نود أن نعترف الدكاترة. روبرت Gereau، هنري لاي، ولارا من الفخار للحصول على مساعدة مفيدة في إعداد هذا النظام. نود أيضا أن نعترف مصادر التمويل لهذا العمل (بيشكتاش - الجمعية الدولية لدراسة الألم في وقت مبكر الوظيفي بحوث غرانت من المسح الضوئي | مؤسسة تصميم إنجر وJENS بروون وصندوق المرض اللعين Hunkele جامعة دوكين).

<ol>
	<li>Merskey, H., Bogduk, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Classification of Chronic Pain. , (1994).</li><li>Gureje, O., Korff, M. V., Simon, G. E., Gater, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peristent+pain+and+well-being.+A+World+Health+Organization+study+in+primary+care.">Peristent pain and well-being. A World Health Organization study in primary care.</a> J. Am. Med. Assoc. 280, 147-151 (1998).</li><li>Gaskin, D. J., Richard, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+economic+costs+of+pain+in+the+United+States.">The economic costs of pain in the United States.</a> J. Pain. 13, 715-754 (2012).</li><li>Berry, S. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prevalence+of+Symptoms+of+Bladder+Pain+Syndrome/Interstitial+Cystitis+Among+Adult+Females+in+the+United+States.">Prevalence of Symptoms of Bladder Pain Syndrome/Interstitial Cystitis Among Adult Females in the United States.</a> J. Urol. 186, 540-544 (2011).</li><li>Tunitsky, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bladder+Pain+Syndrome/Interstitial+Cystitis+in+twin+sisters.">Bladder Pain Syndrome/Interstitial Cystitis in twin sisters.</a> J. Urol. 187, 148-152 (2012).</li><li>Tettamanti, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+smoking,+coffee,+and+tea+consumption+on+Bladder+Pain+Syndrome+in+female+twins.">Influence of smoking, coffee, and tea consumption on Bladder Pain Syndrome in female twins.</a> Urology. 77, 1313-1317 (2011).</li><li>Warren, J. W., et al., Longo, D. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Harrison's Principles of Internal Medicine. , (2012).</li><li>Baldoni, F., Ercolani, M., Baldaro, B., Trombini, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stressful+events+and+psychological+symptoms+in+patients+with+functional+urinary+disorders.">Stressful events and psychological symptoms in patients with functional urinary disorders.</a> Perceptual Motor Skills. 80, 605-606 (1995).</li><li>Tannenbaum, C., Gray, M., Hoffstetter, S., Cardozo, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comorbidities+associated+with+bladder+dysfunction.">Comorbidities associated with bladder dysfunction.</a> Int. J. Clin. Practice. 67, 105-113 (2013).</li><li>Ness, T. J., Elhefni, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reliable+visceromotor+responses+are+evoked+by+noxious+bladder+distention+in+mice.">Reliable visceromotor responses are evoked by noxious bladder distention in mice.</a> J. Urol. 171, 1704-1708 (2004).</li><li>Ness, T. J., Richter, H. E., Varner, R. E., Fillingim, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+psychophysical+study+of+discomfort+produced+by+repeated+filling+of+the+urinary+bladder.">A psychophysical study of discomfort produced by repeated filling of the urinary bladder.</a> Pain. 76, 61-69 (1998).</li><li>Ness, T. J., Gebhart, G. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visceral+pain:+a+review+of+experimental+studies.">Visceral pain: a review of experimental studies.</a> Pain. 41, 167-234 (1990).</li><li>Crock, L. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+amygdala+metabotropic+glutamate+receptor+5+in+the+modulation+of+visceral+pain.">Central amygdala metabotropic glutamate receptor 5 in the modulation of visceral pain.</a> J. Neurosci. 32, 14217-14226 (2012).</li><li>Anderson, R. H., Ness, T. J., Gebhart, G. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+distension+control+device+useful+for+quantitative+studies+of+hollow+organ+sensation.">A distension control device useful for quantitative studies of hollow organ sensation.</a> Physiol. Behav. 41, 635-638 (1987).</li><li>Dimitrakov, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacologic+Management+of+Painful+Bladder+Syndrome/Interstitial+Cystitis:+A+Systematic+Review.">Pharmacologic Management of Painful Bladder Syndrome/Interstitial Cystitis: A Systematic Review.</a> Arch. Intern. Med. 167, 1922-1929 (2007).</li><li>Qin, C., Van Meerveld, B. G. r. e. e. n. w. o. o. d. -., Foreman, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+neuronal+responses+to+urinary+bladder+stimulation+in+rats+with+corticosterone+or+aldosterone+onto+the+amygdala.">Spinal neuronal responses to urinary bladder stimulation in rats with corticosterone or aldosterone onto the amygdala.</a> J. Neurophysiol. 90, 2180-2189 (2003).</li><li>Randich, A., Mebane, H., DeBerry, J. J., Ness, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rostral+ventral+medulla+modulation+of+the+visceromotor+reflex+evoked+by+urinary+bladder+distension+in+female+rats.">Rostral ventral medulla modulation of the visceromotor reflex evoked by urinary bladder distension in female rats.</a> J. Pain. 9, 920-926 (2008).</li></ol>

The authors have nothing to disclose.

