Injeção de sangue autólogo de Modelo hemorragia intracerebral espontânea em ratos
Summary
O modelo de injeção de sangue autólogo de hemorragia intracerebral em camundongos descritos neste protocolo usa a técnica de injecção dupla para minimizar o risco de refluxo de sangue até a faixa de agulha, não anticoagulantes no sistema de bombeamento, e elimina todo o espaço morto e tubos expansíveis no sistema.
Abstract
Investigation of the pathophysiology of injury after intracerebral hemorrhage (ICH) requires a reproducible animal model. While ICH accounts for 10-15% of all strokes, there remains no specific effective therapy. The autologous blood injection model in mice involves the stereotaxic injection of arterial blood into the basal ganglia mimicking a spontaneous hypertensive hemorrhage in man. The response to hemorrhage can then be studied in vivo and the neurobehavioral deficits quantified, allowing for description of the ensuing pathology and the testing of potential therapeutic agents. The procedure described in this protocol uses the double injection technique to minimize risk of blood reflux up the needle track, no anticoagulants in the pumping system, and eliminates all dead space and expandable tubing in the system.
Protocol
Discussion
Este modelo cirúrgico da hemorragia intracerebral em camundongos, utilizando a cauda resultados autólogo de sangue arterial em um modelo reprodutível de hemorragia espontânea gânglios basais. Um modelo em camundongos ICH oferece a vantagem da disponibilidade de animais transgênicos para investigar fisiopatologia, no entanto, seu pequeno tamanho faz com que procedimentos neurocirúrgicos mais tecnicamente difícil do que em animais maiores.
O modelo de colagenase e o modelo de injeção…
Disclosures
The authors have nothing to disclose.
Acknowledgements
O trabalho foi financiado por uma bolsa do Instituto de Medicina Translacional e Terapêutica, e uma bolsa de formação do Instituto de Medicina e Engenharia (T32HL007954) da Universidade da Pensilvânia e da Marlene L. Cohen e H. Jerome Fleisch Scholar Grant no University of Connecticut Health Center (LHS) e NIH NS-029331 (FAW).
Materials
Stereotaxic frame for mouse neurosurgery (Stoelting, 51925)
Microinfusion pump and processor (UMP-3 and Micro4, World Precision Instruments, Sarasota, FL)
Mouse warmer (Stoelting, 50300)
Inhalational mouse anesthesia (Braintree Scientific, EZ-AF9000)
25 μL gastight borosilicate Hamilton syringe with coated plunger and no needle
(Hamilton company, Reno, NV, 1702RN syringe: 765401, ferrule: 30949, spacer: 30946)
fused silica needle cut to 2 cm length (Hamilton, 17739)
*note Hamilton syringe and fused silica needle may be reused for multiple surgeries if sterilized prior to each surgery. These materials are crucial to avoid blood clotting.
Sterile surgical gloves
Surgical gown, bonnet and mask
Betadine
75% ethanol
sterile 27 g needle (single use)
sterile 1 cc syringe (single use)
sterile surgical blade
Cidex
sterile water
buprenorphine and isoflurane
sterile gauze
paraffin wax paper squares
Veterinary surgical glue (Vetbond, 3M, St. Paul, MN)
References
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