Name: intraluminal middle cerebral artery occlusion (mcao) model for ischemic stroke with laser doppler flowmetry guidance in mice
Uploaded: 2011-05-08T14:00:00
Description: Watch this Scientific Journal Video about Intraluminal Middle Cerebral Artery Occlusion MCAO Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice at JoVE.com

This protocol was approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Florida and is in compliance with the 
 "Principle of Laboratory Animal Care" (NIH publication No. 86-23, revised 1985).
 1. Materials
 <ol>
 <li>Animals: Male C57BL/6 mice (Harlan Laboratories, Indianapolis, IN, USA) weighing 20&#x2013;26 grams at the time of surgery.</li>
 <li> Anesthesia: 
 <ol>
 <li>Anesthesia machine: VetEquip inhalation anesthesia system (VetEquip Inc., Pleasan...

<ol>
	<li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recommendations+on+stroke+prevention,+diagnosis,+and+therapy.+Report+of+the+WHO+Task+Force+on+Stroke+and+other+Cerebrovascular+Disorders.">Recommendations on stroke prevention, diagnosis, and therapy. Report of the WHO Task Force on Stroke and other Cerebrovascular Disorders.</a> Stroke; a journal of cerebral circulation. 20, 1407-1431 (1989).</li><li>Lloyd-Jones, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+disease+and+stroke+statistics--2009+update:+a+report+from+the+American+Heart+Association+Statistics+Committee+and+Stroke+Statistics+Subcommittee.">Heart disease and stroke statistics--2009 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee.</a> Circulation. 119, e21-e181 (2009).</li><li>Longa, E. Z., Weinstein, P. R., Carlson, S., Cummins, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversible+middle+cerebral+artery+occlusion+without+craniectomy+in+rats.">Reversible middle cerebral artery occlusion without craniectomy in rats.</a> Stroke; a journal of cerebral circulation. 20, 84-91 (1989).</li><li>Koizumi, J., Yoshida, Y., Nakazawa, T., Ooneda, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+studies+of+ischemic+brain+edema.+I.+A+new+experimental+model+of+cerebral+embolism+in+rats+in+which+recirculation+can+be+introduced+in+the+ischemic+area.">Experimental studies of ischemic brain edema. I. A new experimental model of cerebral embolism in rats in which recirculation can be introduced in the ischemic area.</a> Jpn J Stroke. 8, 1-8 (1986).</li><li>Clark, W. M., Lessov, N. S., Dixon, M. P., Eckenstein, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monofilament+intraluminal+middle+cerebral+artery+occlusion+in+the+mouse.">Monofilament intraluminal middle cerebral artery occlusion in the mouse.</a> Neurol Res. 19, 641-648 (1997).</li><li>Fujii, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strain-related+differences+in+susceptibility+to+transient+forebrain+ischemia+in+SV-129+and+C57black/6+mice.+.">Strain-related differences in susceptibility to transient forebrain ischemia in SV-129 and C57black/6 mice. .</a> Stroke; a journal of cerebral circulation. 28, 1805-1810 (1997).</li><li>Bederson, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+2,3,5-triphenyltetrazolium+chloride+as+a+stain+for+detection+and+quantification+of+experimental+cerebral+infarction+in+rats.">Evaluation of 2,3,5-triphenyltetrazolium chloride as a stain for detection and quantification of experimental cerebral infarction in rats.</a> Stroke; a journal of cerebral circulation. 17, 1304-1308 (1986).</li><li>Qi, X., Hosoi, T., Okuma, Y., Kaneko, M., Nomura, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sodium+4-phenylbutyrate+protects+against+cerebral+ischemic+injury.">Sodium 4-phenylbutyrate protects against cerebral ischemic injury.</a> Mol Pharmacol. 66, 899-908 (2004).</li><li>Barone, F. C., Knudsen, D. J., Nelson, A. H., Feuerstein, G. Z., Willette, 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=Mouse+strain+differences+in+susceptibility+to+cerebral+ischemia+are+related+to+cerebral+vascular+anatomy.">Mouse strain differences in susceptibility to cerebral ischemia are related to cerebral vascular anatomy.</a> J Cereb Blood Flow Metab. 13, 683-692 (1993).</li><li>Tureyen, K., Vemuganti, R., Sailor, K. A., Dempsey, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ideal+suture+diameter+is+critical+for+consistent+middle+cerebral+artery+occlusion+in+mice.">Ideal suture diameter is critical for consistent middle cerebral artery occlusion in mice.</a> Neurosurgery. 56, 196-200 (2005).</li><li>Barber, P. A., Hoyte, L., Colbourne, F., Buchan, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temperature-regulated+model+of+focal+ischemia+in+the+mouse:+a+study+with+histopathological+and+behavioral+outcomes.">Temperature-regulated model of focal ischemia in the mouse: a study with histopathological and behavioral outcomes.</a> Stroke; a journal of cerebral circulation. 35, 1720-1725 (2004).</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></ol>

The MCAO model is an established model that replicates cerebral ischemia in murine species. This model was first used in rats and thereafter 
adapted to mice. Avoiding craniectomy to eliminate the influence of surgical manipulation on blood brain barrier permeability and intracranial pressure is an advantage 
of this technique. However, many variables, such as strain-related differences10, size of suture tip11, duration of occlusion6, 
body temperature12, anesthesia13</su...

