Name: intracavernosal pressure recording to evaluate erectile function in rodents
Uploaded: 2018-06-06T12:30:00
Description: Watch this Scientific Journal Video about Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents at JoVE.com

この作品は、中央大学 (020814380018、020814380077)、中国奨学金議会 (CSC、号 201606195024)、江蘇省の自然科学財団 (BK20160138)、およびキー プロジェクトの基礎的研究資金によって支えられました。科学と技術開発財団、南京医科大学 (2014NJMUZD053) によってサポートされています。

1 歳 3 か月と 18 ヶ月歳 Sprague-dawley ラットは本研究で使用されました。すべての動物は、ケアと実験動物の使用のため NIH ガイドラインに従って処理されていました。動物を対象とする手順は、動物の苦痛を最小限に抑える努力をローカル機関動物のケアと倫理委員会によって承認されました。プロトコル承認機関動物ケアおよび使用委員会 (IACUC) 南京工科大学 (中国南京市) で。
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<ol>
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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+Models+of+Erectile+Dysfunction.">Animal Models of Erectile Dysfunction.</a> Curr Prot Pharmacol. 29, 5.41.1-5.41.22 (2005).</li><li>Giuliano, F., Bernabe, J., Rampin, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Telemetric+monitoring+of+intracavernous+pressure+in+freely+moving+rats+during+copulation.">Telemetric monitoring of intracavernous pressure in freely moving rats during copulation.</a> J Urol. 152 (4), 1271-1274 (1994).</li><li>Bernabe, J., Rampin, O., Giuliano, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracavernous+pressure+changes+during+reflexive+penile+erections+in+the+rat.">Intracavernous pressure changes during reflexive penile erections in the rat.</a> Physiol Behav. 57 (5), 837-841 (1995).</li><li>Bernabe, J., Rampin, O., Sachs, B. 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ICP は、勃起機能の直接測定、信頼性の高い方法14。ICP、勃起と detumescence 時、応答等の期間のピーク ICP、基底 icp データの取得が高原をことができます。他のいくつかのインデックス パラメーターがあります測定パラメーターを直接これらのほか: ICP; ピーク時の 80% に達するとは (1)「T80」、時間(2)「D20」、時間ピーク ICP; の 20% に減少するには(3) の「ΔT80」、の T80; (...

ED は達成または陰茎の勃起を維持することができないこととして定義され、一般的な男性の性的障害1となっています。実験動物は使用され、勃起機能2を調査する再現性のあるモデルを提供します。長い間、勃起機能3,4,5を調査するためいくつかのより大きい動物モデルを採用されています。齧歯動物が他の動物と比較して比較的小さいが、彼らはまた、いくつかの利点6を展示のための男性の勃起不全の研究のため使用します。まず、人間の形態と機能の性的特徴は齧歯動物で締めくくっています。第二に、エドの研究で使用されるより大きい動物と比較して、齧歯動物は、家を購入し、維持するより経済的です。第三に、遺伝子組み換えの齧歯動物モデルは、再現性とその後の行動と神経生理学的研究にメリットをもたらします。したがって、齧歯動物すぐに男性の勃起不全の研究で使用される主な動物となっています。
純粋な遺伝的背景と一貫した培養条件の恩恵、齧歯動物モデルは、一貫して再現性のあるデータ5,6,7、8を提供しています。(APO) アポモルヒネ勃起機能の多くの面に関連する多くの利用可能な研究の間で-誘発勃起反応試験と電気刺激による ICP 応答試験は確実に勃起を反映する最も広く使用されるメソッド9,10、11,12を機能します。ヒートンらによって開発された、勃起機能の APO 誘発テスト13日は、アポモルヒネ ラットへの投与が勃起とあくびを引き出すという現象を利用したバイオアッセイです。容易、非侵襲的、および安定したバイオアッセイ勃起機能を評価するため、として APO 誘発勃起機能テストは広く多くの研究に使用されます。ただし、この試金は勃起または関連付けられて勃起反応14の血流の動的変化の質を十分に反映されません。ICP 測定、クインランらが開発された当初15します。 この方法ではカテーテルは全身の血圧を測定する頸動脈に置かれ、ICP を記録する cru 海綿体に別のカテーテルを挿入。前に、または記録、血管作動性エージェントおよび/または主要な骨盤神経節 (MPG) の電気刺激誘導中に CN もよくラット14に行われたか。この試金は、ED の治療と薬を評価するための信頼性の高いツールをされているし、将来6重要な評価方法として多分使用されます。
APO 誘発勃起機能を簡単に実行テストに比べると、巧妙な誘導録音詳細、実践、そして操作方法への付着に注意が必要。したがって、ここでは、ICP の記録を実行する方法の詳細な説明いたします。

