Name: endothelin-1 induced middle cerebral artery occlusion model for ischemic stroke with laser doppler flowmetry guidance in rat
Uploaded: 2013-02-16T14:00:00
Description: Watch this Scientific Journal Video about Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat at JoVE.com

Dette arbejde blev støttet af tilskud fra American Heart Association Greater Southeast Affiliate (09GRNT2060421), American Medical Association, og fra University of Florida klinisk og translationel Science Institute. Adam Mekka er et NIH / NINDS, NRSA predoctoral fyr (F30 NS-060.335). Robert Regenhardt fik predoctoral stipendium støtte fra University of Florida Multidisciplinary Training Program i Hypertension (T32 HL-083.810).

Denne protokol blev godkendt af Institutional Animal Care og Use Committee (IACUC) ved University of Florida og er i overensstemmelse med &quot;Vejledning for Pleje og anvendelse af forsøgsdyr&quot; (ottende udgave, National Academy of Sciences, 2011). Materialer <ol><li> Dyr: Otte uger gamle, Sprague Dawley hanrotter (Charles River Farms, Wilmington, MA, USA), som vejede 250-300 g på tidspunktet for kirurgi. </li><li> Anæstesi <ol class="lalpha"><li> Inhalationsanæstesi system (VetEqui...

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J., Sumners, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Candesartan+pretreatment+is+cerebroprotective+in+a+rat+model+of+endothelin-1-induced+middle+cerebral+artery+occlusion.">Candesartan pretreatment is cerebroprotective in a rat model of endothelin-1-induced middle cerebral artery occlusion.</a> Exp. Physiol. 94, 937-946 (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>Tomsick, T. 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ET-1-induceret MCAO er en etableret model for eksperimentel iskæmisk slagtilfælde, der anvendes regelmæssigt i flere rottestammer. Mange variable, såsom rotte-stamme, dyrenes alder, kropstemperatur, anæstesi fremgangsmåde, og operatøren ekspertise kan føre til øget variabilitet i infarktvolumener ved brug af denne model 5, 14. Imidlertid har adskillige forskere påvist, at fordelene ved denne model indbefatter den relativt ikke-invasiv metode, dosisrespons af cerebral blodstrøm til ET-1, og evnen t...

<table border="1"><tbody><tr><td></td><td></td><td></td><td><ol><li> Dyr: Otte uger gamle, Sprague Dawley hanrotter (Charles River Farms, Wilmington, MA, USA), som vejede 250-300 g på tidspunktet for kirurgi. </li><li> Anæstesi <ol class="lalpha"><li> Inhalationsanæstesi system (VetEquip Inc., Pleasanton, CA, USA) </li><li> Isofluran bedøvelsesmiddel (Baxter Pharmaceutics, Deerfield, IL, USA) </li></ol></li><li> Stereotaktisk system (David Kopf Instruments, Tujunga, CA, USA) <ol class="lalpha"><li> Små dyr stereotaktisk-system </li><li> Ikke-brud øre stænger for rotter </li><li> Gas anæstesi hoved holder til rotter </li></ol></li><li> Temperatur regulering <ol class="lalpha"><li> BAT-12 mikrosonde termometer (World Precision Instruments, Inc., Sarasota, FL, USA) </li><li> T / PUMP, TP600 Termisk tæppe (Gaymar Industries, Inc., Orchard Park, NY, USA) </li></ol></li><li> Kirurgiske instrumenter <ol class="lalpha"><li> Skalpel håndtag og # 11 klinge, iris pincet, Graefe pincet, bulldog clamp retraktorer, skruetrækker, 10 ul sprøjte med 26 gauge skrå nål (World Precision Instruments, Inc., Sarasota, FL, USA) </li><li> Mikromotorer boremaskine og stereotaktisk holder, Quintessential stereotaktisk Injector (Stoelting, Wood Dale, IL, USA) </li><li> 1,0 mm runde bore bur, 1,0 mm omvendt kegle drill bur (Roboz Surgical Instrument Co, Inc., Gaithersburg, MD, USA) </li></ol></li><li> Kirurgiske Supplies <ol class="lalpha"><li> Monteringsskruer 0-80 x 3/32 med 2,4 mm stav, 21-gauge guide kanyle [4mm lang under soklen] og kanyle dummy (Plast én, Roanoke, VA, USA) </li><li> Jet protese acryl-og væske (Lang Dental Manufacturing Co, Inc., Wheeling, IL, USA) </li><li> 3,0 nylonsutur (Oasis, Mettawa, IL, USA) </li><li> Vatpinde, Puralube øjensalve (Fisher videnskabeligtc, Pittsburg, PA, USA) </li><li> Elektriske hårklippere (Oster, Providence, RI, USA) </li></ol></li><li> Kemikalier <ol class="lalpha"><li> Endothelin-1 (American Peptide, Sunnyvale, CA, USA) </li><li> Chlorhexidin 2% (Agrilabs, St. Joseph, MO, USA) </li><li> Buprenorphin HCI (Hospira Inc., Lake Forest, IL, USA) </li></ol></li><li> Visualisering Udstyr <ol class="lalpha"><li> Kirurgisk mikroskop (Seiler Instrument and Manufacturing, St. Louis, MO, USA) </li><li> Fiber Optic illuminator (TechniQuip Corp, Livermore, CA, USA) </li></ol></li><li> Laser Doppler flowmetry system (ADInstruments, Inc., Colorado Springs, CO, USA) <ol class="lalpha"><li> Standard Pencil Probe </li><li> Probeholderen </li><li> Blood Flowmeter </li><li> Powerlab 4/30 med LabChart 7 </li></ol></li><li> Måling af infarktvolumen <ol class="lalpha"><li> Rottehjernematrix (Zivic-Miller Lab. Inc., Allison Park, PA, USA) </li><li> 2,3,5-triphenyltetrazoliumchlorid (Sigma-Aldrich Co, St Louis, MO, USA) fortyndet til 0,05% i PBS </li><li> Flatbed scanner (Epson Perfection V30, Epson America, Inc., Long Beach, CA, USA) </li><li> Billede J software (ImageJ 1.42q software, US National Institutes of Health, Bethesda, MA, USA) </li></ol></li></ol></td></tr></tbody></table>

