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Neurosci. 27, 7083-7093 (2007).</li><li>Dirnagl, U., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standard+operating+procedures+(SOP)+in+experimental+stroke+research:+SOP+for+middle+cerebral+artery+occlusion+in+the+mouse.">Standard operating procedures (SOP) in experimental stroke research: SOP for middle cerebral artery occlusion in the mouse.</a> Nature Precedings. , (2010).</li><li>Engel, O., Kolodziej, S., Dirnagl, U., Prinz, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+stroke+in+mice+-+middle+cerebral+artery+occlusion+with+the+filament+model.">Modeling stroke in mice - middle cerebral artery occlusion with the filament model.</a> J. Vis. Exp. (47), e2423 (2011).</li><li>Bederson, J. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+middle+cerebral+artery+occlusion:+evaluation+of+the+model+and+development+of+a+neurologic+examination.">Rat middle cerebral artery occlusion: evaluation of the model and development of a neurologic examination.</a> Stroke. 17, 472-476 (1986).</li><li>Lin, T. N., He, Y. Y., Wu, G., Khan, M., Hsu, C. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+brain+edema+on+infarct+volume+in+a+focal+cerebral+ischemia+model+in+rats.">Effect of brain edema on infarct volume in a focal cerebral ischemia model in rats.</a> Stroke. 24, 117-121 (1993).</li><li>Kawano, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=iNOS-derived+NO+and+nox2-derived+superoxide+confer+tolerance+to+excitotoxic+brain+injury+through+peroxynitrite.">iNOS-derived NO and nox2-derived superoxide confer tolerance to excitotoxic brain injury through peroxynitrite.</a> J. Cereb. Blood Flow Metab. 27, 1453-1462 (2007).</li><li>Qi, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sublethal+cerebral+ischemia+inhibits+caspase-3+activation+induced+by+subsequent+prolonged+ischemia+in+the+C57Black/Crj6+strain+mouse.">Sublethal cerebral ischemia inhibits caspase-3 activation induced by subsequent prolonged ischemia in the C57Black/Crj6 strain mouse.</a> Neurosci. Lett. 315, 133-136 (2001).</li><li>Wu, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+forebrain+ischemic+preconditioning+model+established+in+C57Black/Crj6+mice.">A forebrain ischemic preconditioning model established in C57Black/Crj6 mice.</a> J. Neurosci. Methods. 107, 101-106 (2001).</li><li>Liu, S., Zhen, G., Meloni, B. P., Campbell, K., Winn, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+Stroke+Model+Guidelines+for+Preclinical+Stroke+Trials+(1st+Edition).">Rodent Stroke Model Guidelines for Preclinical Stroke Trials (1st Edition).</a> J. Exp. Stroke Transl. Med. 2, 2-27 (2009).</li><li>Nishio, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothermia-induced+ischemic+tolerance.">Hypothermia-induced ischemic tolerance.</a> Ann. N.Y. Acad. Sci. 890, 26-41 (1999).</li><li>DeBow, S., Colbourne, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+temperature+measurement+and+regulation+in+awake+and+freely+moving+rodents.">Brain temperature measurement and regulation in awake and freely moving rodents.</a> Methods. 30, 167-171 (2003).</li><li>Colbourne, F., Sutherland, G. R., Auer, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+automated+system+for+regulating+brain+temperature+in+awake+and+freely+moving+rodents.">An automated system for regulating brain temperature in awake and freely moving rodents.</a> J. Neurosci. Methods. 67, 185-190 (1996).</li><li>Kapinya, K. J., Prass, K., Dirnagl, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoflurane+induced+prolonged+protection+against+cerebral+ischemia+in+mice:+a+redox+sensitive+mechanism?.">Isoflurane induced prolonged protection against cerebral ischemia in mice: a redox sensitive mechanism?.</a> Neuroreport. 13, 1431-1435 (2002).</li><li>Li, L., Zuo, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoflurane+preconditioning+improves+short-term+and+long-term+neurological+outcome+after+focal+brain+ischemia+in+adult+rats.">Isoflurane preconditioning improves short-term and long-term neurological outcome after focal brain ischemia in adult rats.</a> Neuroscience. 164, 497-506 (2009).</li><li>Xiong, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preconditioning+with+isoflurane+produces+dose-dependent+neuroprotection+via+activation+of+adenosine+triphosphate-regulated+potassium+channels+after+focal+cerebral+ischemia+in+rats.">Preconditioning with isoflurane produces dose-dependent neuroprotection via activation of adenosine triphosphate-regulated potassium channels after focal cerebral ischemia in rats.</a> Anesth. Analg. 96, 233-237 (2003).</li><li>Spratt, N. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modification+of+the+method+of+thread+manufacture+improves+stroke+induction+rate+and+reduces+mortality+after+thread-occlusion+of+the+middle+cerebral+artery+in+young+or+aged+rats.">Modification of the method of thread manufacture improves stroke induction rate and reduces mortality after thread-occlusion of the middle cerebral artery in young or aged rats.</a> J. Neurosci. Methods. 155, 285-290 (2006).</li><li>Zarow, G. J., Karibe, H., States, B. A., Graham, S. H., Weinstein, P. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endovascular+suture+occlusion+of+the+middle+cerebral+artery+in+rats:+effect+of+suture+insertion+distance+on+cerebral+blood+flow,+infarct+distribution+and+infarct+volume.">Endovascular suture occlusion of the middle cerebral artery in rats: effect of suture insertion distance on cerebral blood flow, infarct distribution and infarct volume.</a> Neurol. Res. 19, 409-416 (1997).</li></ol>

