XHTW هو كيك WM كلودج مؤسسة الشباب في البحوث الطبية ، وأيضا على دعم من قبل المعاهد الوطنية للصحة / NHLBI المنح R01 - R01 HL089598 وR01HL091947 ، والضمور العضلي جمعية المنحة # 69238. RJvO وهو حاصل على زمالة جمعية الفسيولوجية الأمريكية 2008-2010 بعد الدكتوراه في علم الجينوم الفسيولوجية. كما يدعم هذا العمل في جزء من مؤسسة التحالف من أجل Leducq كناس CaMKII في اشارة الى القلب.

الجزء 1 : إعداد حقل التشغيل <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> وتطهيرها مجال التشغيل مع ايزوبروبيل 75 ٪. </li><li style=";text-align:right;direction:rtl"> تأكد من وسادة التدفئة وعلى الحق في درجة الحرارة. نظام المقترح هو Gaymar ضخ المياه المنتشرة متصلا لوحة العلاج التي تم الاحتفاظ ب 37 درجة مئوية ± 1 درجة مئوية. فمن المهم للحفاظ على درجة حرارة الجسم الطبيعية أثناء الجراحة لتفادي انخفاض سريع في معدل ضربات القلب. </li><li style=";text-align:right;direction:rtl"> يتم تعقيم الأدوات الجراحية في التعقيم حبة الساخن قبل الجراحة. لهذا الإجراء ، سوف تحتاج الصكوك التالية الجراحية : مقص حادة ، وبطبيعة الحال منحني ملقط X 2 ، ودفع غرامة 45 درجة الزاوية ملقط X 2 ، ربيع مقص الزاوية ، مبعاد الصدر وحامل الإبرة. </li><li style=";text-align:right;direction:rtl"> ينبغي تطبيقها سيكون على يد القطن في حالة النزيف. </li></ol> الجزء 2 : إعداد وتركيب الانبوبة من الفئران <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> يتم تخدير الفئران في غرفة التعريفي مع 2 ٪ isoflurane مختلطة مع 0.5 -1.0 لتر / دقيقة 100 ٪ O 2. </li><li style=";text-align:right;direction:rtl"> يتم استخدام الشعر كليبرز حلق الفراء من العنق إلى مستوى وسط الصدر. </li><li style=";text-align:right;direction:rtl"> يتم وضع الماوس في وضع الاستلقاء فوق وسادة التدفئة من أجل الحفاظ على درجة حرارة الجسم. </li><li style=";text-align:right;direction:rtl"> يتم وضع الشريط المطاطي على الأسنان الأمامية للحيوان ليالي لتمديد الرقبة. باستخدام الملقط المنحني في يد واحدة ، يتم التلاعب بها اللسان برفق إلى الجانب. مع من ناحية أخرى ، يتم تنفيذ التنبيب الرغامي استخدام أنابيب PE 90. ثم يتم توصيل الأنبوب الرغامي إلى وحدة تخزين ، تدوير هارفارد القوارض الدراجات التنفس الصناعي في الأنفاس 125-150 / دقيقة وحجم المد والجزر من 0.3ml - 0.1. أثناء العملية الجراحية ، والتخدير في المحافظة 1،5-2 ٪ isoflurane مع 0،5-1،0 لتر / دقيقة 100 ٪ O 2. يتم التحقق من المستوى الصحيح للتخدير عن طريق الضغط على الماوس الظفر (إصبع القدم قرصة منعكس). </li><li style=";text-align:right;direction:rtl"> وتطهيرها والجراحي الميداني مع الحل betadine تليها الكحول 70 ٪. ويتكرر هذا الإجراء ثلاث مرات. </li><li style=";text-align:right;direction:rtl"> لمنع التلوث من الحقل خلال العملية الجراحية ، يتم وضع ثنى العقيمة على الماوس ولم يتبق سوى عملية الميدانية عرضة للخطر. </li><li style=";text-align:right;direction:rtl"> وتستخدم مجموعة من القفازات المعقمة لكل فرد الماوس. </li></ol> الجزء 3 : ربط الشريان الأورطي المستعرض <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> يتم تنفيذ جزئي لبضع الصدر الضلع الثاني تحت المجهر الجراحي والقص تراجع باستخدام مبعاد الصدر. </li><li style=";text-align:right;direction:rtl"> غيض غرامة 45 درجة وتستخدم ملقط الزاوية لفصل بلطف الأنسجة والدهون من الغدة الصعترية قوس الأبهر. </li><li style=";text-align:right;direction:rtl"> بعد التعرف على الأبهر عرضية ، يتم وضع قطعة صغيرة من الحرير 6.0 خياطة الشرايين السباتية بين اللامسمى واليسرى (الشكل 1). </li><li style=";text-align:right;direction:rtl"> وترتبط اثنين عقدة فضفاضة حول الأبهر عرضية وقطعة صغيرة من 27 ½ يتم وضع إبرة حادة موازية لقياس الأبهر عرضية. يرتبط بسرعة عقدة الاولى ضد الإبرة ، تليها الثانية وإزالة الإبرة فورا