Bu iş, Translasyonel Tıp ve Tedavi Enstitüsü dostluk ve Pennsylvania Üniversitesi ve Marlene L. Cohen ve Jerome H. Fleisch Scholar Grant, Tıp ve Mühendislik (T32HL007954) Enstitüsü eğitim bursu tarafından finanse edildi Connecticut Üniversitesi Sağlık Merkezi (LHS) ve NIH NS-029.331 (FAW).

1. Ekipmanların hazırlanması <ol><li> Bakteriyel kontaminasyonu en aza indirmek için,% 75 etanol ile stereotaksik çerçeve ve pompa aşağı doğru silin. </li><li> Hamilton şırınga ve erimiş silika iğne sterilize edin. Not: Eğer kimyasal sterilizasyon kullanılır, kullanmadan önce steril su ile birkaç kez durulama emin olun. </li><li> % 75 etanol ile parafin kağıt yüzeyi silin ve kurumasını bekleyin. </li></ol> 2. Enjeksiyon için fare hazırlanması Not: fareler var, onlara yeni bir çevre gelmesini ve stresi azaltmak için izin hayvan tesis ameliyattan önce en az 7 gün teslim. <ol><li> Ameliyat öncesi bazal için fare tartılır. </li><li> Tepkisiz kadar kuyruk tutam İsofluranın% 30 Oksijen,% 70 oksit ve% 4 ile anestezi neden </li><li> Post-operatif analjezi için buprenorfin 0.1mg/kg intraperitoneal fare enjekte </li><li> Kafa derisi Tıraş </li><li> Steril vazelin ile Coat gözleri </li><li> Betadin x 3 mendil ile kafa derisi hazırlayın, sonra kafa derisi kurumasını bekleyin </li><li> Kafa derisi steril cerrahi neşter kullanılarak 1 cm orta hat sagittal kesi Not: cömert bir kesi kafatası yerlerinden tam maruz sağlayacaktır. </li><li> Tabanından başlayan kuyruk 1 cm ventral yüzeyinin 1 cm Tıraş ve betadin x 3 mendil hazırlamak </li><li> Stereotaksik çerçeve yerleştirin fare Not: fare mükemmel bregma bregma sağ kalma ve en az 3 mm, çerçeve tabanı ile kafatası paralel yüzeyi ile çerçeve güvenli sağlamak için önemlidir. </li></ol> 3. Intraserebral kanama ameliyatı Notlar: tüm ameliyat boyunca sürekli 37 muhafaza İsofluranın% 30 Oksijen,% 70 oksit ve% 1-3, fare ile anestezi ± 0.5 ° C termistör kontrollü ısıtma yastığı ve rektal termometre ile izlenir. <ol><li> 1 cc şırınga çerçeve üzerinde steril 27 g iğne takın. </li><li> Stereotaksik kol tam bregma yerinde iğne kadar ayarlayın. </li><li> Bu iğne 2,5 mm kafatası yüzeyine bregma lateral ve alt böylece kol ayarlayın. </li><li> El tamamen kafatası delinecek çerçeve dikkate alarak hafif aşağı yönlü hareketi uygularken kafatası yüzeyinde burr hole şırınga döndürün. </li><li> Şırınga / iğne ile iğne ve tam burr hole elle çıkarın Not: Eğer kafatası ve beyin parankimi içine iğne yanlışlıkla itme riski en aza indirir iç tablo delikli varken el ile burr hole tamamlanması hemen tanınması için izin verir. </li><li> , Steril cerrahi bıçak kullanarak kuyruk ventral yüzeyinde enine kesi yapın ve arteryel kan 2-3 büyük damla parafin kağıt üzerine düşmesine izin. Sonra hızlı bir şekilde steril gazlı bez kullanarak basıncı ile kanamayı durdurmak. </li><li> 17 mcL kan Hamilton enjektör ve pompa üzerinde yer şırınga içine çekin. </li><li> Point 5 stereotaksik kol ayarlayın ° dikey eksen mediale göreceli. </li><li> Bu iğne ucu kafatasında burr hole üzerinde ve daha sonra alt iğne 3,5 mm kadar dikkatle stereotaksik kol ayarlayın. </li><li> 2 dakika sonra 0.5 mm iğne geri bekleyin (böylece ucu derin 3 mm) </li><li> Beyin iğne etrafında yeniden genişletmek ve enjeksiyon sırasında iğne ekleme parça geri kaçmasını kan riskini en aza indirmek için 5 dakika bekleyin. </li><li> 7.5 mcL kan 1 mcL / dakika enjekte edilir. </li><li> Ilk kan pıhtılaşması izin ve doku vardiya, intrakranial basıncı yükselmeleri en aza indirmek için meydana 5 dakika bekleyin. </li><li> 1 mcL / dakika kalan 7.5 mcL enjekte </li><li> Iğne, kan pıhtılaşması için izin vermek için 25 dakika boyunca yerinde kalması için izin verin Not: Başarısızlık, kan pıhtılaşması, kan iğne ekleme sitesi reflüye neden olacaktır iğne çekilirken beklemek </li><li> Yavaş yavaş geri çekilmesi ve iğne iğne herhangi bir kalıntı kan pıhtılaşma önlemek ve iğne tekrar kullanılabilirliği sağlamak için derhal sıcak su ile durulayın. </li><li> Çerçeveden çıkarın ve fareyi, veteriner cerrahi yapıştırıcı ile kuyruk ve kafa derisi kesiler yakın. </li><li> Anestezi kapalı açın. </li><li> Sürekli gıda nemlendirilmiş için ücretsiz erişim ile ısındı olurken uyandırmak için fare izin verin. </li><li> Littermates tamamen uyanık kafes fare dönün. Gıdaya erişim hayvanlara yardım etmek için kafesin altındaki ıslak gıda pelet yerleştirin. </li></ol> 4. Temsilcisi sonuçları: <img src="/files/ftp_upload/2618/2618fig1.jpg" alt="Şekil 1"/> Şekil 1. Koronal bölümünde, 15 dakika ICH cerrahi sonra fare beyin . Hemen sonra kurban beyin iğne ekleme yerinde bir koronal bölümünde brüt muayene dayalı ICH başarısı için denetlendi. Korpus kallosum geçmiş, ya da ventriküller içine iğne izi kadar, beyin tabanına paletli kanamalar başarısız sayılır ve o fare tüm analizler elendi. Genel ICH success oranları% 0 mortalite ile 50 farelerde% 75-85 idi. <img src="/files/ftp_upload/2618/2618fig2.jpg" alt="Şekil 2"/> Şekil 2 Silindir test sağ bazal ganglion ICH sonra sol hemiparezi göstermektedir. ICH cerrahi sonrası (A) Örnek fare arkada. Sadece sağ ön ayakları, sonra sol bazal ganglion ICH silindir duvara yerleştirilmesi unutmayın. (B) silindir test grafiği sonra farelerin kohortta 1 sonuç ICH cerrahi (n = 5) sahte karşılaştırıldığında (n = 4). Şam farelerin kan enjeksiyon (iğne beyin içine yerleştirilmiş) dışındaki tüm prosedürler vardı. Her fare, 12 cm çapında açık cam silindir yerleştirilir ve 20 rears gözlendi. Silindir duvara ön ayakları ilk yerleşim arka ortalama gol oldu. Fare yere geri döndü ve sonraki arka puan kadar daha sonraki hareketleri (yanal arama gibi) gol değildi. 0 hiçbir ön ayakları tercihi ve 1 sadece sağ ön ayakları nerede, (# + # sağ sol + # iki) / - lateralite indeksi olarak hesaplanır (silindir tarafında # # ön ayakları yerleşimleri sol sağ ön ayakları yerleşimleri) .