יש לנו מה למסור.

במחקר שלנו, שהראינו כי שלושה רצפים של BCCAO, שכל אחד שנמשך לרגע אחד ואחריו reperfusion במשך חמש דקות, הם גירוי מחשב איסכמי מספיק כדי לגרום לתחילת ה-IT. יתר על כן, יש לנו הראינו כי פרוטוקול preconditioning משמש הוא הליך בטוח וקטין פולשני ללא תחלואה ל?...

שבץ איסכמי הוא מחלה עם תמותה גבוהה וניטל כלכלי וחברתי עצום 1. למרות מאמצים מדעיים ניסיוניים וקליניים אינטנסיביים ברחבי העשורים אחרונים, אפשרויות טיפול בחולי שבץ איסכמי חריפים נשארות מוגבלות מאוד 2. בניגוד לכך, עלייה באחוז של קשישים במדינות מפותחות באופן דרמטי להגדיל את שכיחות ושכיחות של חולים עם שבץ מוחי איסכמי בעשורים הקרובים 3. לכן, יש צורך דחוף באסטרטגיות טיפול חדשות בחולים עם שבץ מוחי איסכמי. גישה אחת היא להשיג הבנה נוספת במנגנוני הסתגלות אנדוגני של המוח להתמודד עם גירוי מזיק. neuroprotection המוח המופק מזה ידוע גם בשם נפשית מראש איסכמי (PC), או סובלנות איסכמי (IT), ומתאר תופעה, שבו גירוי מזיק שאינם מזיק להחיל את המוח, גורם להתנגדות חולפתנגד 4 עלבון איסכמי מזיק שלאחר מכן. היא מתרחשת בשני חלונות זמן שונים: מוקדם זה, המתרחש בתוך דקות עד כמה שעות לאחר מחשב ועיכבו את ה-IT, אשר צריך השהיה של כמה שעות להתרחש 4. עד כה, מחקר על זה במוח התמקד בעכב אותו. הרבה פחות ידוע על המנגנונים של תחילת ה-IT. מטרתו של מחקר זה היא לבסס דו צדדי חסימת עורק תרדמה משותפת (BCCAO) שהיא גירוי כדי לגרום לעיכוב זה, כמו גירוי מחשב נאות הוקם כדי לגרום לתחילת ה-IT לאיסכמיה מוחית מוקדי חולפים (הנגרם על ידי חסימה של עורק מוח אמצעי, MCAO) בC57BL6 / עכברי ג&#39;יי. במהלך שני הניתוחים (BCCAO וMCAO), ניטור של זרימת דם במוח (CBF) על ידי לייזר דופלר Flowmetry (LDF) בוצע. BCCAO בוצע בעכברים מורדמים ונושמים באופן ספונטני. שני עורקי התרדמה משותפים (CCA) נחשפווהאפילו על 60 שניות ואחרי 5 דקות של reperfusion. רצף / reperfusion BCCAO זה חזר על עצמו פעמיים. לאחר ניתוח, עכברים התאוששו במהירות ולא הראו שום סימנים של ירידה תפקודית. כדי לשלול, כי פרוטוקול BCCAO מתואר לעיל לא לגרום למוות של תאים במוח מתעכב, ביצענו תיוג TdT בתיווך dUTP-ביוטין ניק סוף (Tunel) מכתים 72 שעות לאחר BCCAO (או ניתוח דמה) ב קבוצה נפרדת. MCAO הוא מודל נרחב הוקם לזירוז של איסכמיה המוחית מוקדי במכרסמים 5-7. הניתוחים בוצעו באמצעות נהלי עבודה אחידים (SOP) 8. לאחר חשיפת המרכז לאמנות עכשווית שמאל, חוט חד סיליקון מכוסה הוכנס המרכז לאמנות העכשווית דיסטלי, התקדמה דרך עורק התרדמה הפנימי (הרשפ&quot;ת) למעגל וויליס ועד עורק המוח הקדמי (ACA) ובכך, מקורו של עורק המוח האמצעי ( MCA) היה האפיל לתקופה מוגדרת של זמן. במחקר זה, שאנובשימוש 45 דקות של MCAO, שתוצאתה בדרך כלל בנגע איסכמי בטריטוריה של MCA אזורים במוח מעורב striatal וקליפת המוח. במחקר שלנו, נתחנו את הפרופיל הזמני של תחילת IT באמצעות BCCAO כגירוי מחשב. הנתונים שלנו מצביעים, כי עיכוב הזמן האופטימלי בין BCCAO וMCAO לגרום מוקדם זה הוא 30 דקות. בסרטון הזה, אנו נותנים הדגמה של שני הניתוחים, כלומר BCCAO וMCAO.