<table border="1">
	<tbody>
		<tr>
			<th>Name of the reagent</th>
			<th>Type</th>
			<th>Company</th>
			<th>Catalogue number</th>
			<th>Comments (optional)</th>
		</tr>
		<tr>
			<td>Male C57BL/6 mice</td>
			<td>Animal</td>
			<td>Harlan Laboratories, Indianapolis, IN, USA</td>
			<td>&nbsp;</td>
			<td>20-26 grs</td>
		</tr>
		<tr>
			<td>Dissection microscope</td>
			<td>Microscope</td>
			<td>Leica stereo Microscope MZ6</td>
			<td>&nbsp;</td>
			<td>6.3:1 zoom</td>
		</tr>
		<tr>
			<td>Fiber optic illuminator</td>
			<td>Illuminator</td>
			<td>TechniQuip Corp., Livermore, CA, USA</td>
			<td>&nbsp;</td>
			<td>FO1&#x2013;150</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Anesthesia</td>
			<td>Baxter Pharmaceutics, Deerfield, IL</td>
			<td>1001936060</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>SuperCut Iris scissor, straight</td>
			<td>Surgical tool</td>
			<td>World Precision Instruments, Inc., Sarasota, FL, USA</td>
			<td>14218</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Vannas micro-scissor, straight</td>
			<td>Surgical tool</td>
			<td>World Precision Instruments, Inc., Sarasota, FL, USA</td>
			<td>14003</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Dumont fine tip forceps</td>
			<td>Surgical tool</td>
			<td>World Precision Instruments, Inc., Sarasota, FL, USA</td>
			<td>503290</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Vessel clip</td>
			<td>Surgical tool</td>
			<td>World Precision Instruments, Inc., Sarasota, FL, USA</td>
			<td>14120</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>High Frequency Desiccator 900</td>
			<td>Surgical tool</td>
			<td>World Precision Instruments, Inc., Sarasota, FL, USA</td>
			<td>500397</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>7-0 surgical Nylon monofilament suture</td>
			<td>Suture</td>
			<td>Ethicon, Inc., Somerville, NJ, USA</td>
			<td>1647G</td>
			<td>Suture for occlusion</td>
		</tr>
		<tr>
			<td>7-0 surgical Silk suture</td>
			<td>Suture</td>
			<td>Ethicon, Inc., Somerville, NJ, USA</td>
			<td>7733G</td>
			<td>Suture for ligation</td>
		</tr>
		<tr>
			<td>Silicone RTV adhesive</td>
			<td>Silicone rubber</td>
			<td>World Precision Instruments, Inc., Sarasota, FL, USA</td>
			<td>1571</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Vetbond</td>
			<td>Tissue adhesive</td>
			<td>3M Company, St Paul, MN, USA</td>
			<td>70200746587</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Puralube Vet Ointment</td>
			<td>Eye ointment</td>
			<td>Pharmaderm, Melville, NY, USA</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Buprenorphine hydrochlorode</td>
			<td>Analgesics</td>
			<td>Hospira Inc., Lake Forest, IL, USA</td>
			<td>NDC 0409-2012-32</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Thermal Cautery Unit</td>
			<td>&nbsp;</td>
			<td>World Precision Instruments, Inc., Sarasota, FL, USA</td>
			<td>501292</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>PeriFlux System 5000</td>
			<td>TCD flowmetry</td>
			<td>Perimed, Stockholm, Sweeden</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>BAT-12</td>
			<td>Rectal thermometer</td>
			<td>World Precision Instruments, Inc., Sarasota, FL, USA</td>
			<td>BAT-12R</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>T/PUMP, TP600</td>
			<td>Thermal blanket</td>
			<td>Gaymar, NY, USA</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>2,3,5-triphenyltetrazolium chloride</td>
			<td>Reagent</td>
			<td>Sigma-Aldrich Co., St Louis, MO, USA</td>
			<td>T8877</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Mice brain matrix slicer</td>
			<td>Brain slicer</td>
			<td>Zivic-Miller Lab., Inc., Allison Park, PA, USA</td>
			<td>BSMAS001-1</td>
			<td>&nbsp;</td>
		</tr>
	</tbody>
</table>

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.

Watch this Scientific Journal Video about Intraluminal Middle Cerebral Artery Occlusion MCAO Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice at JoVE.com

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 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 ...

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

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

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

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