في البشر، والخلالي متلازمة الألم التهاب المثانة / المثانة (IC / BPS) يمثل مشكلة سريرية هامة لأن المرضى قد المنهكة الألم الذي هو في كثير من الأحيان لا تستجيب للعلاج الألم العادية 15. واحدة من التحديات الرئيسية في فهم وعلاج الألم في نهاية المطاف المثانة المزمن هو فهم ا...

ويعرف الألم المزمن رسميا باسم الألم الذي يستمر لثلاثة أشهر تقريبا، أو لمدة أطول من شفاء الأنسجة الطبيعية الوقت 1. هذا النوع من الألم هو واحد من الأسباب الرئيسية أن الناس هي التي تحرك التماس العناية الطبية (2) ويمكن أن تصل تكلفتها 635 مليار دولار سنويا 3. تعتبر استراتيجيات الألم المزمن الحالي التأقلم القديمة؛ بعد عقود من التقدم الطبي، مضادات الالتهاب غير الستيروئيدية (العقاقير المضادة للالتهابات) والمواد الأفيونية لا تزال العلاجات الأولية التي يحددها الأطباء. ولكن هذه العلاجات تستهدف جميع أنواع مختلفة من الألم في نفس الطريق، وتوفير تأثيرات مسكنة في جميع أنحاء الجسم بدلا من التركيز تحديدا على سبب هذا الحادث الألم بالضبط. من أجل مساعدة أولئك الذين يعانون من أفضل الألم المزمن، يجب أن تحول الانتباه البحوث نحو المسببات والعلاج الألم محددة المحتملة المرتبطة الأسباب الأكثر شيوعا لآلام المزمنة. الهدف من هذا manuscriحزب العمال هو لوصف النموذج المستخدم لفهم أفضل لحالة تعرف باسم التهاب المثانة الخلالي / متلازمة المثانة المؤلمة (IC / BPS). IC / BPS هو الشرط المنهكة التي تؤثر على الملايين من الناس، والنساء في المقام الأول فوق سن ال 40 4. ولا تعرف الاسباب الحقيقية لIC / BPS، ومع ذلك ارتبط انتشار لعلم الوراثة 5، والوجبات الغذائية محددة، ومستويات التوتر عالية 6 . وتشمل أعراض IC / BPS ولكن لا تقتصر على: زيادة الحاجة الملحة للتبول، زيادة تواتر التبول، وآلام، وثقب، أو حرق الألم أثناء ملء المثانة ويفرغ، وكثرة التبول اثناء الليل 7. مرضى الذين يعانون من هذه الظروف لديهم مستويات أعلى من التوتر والقلق هي أكثر 8. هذه النتائج زيادة التوتر في زيادة الألم، والذي يؤدي إلى زيادة مستويات التوتر الأصلي. وقد أظهرت الدراسات أن مستويات الاكتئاب والقلق تقلل العلاجات التالية التي تخفف الألم البولية، وبالتالي كسر هذه الدورة بفعالية إيجابية ردود الفعل <sup&gt; 9. كان واحدا من التحديات في اكتشاف علاجات جديدة لIC / BPS قلة فهم العلوم الأساسية من آلام المثانة. من أجل دراسة المساهمات الجهاز العصبي إلى حالة، تم تطوير العديد من النماذج الحيوانية لتقليد الألم والأعراض المرتبطة IC / BPS. تقليديا، وقد تم صوغه التهاب المثانة في الحيوانات مع إدخال المواد الكيميائية المختلفة مثل زيت الخردل، والأسيتون، عديد السكاريد الشحمي، وحامض الهيدروكلوريك، وسيكلوفوسفاميد في المثانة. ومع ذلك لا يوجد عملاء لجهات أجنبية موجودة في البول العقيمة من المرضى IC / BPS، وبالتالي التشكيك في صحة هذه النماذج. آخر نموذج الفئران من آلام المسالك البولية هو انتفاخ المثانة البولية (UBD) 10. في هذا النموذج، يتم تسليم الهواء المضغوط الضغط محددة إلى المثانة حيوان مستيقظا أو تخدير طفيفة على مدى فترة معينة من الزمن. في جميع أنحاء الداخلي، والأسلاك في عضلات البطن المائلة أكتيف الكهربائية سجل متفوقةإيتي من العضلات (مع مخطط كهربية العضل (EMG)). ويعرف هذا النشاط باعتبارها استجابة الحشوي الحركي (VMR) وغير موثوق بها، وقياس استنساخه من حس الألم 10. مماثلة جوفاء الجهاز انتفاخ (مثل المثانة والمستقيم) على متطوعين من البشر صحية تنتج مشاعر الانزعاج وزيادات كبيرة في الألم ذكرت 11،12، والصفات التي غالبا ما تستخدم لتشخيص IC / BPS. وبالتالي، بالتزامن مع أساليب الكهربائية، والدوائية، وoptogenetic من التحفيز أو التثبيط، UBD هو نموذج مفيدة وصالحة لفهم كل من مكونات الجهاز العصبي المحيطي والمركزي من حس الألم المثانة والألم. هنا، نحن تصف الإجراء لUBD كما يستخدم حاليا في المختبر لدينا في غضون عضوات من سلالة الماوس المشتركة، C57BL/6J. بسبب صعوبات في عملية قسطرة للذكور، ويتم تنفيذ هذا الإجراء في المقام الأول في إناث الفئران. وقد تم تكييف هذا البروتوكول من تلك التي وضعتها نيس &lt;م&gt; وآخرون 10. ونشرت سابقا من مختبرنا 13. يصف بروتوكول الأربعة التالية المكونات الأساسية لهذا النموذج في الحيوانات تخدير: (1) القسطرة وجراحة المثانة القطب تسجيل (2) تحريض التخدير الجزئي (3) انتفاخ المثانة وتسجيل VMR، و (4) تحليل البيانات الخام من آثار VMR / EMG . وتناقش الفروق الدقيقة في إجراء UBD-VMR بسبب اختيار الكائن الحي، وعمق التخدير والاستقراء، ودرجة حرارة الجسم أدناه.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Infrared heating blanket and monitoring system</td>
			<td>Kent Scientific</td>
			<td>Right-Temp System&#160;</td>
			<td>This system is setup to monitor two separate temperatures.&#160; This should include the animal and the heating blanket.&#160; In addition, the system can automatically adjust the temperature to maintain a set temp.&#160; However, this automatic function produces electrical interference during EMG recording and must be turned off.&#160; Kent can provide a battery pack for the heating pad for use during the riEMG recording part of the experiment.</td>
		</tr>
		<tr>
			<td>Isoflurane vaporizer</td>
			<td>Draeger</td>
			<td>Vapor 19.1</td>
			<td>Any isoflurane vaporizer will work, but it is helpful to have one that has mutliple notches between 0 and 2% isoflurane.</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>100% Oxygen air tank</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td>For ventilation of animal</td>
		</tr>
		<tr>
			<td>Air breathing grade</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td>For bladder distention</td>
		</tr>
		<tr>
			<td>24GA 0.56inch IV catheter</td>
			<td>BD biosciences&#160;</td>
			<td>381411</td>
