Special thanks to the Gidday lab for their work in developing the RHP protocol, as well as the Neuro-Models Facility (UTSW) for their assistance in the tMCAo surgeries. This work was supported by grants from the American Heart Association (AMS), The Haggerty Center for Brain Injury and Repair (UTSW; AMS), and The Spastic Paralysis Research Foundation of the Illinois-Eastern Iowa District of Kiwanis International (JMG).

The authors have nothing to disclose.

التعرض مرة واحدة لنقص الأكسجة النظامية (أي 2 ساعة من O 2 11٪) في الفئران &quot;عابر&quot; يحمي الدماغ من tMCAo، 29 وهذا يعني أن استجابة جينية لتحدي شروط مسبقة ميتة قصيرة الأمد، ويتم استعادة النمط الظاهري الأساس داخل أيام. العروض المتكررة من التحفيز شروط مسبقة ميت...

باعتبارها السبب الرئيسي للعجز الكبار ورابع سبب رئيسي للوفاة، والسكتة الدماغية هي واحدة من الحالات المرضية الأكثر إضعافا التي تواجه السكان البالغين في الولايات المتحدة. 1 النماذج الحيوانية من السكتة الدماغية تسمح التحقيق التجريبي للطرق جديدة لتقليل الإصابة الدماغية و تحسين الانتعاش بعد السكتة الدماغية. واحد سبيلا جديدا لهذه البحوث متعدية غير شروط مسبقة. شروط مسبقة هو الاستخدام المتعمد للحافزا غير ضارة للحد من الأضرار من لاحقة، وأشد، والإصابة. وقد تبين أن 2 نقص الأوكسجين شروط مسبقة لإحداث تغييرات عديد المظاهر في الدماغ التي توفر الحماية من السكتة الدماغية في كل المجراة في الدراسات المختبرية 3 ومع ذلك، فإن التعرض لنقص الأكسجين واحد لا يقدم سوى العصبية على المدى القصير، وحمل أقل من 72 ساعة من التسامح ضد نقص التروية في فئران بالغة. 4 وحتى بعد أربعة أسابيع من التعرض اليومية 14 ساعة لنقص الأكسجين منخفض الضغط، ولين وآخرون. FOاوند أن العصبية واستمر فقط لمدة أسبوع واحد. 5 شروط مسبقة ميتة المتكرر (رهب) تتميز التغيرات العشوائية في التردد، والمدة، وشدة التعرض ميتة. على النقيض من التحدي شروط مسبقة واحد، RHP يدفع النمط الظاهري cerebroprotective أن تستغرق ما يصل إلى ثمانية أسابيع في الفئران. 6 تخفيض RHP كميات احتشاء، حاجز الدم في الدماغ (BBB) ​​اضطراب، والتهاب الأوعية الدموية، والكريات البيض انسلال لعدة أسابيع بعد التعرض ميتة النهائي . RHP انخفاض تحديدا التهاب في الدماغ الدماغية عن طريق الحد من الخلايا التائية، الوحيدات، والسكان بلعم، مع الحفاظ على السكان الخلية B في نصف الكرة الدماغية. 7 في الواقع، بفعل RHP النمط الظاهري المثبطة للمناعة في الفئران قبل أي إصابة الجهاز العصبي المركزي، بما في ذلك السكتة الدماغية. خلايا B RHP المعاملة معزولة عن رهب معاملة الفئران صحي أظهرت النمط الظاهري المضادة للالتهابات فريدة من نوعها، مع downregulation من كل من العرض مستضد وإنتاج الأجسام المضادة. الانخفاض عام في الآليات المناعية التكيفية الموالية للالتهابات يجعل RHP منهجية ممتازة للحث على المناعة الذاتية للأمراض التهابات الجهاز العصبي المركزي محددة فحسب، ولكن أيضا الإصابة أو المرض النماذج الشاملة التي تشمل أمراض الموالية للالتهابات. RHP يقلل من حيث الحجم واحتشاء BBB اضطراب بعد منتصف انسداد الشريان الدماغي عابر (tMCAo). نماذج حيوانية من السكتة الدماغية، مثل شائعة الاستخدام tMCAo، وتحسين كبير في فهم