<table border="1">
<tbody>
<tr>
	<td>Name of the instrument</td>
 <td>Company</td>
 <td>Catalogue number/model</td>
</tr>
<tr>
	<td>Binocular surgical microscope</td>
 <td>Zeiss</td>
 <td>Stemi 2000 C</td>
</tr>
<tr>
	<td>Light source for microscope</td>
 <td>Zeiss</td>
 <td>SteREO CL 1500 ECO</td>
</tr>
<tr>
	<td>Heating pad with rectal probe</td>
 <td>FST</td>
 <td>21061-10</td>
</tr>
<tr>
	<td>Stereotactic frame</td>
 <td>David Kopf</td>
 <td>Model 930</td>
</tr>
<tr>
	<td>Scissors</td>
 <td>FST</td>
 <td>91460-11</td>
</tr>
<tr>
	<td>Dumont forceps #5</td>
 <td>FST</td>
 <td>11251-10</td>
</tr>
<tr>
	<td>Dumont forceps #7</td>
 <td>FST</td>
 <td>11271-30</td>
</tr>
<tr>
	<td>Mircovascular clamp</td>
 <td>FST</td>
 <td>00398-02</td>
</tr>
<tr>
	<td>Clamp applicator</td>
 <td>FST</td>
 <td>00072-14</td>
</tr>
<tr>
	<td>Springscissors</td>
 <td>FST</td>
 <td>15372-62</td>
</tr>
<tr>
	<td>Needle holder</td>
 <td>FST</td>
 <td>12010-14</td>
</tr>
<tr>
	<td>Needles	</td>
 <td>Feuerstein, Suprama	</td>
 <td>BER 562-20</td>
</tr>
<tr>
	<td>5-0 silk suture</td>
 <td>Feuerstein, Suprama	</td>
 <td></td>
</tr>
<tr>
	<td>7-0 silk suture</td>
 <td>Feuerstein,Suprama	</td>
 <td></td>
</tr>
<tr>
	<td>8-0 silk suture</td>
 <td>Feuerstein, Suprama	</td>
 <td></td>
</tr>
</tbody>
</table>

bilateral common carotid artery occlusion as an adequate preconditioning stimulus to induce early ischemic tolerance to focal cerebral ischemia

יש מצטבר ראיות, שנפשי מראש איסכמי - אתגר איסכמי שאינו מזיק למוח - מעניק הגנה זמנית לעלבון איסכמי מזיק שלאחר מכן. הקמנו חסימת עורק תרדמה משותפת דו צדדית כגירוי נפשית מראש כדי לגרום לסובלנות איסכמי מוקדם לאיסכמיה מוחית חולפת במוקד C57BL6 / עכברי J. בסרטון הזה, נדגים את המתודולוגיה ששמשה למחקר זה.