			<td>For bladder catheterization</td>
		</tr>
		<tr>
			<td>Surgilube (sterile)</td>
			<td>Savage Laboratories</td>
			<td>0281-0205-02</td>
			<td>Any surgical grade lubricate would work fine.</td>
		</tr>
		<tr>
			<td>Mineral oil (sterile)</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Saline (sterile)</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>AG8W Silver Wire, 2 m, 0.20 mm (.008&#34;) D, L, No Insulation&#160;</td>
			<td>Warner Instruments</td>
			<td>W4 64-1318&#160;</td>
			<td>Any silver wire with these specifications will work.&#160; Wire does not need to be &#34;chlorinated.&#34;</td>
		</tr>
		<tr>
			<td>opthalamic ointment</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Small surgical scissors</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Sharp forceps</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>21 Ga needle</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Grass amplifier P511 with 3-lead input cable</td>
			<td>Grass Instruments</td>
			<td>P511 (F-P5IC3/REV1)</td>
			<td>This is the &#34;amplifier&#34; used in the protocol.&#160; Amplifier with the following settings: Calibrator = 1 mV; Lo freq = 300 Hz; Amplification = 20; Hi freq = 10 kHz; Line filter = in.</td>
		</tr>
		<tr>
			<td>Cambridge Electronic Design (CED) 1401 Plus (or equivalent)</td>
			<td>Cambridge Electronic Design</td>
			<td>1401 Plus</td>
			<td>This is the &#34;digitizer&#34; used in the protocol.&#160; Other digitizer systems from WINDAQ or other companies would work fine; Need inputs for pressure signal, EMG, and stimulus signal.</td>
		</tr>
		<tr>
			<td>CED Spike2 software</td>
			<td>Cambridge Electronic Design</td>
			<td>Spike 2</td>
			<td>This is the the &#34;digitizer software&#34; used in the protocol.&#160; Should be from same manufacturer as digitizer.&#160; Program should be setup with 3 channels for pressure (0-100 mmHg scale), EMG signal (typically -5 to +5 V range), and stimulus marker (0-2 V) range.</td>
		</tr>
		<tr>
			<td>Flow chart from air tank to bladder catheter</td>
			<td>n/a</td>
			<td>n/a</td>
			<td>Sequence of connections from pressurized air tank to animal bladder:&#160; Air tank to 1/4 inch tubing to Gilmont flowmeter to Y connector.&#160; Branch 1 of y connector to to sphygmomanometer.&#160; Branch 2 to a single input on the 4-way gang valve to 4-way valve output to the timed pressure regulator to 3/32 tubing from timed pressure regulator to 2nd Y connector (branch 1 to sphygmomanometer) with branch 2 to 3/32 tubing&#160; to a 3rd Y connector.&#160; Branch 1 of Y connector to a 3rd sphygmomanometer and branch 2 to animal bladder catheter.</td>
		</tr>
		<tr>
			<td>Gilmont Flowmeter&#160;</td>
			<td>Gilmont</td>
			<td>GF8321-1401</td>
			<td>Multiple brands of flowmeters will work.&#160; For bladder distention air, flow meter should be able to handle high pressures (such as this Gilmont meter).&#160; For breathing air, flow rate should be adjustable down to 100 cc/min (typical mouse rate is 500-1000 cc/min).&#160;</td>
		</tr>
		<tr>
			<td>4-way gang valve</td>
			<td>Elite</td>
			<td></td>
			<td>This is a specific piece of equipment.&#160; The Elite gang valve is designed for fish tanks at low air pressures.&#160; In the bladder distention setup, this valve acts as a safety valves in case the pressure spikes.&#160; Too high pressure during initial turning on of the tank will ruin the pressure transducer in the Timed pressure regulator and/or the sphygnometers. In addition, by providing a small amount of leak in the system, this valve makes it easier to adjust the pressure between 10-80 mmHg.&#160;&#160;</td>
		</tr>
		<tr>
			<td>1/4 inch tubing</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>3/32 inch tubing</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>&#34;Y&#34; connectors (1/4 and 3/32 inch)</td>
			<td>Multiple</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Sphygmomanometer</td>
			<td>CVS</td>
			<td></td>
			<td>Any analog sphygmomanometer from a drug store will work for this application.&#160;&#160;</td>
		</tr>
		<tr>
			<td>Timed Pressure Regulator</td>
			<td>Custom</td>
			<td></td>
			<td>This is a custom built machine (Washington University in St. Louis Machine shop) that allows for automated pressure delivery including digitization of air pressure, control of trial length, inter-trial interval automation, control of trial number, and stimulus onset digital signal.&#160; However, the basic components of the system (pressure on and off for a given trial period) could be controlled with a simple on/off in-line switch.&#160;&#160; Such analog control of a trial would necessitate additional analysis parameters (see below section 4).&#160; In addition, one would have to manually assign the pressure based on the analog sphygmomanometer during data analysis.&#160;&#160;</td>
		</tr>
		<tr>
			<td>IGOR Pro</td>
			<td>Wavemetrics</td>
			<td>n/a</td>
			<td>For analysis of EMG signal. Many different types of software can be used for data analysis in these experiments.</td>
		</tr>
	</tbody>
</table>