الفيزيولوجيا المرضية من السكتة الدماغية، فضلا عن تصميم neurotherapeutics أكثر فعالية. أول من وضعه كويزومي وآخرون، في عام 1986، 8 الإجراء tMCAo هو الطريقة المستخدمة على نطاق واسع على إحداث سكتة دماغية في القوارض واحدة من الطرق المفضلة للتحقيق في التهاب التالية ضخه. وطرق لtMCAo تتطور، واستخدام أحدث من خيوط المغلفة سيليكون كذلك يقلل من خطر النزف تحت العنكبوتية مقارنة بالطرازات الأخرى 9،10 &lt;/ سوب&gt; وتحسين الاعتمادية، على الرغم من سوء الحظ tMCAo غالبا ما تنتج عن اختلاف كبير في حجم احتشاء 11-13 معظم هذه الدراسات تحدد المناطق احتشاء في أقسام الدماغ الاكليلية التي تلطيخ مع 2،3،5- triphenyltetrazolium كلوريد (TTC)، يعتبر المعيار الذهبي لاحتشاء الكمي لأنه هو وسيلة بسيطة وغير مكلفة لإنتاج حية، نتائج قابلة للتكرار. يقدم TTC باعتباره ركيزة من dehydrogenases الموجودة في الميتوكوندريا. عندما يتعرض شرائح الدماغ إلى حل TTC، يؤخذ TTC بشكل انتقائي في الخلايا الحية حيث المنتج الحد غير قابل للذوبان في، formazan، يترسب إلى لون أحمر عميق في الميتوكوندريا قابلة للحياة. بسبب ضعف الميتوكوندريا في الأنسجة الدماغية، تبقى هذه الأنسجة البيضاء، مما يسمح للتمايز الأنسجة التالفة وصحية. 14 RHP يقلل أيضا من BBB اضطراب في نصف الكرة الدماغية. 6 لذلك القياس الكمي المزدوج للسلامة BBB ضمن نفس بالأمطار كما حجم احتشاء أساس TTC-15 قرارات من شأنها أن توفر معلومات مفيدة حول فعالية كاملة من الحماية الذاتية، والعلاقات السببية المحتملة بين BBB اضطراب واحتشاء في الحيوانات غير المعالجة أو المعالجة. تدفق الدم المحيطي من خلال BBB تعطلت والثانوي إلى السكتة الدماغية، ويزيد من السكان الكريات البيض، السيتوكينات الموالية للالتهابات، الاكسدة، وذمة وعائية المنشأ، والتحول النزفية في نصف الكرة الدماغية، وزيادة في نهاية المطاف معدلات الإصابة والوفيات في المرضى الذين يعانون من السكتة الدماغية . 16،17 وهناك طريقة شائعة لقياس BBB اضطراب في النماذج الحيوانية من خلال تقدير حجم ايفانز الأزرق (EB) تسرب الصبغة في الدماغ. 15،18-21 EB يربط بشكل انتقائي لمصل الزلال، وهو بروتين كروي (MW = 65 كيلو دالتون) لا عبور BBB في الحيوانات لم يصب. 22 وبعد السكتة الدماغية، EB يخترق الدماغ، وتتفلور في 620 نانومتر، مما يسمح لقياس الكثافة البصرية داخلهان لحمة أصيب perfused ل22 الكثافة البصرية يتناسب طرديا مع نفاذية BBB عندما تم غسلها EB من الأوعية الدموية القشرية بعد الوفاة التي كتبها transcardiac الارواء. مع تجهيز الفوري للأدمغة TTC الملطخة في الحيوانات مع الإدارة EB، فإن كلا من حجم احتشاء وBBB اضطراب يمكن قياس فعالية. وتجدر الإشارة إلى أنه، مع ذلك، أن الإصابة العصبية وBBB اضطراب ليست عمليات مصاحبة في المخ بعد السكتة الدماغية، 23،24 حتى اختيار وقت التضحية هو أحد الاعتبارات الهامة. البروتوكول الذي يلي تفاصيل طريقة رهب، وطريقة tMCAo عن الأمر الذي أدى إلى انسداد الشرايين مؤقت نماذج المتوسطة انسداد الشريان الدماغي في المرضى من البشر، والأساليب النسجية المزدوجة لتحديد العصبية والأوعية الدموية النهاية الإصابة بالسكتة الدماغية. TTC يقيس موت الخلايا وتلف الأنسجة التراكمي، والسماح للالكمي من المجلد احتشاء العامأوميا، في حين يوفر EB لتقدير نصف الكرة الغربي من الضرر BBB.

