Abstract
Stroke forskning har tålt mange tilbakeslag i å oversette nevro terapi i klinisk praksis. I motsetning til den virkelige verden terapi (Pa trombolyse) produserer sjelden fordeler i mekaniske okklusjon-baserte eksperimentelle modeller, som dominerer preklinisk hjerneslag forskning. Denne fordelingen mellom benk og nattbord antyder behovet for å ansette Pa-responsive modeller i preklinisk hjerneslag forskning. For dette formål, er en enkel og tPA-reaktive trombotisk slag modell oppfunnet og beskrevet her. Denne modellen består av transient okklusjon av den ensidige arteria carotis communis og levering av 7,5% oksygen gjennom en ansiktsmaske i voksen mus i 30 minutter, under opprettholdelse av dyret rektal temperatur på 37,5 ± 0,5 ° C. Selv reversibel ligering av den ensidige carotid arterie eller hypoksi hver trykkes cerebral blodstrøm bare forbigående, er kombinasjonen av begge fornærmelser forårsaket varig reperfusjon underskudd, fibrin og blodplateavleiring, og store INFARct i midten cerebral arterie-leverte territorium. Viktigere, haleveneinjeksjon av rekombinant tPA ved 0,5, 1, eller fire timer etter Thi (10 mg / kg) gitt tidsavhengig reduksjon av dødelighet og infarktstørrelsen. Denne nye slagmodell, er enkel og kan standardiseres tvers laboratorier for å sammenligne eksperimentelle resultater. Videre induserer det trombose uten craniectomy eller innføre forhånds dannet emboli. Gitt disse unike fordeler, er thi modellen et nyttig tillegg til repertoaret av preklinisk hjerneslag forskning.
Materials
Name | Company | Catalog Number | Comments |
adult male mice | Charles River | C57BL/6 | 10-13 weeks old (22-30 g) |
Mobile Laboratory Animal Anesthesia System | VetEquip | 901807 | anesthesia |
Medical air (Compressed) air tank | Airgas | UN1002 | anesthesia |
Isoflurane | Piramal Healthcare | NDC 66794-013-25 | anesthesia |
Multi-Station Lab Animal AnesthesiaSystem | Surgivet | V703501 | hypoxia system |
7.5% O2 balanced by 92.5% N2 tank | Airgas | UN1956 | hypoxia system |
Temperature Controller with heating lamp | Cole Parmer | EW-89000-10 | temperature controllers |
Rectal probe | Cole Parmer | NCI-00141PG | temperature controllers |
Dissecting microscope | Olympus | SZ40 | surgical setup |
Heat pump with warming pad | Gaymar | TP700 | surgical setup |
Fine curved forceps (serrated) | FST | 11370-31 | surgical instrument |
Fine curved forceps (smooth) | FST | 11373-12 | surgical instrument |
micro scissors | FST | 15000-03 | surgical instrument |
micro needle holders | FST | 12060-01 | surgical instrument |
Halsted-Mosquito hemostats | FST | 13008-12 | surgical instrument |
5-0 silk suture | Harvard Apparatus | 624143 | surgical supplies |
4-0 Nylon monofilament suture | LOOK | 766B | surgical supplies |
Tissue glue | Abbott Laboratories | NC9855218 | surgical supplies |
Puralube Vet ointment | Fisher | NC0138063 | eye dryness prevention |
MoorFLPI-2 blood flow imager | Moor | 780-nm laser source | Laser Speckle Contrast Imaging |
Mannitol | Sigma | M4125 | in vivo TTC |
2,3,5-triphenyltetrazolium chloride (TTC) | Sigma | T8877 | in vivo TTC |
Vibratome | Stoelting | 51425 | brain section for in vivo TTC |
Digital microscope | Dino-Lite | AM2111 | whole-brain imaging |
O.C.T compound | Sakura Finetek | 4583 | |
goat anti-rabbit Alexa Fluro 488 | Invitrogen | A11008 | Immunohistochemistry |
Cryostat | Vibratome | ultrapro 5000 | brain section for IHC |
Evans blue | Sigma | E2129 | Detecting vascular perfusion |
Microtome | Electron Microscopy Sciences | 5000 | brain section for histology |
Avertin (2, 2, 2-Tribromoethanol) | Sigma | T48402 | euthanasia |
Fluorescent microscope | Olympus | DP73 | |
Meloxicam SR | ZooPharm | NSAID analgesia |
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