urinary bladder distention evoked visceromotor responses as a model for bladder pain in mice

ما يقرب من 3-8 ملايين شخص في الولايات المتحدة يعانون من متلازمة التهاب المثانة / المثانة الخلالي ألم (IC / BPS)، وهي حالة تتميز المنهكة زيادة الإلحاح وتكرار التبول، وكذلك كثرة التبول اثناء الليل وآلام الحوض العامة، وخاصة عند ملء المثانة أو يفرغ. على الرغم من سنوات من البحث، وسبب IC / BPS لا يزال بعيد المنال واستراتيجيات العلاج غير قادرين على توفير الإغاثة الكامل للمرضى. من أجل دراسة المساهمات الجهاز العصبي إلى حالة، تم تطوير العديد من النماذج الحيوانية لتقليد الألم والأعراض المرتبطة IC / BPS. واحدة من هذه نموذج الفئران هو البولية المثانة انتفاخ (UBD). في هذا النموذج، يتم تسليم الهواء المضغوط الضغط محددة إلى المثانة من حيوان تخدير طفيفة على مدى فترة معينة من الزمن. في جميع أنحاء الداخلي، والأسلاك في عضلات البطن المائلة العلوية تسجيل النشاط الكهربائي من العضلات. ويعرف هذا النشاط باعتبارها استجابة الحشوي الحركي (VMR) وأناسا مقياس موثوق بها وقابلة للتكرار من حس الألم. هنا، نحن تصف الخطوات اللازمة لتنفيذ هذه التقنية في الفئران بما في ذلك التلاعب الجراحية، وتسجيل الفسيولوجية، وتحليل البيانات. مع استخدام هذا النموذج، يمكن كشف التنسيق بين الخلايا العصبية الحسية الأولية والحبل الشوكي afferents الثانوية، والمناطق الجهاز العصبي المركزي العالي التي تشارك في آلام المثانة. ويمكن بعد هذه المعرفة العلوم الأساسية أن تترجم سريريا لعلاج المرضى الذين يعانون من IC / BPS.