<table border="0" cellpadding="0" cellspacing="0" width="830">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:249px;">Material/ Equipment</td>
			<td style="width:171px;">Company</td>
			<td style="width:127px;">Catalog Number</td>
			<td style="width:283px;">Comments/Description</td>
		</tr>
		<tr height="220">
			<td height="220" style="height:220px;width:249px;">Flowmeters, regulators</td>
			<td style="width:171px;">VetEquip, Inc</td>
			<td style="width:127px;">Specialty order</td>
			<td style="width:283px;">Four flowmeters are attached to 6.0 mm flexible PVC tubing which connects to the inlet port on each induction chamber with a plastic female connector. These flowmeters are bolted to a 6.5&#34; x 1&#34; x 1&#34; metal bar. This metal bar is bolted to a MI-246-P pressure gauge with a DISS outlet. This pressure gauge and flowmeter equipment can be attached to each new gas cylinder with a wrench.</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:249px;">21% O2 tank</td>
			<td style="width:171px;">AirGas</td>
			<td style="width:127px;">OX USP200</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:249px;">11% O2 tank</td>
			<td style="width:171px;">AirGas</td>
			<td style="width:127px;">Specialty order</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:26px;width:249px;">8% O2 tank&#160;</td>
			<td style="width:171px;">AirGas</td>
			<td style="width:127px;">Specialty order</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">15L induction chambers</td>
			<td style="width:171px;">VetEquip</td>
			<td style="width:127px;">941454</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">Moor Laber Dopper Flow&#160;</td>
			<td style="width:171px;">Moor Instruments&#160;</td>
			<td style="width:127px;">moorVMS-LDF1-HP</td>
			<td style="width:283px;">0.8mm diameter probe&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">High Intensity Illuminator&#160;</td>
			<td style="width:171px;">Nikon</td>
			<td style="width:127px;">NI-150</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">Zoom Stereo Microscope&#160;</td>
			<td style="width:171px;">NIkon</td>
			<td style="width:127px;">SMZ800</td>
			<td style="width:283px;">Other surgical microscopes may be used.&#160;</td>
		</tr>
		<tr height="100">
			<td height="100" style="height:100px;width:249px;">Kent Scientific Right Temperature CODA</td>
			<td style="width:171px;">Kent Scientific Corporation</td>
			<td style="width:127px;">Discontinued</td>
			<td style="width:283px;">Recommended replacement is PhysioSuite with RightTemp Temperature Monitoring and Homeothermic Control (Kent Scientific, #PS-RT).</td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:249px;">Hovabator Incubator</td>
			<td style="width:171px;">Stromberg&#39;s</td>
			<td style="width:127px;">2362-E</td>
			<td style="width:283px;">Our model is the 2362N. 2362E is a later model and includes an electronic thermostat.&#160;</td>
		</tr>
		<tr height="120">
			<td height="120" style="height:120px;width:249px;">V010 Anesthesia system&#160;</td>
			<td style="width:171px;">VetEquip</td>
			<td style="width:127px;">901807</td>
			<td style="width:283px;">Includes: ten foot high-pressure oxygen hose, frame, flowmeter, oxygen flush assembly, vaporizer, breathing circuit, chamber, nosecones, waste gas evacuation tubing and two VapoGuard filters</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">250 mL isoflurane&#160;</td>
			<td style="width:171px;">Butler Schein</td>
			<td style="width:127px;">NDC-11695</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">D-6 Vet Trim Animal Cordless Trimmer&#160;</td>
			<td style="width:171px;">Andis&#160;</td>
			<td style="width:127px;">#23905</td>
			<td style="width:283px;">Replacement blades are available from Andis (#23995)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Betadine&#160;</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">19-898-867&#160;</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">Q-tips</td>
			<td style="width:171px;">Multiple sellers&#160;</td>
			<td style="width:127px;">Catalog number not available&#160;</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Gauze Pads</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">67622</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Surgical drape</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">GM300&#160;</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">Silk Sutures&#160;</td>
			<td style="width:171px;">Look/Div Surgical Specialties</td>
			<td style="width:127px;">SP115</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">Nylon Sutures</td>
			<td style="width:171px;">Look/Div Surgical Specialties</td>
			<td style="width:127px;">SP185</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Durmont #5 forceps (2)&#160;</td>
			<td style="width:171px;">Fine Science Tools&#160;</td>
			<td style="width:127px;">11251-35</td>
			<td style="width:283px;">Angled 45&#176;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Surgical Scissors</td>
			<td style="width:171px;">Fine Science Tools&#160;</td>
			<td style="width:127px;">14028-10</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">3mm Vannas</td>
			<td style="width:171px;">Kent Scientific Corporation</td>
			<td style="width:127px;">INS600177</td>
			<td style="width:283px;">Straight blade</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Hartman Hemostats&#160;</td>
			<td style="width:171px;">Fine Scientific Tools</td>
			<td style="width:127px;">13002-10</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:249px;">Occluding filaments</td>
			<td style="width:171px;">Washington University</td>
