Name: photothrombosis-induced focal ischemia as a model of spinal cord injury in mice
Uploaded: 2015-07-16T15:00:00
Description: Watch this Scientific Journal Video about Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice at JoVE.com

Dit werk werd ondersteund door de National Institutes of Health [Grant niet. R01NS069726] en de American Heart Association Grant in Aid Grant [Grant niet. 13GRNT17020004] naar SD.

Opmerking: Muizen (C57BL / 6J, man) leeftijd 10-12 weken werden gebruikt in deze studie. Alle procedures werden uitgevoerd in overeenstemming met de NIH Gids voor de Zorg en gebruik van proefdieren en werden goedgekeurd door de Universiteit van Missouri Institutional Animal Care en gebruik Comite (IACUC). 1. Pre-Surgery <ol><li> De dag voor de operatie autoclaaf en steriliseren van alle chirurgische instrumenten. Wikkel de instrumenten en de autoclaaf bij 121 ° C bij...

<ol>
	<li>Cadotte, D. W., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+injury:+a+systematic+review+of+current+treatment+options.">Spinal cord injury: a systematic review of current treatment options.</a> Clin Orthop Relat Res. 469 (3), 732-741 (2011).</li><li>Cheriyan, 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=Spinal+cord+injury+models:+a+review.">Spinal cord injury models: a review.</a> Spinal Cord. 52 (8), 588-595 (2014).</li><li>Young, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Secondary+injury+mechanisms+in+acute+spinal+cord+injury.">Secondary injury mechanisms in acute spinal cord injury.</a> J Emerg Med. 11, 13-22 (1993).</li><li>Crowe, M. J., Bresnahan, J. C., Shuman, S. L., Masters, J. N., Beattie, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apoptosis+and+delayed+degeneration+after+spinal+cord+injury+in+rats+and+monkeys.">Apoptosis and delayed degeneration after spinal cord injury in rats and monkeys.</a> Nat Med. 3 (1), 73-76 (1997).</li><li>Soubeyrand, M., 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=Effect+of+norepinephrine+on+spinal+cord+blood+flow+and+parenchymal+hemorrhage+size+in+acute-phase+experimental+spinal+cord+injury.">Effect of norepinephrine on spinal cord blood flow and parenchymal hemorrhage size in acute-phase experimental spinal cord injury.</a> Eur Spine J. 23 (3), 658-665 (2014).</li><li>Soubeyrand, M., 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=Real-time+and+spatial+quantification+using+contrast-enhanced+ultrasonography+of+spinal+cord+perfusion+during+experimental+spinal+cord+injury.">Real-time and spatial quantification using contrast-enhanced ultrasonography of spinal cord perfusion during experimental spinal cord injury.</a> Spine (Phila Pa 1976). 37 (22), E1376-E1382 (2012).</li><li>Mautes, A. E., Weinzierl, M. R., Donovan, F., Noble, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+events+after+spinal+cord+injury:+contribution+to+secondary+pathogenesis).">Vascular events after spinal cord injury: contribution to secondary pathogenesis).</a> Phys Ther. 80 (7), 673-687 (2000).</li><li>Liu, X. Z., 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=Neuronal+and+glial+apoptosis+after+traumatic+spinal+cord+injury.">Neuronal and glial apoptosis after traumatic spinal cord injury.</a> J Neurosci. 17 (14), 5395-5406 (1997).</li><li>Liu, 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=An+experimental+study+of+cell+apoptosis+and+correlative+gene+expression+after+tractive+spinal+cord+injury+in+rats.">An experimental study of cell apoptosis and correlative gene expression after tractive spinal cord injury in rats.</a> Zhonghua Wai Ke Za Zhi. 42 (23), 1434-1437 (2004).</li><li>Hirose, K., 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=Activated+protein+C+reduces+the+ischemia/reperfusion-induced+spinal+cord+injury+in+rats+by+inhibiting+neutrophil+activation.">Activated protein C reduces the ischemia/reperfusion-induced spinal cord injury in rats by inhibiting neutrophil activation.