<table border="0" cellpadding="0" cellspacing="0" style="width:1099px;" width="1098">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:128px;">Animal</td>
			<td style="width:289px;"></td>
			<td style="width:336px;"></td>
			<td style="width:345px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rats</td>
			<td></td>
			<td></td>
			<td>Strain: Sprague-Dawley 
			Age: 2-3 month</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rats</td>
			<td></td>
			<td></td>
			<td>Strain: Sprague-Dawley 
			Age: 15-18 month</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Reagents</td>
			<td></td>
			<td></td>
			<td>formula</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:128px;">Saline</td>
			<td style="width:289px;">Sigma-Aldrich, S7653</td>
			<td style="width:336px;"></td>
			<td style="width:345px;">dissolve 8.5 gram sodium chloride in distilled water</td>
		</tr>
		<tr height="68">
			<td height="68" style="height:68px;width:128px;">Pentobarbital sodium solution</td>
			<td style="width:289px;">Sigma-Aldrich, P3761</td>
			<td style="width:336px;"></td>
			<td style="width:345px;">dissolve 1 gram in 100 ml saline</td>
		</tr>
		<tr height="36">
			<td height="36" style="height:36px;width:128px;">Povidone-iodine</td>
			<td>BTP Pharmaceutical Co. Limited</td>
			<td></td>
			<td style="width:345px;">10% (V/V)</td>
		</tr>
		<tr height="36">
			<td height="36" style="height:36px;width:128px;">Ethanol</td>
			<td style="width:289px;">China National Pharmaceutical Group Corporation (SINOPHARM)</td>
			<td style="width:336px;"></td>
			<td style="width:345px;">70% (V/V)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:128px;">Heparin</td>
			<td style="width:289px;">Sigma-Aldrich, H3149</td>
			<td style="width:336px;"></td>
			<td style="width:345px;">dissolve 20000U heparin in 100 ml saline</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Materials</td>
			<td></td>
			<td colspan="2"></td>
		</tr>
		<tr height="35">
			<td height="35" style="height:35px;width:128px;">Hypodermic needle</td>
			<td style="width:289px;">Shandong Weigao Group Medical Polymer Co., Ltd.&#160;</td>
			<td>23G</td>
			<td></td>
		</tr>
		<tr height="35">
			<td height="35" style="height:35px;width:128px;">Syringe</td>
			<td style="width:289px;">Shandong Weigao Group Medical Polymer Co., Ltd</td>
			<td>10 ml</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:128px;">Three-way stopcock</td>
			<td style="width:289px;">Chengdu Instrument factory</td>
			<td>TSK 01</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:128px;">Electrode</td>
			<td style="width:289px;">Chengdu Instrument factory</td>
			<td>JST-1</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:128px;">Catheter tube</td>
			<td style="width:289px;">Chengdu Instrument factory</td>
			<td>PE-10, PE-50</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:128px;">Operating scissors</td>
			<td>Shanghai operation equipment factory</td>
			<td>J22010, J22020</td>
			<td></td>
		</tr>
		<tr height="35">
			<td height="35" style="height:35px;width:128px;">Ophthalmic operating scissors</td>
			<td>Shanghai operation equipment factory</td>
			<td>Y00010, Y00020</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:128px;">Ophthalmic forceps</td>
			<td>Shanghai operation equipment factory</td>
			<td>JD1010, JD1020</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:128px;">MicroScissors</td>
			<td>World Precision Instruments</td>
			<td>WAA260</td>
			<td></td>
		</tr>
		<tr height="35">
			<td height="35" style="height:35px;width:128px;">silk suture</td>
			<td style="width:289px;">Shandong Weigao Group Medical Polymer Co., Ltd.&#160;</td>
			<td>5-0</td>
			<td></td>
		</tr>
		<tr height="35">
			<td height="35" style="height:35px;">Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Equipment</td>
			<td></td>
			<td colspan="2"></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:128px;">Stimulator</td>
			<td style="width:289px;">Nanjing medease science and technology co. ltd (model 4C501H)</td>
			<td style="width:336px;"></td>
			<td style="width:345px;">15 Hz, 5 ms pulse, 5 V, 60 s duration and 5 minutes interval</td>
		</tr>
		<tr height="52">
			<td height="52" style="height:52px;width:128px;">Multichannel signal collection processing system</td>
			<td style="width:289px;">Nanjing medease science and technology co. ltd (model 4C501H)</td>
			<td style="width:336px;"></td>
			<td>Blood pressure model</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:128px;">Pressure transducer</td>
			<td style="width:289px;">Beijing Xin Hang Xing Ye Technology Trading Company Limited (model YP100)</td>
			<td style="width:336px;"></td>
			<td>40KPa</td>
		</tr>
	</tbody>
</table>