			<td style="width:127px;">Specialty order</td>
			<td style="width:283px;">Filaments are silicone coated at Washington Univeristy and provided to UTSW facilities for a fee.&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Evans Blue</td>
			<td style="width:171px;">Sigma Aldritch</td>
			<td style="width:127px;">E2129-10G</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Filter Paper&#160;</td>
			<td style="width:171px;">Sigma Aldritch</td>
			<td style="width:127px;">WHA1001150</td>
			<td style="width:283px;">150 mm, circles, Grade 1&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Weigh Boats</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">02-202-101</td>
			<td style="width:283px;">2.5&#34; diameter</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">0.9% Sodium Chloride Injection USP&#160;</td>
			<td style="width:171px;">Baxter Pharmaceutics&#160;</td>
			<td style="width:127px;">2B1321</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">0.3cc insulin syringe with 29 g needle</td>
			<td style="width:171px;">Becton Dickinson Labware</td>
			<td style="width:127px;">309301</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Flat bottom restrainer&#160;</td>
			<td style="width:171px;">Braintree Scientific&#160;</td>
			<td style="width:127px;">FB&#160;M</td>
			<td style="width:283px;">2.0&#34; diameter</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">TTC</td>
			<td style="width:171px;">Sigma</td>
			<td style="width:127px;">T8877</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">10X PBS, pH 7.4</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">BP399-20</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="80">
			<td height="80" style="height:80px;width:249px;">Water Bath</td>
			<td style="width:171px;">Multiple sellers&#160;</td>
			<td style="width:127px;">Catalog number not available&#160;</td>
			<td style="width:283px;">Scintillation tubes with TTC may be manually held under running warm water as an alternative to the water bath.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">Styrofoam board</td>
			<td style="width:171px;">Multiple sellers&#160;</td>
			<td style="width:127px;">Catalog number not available&#160;</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Large Syringe Kit</td>
			<td style="width:171px;">PumpSystems Inc</td>
			<td style="width:127px;">P-SYRKIT-LG</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Perfusion Pump</td>
			<td style="width:171px;">PumpSystems Inc</td>
			<td style="width:127px;">NE-300&#160;</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">60 cc syringe</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">NC9203256</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">27g winged infusion set</td>
			<td style="width:171px;">Kawasumi Laboratories, Inc</td>
			<td style="width:127px;">D3K1-25G 1</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">20 ml scintillation vial</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">50-367-126</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Stainless steel spatula</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">14-373-25A</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">Alto acrylic 1.0 mm mouse brain, coronal</td>
			<td style="width:171px;">CellPoint Scientific&#160;</td>
			<td style="width:127px;">Catalog number not available&#160;</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">0.21 mm stainless steel blades, 25 pk</td>
			<td style="width:171px;">CellPoint Scientific&#160;</td>
			<td style="width:127px;">Catalog number not available&#160;</td>
			<td style="width:283px;">Reusable cryostat blades are an inexpensive alternative.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">4% paraformaldehyde</td>
			<td style="width:171px;">Santa Cruz Biotechnology&#160;</td>
			<td style="width:127px;">SC-281692</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Superfrost microscope slides&#160;</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">12-550-15</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">HP Scanjet G4050</td>
			<td style="width:171px;">Multiple sellers&#160;</td>
			<td style="width:127px;">Catalog number not available&#160;</td>
			<td style="width:283px;">Other commercial scanners are suitable for this step in the protocol.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">ImageJ&#160;</td>
			<td style="width:171px;">National Institute of Health</td>
			<td style="width:127px;">Catalog number not available&#160;</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Analytical Balance</td>
			<td style="width:171px;">Mettler Toledo&#160;</td>
			<td style="width:127px;">XSE 205U</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Precision Compact Oven&#160;&#160;</td>
			<td style="width:171px;">Thermo Scientific&#160;</td>
			<td style="width:127px;">PR305225M</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">1.7 mL microcentrifuge tubes (Eppendorfs)</td>
			<td style="width:171px;">Denville Scientific&#160;</td>
			<td style="width:127px;">C2170</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">Formamide</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">BP228-100</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:249px;">96-well plates</td>
			<td style="width:171px;">Fisher Scientific</td>
			<td style="width:127px;">07-200-9</td>
			<td style="width:283px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:249px;">Epoch Microplate Spectrophotometer&#160;</td>
			<td style="width:171px;">BioTek&#160;</td>
			<td style="width:127px;">Catalog number not available&#160;</td>
			<td style="width:283px;"></td>
		</tr>
	</tbody>
</table>