</a> Ann Surg. 232 (2), 272-280 (2000).</li><li>Oyinbo, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Secondary+injury+mechanisms+in+traumatic+spinal+cord+injury:+a+nugget+of+this+multiply+cascade.">Secondary injury mechanisms in traumatic spinal cord injury: a nugget of this multiply cascade.</a> Acta Neurobiol Exp (Wars). 71 (2), 281-299 (2011).</li><li>Guerit, J. M., Dion, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=State-of-the-art+of+neuromonitoring+for+prevention+of+immediate+and+delayed+paraplegia+in+thoracic+and+thoracoabdominal+aorta+surgery).">State-of-the-art of neuromonitoring for prevention of immediate and delayed paraplegia in thoracic and thoracoabdominal aorta surgery).</a> Ann Thorac Surg. 74 (5), S1867-S1869 (2002).</li><li>Schepens, M. A., Heijmen, R. H., Ranschaert, W., Sonker, U., Morshuis, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thoracoabdominal+aortic+aneurysm+repair:+results+of+conventional+open+surgery.">Thoracoabdominal aortic aneurysm repair: results of conventional open surgery.</a> Eur J Vasc Endovasc Surg. 37 (6), 640-645 (2009).</li><li>Braeuninger, S., Kleinschnitz, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+of+focal+cerebral+ischemia:+procedural+pitfalls+and+translational+problems.">Rodent models of focal cerebral ischemia: procedural pitfalls and translational problems.</a> Exp Transl Stroke Med. 1, 8 (2009).</li><li>Carmichael, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+of+focal+stroke:+size,+mechanism,+and+purpose.">Rodent models of focal stroke: size, mechanism, and purpose.</a> NeuroRx. 2 (3), 396-409 (2005).</li><li>Dietrich, W. D., Ginsberg, M. D., Busto, R., Watson, B. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photochemically+induced+cortical+infarction+in+the+rat.+1.+Time+course+of+hemodynamic+consequences.">Photochemically induced cortical infarction in the rat. 1. Time course of hemodynamic consequences.</a> J Cereb Blood Flow Metab. 6 (2), 184-194 (1986).</li><li>Zhang, W., 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=Neuronal+protective+role+of+PBEF+in+a+mouse+model+of+cerebral+ischemia.">Neuronal protective role of PBEF in a mouse model of cerebral ischemia.</a> J Cereb Blood Flow Metab. 30 (12), 1962-1971 (2010).</li><li>Li, H., Zhang, N., Sun, G., Ding, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+the+group+I+mGluRs+reduces+acute+brain+damage+and+improves+long-term+histological+outcomes+after+photothrombosis-induced+ischaemia.">Inhibition of the group I mGluRs reduces acute brain damage and improves long-term histological outcomes after photothrombosis-induced ischaemia.</a> ASN Neuro. 5 (3), 195-207 (2013).</li><li>Li, H., 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=Histological,+cellular+and+behavioral+assessments+of+stroke+outcomes+after+photothrombosis-induced+ischemia+in+adult+mice.">Histological, cellular and behavioral assessments of stroke outcomes after photothrombosis-induced ischemia in adult mice.</a> BMC Neurosci. 15, 58 (2014).</li><li>Wang, T., Cui, W., Xie, Y., Zhang, W., Ding, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlling+the+Volume+of+the+Focal+Cerebral+Ischemic+Lesion+through+Photothrombosis.">Controlling the Volume of the Focal Cerebral Ischemic Lesion through Photothrombosis.</a> American Journal of Biomedical Sciences. 2 (1), 33-42 (2010).</li><li>Schroeter, M., Jander, S., Stoll, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+induction+of+focal+cerebral+ischemia+in+mice+by+photothrombosis+of+cortical+microvessels:+characterization+of+inflammatory+responses.">Non-invasive induction of focal cerebral ischemia in mice by photothrombosis of cortical microvessels: characterization of inflammatory responses.