ويمكن رؤية مثال على مجمل الإعداد UBD-VMR في الشكل 1A. زيادة الضغط خطوات لحث زيادة في VMR الخام (أي EMG) (الشكل 1B). وVMR الخام والضغط والتحفيز إشارة علامة التي ينبغي مراعاتها أثناء التسجيل باستخدام برنامج التحويل الرقمي يمكن أن ينظر في الشكل 2. الق...

Watch this Scientific Journal Video about Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice at JoVE.com

Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

ما يقرب من 3-8 ملايين شخص في الولايات المتحدة يعانون من متلازمة ألم التهاب المثانة الخلالي / المثانة (IC / BPS)، وهي حالة تتميز المنهكة في جزء من آلام الحوض. من أجل دراسة المساهمات الجهاز العصبي إلى حالة، يتم استخدام نموذج الفسيولوجية للألم المثانة البولية في الفئران والجرذان.

الردود المثانة البولية انتفاخ مستدعى الحشوي الحركي نموذجا لألم المثانة في الفئران

Approximately 3-8 million people in the United States suffer from interstitial cystitis/bladder pain syndrome (IC/BPS), a debilitating condition ...

urinary-bladder-distention-evoked-visceromotor-responses-as-model-for

Research

JoVE Journal

Medicine

17.6K Views.  Duquesne University.  ما يقرب من 3-8 ملايين شخص في الولايات المتحدة يعانون من متلازمة ألم التهاب المثانة الخلالي / المثانة (IC / BPS)، وهي حالة تتميز المنهكة في جزء من آلام الحوض. من أجل دراسة المساهمات الجهاز العصبي إلى حالة، يتم استخدام نموذج الفسيولوجية للألم المث?...

Video: Urinary Bladder Distention Evoked Visceromotor Responses as a Model for Bladder Pain in Mice

Mouse Bladder Wall Injection

Mouse bladder wall injection is a useful technique to orthotopically study bladder phenomena, including stem cell, smooth muscle, and cancer biology. Before starting injections, the surgical area must be cleaned with soap and water and antiseptic solution. Surgical equipment must be sterilized before use and between each animal. Each mouse is placed under inhaled isoflurane anesthesia (2-5% for induction, 1-3% for maintenance) and its bladder exposed by making a midline abdominal incision with scissors. If the bladder is full, it is partially decompressed by gentle squeezing between two fingers. The cell suspension of interest is intramurally injected into the wall of the bladder dome using a 29 or 30 gauge needle and 1 cc or smaller syringe. The wound is then closed using wound clips and the mouse allowed to recover on a warming pad. Bladder wall injection is a delicate microsurgical technique that can be mastered with practice.

Mouse bladder wall injection is a useful approach to orthotopically study bladder stem cell and cancer biology. This delicate microsurgical method can be mastered with careful technique and practice.

حقن المثانة الماوس الجدار

Mouse bladder wall injection is a useful technique to orthotopically study bladder phenomena, including stem cell, smooth muscle, and cancer biology. ...

An Orthotopic Model of Murine Bladder Cancer

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and subsequent cell adhesion. After their bladders are transurethrally catheterized and drained, animals are again catheterized to permit insertion of a platinum wire into bladders without damaging the urethra or bladder. The catheters are made of Teflon to serve as an insulator for the wire, which will conduct electrical current into the bladder to create a burn injury. An electrocautery unit is used to deliver 2.5W to the exposed end of the wire, burning away extracellular layers and providing attachment sites for carcinoma cells that are delivered in suspension to the bladder through a subsequent catheterization. Cells remain in the bladder for 90 minutes, after which the catheters are removed and the bladders allowed to drain naturally. The development of tumor is monitored via ultrasound. Specific attention is paid to the catheterization technique in the accompanying video.

Bladder tumors are established in female mice in a minimally invasive fashion through catheterization, local cauterization, and subsequent adhesion of carcinoma cells to the burn sites.

نموذج مثلي من سرطان المثانة الفئران

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and ...