quantification of neurovascular protection following repetitive hypoxic preconditioning and transient middle cerebral artery occlusion in mice

النماذج الحيوانية التجريبية من السكتة الدماغية هي أدوات لا تقدر بثمن لفهم السكتة الدماغية علم الأمراض وتطوير استراتيجيات علاجية أكثر فعالية. بروتوكول 2 أسبوع لتكرار شروط مسبقة ميتة (رهب) يدفع حماية طويلة الأمد ضد الجهاز العصبي المركزي (CNS) إصابة في نموذج الفأر من السكتة الدماغية التنسيق. يتكون رهب من 9 التعرض لنقص الأكسجين العشوائية التي تختلف في كل مدة (2 أو 4 ساعات) وكثافة (8٪ و 11٪ O 2). RHP يقلل كميات احتشاء، حاجز الدم في الدماغ (BBB) ​​اضطراب، والاستجابة الالتهابية في مرحلة ما بعد السكتة الدماغية لعدة أسابيع بعد آخر تعرض لنقص الأكسجين، مما يشير إلى الحث على المدى الطويل من النمط الظاهري CNS واقية الذاتية. منهجية لتقدير مزدوج من حجم احتشاء وBBB اضطراب فعالة في تقييم الحماية الوعائية العصبية في الفئران مع RHP أو neuroprotectants المفترض الآخرين. وقد اشترطت الذكور البالغين فئران وبستر السويسرية التي كتبها رهب أو التعرض المدة أي ما يعادل 21٪ O <suب&gt; 2 (أي هواء الغرفة). وكان المستحث A 60 دقيقة عابرة وسط انسداد الشريان الدماغي (tMCAo) 2 أسابيع بعد التعرض ميتة الماضي. وقد أكد كل من انسداد وضخه من قبل عبر الجمجمة دوبلر الليزر flowmetry. اثنان وعشرون ساعة بعد ضخه، ايفانز الأزرق (EB) كانت تدار عن طريق الوريد من خلال حقن الوريد الذيل. في وقت لاحق 2 ساعة، تم التضحية الحيوانات عن طريق أقسام جرعة زائدة والدماغ الأيزوفلورين كانت ملطخة 2،3،5- كلوريد triphenyltetrazolium (TTC). ثم تم كميا كميات عوائق. بعد ذلك، تم استخراج EB من النسيج على مدار 48 ساعة لتحديد BBB اضطراب بعد tMCAo. باختصار، RHP هو بروتوكول بسيط يمكن تكرارها، بأقل تكلفة ممكنة، للحث على حماية الوعائية العصبية الذاتية على المدى الطويل من الاصابة بجلطة في الفئران، مع احتمال متعدية للدول مرض الموالية للالتهابات المستندة إلى الجهاز العصبي المركزي وجهازية أخرى.

<html> وشملت هذه الدراسة 25 من الذكور الفئران وبستر السويسرية التي كانت 10 أسابيع من العمر في بداية العشوائية في RHP (ن = 10) أو 21٪ O 2 (ن = 15) مجموعات. بعد أسبوعين من التعرض RHP النهائي، وأجريت العمليات الجراحية، مع مجموعات أعمى ويوازن بين أيام. بعد tMCAo، توفي 1 الماوس أثناء الا...

Watch this Scientific Journal Video about Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice at JoVE.com

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

الكمي لحماية عائية عصبية بعد المتكرر نقص الأوكسجين شروط مسبقة وعابر الأوسط الشريان الدماغي انسداد في الفئران

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week ...