</a> J Neurosci Methods. 117 (1), 43-49 (2002).</li><li>Boquillon, M., Boquillon, J. P., Bralet, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photochemically+induced,+graded+cerebral+infarction+in+the+mouse+by+laser+irradiation+evolution+of+brain+edema.">Photochemically induced, graded cerebral infarction in the mouse by laser irradiation evolution of brain edema.</a> J Pharmacol Toxicol Methods. 27 (1), 1-6 (1992).</li><li>Kim, G. W., Lewen, A., Copin, J., Watson, B. D., Chan, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+cytosolic+antioxidant,+copper/zinc+superoxide+dismutase,+attenuates+blood-brain+barrier+disruption+and+oxidative+cellular+injury+after+photothrombotic+cortical+ischemia+in+mice.">The cytosolic antioxidant, copper/zinc superoxide dismutase, attenuates blood-brain barrier disruption and oxidative cellular injury after photothrombotic cortical ischemia in mice.</a> Neuroscience. 105 (4), 1007-1018 (2001).</li><li>Schmidt, A., 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=Photochemically+induced+ischemic+stroke+in+rats.">Photochemically induced ischemic stroke in rats.</a> Exp Transl Stroke Med. 4 (1), 13 (2012).</li><li>Lang-Lazdunski, 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=Spinal+cord+ischemia.+Development+of+a+model+in+the+mouse.">Spinal cord ischemia. Development of a model in the mouse.</a> Stroke. 31 (1), 208-213 (2000).</li><li>Wang, Z., 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=Development+of+a+simplified+spinal+cord+ischemia+model+in+mice.">Development of a simplified spinal cord ischemia model in mice.</a> J Neurosci Methods. 189 (2), 246-251 (2010).</li><li>Labat-gest, V., Tomasi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photothrombotic+ischemia:+a+minimally+invasive+and+reproducible+photochemical+cortical+lesion+model+for+mouse+stroke+studies.">Photothrombotic ischemia: a minimally invasive and reproducible photochemical cortical lesion model for mouse stroke studies.</a> J Vis Exp. (76), (2013).</li><li>Lu, H., 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=Induction+and+imaging+of+photothrombotic+stroke+in+conscious+and+freely+moving+rats.">Induction and imaging of photothrombotic stroke in conscious and freely moving rats.</a> J Biomed Opt. 19 (9), 96013 (2014).</li><li>Seto, A., 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=Induction+of+ischemic+stroke+in+awake+freely+moving+mice+reveals+that+isoflurane+anesthesia+can+mask+the+benefits+of+a+neuroprotection+therapy.">Induction of ischemic stroke in awake freely moving mice reveals that isoflurane anesthesia can mask the benefits of a neuroprotection therapy.</a> Front Neuroenergetics. 6 (1), (2014).</li><li>Bell, M. 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=Toll-like+receptor+4-dependent+microglial+activation+mediates+spinal+cord+ischemia-reperfusion+injury.">Toll-like receptor 4-dependent microglial activation mediates spinal cord ischemia-reperfusion injury.</a> Circulation. 128 (11 Suppl 1), S152-S156 (2013).</li><li>Smith, P. D., 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=The+evolution+of+chemokine+release+supports+a+bimodal+mechanism+of+spinal+cord+ischemia+and+reperfusion+injury.">The evolution of chemokine release supports a bimodal mechanism of spinal cord ischemia and reperfusion injury.</a> Circulation. 126 (11 Suppl 1), S110-S117 (2012).</li><li>Jia, Z., 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=Oxidative+stress+in+spinal+cord+injury+and+antioxidant-based+intervention.">Oxidative stress in spinal cord injury and antioxidant-based intervention.</a> Spinal Cord. 50 (4), 264-274 (2012).</li></ol>