intracavernosal pressure recording to evaluate erectile function in rodents

勃起不全 (ED) は達成するか、陰茎の勃起を維持することができないこととして定義され、この流行している男性の性的障害となっています。齧歯動物は、勃起機能の生理学・病理学を研究する多くの研究により雇われています。齧歯動物で勃起機能は intracavernosal 圧力 (ICP) を測定することによって評価できます。実習では、次の (CNs) の海綿状の神経への電気刺激誘導を監視できます。(平均動脈圧) 頸動脈の動脈の圧は、誘導のための参照として使用されます。ICP プロトコルを記録を使用して、ICP 応答曲線から勃起機能の多くの重要なパラメーターを測定できます。ICP 測定アポモルヒネ誘発勃起テストより多くの情報を提供しています、このメソッドに勃起機能を評価する最も人気のある 1 つを作る、海綿体テレメトリ監視よりも安いです。巧妙な誘導録音 APO 誘発勃起機能を簡単に実行テストと比較して、しかし、詳細、実践、そして操作方法への付着に注意が必要です。この作品は、ラットにおける ICP 記録への入門は効率的に手順を補完するために提供しています。

数多くの研究は、高齢者でその勃起不全を示している男性共通の問題となっています。しかし、医療は加 ED16の管理に制限されます。加 ED の齧歯動物モデルは、多くの治療法は高齢ラットの勃起機能にテストされます。我々 が上記で紹介した、ICP 録音テスト勃起機能の潜在的な治療や薬の効果を定量化する貴重なもである実験動物の総人口から ED ?...

Watch this Scientific Journal Video about Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents at JoVE.com

Intracavernosal Pressure Recording to Evaluate Erectile Function in Rodents

齧歯動物で勃起機能評価 Intracavernosal 圧記録

Intracavernosal 圧記録 (ICP) は、実験動物の勃起機能を評価する重要な方法です。ここでは、ICP の記録手順の crura ペニスを catheterizing とラットの海綿状の神経を電気刺激により詳細なプロトコルを示す.

Erectile dysfunction (ED) is defined as the inability to attain or keep an erection of the penis, and this has become a prevalent male sexual disorder. ...