<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>

Çıkar çatışması ilan etti.

Intraserebral kanama, spontan bazal ganglia kanama tekrarlanabilir bir model otolog kuyruk arter kan sonuçlarını kullanarak farelerde Bu cerrahi modeli. Farelerde bir ICH modeli patofizyolojisi araştırmak amacıyla transgenik hayvanların durumu avantaj sunar; Ancak, bunların küçük boyutlu büyük hayvanların daha nöroşirürjikal işlemler teknik açıdan daha zor hale getirir. Kollajenaz model ve otolog kan enjeksiyonu modeli deneysel ICH iki köklü modelleri. Kollajenaz modeli...

Stereotaksik fare nöroşirürji için bir çerçeve (Stoelting, 51.925) Mikroinfüzyon pompa ve işlemci (UMP-3 ve Micro4, Dünya Hassas Aletler, Sarasota, FL) Fare sıcak (Stoelting, 50300) İnhalasyon fare anestezi (Braintree Bilimsel, EZ-AF9000) 25 mcL gaz geçirmez borosilikat Hamilton kaplı piston enjektör ve herhangi bir iğne (Hamilton şirket, Reno, NV, 1702RN şırınga: 765.401, halkalı: 30.949, mesafe: 30.946) erimiş silika iğne, 2 cm uzunluğunda (Hamilton, 17739) kesilmiş * Not Hamilton şırınga ve erimiş silika iğne eğer her ameliyat öncesi sterilize birden fazla ameliyatları için yeniden olabilir. Bu malzemeler, kan pıhtılaşmasını önlemek için çok önemlidir. Steril cerrahi eldiven Cerrahi önlük, bone ve maske Betadine % 75 etanol steril 27 g iğne (tek kullanımlık) steril 1 cc şırınga (tek kullanımlık) steril cerrahi bıçak Cidex steril su buprenorfin ve izofluran steril gazlı bez parafin kağıt kareler Veteriner cerrahi yapıştırıcı (Vetbond, 3M, St Paul, 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

Bu protokolü açıklanan farelerde intraserebral kanama otolog kan enjeksiyonu modeli, iğne izi kadar kan reflü, pompa sistemi hiçbir antikoagülanlar riskini en aza indirmek için çift enjeksiyon tekniği kullanır ve sistemdeki tüm ölü boşluk ve genişletilebilir boru ortadan kaldırır.