פרוטוקול BCCAO שלנו הסתיים בירידות CBF מיידיות ועמוקות (&gt; 90%), כאשר שניהם היו המרכז לאמנות עכשווית של האפיל (איור 1). פתיחתו מחדש של המרכז לאמנות העכשווית שהובילה להשלמת reperfusion כפיקוח על ידי LDF (איור 1). לאחר מחשב, בעלי חיים לא הראו סימנים של נכות תפקודית. ית...

Watch this Scientific Journal Video about Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia at JoVE.com

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

יש מצטבר ראיות, שנפשי מראש איסכמי (PC) - אתגר איסכמי שאינו מזיק למוח - מעניק הגנה זמנית לעלבון איסכמי מזיק שלאחר מכן. הקמנו הבילטרלי חסימת עורק תרדמה משותפת (BCCAO) כגירוי נפשית מראש כדי לגרום לסובלנות איסכמי מוקדם (IT) לאיסכמיה מוחית מוקדי חולפים (הנגרם על ידי חסימה של עורק מוח אמצעי, MCAO) בC57BL6 / י עכברים.

ספיגה דו צדדי משותף עורק תרדמה כגירוי נפשית מראש מספיק כדי לגרום סובלנות איסכמי מוקדמת לאיסכמיה מוחית מיקוד

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

bilateral-common-carotid-artery-occlusion-as-an-adequate

Research

JoVE Journal

Medicine

28.8K Views.  Charité - Universitätsmedizin Berlin, Germany. יש מצטבר ראיות, שנפשי מראש איסכמי (PC) - אתגר איסכמי שאינו מזיק למוח - מעניק הגנה זמנית לעלבון איסכמי מזיק שלאחר מכן. הקמנו הבילטרלי חסימת עורק תרדמה משותפת (BCCAO) כגירוי נפשית מראש כדי לגרום לסובלנות איסכמי מוקדם (IT) לאיסכמיה מוחית מוקדי חולפים (הנגרם על ידי חסימה של ?...

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

Mouse Model of Middle Cerebral Artery Occlusion

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the brain (ischemic stroke) 2. Since most ischemic strokes (~80%) occur in the territory of middle cerebral artery (MCA) 3, many animal stroke models that have been developed have focused on this artery. The intraluminal monofilament model of middle cerebral artery occlusion (MCAO) involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the MCA territory 4. The technique can be used to model permanent or transient occlusion 5. If the suture is removed after a certain interval (30 min, 1 h, or 2 h), reperfusion is achieved (transient MCAO); if the filament is left in place (24 h) the procedure is suitable as a model of permanent MCAO. This technique does not require craniectomy, a neurosurgical procedure to remove a portion of skull, which may affect intracranial pressure and temperature 6. It has become the most frequently used method to mimic permanent and transient focal cerebral ischemia in rats and mice 7,8. To evaluate the extent of cerebral infarction, we stain brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) to identify ischemic brain tissue 9. In this video, we demonstrate the MCAO method and the determination of infarct size by TTC staining.

We demonstrate in the video a method for producing a middle cerebral artery occlusion in adult mice using an intraluminal monofilament. We also show how to evaluate the extent of cerebral infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining.

עכבר דגם ספיגה התיכון עורק מוחי

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the ...

Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United States in 20091,2. The intraluminal middle cerebral artery occlusion (MCAO) model was developed by Koizumi4 in 1986 to simulate this 
impactful human pathology in the rat. A modification of the MCAO method was later presented by Longa3. Both techniques have been widely used to identify 
molecular mechanisms of brain injury resulting from ischemic stroke and potential therapeutic modalities5. This relatively noninvasive method in rats has been 
extended to use in mice to take advantage of transgenic and knockout strains6,7. To model focal cerebral ischemia, an intraluminal suture is advanced via 
the internal carotid artery to occlude the base of the MCA. Retracting the suture after a specified period of time mimics spontaneous reperfusion, but the 
suture can also be permanently retained. This video will be demonstrating the two major approaches for performing intraluminal MCAO procedure in mice in a 
stepwise fashion, as well as providing insights for potential drawbacks and pitfalls. The ischemic brain tissue will subsequently be stained by 
2,3,5-triphenyltetrazolium chloride (TTC) to evaluate the extent of cerebral infarction8.

Intraluminal Middle Cerebral Artery Occlusion MCAO Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

The intraluminal middle cerebral artery occlusion (MCAO) model is the most frequent used model among experimental ischemic stroke models. Here we will demonstrate the entire model in detail with the guide of Laser Doppler flowmetry, and its representative results.

Intraluminal העורקים התיכון מוחין ספיגה (MCAO) מודל שבץ איסכמי עם הדרכה דופלר Flowmetry לייזר עכברים

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United ...

Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in 20091, 2. Several models of cerebral ischemia have been developed to mimic the human condition of stroke. It has been suggested that up to 80% of all strokes result from ischemic damage in the middle cerebral artery (MCA) area3. In the early 1990s, endothelin-1 (ET-1) 4 was used to induce ischemia by applying it directly adjacent to the surface of the MCA after craniotomy. Later, this model was modified 5 by using a stereotaxic injection of ET-1 adjacent to the MCA to produce focal cerebral ischemia. The main advantages of this model include the ability to perform the procedure quickly, the ability to control artery constriction by altering the dose of ET-1 delivered, no need to manipulate the extracranial vessels supplying blood to the brain as well as gradual reperfusion rates that more closely mimics the reperfusion in humans5-7. On the other hand, the ET-1 model has disadvantages that include the need for a craniotomy, as well as higher variability in stroke volume8. This variability can be reduced with the use of laser Doppler flowmetry (LDF) to verify cerebral ischemia during ET-1 infusion. Factors that affect stroke variability include precision of infusion and the batch of the ET-1 used6. Another important consideration is that although reperfusion is a common occurrence in human stroke, the duration of occlusion for ET-1 induced MCAO may not closely mimic that of human stroke where many patients have partial reperfusion over a period of hours to days following occlusion9, 10. This protocol will describe in detail the ET-1 induced MCAO model for ischemic stroke in rats. It will also draw attention to special considerations and potential drawbacks throughout the procedure.

Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed.

אנדותלין-1 דגם מושרה מזרח מוחות עורקים ספיגים לשבץ איסכמי עם הדרכת Flowmetry ליזר דופלר בעכברוש

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in ...

Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be performed in many locations overlying the rat cerebrum, but is most easily carried out by creating a craniectomy over the readily accessible frontal or parietal bones. Most frequently, we have used this technique in combination with the endothelin-1 middle cerebral artery occlusion model of ischemic stroke to quantify the changes in middle cerebral artery vessel diameter that occur with injection of endothelin-1 into the brain parenchyma adjacent to the proximal MCA4, 5. In order to visualize the proximal portion of the MCA during endothelin -1 induced MCAO, we use a technique to create a cranial window through the temporal bone on the lateral aspect of the rat skull (Figure 1). Cerebral arteries can be visualized either with the dura intact or with the dura incised and retracted. Most commonly, we leave the dura intact during visualization since endothelin-1 induced MCAO involves delivery of the vasoconstricting peptide into the brain parenchyma. This bypasses the need to incise the dura directly over the visualized vessels for drug delivery. This protocol will describe how to create a cranial window to visualize cerebral arteries in a step-wise fashion, as well as how to avoid many of the potential pitfalls pertaining to this method.

This article describes a method for visualizing rat cerebral arteries through a cranial window using temporal craniectomy in order to view proximal portions of the middle cerebral artery (Figure 1). This versatile method can be combined with various techniques of drug delivery to measure cerebral artery reactivity in vivo.

ניצול חלון גולגולת לדמיין עורק המוח האמצעי במהלך ספיג אנדותלין-1 מושרה מזרח מוחות עורקים

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be ...

