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

This work was supported by the Fundamental Research Funds for the Central Universities (020814380018, 020814380077), The China Scholarship Council (CSC, No. 201606195024), Natural Science Foundation of Jiangsu Province (BK20160138), and Key Project supported by Science and Technology Development Foundation, Nanjing Medical University (2014NJMUZD053).

Three-month-old and 18-month-old Sprague-Dawley rats were used in the present study. All animals were handled in accordance with the NIH Guidelines for the Care and Use of Laboratory Animals. Procedures involving animal subjects were approved by the local Institutional Animal Care and Ethics Committee, with an effort to minimize animal suffering. The protocols were approved by the Institutional Animal Care and Use Committee (IACUC) at Nanjing Tech University (Nanjing, China).
The rats were div...

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As a direct measure of erectile function, ICP is a reliable method14. It allows for the acquisition of data on basal ICP, peak ICP, plateau ICP, time to erection and detumescence time, duration of response, etc. Besides these direct measured parameters, there are some other index parameters: (1) &#34;T80&#34;, the time to reach 80% of peak ICP; (2) &#34;D20&#34;, the time to decrease to 20% of peak ICP; (3) &#34;&#916;T80&#34;, the rate of increase in pressure (per second) at T80; and (4)...

ED is defined as the inability to attain or keep a penile erection, and has become a common male sexual disorder1. Experimental animals are used and provide reproducible models to investigate erectile function2. For a long time, several larger animal models have been employed for investigating erectile function3,4,5. Although rodents are relatively small compared to other animals, they are also used for the study of male erectile dysfunction due to exhibiting several advantages6. First, the morphological and functional sexual characteristics of humans are recapitulated in rodents. Second, compared to larger animals used in ED studies, rodents are more economical to purchase, house, and maintain. Third, genetically modified rodent models provide advantages in reproducible and subsequent behavioral as well as neurophysiological studies. Therefore, rodents have quickly become the primary animals used in the study of male erectile dysfunction.
Benefiting from a pure genetic background and consistent culture conditions, rodent models have provided consistently reproducible data5,6,7,8. Among the numerous available studies related to many aspects of erectile functions, the apomorphine (APO)-induced erectile response test and the electric-stimulation-induced ICP response test are the most widely used methods that reliably reflect erectile function9,10,11,12. The APO-induced erectile function test, developed by Heaton et al.13, is a bio-assay that utilizes the phenomenon that administration of apomorphine to rats elicits erections and yawns. As an easy, noninvasive, and stable bio-assay to evaluate erectile function, the APO-induced erectile function test is widely used in many studies. However, this assay does not adequately reflect the quality of erections or the dynamic changes in blood flow associated with an erectile response14. ICP measurements were initially developed by Quinlan et al.15. In this method, a catheter is placed into the carotid artery to measure systemic blood pressure, and another catheter is inserted into the crus corpus cavernosum to record the ICP. Before or during the ICP recording, a vasoactive agent and/or electrical field stimulation of the major pelvic ganglion (MPG) or CN were often given to the rats14. This assay has been a reliable tool for evaluating the therapies and medicines for ED, and will likely be used as a vital evaluation method in the future6.
Compared to the easily-performed APO-induced erectile function test, successful ICP recordings require attention to detail, practice, and adherence to the operation method. Therefore, here, we provide a detailed description of how to perform ICP recording.

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		<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

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. Rodents are employed by many studies to research the physiology/pathology of erectile function. Erectile function in rodents can be evaluated by measuring the intracavernosal pressure (ICP). In practice, ICP can be monitored following electrical stimulation of the cavernous nerves (CNs). The arterial pressure of the carotid artery (the mean arterial pressure) is used as the reference for ICP. Using ICP recording protocols, many key parameters of erectile function can be measured from the ICP response curve. The ICP measurement provides more information than the apomorphine-induced penile erection test, and is cheaper than telemetric monitoring of the corpus spongiosum penis, making this method the most popular one to evaluate erectile function. However, compared to the easily-performed APO-induced erectile function test, successful ICP recordings require attention to detail, practice, and adherence to the operation method. In this work, an introduction to ICP recording in rats is provided to complement the procedure efficiently.

Numerous studies have shown that erectile dysfunction in aged males is becoming a common problem. However, medical treatment is limited in the management of aging-related ED16. In rodent models of aging-related ED, many therapies are tested on the erectile function of aged rats. As we have introduced above, the ICP recording test could be used to distinguish the ED animals from the total population of experimental animals, which is also valuable to quantify the eff...

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

Intracavernosal pressure recording (ICP) is an important method to evaluate the erectile function of experimental animals. Here, a detailed protocol is demonstrated for the recording procedure of ICP by catheterizing the crura penis and then electrically stimulating the cavernous nerves in rats.

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 

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

Pressure-polishing Pipettes for Improved Patch-clamp Recording

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording.  We first developed this method for fabricating pipettes suitable for ...

Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method

The CODA 8-Channel High Throughput Non-Invasive Blood Pressure system measures the blood pressure in up to 8 mice or rats simultaneously. The CODA ...

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 ...

Exploring Arterial Smooth Muscle Kv7 Potassium Channel Function using Patch Clamp Electrophysiology and Pressure Myography

Contraction  or relaxation of smooth muscle cells within the walls of resistance arteries  determines the artery diameter and thereby controls flow of ...

Assessing Murine Resistance Artery Function Using Pressure Myography

Pressure myograph systems are  exquisitely useful in the functional assessment of small arteries, pressurized  to a suitable transmural pressure. The  ...

Force Measurement During Contraction to Assess Muscle Function in Zebrafish Larvae

Zebrafish larvae provide models of muscle development, muscle disease and muscle-related chemical toxicity, but related studies often lack functional ...

One-channel Cell-attached Patch-clamp Recording

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate ...

Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

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

Photodiode-Based Optical Imaging for Recording Network Dynamics with Single-Neuron Resolution in Non-Transgenic Invertebrates

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