The Use of Cystometry in Small Rodents: A Study of Bladder Chemosensation

The lower urinary tract (LUT) functions as a dynamic reservoir that is able to store urine and to efficiently expel it at a convenient time. While storing urine, however, the bladder is exposed for prolonged periods to waste products. By acting as a tight barrier, the epithelial lining of the LUT, the urothelium, avoids re-absorption of harmful substances. Moreover, noxious chemicals stimulate the bladder's nociceptive innervation and initiate voiding contractions that expel the bladder's contents. Interestingly, the bladder's sensitivity to noxious chemicals has been used successfully in clinical practice, by intravesically infusing the TRPV1 agonist capsaicin to treat neurogenic bladder overactivity1. This underscores the advantage of viewing the bladder as a chemosensory organ and prompts for further clinical research. However, ethical issues severely limit the possibilities to perform, in human subjects, the invasive measurements that are necessary to unravel the molecular bases of LUT clinical pharmacology. A way to overcome this limitation is the use of several animal models2. Here we describe the implementation of cystometry in mice and rats, a technique that allows measuring the intravesical pressure in conditions of controlled bladder perfusion.
	After laparotomy, a catheter is implanted in the bladder dome and tunneled subcutaneously to the interscapular region. Then the bladder can be filled at a controlled rate, while the urethra is left free for micturition. During the repetitive cycles of filling and voiding, intravesical pressure can be measured via the implanted catheter. As such, the pressure changes can be quantified and analyzed. Moreover, simultaneous measurement of the voided volume allows distinguishing voiding contractions from non-voiding contractions3.
	Importantly, due to the differences in micturition control between rodents and humans, cystometric measurements in these animals have only limited translational value4. Nevertheless, they are quite instrumental in the study of bladder pathophysiology and pharmacology in experimental pre-clinical settings. Recent research using this technique has revealed the key role of novel molecular players in the mechano- and chemo-sensory properties of the bladder.

Cystometry is an efficient technique to measure bladder function of small animals in vivo. The bladder is continuously infused at rates controlled through an intravesical catheter, whereas the urethra is left free for micturition. This allows for repetitive filling and emptying of the bladder, while intravesical pressure and voided volume are recorded.

استعمال قياس المثانة في القوارض الصغيرة: دراسة في Chemosensation المثانة

	The lower urinary tract (LUT) functions as a  dynamic reservoir that is able to store urine and to efficiently expel it at a  convenient time. While ...

An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder tumor model. The mouse model is established by urethral catheterization under inhaled general anesthetic. Chemical burn is then introduced to the bladder mucosa using intravesical silver nitrate solution to disrupt the bladder glycosaminoglycan layer and allows cells to attach. Following several washes with sterile water, human bladder cancer KU-7-luc2-GFP cells are instilled through the catheter into the bladder to dwell for 2 hours. Subsequent growth of bladder tumors is confirmed and monitored by in vivo bladder ultrasound and bioluminescent imaging. The tumors are then treated intravesically with saRNA formulated in lipid nanoparticles (LNPs). Tumor growth is monitored with ultrasound and bioluminescence. All steps of this procedure are demonstrated in the accompanying video.

Establishing an orthotopic bladder tumor model to evaluate antitumor effects of intravesically delivered saRNA and monitoring tumor growth by ultrasound and bioluminescent imaging.

ورم في المثانة Orthotopic النموذجي وتقييم المعالجة سارنا داخل المثانة

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder ...

An Orthotopic Bladder Cancer Model for Gene Delivery Studies

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are needed. The tumor microenvironment can significantly influence tumor development and therapy response. It is therefore often desirable to grow tumor cells in the organ from which they originated. This protocol describes an orthotopic model of bladder cancer, in which MB49 murine bladder carcinoma cells are instilled into the bladder via catheterization. Successful tumor cell implantation in this model requires disruption of the protective glycosaminoglycan layer, which can be accomplished by physical or chemical means. In our protocol the bladder is treated with trypsin prior to cell instillation. Catheterization of the bladder can also be used to deliver therapeutics once the tumors are established. This protocol describes the delivery of an adenoviral construct that expresses a luciferase reporter gene. While our protocol has been optimized for short-term studies and focuses on gene delivery, the methodology of mouse bladder catheterization has broad applications.

Implantation of cancer cells into the organ of origin can serve as a useful preclinical model to evaluate novel therapies. MB49 bladder carcinoma cells can be grown within the bladder following intravesical instillation. This protocol demonstrates catheterization of the mouse bladder for the purpose of tumor implantation and adenoviral delivery.

وسرطان المثانة نموذج مثلي للدراسات توصيل الجينات

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are ...