من أجل تسفر عن انقباض 0.4mm في القطر. التحكم في الفئران صورية ، فإن الإجراء بأكمله متطابقة باستثناء ربط الشريان الأورطي. </li><li style=";text-align:right;direction:rtl"> تتم إزالة مبعاد الصدر وتدفق للخارج التنفس الصناعي لمدة 2 ثانية مقروص لإعادة تضخيم الرئتين. </li><li style=";text-align:right;direction:rtl"> إغلاق القفص الصدري باستخدام خياطة prolene 6.0 مع نمط خياطة توقف. </li><li style=";text-align:right;direction:rtl"> إغلاق خياطة الجلد باستخدام prolene 6.0 مع وجود نمط مستمر خياطة. </li></ol> الجزء 4 : في مرحلة ما بعد المنطوق الاسترداد <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> لتسكين بعد العملية ، يتم حقن الفأر مع البوبرينورفين (0،1 ملغ / كلغ) intraperitoneally. إذا كانت هناك علامات الجفاف بعد الجراحة ، ويتم إعطاء المالحة intraperitoneally العقيمة. </li><li style=";text-align:right;direction:rtl"> وخفضت تدريجيا تخدير للموقف قبالة وإزالة الأنبوب الرغامي عند ظهور علامات التنفس العفوي يحدث. </li><li style=";text-align:right;direction:rtl"> يتم نقل الماوس إلى وضعية الانبطاح وسمح لاسترداد على وسادة التدفئة. </li></ol> الجزء 5 : تأكيد نجاح عملية ربط الشريان الأورطي المستعرض <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> بعد أسبوع واحد تاك ، والفأر هو إعادة تخدير لتحديد درجة من الضغط الزائد الناتج عن ربط الشريان الأورطي عرضية. كما هو موضح أعلاه ، يتم الاحتفاظ التخدير في 1،5-2 ٪ isoflurane مع 0،5-1،0 لتر / دقيقة 100 ٪ O 2 والحفاظ على درجة حرارة الجسم عند 37 درجة مئوية ± 1 درجة مئوية. </li><li style=";text-align:right;direction:rtl"> يتم وضع مسبار دوبلر 20 ميغاهيرتز على الجانبين الأيسر والأيمن من الرقبة بزاوية 45 درجة للكشف عن تدفق السرعات. تم استخدام الكمبيوتر المستندة إلى معالج الاشارات دوبلر (اندوس الآلات ، وهيوستن ، تكساس) لعرض وتخزين الاشارات دوبلر 5 </li><li style=";text-align:right;direction:rtl"> اعتمادا على درجة من الضغط الزائد اللازم لبروتوكول تجريبي ، يتم تضمين فقط مع الفئران السباتي الأيمن (RC) / يسار السباتي (LC) نسبة التدفق داخل نطاق معين لمزيد من التحليل. على سبيل المثال ، وعلى درجة معتدلة من الضغط الزائد يؤدي إلى نسبة 5-8 ، في حين أن أكثر تشددا مما أدى إلى تقليص عبء ضغط شديد يؤدي رOA نسبة 80-10 (الشكل 5.3A). حيوان صورية (لا تعمل إلا ligated) ، ومع ذلك ، فمن المتوقع أن يكون لها نسبة من ~ 1 (الشكل 5.3B). </li></ol> 6 : نتائج الممثل بقاء نموذجية الجراحية التالية TAC في البرية من نوع الفئران حوالي 80-90 ٪. وربط جراحية ناجحة في الشريان الأورطي عرضية يؤدي إلى سرعة تدفق دوبلر نسبة بين الشريان السباتي الأيمن والأيسر (RC / LC) من 50-10 (الشكل 1). بالمقارنة مع الفئران التي تديرها صورية (الشكل 2A) ، ومن المتوقع أن الفئران مع الضغط الزائد على وضع تضخم القلب في غضون 1-2 أسابيع ، وبعد توسع القلب 6-8 أسابيع ، تبعا لضيق للانقباض (الشكل 2B). الوفيات خلال الشهر الأول بعد TAC منخفضة عادة (&lt;20 ٪) في الفئران من النوع البري ، على الرغم من الفئران المعدلة وراثيا قد يحمل معدلات البقاء على قيد الحياة المختلفة. 6،7 <img src="/files/ftp_upload/1729/1729fig1tmb.jpg" alt="الشكل 1" border="0" /> الشكل 1. تأكيدا لربط الأورطي باستخدام مستعرض دوبلر قياس السرعة. إشارات الممثل سرعة دوبلر للحق (RC) وترك الشرايين السباتية (LC) 6 أيام ما بعد معاهدة الصداقة والتعاون. هذه التسجيلات يبرهن على وجود ربط ناجحة مع نسبة تدفق RC / LC من ~ 6.8. الرجاء <a href="https://www.jove.com/files/ftp_upload/1729/1729fig1.jpg">انقر هنا</a> لرؤية نسخة أكبر من الرقم 1. <img src="/files/ftp_upload/1729/1729fig2.jpg" alt="الشكل 2" /> الشكل 2. ممثل صور القلب الجامع عقب تاك. ألف وباء. الشام التي تديرها الماوس نوع البرية القلب. نوع البرية قلب فأر بعد 16 أسبوعا تاك. كل سطر = 1MM.