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13.7K Views.  University of Texas Southwestern Medical Center. This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

Video: Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

Modeling Stroke in Mice - Middle Cerebral Artery Occlusion with the Filament Model

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are very limited. To meet the need for novel therapeutic approaches, experimental stroke research frequently employs rodent models of focal cerebral ischaemia. Most researchers use permanent or transient occlusion of the middle cerebral artery (MCA) in mice or rats.
Proximal occlusion of the middle cerebral artery (MCA) via the intraluminal suture technique (so called filament or suture model) is probably the most frequently used model in experimental stroke research. The intraluminal MCAO model offers the advantage of inducing reproducible transient or permanent ischaemia of the MCA territory in a relatively non-invasive manner. Intraluminal approaches interrupt the blood flow of the entire territory of this artery. Filament occlusion thus arrests flow proximal to the lenticulo-striate arteries, which supply the basal ganglia. Filament occlusion of the MCA results in reproducible lesions in the cortex and striatum and can be either permanent or transient. In contrast, models inducing distal (to the branching of the lenticulo-striate arteries) MCA occlusion typically spare the striatum and primarily involve the neocortex. In addition these models do require craniectomy. In the model demonstrated in this article, a silicon coated filament is introduced into the common carotid artery and advanced along the internal carotid artery into the Circle of Willis, where it blocks the origin of the middle cerebral artery. In patients, occlusions of the middle cerebral artery are among the most common causes of ischaemic stroke. Since varying ischemic intervals can be chosen freely in this model depending on the time point of reperfusion, ischaemic lesions with varying degrees of severity can be produced. Reperfusion by removal of the occluding filament at least partially models the restoration of blood flow after spontaneous or therapeutic (tPA) lysis of a thromboembolic clot in humans.
In this video we will present the basic technique as well as the major pitfalls and confounders which may limit the predictive value of this model.

Filamentous occlusion of the Middle cerebral artery is a common model for studying ischemic stroke in mice.

السكتة الدماغية النمذجة في الفئران -- الشرق انسداد الشريان الدماغي مع نموذج الشعيرة

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are ...

Mouse Model of Middle Cerebral Artery Occlusion

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the brain (ischemic stroke) 2. Since most ischemic strokes (~80%) occur in the territory of middle cerebral artery (MCA) 3, many animal stroke models that have been developed have focused on this artery. The intraluminal monofilament model of middle cerebral artery occlusion (MCAO) involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the MCA territory 4. The technique can be used to model permanent or transient occlusion 5. If the suture is removed after a certain interval (30 min, 1 h, or 2 h), reperfusion is achieved (transient MCAO); if the filament is left in place (24 h) the procedure is suitable as a model of permanent MCAO. This technique does not require craniectomy, a neurosurgical procedure to remove a portion of skull, which may affect intracranial pressure and temperature 6. It has become the most frequently used method to mimic permanent and transient focal cerebral ischemia in rats and mice 7,8. To evaluate the extent of cerebral infarction, we stain brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) to identify ischemic brain tissue 9. In this video, we demonstrate the MCAO method and the determination of infarct size by TTC staining.

We demonstrate in the video a method for producing a middle cerebral artery occlusion in adult mice using an intraluminal monofilament. We also show how to evaluate the extent of cerebral infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining.

نموذج الفأر من انسداد الشريان الدماغي الأوسط

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the ...

Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United States in 20091,2. The intraluminal middle cerebral artery occlusion (MCAO) model was developed by Koizumi4 in 1986 to simulate this 
impactful human pathology in the rat. A modification of the MCAO method was later presented by Longa3. Both techniques have been widely used to identify 
molecular mechanisms of brain injury resulting from ischemic stroke and potential therapeutic modalities5. This relatively noninvasive method in rats has been 
extended to use in mice to take advantage of transgenic and knockout strains6,7. To model focal cerebral ischemia, an intraluminal suture is advanced via 
the internal carotid artery to occlude the base of the MCA. Retracting the suture after a specified period of time mimics spontaneous reperfusion, but the 
suture can also be permanently retained. This video will be demonstrating the two major approaches for performing intraluminal MCAO procedure in mice in a 
stepwise fashion, as well as providing insights for potential drawbacks and pitfalls. The ischemic brain tissue will subsequently be stained by 
2,3,5-triphenyltetrazolium chloride (TTC) to evaluate the extent of cerebral infarction8.

Intraluminal Middle Cerebral Artery Occlusion MCAO Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

The intraluminal middle cerebral artery occlusion (MCAO) model is the most frequent used model among experimental ischemic stroke models. Here we will demonstrate the entire model in detail with the guide of Laser Doppler flowmetry, and its representative results.

الشريان الدماغي الأوسط انسداد داخل اللمعة (MCAO) النموذجي للاصابة بالسكتة الدماغية مع توجيه الليزر دوبلر Flowmetry في الفئران

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United ...

Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in 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 20091, 2. Several models of cerebral ischemia have been developed to mimic the human condition of stroke. It has been suggested that up to 80% of all strokes result from ischemic damage in the middle cerebral artery (MCA) area3. In the early 1990s, endothelin-1 (ET-1) 4 was used to induce ischemia by applying it directly adjacent to the surface of the MCA after craniotomy. Later, this model was modified 5 by using a stereotaxic injection of ET-1 adjacent to the MCA to produce focal cerebral ischemia. The main advantages of this model include the ability to perform the procedure quickly, the ability to control artery constriction by altering the dose of ET-1 delivered, no need to manipulate the extracranial vessels supplying blood to the brain as well as gradual reperfusion rates that more closely mimics the reperfusion in humans5-7. On the other hand, the ET-1 model has disadvantages that include the need for a craniotomy, as well as higher variability in stroke volume8. This variability can be reduced with the use of laser Doppler flowmetry (LDF) to verify cerebral ischemia during ET-1 infusion. Factors that affect stroke variability include precision of infusion and the batch of the ET-1 used6. Another important consideration is that although reperfusion is a common occurrence in human stroke, the duration of occlusion for ET-1 induced MCAO may not closely mimic that of human stroke where many patients have partial reperfusion over a period of hours to days following occlusion9, 10. This protocol will describe in detail the ET-1 induced MCAO model for ischemic stroke in rats. It will also draw attention to special considerations and potential drawbacks throughout the procedure.

Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed.

Endothelin-1 المستحثة انسداد الشريان الدماغي الأوسط نموذج للالسكتة الدماغية مع التوجيه بالليزر Flowmetry دوبلر في الجرذ

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 ...

Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be performed in many locations overlying the rat cerebrum, but is most easily carried out by creating a craniectomy over the readily accessible frontal or parietal bones. Most frequently, we have used this technique in combination with the endothelin-1 middle cerebral artery occlusion model of ischemic stroke to quantify the changes in middle cerebral artery vessel diameter that occur with injection of endothelin-1 into the brain parenchyma adjacent to the proximal MCA4, 5. In order to visualize the proximal portion of the MCA during endothelin -1 induced MCAO, we use a technique to create a cranial window through the temporal bone on the lateral aspect of the rat skull (Figure 1). Cerebral arteries can be visualized either with the dura intact or with the dura incised and retracted. Most commonly, we leave the dura intact during visualization since endothelin-1 induced MCAO involves delivery of the vasoconstricting peptide into the brain parenchyma. This bypasses the need to incise the dura directly over the visualized vessels for drug delivery. This protocol will describe how to create a cranial window to visualize cerebral arteries in a step-wise fashion, as well as how to avoid many of the potential pitfalls pertaining to this method.

This article describes a method for visualizing rat cerebral arteries through a cranial window using temporal craniectomy in order to view proximal portions of the middle cerebral artery (Figure 1). This versatile method can be combined with various techniques of drug delivery to measure cerebral artery reactivity in vivo.

الاستفادة من وجود نافذة القحف لتصور الشريان الدماغي الأوسط خلال Endothelin-1 انسداد الشريان الدماغي الأوسط المستحثة

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be ...

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

الأمثل نظام لرصد الإرواء الدماغي في نموذج الفئران من السكتة الدماغية داخل اللمعة انسداد الشريان الدماغي الأوسط

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

Modeling Stroke in Mice: Permanent Coagulation of the Distal Middle Cerebral Artery

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical options are available for a small proportion of stroke patients in the acute phase. Current research is intensively searching for novel therapeutic strategies and is increasingly focusing on the sub-acute and chronic phase after stroke because more patients might be eligible for therapeutic interventions in a prolonged time window. These delayed mechanisms include important pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity and regeneration. In order to analyze these mechanisms and to subsequently evaluate novel drug targets, experimental stroke models with clinical relevance, low mortality and high reproducibility are sought after. Moreover, mice are the smallest mammals in which a focal stroke lesion can be induced and for which a broad spectrum of transgenic models are available. Therefore, we describe here the mouse model of transcranial, permanent coagulation of the middle cerebral artery via electrocoagulation distal of the lenticulostriatal arteries, the so-called &#8220;coagulation model&#8221;. The resulting infarct in this model is located mainly in the cortex; the relative infarct volume in relation to brain size corresponds to the majority of human strokes. Moreover, the model fulfills the above-mentioned criteria of reproducibility and low mortality. In this video we demonstrate the surgical methods of stroke induction in the &#8220;coagulation model&#8221; and report histological and functional analysis tools.

Various murine models of middle cerebral artery occlusion (MCAO) are widely used in experimental brain research. Here, we demonstrate the model of transcranial permanent distal MCAO which produces consistent cortical infarction of a size corresponding to damage imposed by the majority of human ischemic strokes.

