 주입에 의해 유도된 광범위한 대뇌 피질 탈량의 쥐 모델

Multiple sclerosis (MS) is the most common immune-mediated disease of the central nervous system (CNS) and progressively leads to physical disability and ...

A Volumetric Method for Quantification of Cerebral Vasospasm in a Murine Model of Subarachnoid Hemorrhage

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor determining patient outcome and is therefore frequently taken as a study endpoint. However, in small animal studies on SAH, quantification of cerebral vasospasm is a major challenge. Here, an ex vivo method is presented that allows quantification of volumes of entire vessel segments, which can be used as an objective measure to quantify cerebral vasospasm. In a first step, endovascular casting of the cerebral vasculature is performed using a radiopaque casting agent. Then, cross-sectional imaging data are acquired by micro computed tomography. The final step involves 3-dimensional reconstruction of the virtual vascular tree, followed by an algorithm to calculate center lines and volumes of the selected vessel segments. The method resulted in a highly accurate virtual reconstruction of the cerebrovascular tree shown by a diameter-based comparison of anatomical samples with their virtual reconstructions. Compared with vessel diameters alone, the vessel volumes highlight the differences between vasospastic and non-vasospastic vessels shown in a series of SAH and sham-operated mice.

The goal of this article is to present a method that allows a 3-dimensional reconstruction of the cerebrovascular tree in mice after micro computed tomography and determination of volumes of entire vessel segments that can be used to quantify cerebral vasospasm in murine models of subarachnoid hemorrhage.

거미 막 밑 출혈의 Murine 모델에 대뇌 Vasospasm의 정량화에 대 한 체적 메서드

Subarachnoid hemorrhage (SAH) is a subtype of hemorrhagic stroke. Cerebral vasospasm that occurs in the aftermath of the bleeding is an important factor ...

Minimally Invasive Endoscopic Intracerebral Hemorrhage Evacuation

Intracerebral hemorrhage (ICH) is a subtype of stroke with high mortality and poor functional outcomes, largely because there are no evidence-based treatment options for this devastating disease process. In the past decade, a number of minimally invasive surgeries have emerged to address this issue, one of which is endoscopic evacuation. Stereotactic ICH Underwater Blood Aspiration (SCUBA) is a novel endoscopic evacuation technique performed in a fluid-filled cavity using an aspiration system to provide an additional degree of freedom during the procedure. The SCUBA procedure utilizes a suction device, endoscope, and sheath and is divided into two phases. The first phase involves maximal aspiration and minimal irrigation to decrease clot burden. The second phase involves increasing irrigation for visibility, decreasing aspiration strength for targeted aspiration without disturbing the cavity wall, and cauterizing any bleeding vessels. Using the endoscope and aspiration wand, this technique aims to maximize hematoma evacuation while minimizing collateral damage to the surrounding brain. Advantages of the SCUBA technique include the use of a low-profile endoscopic sheath minimizing brain disruption and improved visualization with a fluid-filled cavity rather than an air-filled one.

This paper details the surgical protocol for minimally invasive endoscopic intracerebral hemorrhage evacuation using the SCUBA technique.

최소 침습적 내시경 출혈 대피

Intracerebral hemorrhage (ICH) is a subtype of stroke with high mortality and poor functional outcomes, largely because there are no evidence-based ...

In Vivo Direct Reprogramming of Resident Glial Cells into Interneurons by Intracerebral Injection of Viral Vectors

Converting resident glia in the brain into functional and subtype-specific neurons in vivo provides a step forward towards the development of alternative cell replacement therapies while also creating tools to study cell fate in situ. To date, it has been possible to obtain neurons via in vivo reprogramming, but the precise phenotype of these neurons or how they mature has not been analyzed in detail. In this protocol, we describe a more efficient conversion and cell-specific identification of the in vivo reprogrammed neurons, using an AAV-based viral vector system. We also provide a protocol for functional assessment of the reprogrammed cells&#8217; neuronal maturation. By injecting flip-excision (FLEX) vectors, containing the reprogramming and synapsin-driven reporter genes&#160;to specific cell types in the brain that serve as the target for cell reprogramming. This technique allows for the easy identification of newly reprogrammed neurons. Results show that the obtained reprogrammed neurons functionally mature over time, receive synaptic contacts and show electrophysiological properties of different types of interneurons. Using the transcription factors Ascl1, Lmx1a and Nurr1, the majority of the reprogrammed cells have properties of fast-spiking, parvalbumin-containing interneurons.