<ol>
	<li>Rockman, H. A., Ross, R. S., Harris, A. N., Knowlton, K. U., Steinhelper, M. E., Field, L. J., Ross, J., Chein, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Segregation+of+atrial-specific+and+inducible+expression+of+an+atrial+natriuretic+factor+transgene+in+an+in+vivo+murine+model+of+cardiac+hypertrophy.">Segregation of atrial-specific and inducible expression of an atrial natriuretic factor transgene in an in vivo murine model of cardiac hypertrophy.</a> Proc. Natl. Acad. Sci. USA. 88, 8277-8281 (1991).</li><li>Heineke, J., Molkentin, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+cardiac+hypertrophy+by+intracellular+signaling+pathways.">Regulation of cardiac hypertrophy by intracellular signaling pathways.</a> Nat. Rev. Mol. Cell. Biol. 7, 589-600 (2006).</li><li>Hartley, C. J., Reddy, A. K., Madala, S., Michael, L. H., Entman, M. L., Taffett, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doppler+estimation+of+reduced+coronary+flow+reserve+in+mice+with+pressure+overload+cardiac+hypertrophy.">Doppler estimation of reduced coronary flow reserve in mice with pressure overload cardiac hypertrophy.</a> Ultrasound Med. Biol. 34, 892-901 (2008).</li><li>Reddy, A. K., Taffett, G. E., Li, Y. -. H., Lim, S. -. W., Pham, T. T., Pocius, J. S., Entman, M. L., Michael, L. H., Hartley, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doppler+signal+processing+for+use+in+mice:+applications.">Doppler signal processing for use in mice: applications.</a> IEEE Trans. Biomed. Eng. 52, 1771-1783 (2005).</li><li>Li, Y. -. H., Reddy, A. K., Ochoa, L. N., Pham, T. T., Hartley, C. J., Micheal, L. H., Entman, M. L., Taffett, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+age+on+peripheral+vascular+response+to+transverse+aortic+banding+in+mice.">Effect of age on peripheral vascular response to transverse aortic banding in mice.</a> J. Gerontol. 58A, 895-899 (2003).</li><li>Bourajjaj, M., Armand, A. S., Martins, d. a. C. o. s. t. a., Weijts, P. A., Nagel, B. V. a. n. d. e. r., Heeneman, R., Wehrens, S., Windt, X. H. D. e., J, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NFATc2+is+a+necessary+mediator+of+calcineurin-dependent+cardiac+hypertrophy+and+heart+failure.">NFATc2 is a necessary mediator of calcineurin-dependent cardiac hypertrophy and heart failure.</a> J. Biol. Chem. 28, 22295-22303 (2008).</li><li>Oort, R. J. v. a. n., Respress, J. L., Li, N., Reynolds, C., Almeida, A. C. D. e., Skapura, D. G., Windt, L. J. D. e., Wehrens, X. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+development+of+pressure+overload+induced+cardiac+hypertrophy+and+dysfunction+in+an+RyR2-R176Q+knockin+mouse+model.">Accelerated development of pressure overload induced cardiac hypertrophy and dysfunction in an RyR2-R176Q knockin mouse model.</a> Hypertension. 55, (2010).</li><li>Tarnavski, O., McMullen, J. R., Schinke, M., Nie, Q., Kong, S., Izumo, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+cardiac+surgery:+comprehensive+techniques+for+the+generation+of+mouse+models+of+human+diseases+and+their+application+for+genomic+studies.">Mouse cardiac surgery: comprehensive techniques for the generation of mouse models of human diseases and their application for genomic studies.</a> Physiol Genomics. 16, 349-360 (2004).</li></ol>