The overall goal of this procedure is to test erectile function of rats by catheterizing the crus corpus cavernosum and recording intracavernous pressure after stimulation of the cavernous nerves. This method can help answer key questions in the male erectile dysfunction field, such as, for evaluating the therapies and medicines for erectile dysfunction. So one advantage of this technique is that it provides a more accurate evaluation of erection function and reflexes, the dynamic change in blood flow associated with erectile response.Visual demonstration of this method is critical as catheterization is difficult to learn because the small area of the white tunica albuginea of crus corpus cavernosum makes it difficult to insert the needle into the penis crura. To prepare for the surgery, make a pair of bipolar electrodes for intracavernosal pressure recordings. Slightly bend the ends of the electrodes and adjust the distance between them to one to two millimeters.Then connect the electrodes to the stimulator using two crocodile clamps. Next, assemble the catheter system. First, use tubing to connect a hypodermic 23-gauge needle to a three-way stopcock.Then connect the stopcock to a pressure transducer. Next attach a 10 milliliter syringe loaded with heparin saline to the third position of the stopcock. Then fill the whole system with heparin saline and check for leaks.Next lift the needle 20 centimeters and calibrate the recording system to 20 centimeters of water. After the calibration, adjust the height of the needle and check the accuracy of the recording system. As needed, repeat the calibration until the accuracy is confirmed.Next, transfer the rats from the animal facility to the surgery room and allow them to acclimate to the room for at least 30 minutes. Meanwhile, sterilize the surgical tools with 70%ethanol and prepare the necessary surgical materials. After 30 minutes prepare the first rat for surgery.Anesthetize it with sodium pentobarbital and wait for five to 10 minutes. Then pinch the rats toes and check for an absence of reflex to confirm a proper anesthetization. Next, shave the fur from the abdomen and neck and place the rat supine on a surgical board.Now wipe the exposed skin with three alternating scrubs with 10%povidone iodine followed by 70%ethanol. Also, apply ophthalmic ointment to prevent the eyes from drying out. Then proceed with the surgery.Before isolating the cavernous nerve, catheterize the left carotid artery according to the text protocol. Start the nerve isolation by lifting the skin and muscle of the abdomen with a pair of forceps. Then, with scissors, make a midline incision from the lower abdomen to the penis.Next, gently move aside the intestines into the upper part of the abdominal cavity. Then grasp the bladder with a pair of forceps and pull it out of the abdominal cavity. Now find the ventral lobes of the prostate which are located on the ventral portion of the urethra.Next, expose the dorsal lobe of the prostate by pulling out the ventral lobes, seminal vesicle, and vas deferens. Now find the point of adhesion of the vas deferens and prostate. Then separate the space between the prostate and vas deferens.Proceed by carefully exposing the fibrous capsule located posterior to the junction point of the prostate and vas deferens. Then find the major pelvic ganglion and cavernous nerves located on the surface of the prostate. Now carefully dry the cavernous nerves area with a swab or tissue paper, and then carefully isolate and hook the right cavernous nerve with the bipolar electrodes.To continue the surgery, cut a small incision in the skin of the penis with dissecting scissors and then carefully denude the skin of the penis shaft. Then expose enough tissue to dissect the striated penile musculature. Now find the upper branch of pubis bone.Then expose the bulbospongiosus muscle which covers the spongios bulb. Next, using curved forceps, divide the bulbospongiosus muscle from the ischiocavernosus muscle. Proceed by carefully isolating the ischiocavernosus muscle.Then cut the ischiocavernosus muscle to expose the white tunica albuginea of the crus corpus cavernosum. Now perform the catheterization. Carefully follow the anatomical direction of the crus corpus cavernosum and gently insert the needle through the white tunica albuginea into the cavernosum.The location of the needle is crucial to the operation's success, so as a test, inject a small amount of heparinized saline. This should result in a slight penile tumescence if the needle has been inserted correctly. Next, without repositioning the needle, or disrupting the connecting tube, carefully release the needle.Then inspect the area for any leakage. If none is found, proceed with stimulating the nerve and analyzing the data. The described ICP recording test was used to distinguished aged rats with erectile disfunction to help quantify the effect of potential treatment or drugs on erectile function.A typical ICP response curve of the erectile disfunction group was much lower than the control group's curve. Usually the highest ICP was chosen for statistical analysis. After calculating the ratio of ICP to mean arterial pressure, the pooled data from five rats showed that the ratio in the aged group decreased significantly compared to that of the control groups.Besides these two key parameters, the peak ICP, the plateau ICP, the detumescence time, the duration of response, and the area under the curve, all significantly decreased in aged rats, thus, data is full of quantitative measurements that reflect erectile function. After watching this video, you should have a good understanding of how to efficiently perform the ICP recording on animals. Don't forget that while attempting this procedure, it's important to remember to use heparin in order to ensure the canula tube remains unobstructed.Once mastered, this technique can be done in about two hours if it is performed properly.

Research

JoVE Journal

Biology

Survivable Stereotaxic Surgery in Rodents 

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The ability to measure extracellular basal levels of neurotransmitters in the brain of awake animals allows for the determination of effects of different ...

生物学

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Ambulatory ECG Recording in Mice

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Force Measurement During Contraction to Assess Muscle Function in Zebrafish Larvae

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One-channel Cell-attached Patch-clamp Recording

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The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a unique channel-forming member of the ATP Binding Cassette (ABC) superfamily of ...

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The development of transgenic invertebrate preparations in which the activity of specifiable sets of neurons can be recorded and manipulated with light ...