This protocol aims at generating directly reprogrammed interneurons in vivo, using an AAV-based viral system in the brain and a FLEX synapsin-driven GFP reporter, which allows for cell identification and further analysis in vivo.

바이러스 벡터의 intracerebral 주입에 의해 인터뉴런으로 상주 신경교 세포의 생체 내 직접 재프로그래밍

Converting resident glia in the brain into functional and subtype-specific neurons in vivo provides a step forward towards the development of alternative ...

Double Direct Injection of Blood into the Cisterna Magna as a Model of Subarachnoid Hemorrhage

Among strokes, subarachnoid hemorrhage (SAH) consecutive to the rupture of a cerebral arterial aneurysm represents 5-9% but is responsible for about 30% of the total stroke-related mortality with an important morbidity in terms of neurological outcome. A delayed cerebral vasospasm (CVS) may occur most often in association with a delayed cerebral ischemia. Different animal models of SAH are now being used including endovascular perforation and direct injection of blood into the cisterna magna or even the prechiasmatic cistern, each exhibiting distinct advantages and disadvantages. In this article, a standardized mouse model of SAH by double direct injection of determined volumes of autologous whole blood into the cisterna magna is presented. Briefly, mice were weighed and then anesthetized by isoflurane inhalation. Then, the animal was placed in a reclining position on a heated blanket maintaining a rectal temperature of 37 &#176;C and positioned in a stereotactic frame with a cervical bend of about 30&#176;. Once in place, the tip of an elongated glass micropipette filled with the homologous arterial blood taken from carotid artery of another mouse of the same age and gender (C57Bl/6J) was positioned at a right angle in contact with the atlanto-occipital membrane by means of a micromanipulator. Then 60 &#181;L of blood was injected in the cisterna magna followed by a 30&#176; downward tilt of the animal for 2 minutes. The second infusion of 30 &#181;L of blood into the cisterna magna was performed 24 h after the first one. The individual follow-up of each animal is carried out daily (careful evaluation of weight and well-being). This procedure allows a predictable and highly reproducible distribution of blood, likely accompanied by intracranial pressure elevation that can be mimicked by an equivalent injection of an artificial cerebral spinal fluid (CSF), and represents an acute to mild-model of SAH inducing low mortality.

We described in this protocol a standardized subarachnoid hemorrhage (SAH) mouse model by a double injection of autologous whole-blood into the cisterna magna. The high degree of standardization of the double-injection procedure represents a middle-to-acute model of SAH with relative safety regarding mortality.

수브라크누드 출혈의 모델로 시스테나 마그나에 혈액을 이중 직접 주입

Among strokes, subarachnoid hemorrhage (SAH) consecutive to the rupture of a cerebral arterial aneurysm represents 5-9% but is responsible for about 30% ...

Massive Pontine Hemorrhage by Dual Injection of Autologous Blood

We provide a protocol to establish a massive pontine hemorrhage model in a rat. Rats weighing about 250 grams were used in this study. One hundred microliters of autologous blood was taken from the tail vein and stereotaxically injected into the pons. The injection process was divided into 2 steps: First, 10 &#181;L of blood was injected into a specific location, anteroposterior position (AP) -9.0 mm; lateral (Lat) 0 mm; vertical (Vert) -9.2 mm, followed by a second injection of the residual blood located at AP -9.0 mm; Lat 0 mm; Vert -9.0 mm with a 20-minute interval. The balance beam test, limb placement test, and the modified Voestch neuroscore were used to evaluate neurological function. Magnetic Resonance Imaging (MRI) was used to assess the volume of hemorrhage in vivo. The symptoms of this model were in line with patients with massive pontine hemorrhage.