تاك ، الذي يحاكي الإنسان تضيق الأبهر ، هو طريقة شائعة للحث على تضخم القلب وقصور في القلب لدى الفئران. مواقع بديلة لانقباض الأبهر تشمل الأبهر الصاعد والبطن. تصاعدي انقباض الأبهر يوفر الزائد المدقع وأكثر سرعة في البطين الأيسر (LV). في المقابل ، انقباض البطن الأبهر يترك ا?...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Gaymar T/pump</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>Model TP-500</td>
<td>Circulating water pump</td>
</tr>
<tr>
<td>Temp. therapy pads</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>60-3414</td>
<td>&nbsp;</td>
<tr>
<td>Hot bead sterilizer</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>18000-45</td>
<td>&nbsp;</td>
<tr>
<td>Blunt scissors</td>
<td>&nbsp;</td>
<td>Roboz</td>
<td>RS-5980</td>
<td>&nbsp;</td>
<tr>
<td>Angled spring scissors</td>
<td>&nbsp;</td>
<td>Roboz</td>
<td>RS-5668</td>
<td>&nbsp;</td>
<tr>
<td>Chest retractor</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>17002-02</td>
<td>&nbsp;</td>
<tr>
<td>Course curved forceps</td>
<td>&nbsp;</td>
<td>Roboz</td>
<td>RS-5138</td>
<td>&nbsp;</td>
<tr>
<td>Dumont fine 45&deg; angled forceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11253-25</td>
<td>&nbsp;</td>
<tr>
<td>Needle holder</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>12565-14</td>
<td>&nbsp;</td>
<tr>
<td>Electric shaver</td>
<td>&nbsp;</td>
<td>General Electric</td>
<td>&nbsp;</td>
<td>For shaving mouse fur</td>
</tr>
<tr>
<td>PE 90 tubing</td>
<td>&nbsp;</td>
<td>BD</td>
<td>427420</td>
<td>For intubation</td>
</tr>
<tr>
<td>MiniVent (ventilator)</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>Type 845</td>
<td>&nbsp;</td>
<tr>
<td>Betadine</td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>19-027132</td>
<td>May be purchased at medical supply store</td>
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<tr>
<td>Sterile gloves</td>
<td>&nbsp;</td>
<td>Mckesson</td>
<td>20-1565</td>
<td>&nbsp;</td>
<tr>
<td>Stereo microscope</td>
<td>&nbsp;</td>
<td>Unico</td>
<td>ZM186</td>
<td>&nbsp;</td>
<tr>
<td>6-0 silk suture</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>18020-60</td>
<td>&nbsp;</td>
<tr>
<td>27 &frac12; gauge needle</td>
<td>&nbsp;</td>
<td>BD</td>
<td>305109</td>
<td>&nbsp;</td>
<tr>
<td>6-0 prolene suture</td>
<td>&nbsp;</td>
<td>Mckesson</td>
<td>3286</td>
<td>&nbsp;</td>
<tr>
<td>Buprenorphine</td>
<td>&nbsp;</td>
<td>Institutional Animal Facility</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
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transverse aortic constriction in mice

