Intracavernous pressure (ICP) elicited by electrical stimulation was previously described.34 In brief, mice are anesthetizedby 4% isoflurane for induction, and 1.5–2.0% isoflurane for maintenance using an inhalationanesthesia apparatus (Nakazawa Seisaku‐sho, Funabashi, Japan). The left crus of the corpuscavernosum (CC) was cannulated using a 23‐G needle for continuous ICP monitoring. The pressure transducer was connected through a transducer amplifier to a data acquisition board (PowerLab 2/26, AD Instruments Pty). Stainless steel bipolar wire electrodes (Unique Medical) and a pulse generator (Nihon Kohden) were used for cavernous nerve stimulations to electrically stimulate penile erections. The stimulation parameters were as follows: 5 V, 16 Hz, and duration time: 1 min.
Pulse generator
Manufactured by Nihon Kohden
Sourced in Japan
The Nihon Kohden Pulse Generator is a device that generates electrical pulses. It is designed to provide precise and reliable pulse signals for various applications in a laboratory setting.
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8 protocols using pulse generator
1
Electrical Stimulation-Induced Penile Erection
2
In Vivo Optogenetic Manipulation of Sleep
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Fourteen days after the surgery for implanting the EEG/EMG electrodes and optic guide cannula, the EEG/EMG recording cables were connected to the amplifier and fiber optic cables (1-m long, 200-μm diameter, Newdoon Inc., Hangzhou, China) were placed inside the implanted cannula simultaneously, and fiber-optic rotary joints (Doric Lenses, Québec, Canada) were used for unrestricted in vivo illumination. Rats were acclimatized for 2 days before the photostimulation sessions. Light pulse trains were programmed using a pulse generator (Nihon Kohden, Tokyo, Japan) that provided simultaneous input into 2 blue light lasers (473 nm, 100 mW intensity; SLOC, Shanghai, China). For acute optogenetic procedure, each stimulation train was applied 60 s after a stable NREM or REM sleep event as detected by real-time online polysomnographic recording. For the chronic photostimulation experiments, blue light stimulation (5-ms pulses, 50 trains of 20 Hz for MPB, 25 trains of 40 Hz for BF, LH and VM, main interval 30 s) was applied for 1 h during 9:00-10:00.
3
Evaluating Erectile Function in Rats
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Intracavernous pressure (ICP) was measured by electrical stimulation as previously reported.9 (link), 10 (link), 11 (link), 12 (link), 13 (link) Briefly, rats from each group (n = 8) were anaesthetized with isoflurane using inhalation anesthesia apparatus. The carotid artery was cannulated for continuous monitoring of the mean arterial pressure (MAP) and the left crus of the CC was cannulated using a 23-G needle for continuous ICP monitoring. The pressure transducer was connected through an amplifier to a data acquisition board (PowerLab 2/26, ADInstruments Pty. Ltd., New South Wales, Australia). Stainless steel bipolar wire electrodes (Unique Medical, Osaka, Japan) and a pulse generator (Nihon Kohden, Tokyo, Japan) were used for the penile stimulations with the following parameters: 1 min at 5 V, 1–16 Hz, and a square wave duration of 5 microseconds. Erectile function was evaluated using the maximum ICP/MAP ratio.
4
Penile Erectile Function Assessment
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ICP was measured using electrical stimulation, as previously reported.22 , 23 , 24 Briefly, the rats from each group (n = 6–12) were anesthetized using isoflurane (Mylan, Canonsburg, PA, USA). The carotid artery was cannulated for the continuous monitoring of the mean arterial pressure (MAP), and the left crus of the corpus cavernosum was cannulated using a 23‐G needle for continuous ICP monitoring. The pressure transducer was connected via an amplifier to a data acquisition board (PowerLab 2/26; ADInstruments Pty. Ltd., Bella Vista, Australia). Stainless steel bipolar wire electrodes (Unique Medical, Osaka, Japan) and a pulse generator (Nihon Kohden, Tokyo, Japan) were used for penile stimulation under the following conditions: 1 min at 5 V, 1–16 Hz, and a square wave duration of 5 ms. Erectile function was evaluated using the maximum ICP/MAP ratio, as ICP is influenced by systemic arterial pressure.25
5
Intracavernous Pressure Measurement by Electrical Stimulation
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Intracavernos pressure (ICP) was measured by electrical stimulation as previously reported.3 (link),10 (link), 11 (link), 12 (link), 13 , 14 Rats from each group were briefly anesthetized using isoflurane (Mylan, Canonsburg, PA, USA) using inhalation anesthesia apparatus (Nakazawa Seisaku-sho, Funabashi, Japan). The carotid artery was cannulated for continuous monitoring of the mean arterial pressure (MAP), and the left crus of the corpus cavernosum was cannulated using a 23-G needle for continuous ICP monitoring. The pressure transducer was connected through an amplifier to a data acquisition board (PowerLab 2/26; ADInstruments Pty. Ltd., Bella Vista, Australia). Stainless steel bipolar wire electrodes (Unique Medical, Osaka, Japan) and a pulse generator (Nihon Kohden, Tokyo, Japan) were used for penile stimulation with the following parameters: 1 minute at 5 V, 1–16 Hz, and a square wave duration of 5 minutes. Erectile function was evaluated using the maximum ICP/MAP ratio, as ICP is influenced by systemic arterial pressure.