We present a protocol to establish a massive pontine hemorrhage model in a rat via dual injection of autologous blood.

자동 혈액의 이중 주입에 의한 대규모 폰틴 출혈

We provide a protocol to establish a massive pontine hemorrhage model in a rat. Rats weighing about 250 grams were used in this study. One hundred ...

Pre-Chiasmatic, Single Injection of Autologous Blood to Induce Experimental Subarachnoid Hemorrhage in a Rat Model

Despite advances in treatment over the last decades, subarachnoid hemorrhage (SAH) continues to carry a high burden of morbidity and mortality, largely afflicting a fairly young population. Several animal models of SAH have been developed to investigate the pathophysiological mechanisms behind SAH and to test pharmacological interventions. The pre-chiasmatic, single injection model in the rat presented in this article is an experimental model of SAH with a predetermined blood volume. Briefly, the animal is anesthetized, intubated, and kept under mechanical ventilation. Temperature is regulated with a heating pad. A catheter is placed in the tail artery, enabling continuous blood pressure measurement as well as blood sampling. The atlantooccipital membrane is incised and a catheter for pressure recording is placed in the cisterna magna to enable intracerebral pressure measurement. This catheter can also be used for intrathecal therapeutic interventions. The rat is placed in a stereotaxic frame, a burr hole is drilled anteriorly to the bregma, and a catheter is inserted through the burr hole and placed just anterior to the optic chiasm. Autologous blood (0.3 mL) is withdrawn from the tail catheter and manually injected. This results in a rise of intracerebral pressure and a decrease of cerebral blood flow. The animal is kept sedated for 30 min and given subcutaneous saline and analgesics. The animal is extubated and returned to its cage. The pre-chiasmatic model has a high reproducibility rate and limited variation between animals due to the pre-determined blood volume. It mimics SAH in humans making it a relevant model for SAH research.

Subarachnoid hemorrhage continues to carry a high burden of mortality and morbidity in man. To facilitate further research into the condition and its pathophysiology, a pre-chiasmatic, single injection model is presented.

쥐 모델에서 실험적 지주막하 출혈을 유도하기 위한 자가 혈액의 Pre-chiasmatic, 단일 주사

Despite advances in treatment over the last decades, subarachnoid hemorrhage (SAH) continues to carry a high burden of morbidity and mortality, largely ...

Intracranial Pressure Monitoring In Nontraumatic Intraventricular Hemorrhage Rodent Model

Survivors of intraventricular hemorrhage are often left with significant long-term memory impairment; thus, research utilizing intraventricular hemorrhage animal models is essential. In this study, we sought out ways to measure intracranial pressure, mean arterial pressure, and cerebral perfusion pressure during nontraumatic intraventricular hemorrhage in rats. The experimental design included three Sprague Dawley groups: sham, standard 200 &#181;l intraventricular hemorrhage, and vehicle control groups. By introducing an intraparenchymal fiberoptic pressure sensor, precise intracranial pressure measurements were obtained in all groups. Cerebral perfusion pressures were calculated with the knowledge of intracranial pressure and mean arterial pressure values. As expected, the intraventricular hemorrhage and vehicle control groups both experienced a rise in the intracranial pressure and subsequent decline in cerebral perfusion pressure during intraventricular injection of autologous blood and artificial cerebrospinal fluid, respectively. The addition of an intraparenchymal fiberoptic pressure sensor is beneficial in monitoring precise intracranial pressure changes.

Monitoring intracranial pressure in rodent&#160;models of nontraumatic intraventricular hemorrhage is not common in the current literature. Herein, we demonstrate a technique for measuring intracranial pressure, mean arterial pressure, and cerebral perfusion pressure during intraventricular hemorrhage in a rat&#160;animal model.

비외상성 뇌실내 출혈 설치류 모델의 두개내압 모니터링

Survivors of intraventricular hemorrhage are often left with significant long-term memory impairment; thus, research utilizing intraventricular hemorrhage ...