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

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

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

למבוגרים עכבר ורידי יתר לחץ דם דגם: עורק תרדמה משותף לחיצוני הצוואר וריד השקה.

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

Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic stroke. However, the use of tPA is limited by its narrow therapeutic window and by increased risk of hemorrhagic transformation. There is an urgent need to develop suitable stroke models to study new thrombolytic agents and strategies for treatment of ischemic stroke. At present, two major types of ischemic stroke models have been developed in rats and mice: intraluminal suture MCAO and embolic MCAO. Although MCAO models via the intraluminal suture technique have been widely used in mechanism-driven stroke research, these suture models do not mimic the clinical situation and are not suitable for thrombolytic studies. Among these models, the embolic MCAO model closely mimics human ischemic stroke and is suitable for preclinical investigation of thrombolytic therapy. This embolic model was first developed in rats by Overgaard et al.1 in 1992 and further characterized by Zhang et al. in 19972. Although embolic MCAO has gained increasing attention, there are technical problems faced by many laboratories. To meet increasing needs for thrombolytic research, we present a highly reproducible model of embolic MCAO in the rat, which can develop a predictable infarct volume within the MCA territory. In brief, a modified PE-50 tube is gently advanced from the external carotid artery (ECA) into the lumen of the internal carotid artery (ICA) until the tip of the catheter reaches the origin of the MCA. Through the catheter, a single homologous blood clot is placed at the origin of the MCA. To identify the success of MCA occlusion, regional cerebral blood flow was monitored, neurological deficits and infarct volumes were measured. The techniques presented in this paper should help investigators to overcome technical problems for establishing this model for stroke research.

Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats

In this paper, we demonstrate a standard method for producing an embolic middle cerebral artery occlusion with homologous blood clots (fibrin-rich) in adult rat. This model closely mimics human ischemic stroke and is suitable for preclinical study of thrombolytic therapy for ischemic stroke.

עורקי תסחיפים התיכון מוחין ספיגה (MCAO) לשבץ איסכמי עם קרישי דם הומולוגיים בחולדות

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic ...

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week protocol for repetitive hypoxic preconditioning (RHP) induces long-term protection against central nervous system (CNS) injury in a mouse model of focal ischemic stroke. RHP consists of 9 stochastic exposures to hypoxia that vary in both duration (2 or 4 hr) and intensity (8% and 11% O2). RHP reduces infarct volumes, blood-brain barrier (BBB) disruption, and the post-stroke inflammatory response for weeks following the last exposure to hypoxia, suggesting a long-term induction of an endogenous CNS-protective phenotype. The methodology for the dual quantification of infarct volume and BBB disruption is effective in assessing neurovascular protection in mice with RHP or other putative neuroprotectants. Adult male Swiss Webster mice were preconditioned by RHP or duration-equivalent exposures to 21% O2 (i.e. room air). A 60 min transient middle cerebral artery occlusion (tMCAo) was induced 2 weeks following the last hypoxic exposure. Both the occlusion and reperfusion were confirmed by transcranial laser Doppler flowmetry. Twenty-two hr after reperfusion, Evans Blue (EB) was intravenously administered through a tail vein injection. 2 hr later, animals were sacrificed by isoflurane overdose and brain sections were stained with 2,3,5- triphenyltetrazolium chloride (TTC). Infarcts volumes were then quantified. Next, EB was extracted from the tissue over 48 hr to determine BBB disruption after tMCAo. In summary, RHP is a simple protocol that can be replicated, with minimal cost, to induce long-term endogenous neurovascular protection from stroke injury in mice, with the translational potential for other CNS-based and systemic pro-inflammatory disease states.

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

כימות של הגנת עצבים וכלי דם בעקבות חסימת עורק חוזר חוסר חמצן אכשור קד וחלוף התיכון מוחין של עכברים

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week ...

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.

Performing מוחות תיכונים דיסטלי קבוע עם ספיגה עורקת Common הראש אצל חולדות בנים לחקר קורטיקלי איסכמיה עם נכות קבועה זה

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.

אין ויוו העברה גנטית כדי אנדותל העורק נפוצות הארנב

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.

דגם ארנב של ביטוי Transgene עמיד ומוחלף כדי שתלי Interposition נפוצים של העורק הראשי

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