السكتة الدماغية النمذجة في الفئران: التخثر الدائم من الشريان الدماغي الأوسط القاصي

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical ...

Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic stroke. However, the use of tPA is limited by its narrow therapeutic window and by increased risk of hemorrhagic transformation. There is an urgent need to develop suitable stroke models to study new thrombolytic agents and strategies for treatment of ischemic stroke. At present, two major types of ischemic stroke models have been developed in rats and mice: intraluminal suture MCAO and embolic MCAO. Although MCAO models via the intraluminal suture technique have been widely used in mechanism-driven stroke research, these suture models do not mimic the clinical situation and are not suitable for thrombolytic studies. Among these models, the embolic MCAO model closely mimics human ischemic stroke and is suitable for preclinical investigation of thrombolytic therapy. This embolic model was first developed in rats by Overgaard et al.1 in 1992 and further characterized by Zhang et al. in 19972. Although embolic MCAO has gained increasing attention, there are technical problems faced by many laboratories. To meet increasing needs for thrombolytic research, we present a highly reproducible model of embolic MCAO in the rat, which can develop a predictable infarct volume within the MCA territory. In brief, a modified PE-50 tube is gently advanced from the external carotid artery (ECA) into the lumen of the internal carotid artery (ICA) until the tip of the catheter reaches the origin of the MCA. Through the catheter, a single homologous blood clot is placed at the origin of the MCA. To identify the success of MCA occlusion, regional cerebral blood flow was monitored, neurological deficits and infarct volumes were measured. The techniques presented in this paper should help investigators to overcome technical problems for establishing this model for stroke research.

Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats

In this paper, we demonstrate a standard method for producing an embolic middle cerebral artery occlusion with homologous blood clots (fibrin-rich) in adult rat. This model closely mimics human ischemic stroke and is suitable for preclinical study of thrombolytic therapy for ischemic stroke.

صمي الأوسط انسداد الشريان الدماغي (MCAO) لالدماغية السكتة الدماغية مع مثلي تجلط الدم في الجرذان

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic ...

Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ischemic in origin and occur in the territory of the middle cerebral artery (MCA). Currently the only Food and Drug Administration (FDA) approved drug for the treatment of this devastating disease is tissue plasminogen activator (tPA). However, tPA has a small therapeutic window for administration (3 - 6 hr), and is only effective in 4% of the patients who actually receive it. Current research focuses on understanding the pathophysiology of stroke in order to find potential therapeutic targets. Thus, reliable models are crucial, and the MCA occlusion (MCAo) model (also termed the intraluminal filament or suture model) is deemed to be the most clinically relevant surgical model of ischemic stroke, and is fairly non-invasive and easily reproducible. Typically the MCAo model is used with rodents, especially with mice due to all the genetic variations available for this species. Here we describe (and present in the video) how to successfully perform the MCAo model (with reperfusion) in mice to generate reliable and reproducible data.

In order to understand the pathophysiology of stroke, it is important to use reliable models. This paper will describe one of the most frequently used stroke models in mice, termed the middle cerebral artery occlusion (MCAo) model (also termed the intraluminal filament or suture model) with reperfusion.

نهج الجراحي لانسداد الشريان الدماغي الأوسط وضخه المستحثة السكتة الدماغية في الفئران

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature and mature brain. Stroke occurs commonly in the perinatal period in humans, and transient ischemia-reperfusion is the most common form of stroke in neonates. The reperfusion phase is a critical component of injury progression, which occurs over a period of days to weeks, and of the endogenous response to injury. This postnatal day 10 (p10) rat model of transient middle cerebral artery occlusion (tMCAO) creates a unilateral, non-hemorrhagic focal ischemia-reperfusion injury that can be utilized to study the mechanisms of focal injury and repair in the full-term-equivalent brain. The injury pattern that is produced by tMCAO is consistent and highly reproducible and can be confirmed with MRI or histological analyses. The severity of injury can be manipulated through changes in occlusion time and other methods that will be discussed.

Neonatal stroke is a significant cause of early brain injury requiring a translational model with consistent focal injury patterns and high reproducibility in order to enable study. This study describes the detailed surgical procedure for creating a non-hemorrhagic, unilateral focal ischemia-reperfusion injury in full-term-equivalent rodents.

عابر الدماغي الأوسط الشريان انسداد نموذج من السكتة الدماغية حديثي الولادة في الفئران P10

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...