مستعرضة انقباض الأبهر (TAC) في الماوس هو نموذج تجريبي استخداما للضغط الزائد الناجم عن تضخم القلب وقصور في القلب.<sup&gt; 1</sup&gt; TAC يؤدي الى تضخم في البداية تعويض للقلب ، والتي غالبا ما يترافق مع تعزيز مؤقت لانقباض القلب. بمرور الوقت ، ومع ذلك ، استجابة لزيادة الحمل على الدورة الدموية المزمنة يصبح التأقلم ، مما أدى إلى اتساع القلب وقصور في القلب.<sup&gt; 2</sup&gt; تم التحقق من صحة النموذج لأول مرة TAC الفئران التي Rockman<em&gt; وآخرون</em&gt;.<sup&gt; 1</sup&gt; ، ومنذ ذلك الحين يستخدم على نطاق واسع باعتبارها أداة قيمة لتقليد أمراض القلب والشرايين البشرية وتوضيح العمليات الأساسية إشارات تشارك في الاستجابة ضخامي القلب والقلب التنمية الفشل. بالمقارنة مع غيرها من نماذج تجريبية من قصور في القلب ، مثل انسداد كامل للالأمامي الأيسر النازل (LAD) الشريان التاجي ، ومعاهدة الصداقة والتعاون يقدم نموذجا أكثر استنساخه من تضخم القلب ودوام أكثر تدرجا في تطوير بقصور في القلب. هنا ، نحن تصف الإجراء خطوة بخطوة لتنفيذ معاهدة الصداقة والتعاون الجراحية في الفئران. لتحديد مستوى الضغط الزائد التي تنتجها عملية ربط الشريان الأورطي ، يتم استخدام مسبار دوبلر عالية التردد لقياس النسبة بين سرعات تدفق الدم في الشرايين السباتية في اليمين واليسار.<sup&gt; 3 و 4</sup&gt; مع بقاء معدلات العمليات الجراحية في 80-90 ٪ ، الأبهر عرضية النطاقات هي تقنية فعالة لحفز تضخم البطين الأيسر وفشل القلب في الفئران.

Watch this Scientific Journal Video about Transverse Aortic Constriction in Mice at JoVE.com

Transverse Aortic Constriction in Mice

مستعرضة انقباض الأبهر (TAC) في الفأر نموذج استخداما تجريبيا لدراسة الآليات الكامنة وراء تضخم القلب والقلب التنمية الفشل. هنا ، نحن تصف إجراءات لتقليص الأبهر لخلق درجة استنساخه من تضخم القلب في الفئران.

المستعرضة في انقباض الأورطي الفئران

Transverse aortic constriction (TAC) in the mouse is a commonly used experimental model for pressure overload-induced cardiac hypertrophy and heart ...

Research

JoVE Journal

Biology

66.6K Views.  Baylor College of Medicine (BCM). مستعرضة انقباض الأبهر (TAC) في الفأر نموذج استخداما تجريبيا لدراسة الآليات الكامنة وراء تضخم القلب والقلب التنمية الفشل. هنا ، نحن تصف إجراءات لتقليص الأبهر لخلق درجة استنساخه من تضخم القلب في الفئران.

Video: Transverse Aortic Constriction in Mice

Heterotopic Heart Transplantation in Mice

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable for studying rejection and immune response now that newer transgenic and gene knockout mice are available, and a large number of immunologic reagents have been developed. The heart transplant model is less stringent than the skin transplant models, although technically more challenging. We have developed a modified technique and have completed over 1000 successful cases of heterotopic heart transplantation in mice. When making anastomosis of the ascending aorta and abdominal aorta, two stay sutures are placed at the proximal and distal apexes of recipient abdominal aorta with the donor s ascending aorta, then using 11-0 suture for anastomosis on both side of aorta with continuing sutures. The stay sutures make the anastomosis easier and 11-0 is an ideal suture size to avoid bleeding and thrombosis.
When making anastomosis of pulmonary artery and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s pulmonary artery. The left wall of the inferior vena cava and donor s pulmonary artery is closed with continuing sutures in the inside of the inferior vena cava after, one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s pulmonary artery are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis. In this article, we provide details of the technique to supplement the video.

The mouse heterotopic heart transplantation model has been proven by many investigators to be an important method for studying mechanisms of rejection and immune response. However, the techniques involved are still challenging. By modifying standard techniques we have had success with more than 1000 transplants.

منتبذ زرع القلب في الفئران

The mouse heterotopic heart transplantation has been used widely since it was introduced by Drs. Corry and Russell in 1973. It is particularly valuable ...