Farelerde Spontan İntraserebral Kanama Model Otolog Kan Enjeksiyon

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 Görüntüleme.  University of Connecticut Health Center. Bu protokolü açıklanan farelerde intraserebral kanama otolog kan enjeksiyonu modeli, iğne izi kadar kan reflü, pompa sistemi hiçbir antikoagülanlar riskini en aza indirmek için çift enjeksiyon tekniği kullanır ve sistemdeki tüm ölü boşluk ve genişletilebilir boru ortadan kaldırır.

Video: 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'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.

Yöntem Spontan Karakterizasyonu Ürkütme kaynaklı hareket yeteneği bir Rotenon ile uyarılan Parkinson Hastalığı Modeli için<em&gt; Drosophila</em

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

Nörobilim

Intracerebroventricular Injection of Amyloid-&#946; Peptides in Normal Mice to Acutely Induce Alzheimer-like Cognitive Deficits

Amyloid-&#946; (A&#946;) is a major pathological mediator of both familial and sporadic Alzheimer's disease (AD). In the brains of AD patients, progressive accumulation of A&#946; oligomers and plaques is observed. Such A&#946; abnormalities are believed to block long-term potentiation, impair synaptic function, and induce cognitive deficits. Clinical and experimental evidences have revealed that the acute increase of A&#946; levels in the brain allows development of Alzheimer-like phenotypes. Hence, a detailed protocol describing how to acutely generate an AD mouse model via the intracerebroventricular (ICV) injection of A&#946; is necessary in many cases. In this protocol, the steps of the experiment with an A&#946;-injected mouse are included, from the preparation of peptides to the testing of behavioral abnormalities. The process of preparing the tools and animal subjects before the injection, of injecting the A&#946; into the mouse brain via ICV injection, and of assessing the degree of cognitive impairment are easily explained throughout the protocol, with an emphasis on tips for effective ICV injection of A&#946;. By mimicking certain aspects of AD with a designated injection of A&#946;, researchers can bypass the aging process and focus on the downstream pathology of A&#946; abnormalities.

The amyloid-&#946; (A&#946;)-injected animal model enables the administration of a defined quantity and species of A&#946; fragments and reduces individual differences within each study group. This protocol describes the intracerebroventricular (ICV) injection of A&#946; without stereotactic instruments, enabling the production of Alzheimer-like behavioral abnormalities in normal mice.

Normal Farelerde Amiloid-β Peptitler intraserebroventriküler enjeksiyon Akut Alzheimer gibi Bilişsel Açıkları tetikleyebilecek

Amyloid-&#946; (A&#946;) is a major pathological mediator of both familial and sporadic Alzheimer's disease (AD). In the brains of AD patients, ...

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.

İntraserebral enjeksiyon stratejileri kullanarak Zika virüs kaynaklı nörolojik bozukluklar için fare modelleri oluşturma: embriyonik, Yenidoğan ve yetişkin

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.

Pro-enflamatuar Sitokinlerin İntraserebral Enjeksiyonu ile İndüklenen Yaygın Serebral Kortikal Demiyelinasyonun Sıçan Modeli

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.

Subaraknoid kanama, bir fare modeli serebral Vasospasm miktar hacimsel bir yöntemi

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.

Minimal İnvaziv Endoskopik İntraserebral Kanama Tahliyesi

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

Direct Intrathecal Injection of Recombinant Adeno-associated Viruses in Adult Mice

Intrathecal (IT) injection of adeno-associated virus (AAV) has drawn considerable interest in CNS gene therapy by virtue of its safety, noninvasiveness, and excellent transduction efficacy in the CNS.&#160;Previous studies have demonstrated the therapeutic potency of AAV-delivered gene therapy in neurodegenerative disorders by IT administration. However, high rates of unpredictable failure due to the technical limitation of IT administration in small animals have been reported. Here, we established a scoring system to indicate the success extent of lumbar puncture in small animals by adding 1% lidocaine hydrochloride into the injection solution. We further show that the extent of transient weakness following injection can predict the transduction efficiency of AAV. Thus, this IT injection method can be used to optimize therapeutic trials in mouse models of CNS diseases that afflict wide regions of the CNS.

Here we present a direct intrathecal injection technique using 1% lidocaine hydrochloride in a viral solution to ensure efficient adeno-associated virus delivery to small animals and establish a scoring system to predict transduction efficiency in the central nervous system according to the degree of transient weakness induced by lidocaine.

Rekombinant Adeno ilişkili virüs yetişkin farelerde doğrudan intratekal enjeksiyon

Intrathecal (IT) injection of adeno-associated virus (AAV) has drawn considerable interest in CNS gene therapy by virtue of its safety, noninvasiveness, ...

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.

In vivo doğrudan reprogramming Resident glial hücreler Interneurons viral vektörler ıntracerebral enjeksiyon ile

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.

Subaraknoid Kanama Modeli olarak Cisterna Magna içine Kan Çift Doğrudan Enjeksiyon

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.