endothelin-1 induced middle cerebral artery occlusion model for ischemic stroke with laser doppler flowmetry guidance in rat

Slagtilfælde er den største årsag til invaliditet og tredje hyppigste dødsårsag i verden, koster en anslået 70 milliarder dollar i USA i 2009 1, 2. Mange modeller af cerebral iskæmi er blevet udviklet til at efterligne den menneskelige tilstand af slagtilfælde. Det er blevet foreslået, at op til 80% af alle slagtilfælde skyldes iskæmisk skade i den midterste cerebrale arterie (MCA) område 3. I begyndelsen af 1990&#39;erne, 4 endothelin-1 (ET-1) blev anvendt til at inducere iskæmi ved at anvende det direkte op til overfladen af MCA efter kraniotomi. Senere blev denne model modificeret 5 ved anvendelse af en stereotaksisk injektion af ET-1 ved siden af MCA at producere fokal cerebral iskæmi. Hovedfordelene ved denne model omfatter evnen til at udføre proceduren hurtigt, evnen til at kontrollere arterie konstriktion ved at ændre dosis af ET-1 afgivet, ikke nødvendigt at bearbejde de ekstrakraniale kar, der forsyner blod til hjernen og gradvis reperfusiom satser der i højere grad efterligner reperfusion hos mennesker 5-7. På den anden side har den ET-1 model ulemper, der omfatter behovet for en kraniotomi, samt højere variation i slagvolumen 8. Denne variabilitet kan reduceres ved anvendelse af laser-doppler-flowmetri (LDF) at kontrollere, cerebral iskæmi under ET-1 infusion. Faktorer, der påvirker slagtilfælde variabilitet omfatter præcision af infusion og det parti af ET-1 brugt 6. En anden vigtig overvejelse er, at selv om reperfusion er en almindelig forekomst i humant slagtilfælde, kan varigheden af okklusionen for ET-1-induceret MCAO ikke nøje efterligner den for humant slagtilfælde, hvor mange patienter har partiel reperfusion i løbet af timer til dage efter okklusion 9, 10. Denne protokol vil beskrive i detaljer ET-1-induceret MCAO model for iskæmisk slagtilfælde hos rotter. Det vil også henlede opmærksomheden på særlige hensyn og potentielle ulemper hele proceduren.

1. Post-Op neurologisk evaluering Efter at dyret kommer til bevidsthed, kan en lang række metoder anvendes til at evaluere neurologiske defekter, herunder balance, gribestyrke, paw placering, postural asymmetri og trappe klatring. Den solsikkefrø opgave er en grov vurdering af motorisk og sensorisk funktion, der har signifikant sammenhæng med infarktvolumen 7, 12. I løbet af denne opgave, er rotter timet under åbning og forbruge 5 solsikkefrø. De fem frø anbringes i et hjørne...

Watch this Scientific Journal Video about Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat at JoVE.com

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

Adskillige dyremodeller for cerebral iskæmi er blevet udviklet til at simulere den menneskelige tilstand af slagtilfælde. Denne protokol beskrives endothelin-1 (ET-1) inducerede mellem-cerebral arterieokklusion (MCAO) model for iskæmisk slagtilfælde i rotter. Desuden er vigtige overvejelser, fordele og ulemper ved denne diskuteret.

Endothelin-1-induceret mellem-cerebral arterieokklusion Model for iskæmisk slagtilfælde med laser-doppler-flowmetri Vejledning i 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 ...

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