In deze studie, beschreven we een fototrombotisch model van SC ischemie. Door de vooruitgang in gentechnologie is er een uitgesproken in de handel verkrijgbare transgene muizen die het mogelijk heeft gemaakt om het effect van specifieke genen die betrokken zijn bij de pathofysiologie van ischemische SC kunnen bestuderen. Het doel van de studie was om een ​​reproduceerbaar muismodel van ruggemerg ischemie ontwikkelen. Hier aangepast we een corticale PT model aan SCI induceren in muizen. Na de operati...

Traumatisch ruggenmergletsel (SCI) is een verwoestende klinische aandoening van de sensomotorische en autonome functies van de SC. Patiënten overleven SCI worden vaak achter met slopende dwarslaesie die een aanzienlijke invloed op hun dagelijkse activiteiten en de kwaliteit van het leven 1. Experimentele SCI modellen zijn een onmisbaar instrument in het wetenschappelijk onderzoek naar de pathofysiologie van SCI en bijbehorende neurale herstelprocessen begrijpen geweest. Deze modellen zijn ook gebruikt om de preklinische effectiviteit van verschillende experimentele neuroprotectieve interventies die zijn gericht op functioneel herstel testen. Momenteel is de meerderheid van de SCI-modellen in de praktijk in dienst van het gebruik van fysieke stomp mechanisch verstoren en verwonden van de SC. Deze werkwijzen omvatten kneuzing, compressie, dislocatie en doorsnijding van de SC 2. Gesuggereerd is dat na de primaire mechanische insult secundaire beschadiging in de vorm van ischemie sets in de gewonde SC<html> 3,4. De etiologie van secundaire ischemie bevat uitgebreide weefsel degeneratie, parenchymale bloedingen en soms door verstopping van bloedvaten door weefseloedeem 5-7. Door de secundaire schade de integriteit van SC wordt verder beïnvloed worden neuronen en gliacellen ernstig verminderde functie en levensvatbaarheid en apoptose ondergaan die leidt tot groei infarct in de chronische fase van letsel, analoog aan de groei van ischemische penumbra na beroerte 8,9. Verschillende mechanismen als excitotoxiciteit, vrije radicalen en ontstekingen zijn gerapporteerd verantwoordelijk voor ischemische celdood na SCI 10,11 te zijn. Bovendien, SC ischemie is een ernstige complicatie van thoraco-abdominale aorta aneurysma chirurgie die vaak leiden tot paraplegie bij de patiënten 12,13. Ondanks deze grote klinische effect weinig modellen van ruggemerg ischemie hoge reproduceerbaarheid beschikbaar. nt &quot;&gt; Photothrombosis (PT) is een algemeen gebruikte werkwijze voor de inductie van focale ischemie in de hersenen 14-20. De techniek is tamelijk non-invasieve, uiterst reproduceerbaar en levert een nauwkeurige focale ischemische laesie op het blootgestelde oppervlak van de hersenen 17 -21. Dit wordt bereikt door de systemische toediening van de fotoactieve kleurstoffen zoals Rose Bengal (RB) 16-20,22 of erythrosine B 23 gevolgd door lokale bestraling van bloedvaten met de juiste lichtbron. Fotoactivatie van de kleurstof zorgt ervoor dat de vorming van vrije radicalen die verstoort de integriteit van de gladde vasculaire endotheel, waardoor de bloedplaatjes te verzamelen, die vervolgens vormt een thrombus. De obstructie van de bloedstroom door de thrombus resulteert in een infarct in de door het vat 24 toegevoerd regio. wegens gemak van controle van de intensiteit en duur van de bestraling procedure levert een zeer gelijkmatige en reproduceerbare infarct. Bovendien kan deze methode worden toegepast om een ​​infarc inducerent op verschillende anatomische locaties waardoor ruimtelijke (bv grijze stof versus witte stof) inzicht in het effect van ischemie. Het doel van dit onderzoek is het eenvoudig en zeer reproduceerbaar model van SC ischemie in muizen ontwikkelen. We beschreven procedure van een PT model SC ischemie in muizen. Resultaten van histologie en immunokleuring aangetoond dat PT effectief induceren SC infarct, neuronaal verlies en reactieve gliosis.