Small Bowel Transplantation In Mice

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive medications, infection prophylaxis, and suitable patient selection helped improve actuarial graft and patient survival rates for all types of intestine transplantation. Patients with irreversible intestinal failure and complications of parenteral nutrition should now be routinely considered for small intestine transplantation. However, Survival rates for small intestinal transplantation have been slow to improve compares increasingly favorably with renal, liver, heart and lung. The small bowel transplantation is still unsatisfactory compared with other organs. Further progress may depend on better understanding of immunology and physiology of the graft and can be greatly facilitated by animal models. A wider use of mouse small bowel transplantation model is needed in the study of immunology and physiology of the transplantation gut as well as efficient methods in diagnosing early rejection. However, this model is limited to use because the techniques involved is an extremely technically challenging. We have developed a modified technique. When making anastomosis of portal vein and inferior vena cava, two stay sutures are made at the proximal apex and distal apex of the recipient s inferior vena cava with the donor s portal vein. The left wall of the inferior vena cava and donor s portal vein is closed with continuing sutures in the inside of the inferior vena cava after, after one knot with the proximal apex stay suture the right wall of the inferior vena cava and the donor s portal vein are closed with continuing sutures outside the inferior vena cave with 10-0 sutures. This method is easier to perform because anastomosis is made just on the one side of the inferior vena cava and 10-0 sutures is the right size to avoid bleeding and thrombosis.  In this article, we provide details of the technique to supplement the video.

The mouse small bowel transplantation model has been recognized as an important tool to study mechanismes of immune rejection and screen new immunosuppressive drugs. However, this model is limited to use because the techniques involved is an extremely technically challenge. Now we introduce the modified technique.

زرع الأمعاء الصغيرة في الفئران

Since 1990, the development of tacrolimus-based immunosuppression and improved surgical techniques, the increased array of potent immunosuppressive ...

Aortic Ring Assay

Angiogenesis, the sprouting of blood vessels from preexisting vasculature is associated with both natural and pathological processes. Various angiogenesis assays involve the study of individual endothelial cells in culture conditions (1). The aortic ring assay is an angiogenesis model that is based on organ culture. In this assay, angiogenic vessels grow from a segment of the aorta (modified from (2)). Briefly, mouse thoracic aorta is excised, the fat layer and adventitia are removed, and rings approximately 1 mm in length are prepared. Individual rings are then embedded in a small solid dome of basement matrix extract (BME), cast inside individual wells of a 48-well plate. Angiogenic factors and inhibitors of angiogenesis can be directly added to the rings, and a mixed co-culture of aortic rings and other cell types can be employed for the study of paracrine angiogenic effects. Sprouting is observed by inspection under a stereomicroscope over a period of 6-12 days. Due to the large variation caused by the irregularities in the aortic segments, experimentation in 6-plicates is strongly advised. Neovessel outgrowth is monitored throughout the experiment and imaged using phase microscopy, and supernatants are collected for measurement of relevant angiogenic and anti-angiogenic factors, cell death markers and nitrite.

Angiogenesis, the sprouting of blood vessels from pre-existing vasculature, is associated with both natural and pathological processes. Here we demonstrate an aortic ring assay that allows angiogenic potentiators and inhibitors to be directly added to aortic rings in culture. Sprouting and neovessel outgrowth can be determined by inspecting the aortic rings over a period of 6-12 days.

الأبهر خاتم الفحص

Angiogenesis, the sprouting of blood vessels from preexisting vasculature is associated with both natural and pathological processes. Various angiogenesis ...

Ambulatory ECG Recording in Mice

Telemetric ECG recording in mice is essential to understanding the mechanisms behind arrhythmias, conduction disorders, and sudden cardiac death. Although the surface ECG is utilized for short-term measurements of waveform intervals, it is not practical for long-term studies of heart rate variability or the capture of rare episodes of arrhythmias. Implantable ECG telemeters offer the advantages of simple surgical implantation, long-term recording of electrograms in ambulatory mice, and scalability with simultaneous recordings of multiple animals. Here, we present a step-by-step guide to the implantation of telemeters for ambulatory ECG recording in mice. Careful adherence to aseptic technique is required for favorable survival results with the possibility of implantation and recording from weeks to months. Thus, implantable ECG telemetry is a valuable tool for detection of critical information on cardiac electrophysiology in ambulatory animal models such as the mouse. 

Telemetric ECG has emerged as an essential tool in evaluating animal models for cardiac arrhythmias and sudden cardiac death. Here, we present a stepwise guide to telemetric ECG recordings for application in long-term ambulatory ECG monitoring in mice.

الإسعافية ECG التسجيل في الفئران

Telemetric ECG recording in mice is essential to understanding the mechanisms behind arrhythmias, conduction disorders, and sudden cardiac death. Although ...

