Name: cavernous nerve stimulation and recording of intracavernous pressure in a rat
Uploaded: 2018-04-23T12:30:00
Description: Watch this Scientific Journal Video about Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat at JoVE.com

The authors have no acknowledgements.

Animals were housed in the University of Southern Denmark Animal Care Facility as per institutional guidelines. All animal experiments were carried out in accordance with the National Institutes of Health guide for the care and use of laboratory animals. This is an acute, non-survival surgery procedure.&#160;
1. Preparation of Tubing, Electrode, and Instruments for the Surgical Procedure
<ol>
	<li>Use the following microsurgical instruments: surgical scissors, angled micro scissors, a tissue...

<ol>
	<li>Eckhard, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Untersuchungen &#252;ber die Erektion des Hundes In: Beitr&#228;ge zur Anatomie und Physiologie. , 123-166 (1863).</li><li>Quinlan, D. M., Nelson, R. J., Partin, A. W., Mostwin, J. L., Walsh, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+rat+as+a+model+for+the+study+of+penile+erection.">The rat as a model for the study of penile erection.</a> J Urol. 141 (3), 656-661 (1989).</li><li>Heaton, J. P., Varrin, S. 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L., Becker, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunophilin+ligands+promote+penile+neurogenesis+and+erection+recovery+after+cavernous+nerve+injury.">Immunophilin ligands promote penile neurogenesis and erection recovery after cavernous nerve injury.</a> J Urol. 171 (1), 495-500 (2004).</li><li>Yamashita, S., 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=Nerve+injury-related+erectile+dysfunction+following+nerve-sparing+radical+prostatectomy:+a+novel+experimental+dissection+model.">Nerve injury-related erectile dysfunction following nerve-sparing radical prostatectomy: a novel experimental dissection model.</a> Int J Urol. 16 (11), 905-911 (2009).</li><li>Burnett, A. 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=GGF2+is+neuroprotective+in+a+rat+model+of+cavernous+nerve+injury-induced+erectile+dysfunction.">GGF2 is neuroprotective in a rat model of cavernous nerve injury-induced erectile dysfunction.</a> J Sex Med. 12 (4), 897-905 (2015).</li><li>Lin, 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=Nanoparticle+Improved+Stem+Cell+Therapy+for+Erectile+Dysfunction+in+a+Rat+Model+of+Cavernous+Nerve+Injury.">Nanoparticle Improved Stem Cell Therapy for Erectile Dysfunction in a Rat Model of Cavernous Nerve Injury.</a> J Urol. 195 (3), 788-795 (2016).</li><li>Kapoor, M. S., Khan, S. A., Gupta, S. K., Choudhary, R., Bodakhe, S. 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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+sequelae+of+cavernous+nerve+injury+in+the+rat:+is+there+model+dependency.">Functional sequelae of cavernous nerve injury in the rat: is there model dependency.</a> J Sex Med. 3 (1), 77-83 (2006).</li><li>Albersen, M., 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=Injections+of+adipose+tissue-derived+stem+cells+and+stem+cell+lysate+improve+recovery+of+erectile+function+in+a+rat+model+of+cavernous+nerve+injury.">Injections of adipose tissue-derived stem cells and stem cell lysate improve recovery of erectile function in a rat model of cavernous nerve injury.</a> J Sex Med. 7 (10), 3331-3340 (2010).</li></ol>

The principal goal of this study was to describe a simplified surgical technique of penile crus cannulation for ICP recording and isolation of the CN for electrostimulation. We introduced modifications to the dissection of the cavernous body to simplify the surgery and provide reproducible recordings of the increase of the ICP with CN stimulation. With a 1 cm vertical skin incision, lateral to the base of the penis, using the palpable ischial tuberosity as guidance, we achieved good exposure of the ischiocavernosus muscl...

In vivo study of erectile function in experimental animals started in 1863 with the pioneering experimental work of Eckhard1. Electrostimulation of the pelvic nerves was used to induce the increased ICP in dogs. Throughout the 20th century, similar experimental protocols were used in larger animals such as dogs, monkeys, cats and rabbits. Evaluating erectile function in a rat was first developed by Quinlan et al. in 19892. The method has since been modified and updated by several other groups34. Today, the rat is the most widely used animal model for studying the pathology of erectile dysfunction and evaluating emerging treatment options. The main steps of the procedure include, recording systemic blood pressure using a line in the carotid artery, cannulation of the penile crus to measure ICP, and stimulation of the CN to induce an increase in the ICP. Although several researchers have refined the model, its reproducibility remains a problem, and variable results have been reported by different laboratories. Several pitfalls still persist.
Previous articles5,6,7,8,9,10 describe the use of full penile exposure with degloving of the penis for cavernous body cannulation. This is not an optimal approach as manipulation and disruptive dissection causes injury to structures, which are essential to erectile function. The dissection of the CN has been well described10,11, but stimulation of the nerve is not optimal due to multiple factors that could affect experimental results. The technique of CN stimulation includes lifting the nerve from the surrounding tissue by pulling on the bipolar hook electrode, which is positioned around the nerve, and drying the nerve before each stimulation. This can lead to various degrees of nerve damage and electrical current leakage, resulting in a diminished response or false increase in the ICP through stimulation of alternative pathways e.g., pelvic floor muscles, bladder and gastrointestinal tract12. All of these factors limit reproducibility.
During our study, we observed that both the depth and type of anesthesia have a profound effect on the ICP. The anesthetics used are sodium pentobarbital, ketamine/xylazine or ketamine/midazolam injection, or isoflurane/oxygen inhalation.
Here we describe a simplified surgical method and provide data in support of standardization of the experimental protocol.

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cavernous nerve stimulation and recording of intracavernous pressure in a rat

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for erectile dysfunction. However, the methods used vary between laboratories, and pitfalls still exist. The goal of this study was to describe a surgical technique that would provide a reliable and reproducible model. By exposing the ischiocavernosus muscle at its point of insertion on the ischial tuberosity, the penile crus could be cannulated with minimal dissection and injury to the structures involved in erectile function. Repeated stimulation of the CN, without the need for lifting and drying, was achieved by using a 125 &#181;m bipolar silver electrode and biocompatible silicon glue to isolate the electrode-nerve complex. This method prevents neuropraxia by reducing stretching and drying the nerve and provides complete isolation of the nerve, negating electrical leakage and preventing stimulation of alternative pathways.

The use of this protocol with the recommended stimulation settings, under inhalation anesthesia with isoflurane 2.0% oxygen 0.8 L/min, should produce results as shown in Figure 11 and Figure12, where there is multiple back to back stimulations between 75-80 mm Hg. Figure 13 shows the same stable response over a 20-min stimulation with the response stable at 73-77 mm Hg. Test the line for ICP meas...

Watch this Scientific Journal Video about Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat at JoVE.com

Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat

This study describes a simplified surgical procedure and technique for performing cavernous nerve stimulation with the isolation of the nerve-electrode complex using silicone glue and intracavernous pressure measurement.

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for ...

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