Yanal Kronik Kranial Pencere Hazırlık sağlar<em> İn Vivo</em> Farelerde Distal Orta Serebral Arter Tıkanıklığı takiben Gözlem
Summary
distal orta serebral arter dalı (MCAO) Cerrahi tıkanması, deneysel inme araştırmalarında sıkça kullanılan bir modeldir. Bu el yazması farelerde boyuna intravital mikroskopi için bir fırsat sunuyor yanal kafatası pencere takılması, birlikte kalıcı MCAO temel tekniğini açıklar.
Abstract
Fokal serebral iskemi (yani, iskemik inme) nöronal fonksiyon ciddi bir kaybı ve dolayısıyla motor ve zihinsel engelli bir dizi yol açan başlıca beyin yaralanmaya neden olabilir. Inme 1 dünya çapında uzun süreli sakatlık ve ölüm başlıca nedenleri arasında olduğu gibi yüksek prevalans, ciddi bir sağlık yükünü oluşturmaktadır. nöronal fonksiyonu geri kazanımı, çoğu durumda, sadece kısmi vardır. Şimdiye kadar, tedavi seçenekleri oldukça nedeniyle trombolitik 2,3 dar zaman penceresi, özellikle de sınırlıdır. inme kurtarma hızlandırmak için yöntemlerin belirlenmesi birinci sınıf bir tıbbi hedef olmaya devam; Ancak, bu toparlanma sürecine yetersiz mekanistik anlayışlar tarafından engellenmiştir. Deneysel inme araştırmacılar sıklıkla fokal serebral iskemi kemirgen modelleri kullanır. Akut fazda ötesinde, inme araştırma giderek serebral iskemi aşağıdaki-sub akut ve kronik fazda odaklanmıştır. Çoğu inme araştırmacılar uygulamak kalıcı veya tranfarelerde ya da sıçanlarda MCA'nın SIENT oklüzyonu. Hastalarda, MCA tıkanıklıklar iskemik inme 4 en sık nedenleri arasındadır. Filament modelini kullanarak MCA proksimal oklüzyon yanı sıra, uzak MCA cerrahi tıkanması, deneysel inme araştırma 5. en sık kullanılan model muhtemelen. MCA dalı (lenticulo-striat arterlerin dallanma kadar) distalinde Tıkanıklığı tipik striatum yedek ve öncelikle neokorteks etkiler. Gemi tıkanıklığı kalıcı veya geçici olabilir. uzun dönem sonuçları açısından lezyon hacmi ve çok düşük mortalite oranları yüksek tekrarlanabilirlik bu modelin başlıca avantajları vardır. Burada, sagital sinüs kronik kafa penceresi (CW) hazırlık yanal gerçekleştirmek için, ve daha sonra nasıl bir kraniotomi yaklaşımı kullanarak pencerenin altında bir uzak inme neden cerrahi olarak nasıl gösterilmektedir. Bu yaklaşım ile iskemi sonrası akut ve kronik değişikliklerin sıralı görüntüleme için uygulanabilirKonfokal lazer tarama ve iki foton intravital mikroskopi epi-aydınlatıcı.
Introduction
Stroke is among the principal causes of long-term disability and death worldwide1, coming second after coronary heart disease. In addition, stroke is the primary cause of long-term disability, underscoring its tremendous socioeconomic impact6-8. Beyond acute treatment, investigating new approaches and mechanisms to accelerate and enhance recovery after stroke remains a prime medical goal7.
In the last few decades, data from experimental stroke research has contributed substantially to understanding the complex pathophysiological cascades triggered by ischemia9,10. Excitotoxicity, apoptosis, peri-infarct depolarization, and inflammation have been identified as the most relevant mediators of cell death following focal cerebral ischemia. Moreover, using animal models of cerebral ischemia, important concepts, diagnostic modalities, and therapeutic approaches have been developed and validated (e.g., “penumbra” and thrombolysis)11.
The availability of experimental stroke models, combined with non-invasive imaging modalities (e.g., magnetic resonance imaging (MRI), computed tomography, or laser speckle contrast analysis), enables the researcher to investigate hyperacute and chronic pathophysiological changes induced by the ischemic insult in a longitudinal manner12. Along with studying the spatiotemporal profile of the evolving lesion, changes resembling neuronal plasticity can be investigated and correlated to functional outcomes and histological findings. Within the last few years, further methodological advances have been made using the combination of cerebral ischemia models and in vivo microscopy via cranial windows13. These new techniques allow investigators to analyze the neurovascular unit at the cellular and molecular level, with great analytic power in the acute, subacute, and chronic phases following focal cerebral ischemia14. Moreover, in vivo microscopy imaging of microcirculatory dynamics has revealed novel aspects of cerebral microvasculature function and angioarchitecture, with significant pathophysiological relevance15-17.
In this protocol, we present how to perform a chronic CW preparation lateral to the sagittal sinus and how to surgically induce a distal stroke underneath the window. This mouse model can be applied to sequential imaging of acute, subacute, and chronic changes following focal cerebral ischemia via epi-illuminating, confocal laser scanning, and two-photon intravital microscopy.
Protocol
Representative Results
Discussion
İnme 1 dünya çapında uzun süreli sakatlık ve ölüm başlıca nedenleri arasındadır. Akut tedavi, hızlandırmak ve inme sonrası iyileşmeyi artırmak için yeni yaklaşımlar ve mekanizmaları araştıran ötesinde bir asal tıbbi gol 7 kalır. Deneysel inme araştırmacılar sıklıkla fokal serebral iskemi kemirgen modelleri kullanır. Aslında, geçici veya kalıcı MCAO uyaran modelleri hastalarda 4 fokal serebral iskemi en yaygın türlerinden biri taklit. MCA proksimal ok…
Disclosures
The authors have nothing to disclose.
Acknowledgements
VP is a participant in the Charité Clinical Scientist Program, funded by the Charité – Universitätsmedizin Berlin and the Berlin Institute of Health. TB is an SNSF PostDoc Mobility fellow. The authors receive grant support from EinsteinStiftung/A-2012-153 to PV.
Materials
| Binocular surgical microscope | Zeiss | Stemi 2000 C | |
| Light source for microscope | Zeiss | CL 6000 LED | |
| Heating pad with rectal probe | FST | 21061-10 | |
| Stereotactic frame | Kopf | Model 930 | |
| Anaethesia system for isoflurane | Draeger | ||
| Isoflurane | Abott | ||
| Dumont forceps #5 | FST | 11251-10 | |
| Dumont forceps #7 | FST | 11271-30 | |
| Bipolar Forceps | Erbe | 20195-501 | |
| Bipolar Forceps | Erbe 20195-022 | ||
| Microdrill | FST 18000-17 | ||
| Needle holder | FST | 12010-14 | |
| 5-0 silk suture | Feuerstein, Suprama | ||
| 7-0 silk suture | Feuerstein,Suprama | ||
| 8-0 silk suture | Feuerstein, Suprama | ||
| Veterinary Recovery Chamber | Peco Services | V1200 | |
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