Intravenous Injections in Neonatal Mice

Intravenous injection is a clinically applicable manner to deliver therapeutics. For adult rodents and larger animals, intravenous injections are technically feasible and routine. However, some mouse models can have early onset of disease with a rapid progression that makes administration of potential therapies difficult. The temporal (or facial) vein is just anterior to the ear bud in mice and is clearly visible for the first two days after birth on either side of the head using a dissecting microscope. During this window, the temporal vein can be injected with volumes up to 50 &#956;l. The injection is safe and well tolerated by both the pups and the dams. A typical injection procedure is completed within 1-2 min, after which the pup is returned to the home cage. By the third postnatal day the vein is difficult to visualize and the injection procedure becomes technically unreliable. This technique has been used for delivery of adeno-associated virus (AAV) vectors, which in turn can provide almost body-wide, stable transgene expression for the life of the animal depending on the viral serotype chosen.

Animal models of pediatric disease can experience early onset and aggressive disease progression. Clinically relevant therapy delivery to young mouse models can be technically difficult. This protocol describes a non-invasive intravenous injection method for newborn mice within the first two postnatal days of life.

الحقن في الوريد في الفئران حديثي الولادة

Intravenous injection is a clinically applicable manner to deliver therapeutics. For adult rodents and larger animals, intravenous injections are ...

Isolation and Primary Culture of Mouse Aortic Endothelial Cells

The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. The mouse is an important model for cardiovascular disease research. This study demonstrates a simple method to isolate and culture endothelial cells from the mouse aorta without any special equipment. To isolate endothelial cells, the thoracic aorta is quickly removed from the mouse body, and the attached adipose tissue and connective tissue are removed from the aorta. The aorta is cut into 1 mm rings. Each aortic ring is opened and seeded onto a growth factor reduced matrix with the endothelium facing down. The segments are cultured in endothelial cell growth medium for about 4 days. The endothelial sprouting starts as early as day 2. The segments are then removed and the cells are cultured continually until they reach confluence. The endothelial cells are harvested using neutral proteinase and cultured in endothelial cell growth medium for another two passages before being used for experiments. Immunofluorescence staining indicated that after the second passage the majority of cells were double positive for Dil-ac-LDL uptake, Lectin binding, and CD31 staining, the typical characteristics of endothelial cells. It is suggested that cells at the second to third passages are suitable for in vitro and in vivo experiments to study the endothelial biology. Our protocol provides an effective means of identifying specific cellular and molecular mechanisms in endothelial cell physiopathology.

The vascular endothelial cells play a significant role in many important cardiovascular disorders. This article describes a simple method to isolate and expand endothelial cells from the mouse aorta without using any special equipment. Our protocol provides an effective means of identifying mechanisms in endothelial cell physiopathology.

العزلة والثقافة الابتدائية من خلايا الفأر الأورطي غشائي

The vascular endothelium is essential to normal vascular homeostasis. Its dysfunction participates in various cardiovascular disorders. The mouse is an ...

Induction of Right Ventricular Failure by Pulmonary Artery Constriction and Evaluation of Right Ventricular Function in Mice

The mechanism of right ventricular failure (RVF) requires clarification due to the uniqueness, high morbidity, high mortality, and refractory nature of RVF. Previous rat models imitating RVF progression have been described. Compared with rats, mice are more accessible, economical, and widely used in animal experiments. We developed a pulmonary artery constriction (PAC) approach which is comprised of banding the pulmonary trunk in mice to induce right ventricular (RV) hypertrophy. A special surgical latch needle was designed that allows for easier separation of the aorta and the pulmonary trunk. In our experiments, the use of this fabricated latch needle reduced the risk of arteriorrhexis and improved the surgical success rate to 90%. We used different padding needle diameters to precisely create quantitative constriction, which can induce different degrees of RV hypertrophy. We quantified the degree of constriction by evaluating the blood flow velocity of the PA, which was measured by noninvasive transthoracic echocardiography. RV function was precisely evaluated by right heart catheterization at 8 weeks after surgery. The surgical instruments made inhouse were composed of common materials using a simple process that is easy to master. Therefore, the PAC approach described here is easy to imitate using instruments made in the lab and can be widely used in other labs. This study presents a modified PAC approach that has a higher success rate than other models and an 8-week postsurgery survival rate of 97.8%. This PAC approach provides a useful technique for studying the mechanism of RVF and will enable an increased understanding of RVF.

Here, we provide a useful approach for studying the mechanism of right ventricular failure. A more convenient and efficient approach to pulmonary artery constriction is established using surgical instruments made inhouse. In addition, methods to evaluate the quality of this approach by echocardiography and catheterization are provided.

تحريض الفشل البطيني الأيمن عن طريق انقباض الشريان الرئوي وتقييم وظيفة البطين الأيمن في الفئران

The mechanism of right ventricular failure (RVF) requires clarification due to the uniqueness, high morbidity, high mortality, and refractory nature of ...