6
Cavernous Nerve Electrostimulation for Erectile Function
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Erectile function was assessed based on changes in intracavernous pressure (ICP), followed by electrostimulation of the cavernous nerves, as previously reported (Hotta et al., 2014 (link)). Briefly, under anesthesia with isoflurane (Pfizer Inc.), body temperature was maintained for each animal. First, polyethylene tubing (PE‐50) was inserted into the left carotid artery in order to record blood pressure. Next, the area posterolateral to the prostate was explored on both sides, and the major pelvic ganglions and cavernous nerves were identified and exposed. The skin overlying the penis was incised, the left penile crus was exposed, and a 23‐gauge needle connected to PE‐50 tubing filled with heparinized saline (5 U heparin/ml) was inserted into the crus for ICP measurement. Next, electrostimulation was performed using stainless steel bipolar wire electrodes (Unique Medical) and a pulse generator (Nihon Kohden) for cavernous nerve stimulation with the following parameters: 1 min at 5 V, 16 Hz, and a square wave duration of 5 ms. Systemic arterial pressure and ICP were recorded and analyzed with LabChart8 software (ADInstruments). Erectile function was evaluated using the maximum ICP to mean arterial pressure (MAP) ratio because ICP is influenced by systemic arterial pressure.
7
Erectile Function Evaluation via ICP
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Erectile function was evaluated using ICP measurement as previously reported.20 (link),21 (link) The rats from each group were anaesthetized briefly using isoflurane using an inhalation anesthesia apparatus. The carotid artery was cannulated for continuous monitoring of the arterial pressure, and the left crus of the CC were cannulated using a 23-G needle for continuous ICP monitoring. The pressure transducer was connected through an amplifier to a data acquisition board (PowerLab 2/26, AD Instruments Pty., Ltd., New South Wales, Australia). Stainless steel bipolar wire electrodes (Unique Medical, Osaka, Japan) and a pulse generator (Nihon Kohden, Tokyo, Japan) were used for cavernous nerve stimulations with the following parameters: 1 minutes at 10 V, 20 Hz, and a square wave duration of 5 milliseconds or 1 minute at 5 V, 1–16 Hz, and a square wave duration of 5 milliseconds. As ICP is influenced by systemic arterial pressure, erectile function was evaluated using the maximum ICP/mean arterial pressure (MAP) ratio. The MAP was calculated from blood pressure measurements during electrical stimulation using LabChart ver.7 (AD Instruments Pty).
8
Intracavernous Pressure Evaluation in Rats
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Intracavernous pressure (ICP) was measured after electrical stimulation as reported previously16 (link),17 (link). Briefly, rats from each group were anesthetized using isoflurane (Mylan, Canonsburg, PA, USA). Under inhalational anesthesia, with 4% isoflurane for induction and 1.0%–1.5% isoflurane for maintenance, the carotid artery was cannulated for continuous monitoring of the mean arterial pressure (MAP) and the left crus of the CC was cannulated using a 23-G needle for continuous monitoring of the ICP. The pressure transducer was connected through an amplifier to a data acquisition board (PowerLab 2/26, ADInstruments Pty. Ltd., New South Wales, Australia). Stainless steel bipolar wire electrodes (Unique Medical, Osaka, Japan) and a pulse generator (Nihon Kohden, Tokyo, Japan) were used for penile stimulations with the following parameters: 1 minute at 10 V, 20 Hz, and a square wave duration of 5 ms. Erectile function was evaluated using the maximum ICP/MAP ratio.
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