Induction of Invasive Transitional Cell Bladder Carcinoma in Immune Intact Human MUC1 Transgenic Mice:  A Model for Immunotherapy Development

A preclinical model of invasive bladder cancer was developed in human mucin 1 (MUC1) transgenic (MUC1.Tg) mice for the purpose of evaluating immunotherapy and/or cytotoxic chemotherapy. To induce bladder cancer, C57BL/6 mice (MUC1.Tg and wild type) were treated orally with the carcinogen N-butyl-N-(4-hydroxybutyl)nitrosamine (OH-BBN) at 3.0 mg/day, 5 days/week for 12 weeks. To assess the effects of OH-BBN on serum cytokine profile during tumor development, whole blood was collected via submandibular bleeds prior to treatment and every four weeks. In addition, a MUC1-targeted peptide vaccine and placebo were administered to groups of mice weekly for eight weeks. Multiplex fluorometric microbead immunoanalyses of serum cytokines during tumor development and following vaccination were performed. At termination, interferon gamma (IFN-&#947;)/interleukin-4 (IL-4) ELISpot analysis for MUC1 specific T-cell immune response and histopathological evaluations of tumor type and grade were performed. The results showed that: (1) the incidence of bladder cancer in both MUC1.Tg and wild type mice was 67%; (2) transitional cell carcinomas (TCC) developed at a 2:1 ratio compared to squamous cell carcinomas (SCC); (3) inflammatory cytokines increased with time during tumor development; and (4) administration of the peptide vaccine induces a Th1-polarized serum cytokine profile and a MUC1 specific T-cell response. All tumors in MUC1.Tg mice were positive for MUC1 expression, and half of all tumors in MUC1.Tg and wild type mice were invasive. In conclusion, using a team approach through the coordination of the efforts of pharmacologists, immunologists, pathologists and molecular biologists, we have developed an immune intact transgenic mouse model of bladder cancer that expresses hMUC1.

An N-butyl-N-(4-hydroxybutyl)nitrosamine-induced bladder cancer model was developed in human mucin 1 (MUC1) transgenic mice for the purpose of testing MUC1-directed immunotherapy. After administering a MUC1-targeted peptide vaccine, a cytotoxic T lymphocyte response to MUC1 was confirmed by measuring serum cytokine levels and T-cell specific activity.

تحريض الغازية سرطان المثانة الانتقالية الخليوي في المناعة السليمة الإنسان MUC1 الفئران المعدلة وراثيا: نموذج للتنمية الخلايا الجذعية

A preclinical model of invasive bladder cancer was developed in human mucin 1 (MUC1) transgenic (MUC1.Tg) mice for the purpose of evaluating immunotherapy ...

Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable cell lines are inoculated either by intravesical instillation (model of nonmuscle invasive growth) or intramural injection into the bladder wall (model of invasive growth). Both procedures are complex and highly time-consuming. Additionally, the superficial model has its shortcomings due to the lack of cell lines that are tumorigenic following instillation. Intramural injection, on the other hand, is marred by the invasiveness of the procedure and the associated morbidity for the host mouse.
With these shortcomings in mind, we modified previous methods to develop a minimally invasive approach for creating orthotopic bladder cancer xenografts. Using ultrasound guidance we have successfully performed percutaneous inoculation of the bladder cancer cell lines UM-UC1, UM-UC3 and UM-UC13 into 50 athymic nude. We have been able to demonstrate that this approach is time efficient, precise and safe. With this technique, initially a space is created under the bladder mucosa with PBS, and tumor cells are then injected into this space in a second step. Tumor growth is monitored at regular intervals with bioluminescence imaging and ultrasound. The average tumor volumes increased steadily in in all but one of our 50 mice over the study period.
In our institution, this novel approach, which allows bladder cancer xenograft inoculation in a minimally-invasive, rapid and highly precise way, has replaced the traditional model.

The established technique to inoculate primary invasive orthotopic bladder cancer xenografts requires laparotomy and mobilization of the bladder. This procedure inflicts significant morbidity on the mice, is technically challenging and time-consuming. We therefore developed a high-precision, percutaneous approach utilizing ultrasound guidance.

مؤسسة المناظير الفئران مثلي المثانة Xenografts

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable ...

The Sciatic Nerve Cuffing Model of Neuropathic Pain in Mice

Neuropathic pain arises as a consequence of a lesion or a disease affecting the somatosensory system. This syndrome results from maladaptive changes in injured sensory neurons and along the entire nociceptive pathway within the central nervous system. It is usually chronic and challenging to treat. In order to study neuropathic pain and its treatments, different models have been developed in rodents. These models derive from known etiologies, thus reproducing peripheral nerve injuries, central injuries, and metabolic-, infectious- or chemotherapy-related neuropathies. Murine models of peripheral nerve injury often target the sciatic nerve which is easy to access and allows nociceptive tests on the hind paw. These models rely on a compression and/or a section. Here, the detailed surgery procedure for the &#34;cuff model&#34; of neuropathic pain in mice is described. In this model, a cuff of PE-20 polyethylene tubing of standardized length (2 mm) is unilaterally implanted around the main branch of the sciatic nerve. It induces a long-lasting mechanical allodynia, i.e., a nociceptive response to a normally non-nociceptive stimulus that can be evaluated by using von Frey filaments. Besides the detailed surgery and testing procedures, the interest of this model for the study of neuropathic pain mechanism, for the study of neuropathic pain sensory and anxiodepressive aspects, and for the study of neuropathic pain treatments are also discussed.