<table border="0" cellpadding="0" cellspacing="0" width="1050">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">&#160;Rose Bengal</td>
			<td style="width:233px;">Sigma-Aldrich</td>
			<td style="width:180px;">330000</td>
			<td style="width:421px;">20 mg/ml in sterile saline</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">C57Bl6/J</td>
			<td>Jackson lab</td>
			<td>664</td>
			<td>22-25g</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ketamine&#160;</td>
			<td>VEDCO</td>
			<td>NDC-50989-996-06</td>
			<td>100 mg/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Xylazine&#160;</td>
			<td>VEDCO</td>
			<td>NDC-50989-234-11</td>
			<td>100 mg/ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Betadine solution</td>
			<td>Purdue</td>
			<td>NDC-67618-150-01</td>
			<td>10% povidone iodine topical solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Normal saline</td>
			<td>Abott Laboratories</td>
			<td>04930-04-10</td>
			<td>For diluting RB, anaesthesia and for preventing tissue from drying</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Artificial tears ointment&#160;</td>
			<td>Rugby</td>
			<td>NDC-0536-6550-91</td>
			<td>83% white petrolatum</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ethanol</td>
			<td>Decon labs.Inc</td>
			<td>2716</td>
			<td>70% ethanol for disinfection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Metal halide lamp</td>
			<td>EXFO, Canada</td>
			<td>X-Cite 120 PC&#160;</td>
			<td>Set power at 12%</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Spring scissors&#160;</td>
			<td>Fine Science Tool</td>
			<td>15000-10</td>
			<td>for minor dissection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scissors (angled to side)</td>
			<td>Fine Science Tool</td>
			<td>14063-011</td>
			<td>No. 3 handle</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Standard scalpel</td>
			<td>Fine Science Tool</td>
			<td>10003-12</td>
			<td>for removing muscle</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Scalpel blade</td>
			<td>Feather</td>
			<td>2976</td>
			<td>No. 10</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Forceps (curved)</td>
			<td>Fine Science Tool</td>
			<td>11150-10</td>
			<td>for holding tissue</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Forceps (straight)</td>
			<td>Fine Science Tool</td>
			<td>11151-10</td>
			<td>for holding tissue</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Needle holder&#160;</td>
			<td>Fine Science Tool</td>
			<td>12002-12</td>
			<td>for suturing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tissue adhesive glue</td>
			<td>3M Vetbond</td>
			<td>1469SB</td>
			<td>to adhere to edges of the cut skin</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Monofilament polypropylene&#160;</td>
			<td>USSC Sutures</td>
			<td>VP-521</td>
			<td>Size = 4-0 (for fascia)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Perma-hand silk</td>
			<td>Ethicon</td>
			<td>683G</td>
			<td>Size = 4-0 (for skin)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Micro drill</td>
			<td>Roboz Surgical Instrument Co. Inc.</td>
			<td>RS-6300</td>
			<td>with bone polishing drill bit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Laser doppler flowmeter</td>
			<td>Moor Instruments</td>
			<td>moorVMS-LDF1</td>
			<td>for monitoring change in blood flow</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Heating pad</td>
			<td>Fine Science Tool</td>
			<td>21052-00</td>
			<td>to prevent hypothermia</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Lab-Jack</td>
			<td>Fisher scientific&#160;</td>
			<td>14-673-50</td>
			<td>4x4 in plate to adjust the height of the animal</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">X-Y gliding stage&#160;</td>
			<td>Amscope</td>
			<td>GT100</td>
			<td>for positioning the animal under microscope&#160;&#160;</td>
		</tr>
	</tbody>
</table>

photothrombosis-induced focal ischemia as a model of spinal cord injury in mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Het doel van deze studie was het ruggemerg ischemie in muizen met een model PT produceren. Na het gewenste gebied van het bot boven het ruggenmerg (T10 - T12) werd uitgedund, werd Bengaals geïnjecteerd door retro-orbitale sinus route, en ischemie werd geïnduceerd door PT Figuur 1A, B tonen de muis geplaatst in een op maat gemaakte chirurgische. platform tijdens de operatie. De muis werd op zijn plaats gehouden door een klem snuit en twee verstelbare gewervelde klemmen om het ruggenmerg stabiliseren <s...

Watch this Scientific Journal Video about Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice at JoVE.com

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

Photothrombosis-geïnduceerde focale ischemie als een model van Spinal Cord Injury in Muizen

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

Research

JoVE Journal

Medicine

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Acute en chronische Tactile Sensory Testing na ruggenmergletsel bij ratten

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

Controlled Cervical Laceration Injury in Mice

Gecontroleerde Cervicale Laceration Letsel in Muizen

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

A Contusion Model of Severe Spinal Cord Injury in Rats

Een Kneuzing Model van Severe ruggenmergletsel bij ratten

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemie-reperfusie model van Acute Kidney Injury en Post Letsel Fibrosis in Muizen

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion

Reproduceerbaar Paraplegie door thoracale aorta-occlusie in een muizenmodel van Spinal Cord ischemie-reperfusie

Background
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Gecontroleerde Corticale Impact Model voor traumatisch hersenletsel

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

Een radio-telemetrische systeem om de cardiovasculaire functie bij ratten met Spinal Cord doorsnijding en embryonale neurale stamcellen Enten Monitor

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Neural Stem Cell Transplantation in Experimental Contusive Model of Spinal Cord Injury

Neurale stamceltransplantatie bij Experimentele Contusive Model van Spinal Cord Injury

Spinal cord injury is a devastating clinical condition, characterized by a complex of neurological dysfunctions. Animal models of spinal cord injury can ...

Calibrated Forceps Model of Spinal Cord Compression Injury

Gekalibreerde tang Model van Spinal Cord Injury Compression

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Een Neonatale Mouse Spinal Cord Injury Compressie Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...