Inducing Apical Periodontitis in Mice

The mechanisms involved in local induced inflammation can be studied using several available animal models. One of these is the induction of apical periodontitis (AP). Apical periodontitis is a common pathology of an inflammatory nature in the periodontal tissues surrounding the tooth root. In order to better understand the nature and mechanism of this pathology it is advantageous to perform the procedure in mice. The induction of this odontogenic inflammation is achieved by drilling into the mouse tooth until the dental pulp is exposed. Next, the tooth pulp remains exposed to be contaminated by the natural oral flora over time, causing apical periodontitis. After this time period, the animal is sacrificed, and the tooth and the jaw bone can be analyzed in various ways. Typical analyses include micro-CT imaging (to evaluate bone resorption), histological staining, immunohistochemistry, and RNA expression. This protocol is useful for research in the field of oral biology to better understand this inflammatory process in an in vivo experimental setting with uniform conditions. The procedure requires a careful handling of the mice and the isolated jaw, and a visual demonstration of the technique is useful. All technical aspects of the procedures leading to induced apical periodontitis and its characterization in a mouse model are demonstrated.

Here, we present a protocol to locally induce apical periodontitis in mice. We show how to drill a hole in the mouse&#39;s tooth and expose its pulp, in order to cause local inflammation. Analysis methods to investigate the nature of this inflammation, such as micro-CT and histology, are also demonstrated.

تحفيز أمراض اللثة المثلى في الفئران

The mechanisms involved in local induced inflammation can be studied using several available animal models. One of these is the induction of apical ...

Transverse Fracture of the Mouse Femur with Stabilizing Pin

Fracture repair is an essential function of the skeleton that cannot be reliably modeled in vitro. A mouse injury model is an efficient approach to test whether a gene, gene product or drug influences bone repair because murine bones recapitulate the stages observed during human fracture healing. When a mouse or human breaks a bone, an inflammatory response is initiated, and the periosteum, a stem cell niche surrounding the bone itself, is activated and expands. Cells residing in the periosteum then differentiate to form a vascularized soft callus. The transition from the soft callus to a hard callus occurs as the recruited skeletal progenitor cells differentiate into mineralizing cells, and the bridging of the fractured ends results in the bone union. The mineralized callus then undergoes remodeling to restore the original shape and structure of the healed bone. Fracture healing has been studied in mice using various injury models. Still, the best way to recapitulate this entire biological process is to break through the cross-section of a long bone that encompasses both cortices. This protocol describes how a stabilized, transverse femur fracture can be safely performed to assess healing in adult mice. A surgical protocol including detailed harvesting and imaging techniques to characterize the different stages of fracture healing is also provided.

This protocol describes a method to perform fractures on adult mice and monitor the healing process.

كسر عرضي لعظم عظم الفخذ مع دبوس التثبيت

Fracture repair is an essential function of the skeleton that cannot be reliably modeled in vitro. A mouse injury model is an efficient approach to test ...

A Modified Technique for Transverse Aortic Constriction in Mice

Transverse aortic constriction (TAC) is a frequently used surgery in research regarding heart failure and cardiac hypertrophy based on the formation of pressure overload in mouse models. The main challenge of this procedure is to clearly visualize the transverse aortic arch and precisely band the target vessel. Classical approaches perform a partial thoracotomy to expose the transverse aortic arch. However, it is an open-chest model that causes a rather large surgical trauma and requires a ventilator during the surgery. To prevent unnecessary trauma and simplify the operating procedure, the aortic arch is approached via the proximal proportion of the sternum, reaching and binding the target vessel using a small self-made retractor that contains a snare. This procedure can be conducted without entering the pleura cavity and does not need a ventilator or microsurgical operation, which leaves the mice with physiological breathing patterns, simplifies the procedure, and significantly reduces operation time. Due to the less invasive approach and less operation time, mice can undergo fewer stress reactions and recover rapidly.

The present protocol describes a modified and simplified technique with a minimally invasive transverse aortic constriction (TAC) procedure using a self-made retractor. This procedure can be conducted without a ventilator or microscope and introduces pressure overload, eventually leading to cardiac hypertrophy or heart failure.

تقنية معدلة لتضيق الأبهر المستعرض في الفئران

Transverse aortic constriction (TAC) is a frequently used surgery in research regarding heart failure and cardiac hypertrophy based on the formation of ...