Neuropathic pain is a consequence of a lesion or disease affecting the somatosensory system. The &#8220;cuff model&#8221; of neuropathic pain in mice consists of the implantation of a polyethylene cuff around the main branch of the sciatic nerve. Mechanical allodynia is tested using von Frey filaments.

العصب الوركي استكفاف نموذج من ألم الأعصاب في الفئران

Neuropathic pain arises as a consequence of a lesion or a disease affecting the somatosensory system. This syndrome results from maladaptive changes in ...

Bladder Smooth Muscle Strip Contractility as a Method to Evaluate Lower Urinary Tract Pharmacology

We describe an in vitro method to measure bladder smooth muscle contractility, and its use for investigating physiological and pharmacological properties of the smooth muscle as well as changes induced by pathology. This method provides critical information for understanding bladder function while overcoming major methodological difficulties encountered in in vivo experiments, such as surgical and pharmacological manipulations that affect stability and survival of the preparations, the use of human tissue, and/or the use of expensive chemicals. It also provides a way to investigate the properties of each bladder component (i.e. smooth muscle, mucosa, nerves) in healthy and pathological conditions.
The urinary bladder is removed from an anesthetized animal, placed in Krebs solution and cut into strips. Strips are placed into a chamber filled with warm Krebs solution. One end is attached to an isometric tension transducer to measure contraction force, the other end is attached to a fixed rod. Tissue is stimulated by directly adding compounds to the bath or by electric field stimulation electrodes that activate nerves, similar to triggering bladder contractions in vivo. We demonstrate the use of this method to evaluate spontaneous smooth muscle contractility during development and after an experimental spinal cord injury, the nature of neurotransmission (transmitters and receptors involved), factors involved in modulation of smooth muscle activity, the role of individual bladder components, and species and organ differences in response to pharmacological agents. Additionally, it could be used for investigating intracellular pathways involved in contraction and/or relaxation of the smooth muscle, drug structure-activity relationships and evaluation of transmitter release.
The in vitro smooth muscle contractility method has been used extensively for over 50 years, and has provided data that significantly contributed to our understanding of bladder function as well as to pharmaceutical development of compounds currently used clinically for bladder management.

This manuscript presents a simple, yet powerful, in vitro method for evaluating smooth muscle contractility in response to pharmacological agents or nerve stimulation. Main applications are drug screening and understanding tissue physiology, pharmacology,&#160;and pathology.

المثانة العضلات الملساء قطاع انقباض كوسيلة لتقييم المسالك البولية السفلى الصيدلة

We describe an in vitro method to measure bladder smooth muscle contractility, and its use for investigating physiological and pharmacological properties ...

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Bladder cancer is one of the most common malignancies of the urogenital tract. Intravesical injection of Bacillus Calmette-Gu&#233;rin (BCG) is the gold standard treatment for the high-grade non-muscle invasive bladder cancer (NMIBC). However, since the treatment-related side effects are relevant, newer biological response modifiers with a better benefit/side effects ratio are needed.
The tumour microenvironment can influence both tumour development and therapy efficacy. In order to obtain a good model, it is desirable to implant tumour cells in the organ from which the cancer originates.
In this protocol, we describe a method for establishing a tumour in the bladder cavity of female mice and subsequent delivery of therapeutic agents; the latter are exemplified by our use of Helicobacter pylori neutrophil activating protein (HP-NAP). A preliminary chemical burn of the mucosa, followed by the injection of mouse urothelial carcinoma cell line MB49 via urethral catheterization, enables the cells to attach to the bladder mucosa. After a period, required to allow an initial proliferation of the cells, mice are treated with HP-NAP, administrated again via catheterization. The anti-tumour activity of HP-NAP is evaluated comparing the tumour volume, the extent of necrosis and the degree of vascularization between vehicle- and HP-NAP-treated animals.

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Here the method to establish a syngeneic mouse model of orthotopic bladder tumour to evaluate the anti-tumour efficacy of the bacterial protein HP-NAP is described.

تقييم فعالية<em&gt; H. بيلوري</em&gt; البروتين HP-NAP كأداة علاجية لعلاج سرطان المثانة في مثلي فأري موديل

Bladder cancer is one of the most common malignancies of the urogenital tract. Intravesical injection of Bacillus Calmette-Gu&#233;rin (BCG) is the gold ...