Korean J Urol.  2010 Aug;51(8):572-578. 10.4111/kju.2010.51.8.572.

Direct Effect of Carbon Monoxide on Relaxation Induced by Electrical Field Stimulation in Rat Corpus Cavernosum

• Kim DW
• Zhao C
• Kim MK
• Park JK

• KMID: 1997027
• DOI: http://doi.org/10.4111/kju.2010.51.8.572

Abstract

PURPOSE
Carbon monoxide (CO) may mediate smooth muscle relaxation in the rat corpus cavernosum smooth muscle (CCSM). We hypothesized that CO plays a role in neurally derived, frequency-dependent relaxation of rat CCSM.
MATERIALS AND METHODS
To study the effect of CO on CCSM relaxation induced by electrical field stimulation (EFS), a CCSM bundle was mounted on a force transducer and perfused with Hanks' balanced salt solution at 37degrees C with 95% O2 and 5% CO2. After 1 hour equilibration with -500 mg of passive tension, contraction of the CCSM bundle was elicited by 10(-5) M phenylephrine, which was continuously added with different concentrations of CO (1%, 2%, and 5%). Frequency-dependent relaxation was induced by EFS trains (0.2 ms at 0.5-32 Hz, for 10 s) repeated at 2 min intervals over 15 min in the presence of adrenergic and muscarinic receptor blocking agents (guanethidine and atropine, respectively). To study the distribution of heme oxygenase-2 (HO-2) in the rat CCSM, we performed immunohistochemical evaluation.
RESULTS
CO produced a dose-dependent enhancement of EFS-induced relaxation. Pretreatment with N(G)-nitro-L-arginine (a nitric oxide synthase blocker) greatly reduced the EFS-induced relaxation in the presence of CO (-45%). Pretreatment with zinc protoporphyrin-IX (ZnPP-9, a heme oxygenase inhibitor) had no significant effect on EFS-induced relaxation in the absence or the presence of CO. We found immunoreactivity for HO-2 in CCSM and immunoreactivity for protein gene product 9.5 (PGP 9.5) in nerve fibers.
CONCLUSIONS
We conclude that CO produced a dose-dependent enhancement of EFS-induced relaxation in rat CCSM bundles, but neurally derived, frequency-dependent relaxation in the rat CCSM depended mostly on nitric oxide in response to nonadrenergic noncholinergic neurotransmission. Immunoreactivity for HO-2 was found in rat CCSM but not nerve fibers.

Keyword

Carbon monoxide; Cyclic GMP; Electric stimulation; Muscle relaxation; Penile erection

MeSH Terms

Animals
Atropine
Carbon
Carbon Monoxide
Contracts
Cyclic GMP
Electric Stimulation
Heme
Heme Oxygenase (Decyclizing)
Isotonic Solutions
Male
Muscle Relaxation
Muscle, Smooth
Nerve Fibers
Nitric Oxide
Nitric Oxide Synthase
Penile Erection
Phenylephrine
Rats
Receptors, Muscarinic
Relaxation
Synaptic Transmission
Transducers
Zinc
Atropine
Carbon
Carbon Monoxide
Cyclic GMP
Heme
Heme Oxygenase (Decyclizing)
Isotonic Solutions
Nitric Oxide
Nitric Oxide Synthase
Phenylephrine
Receptors, Muscarinic
Zinc

Figure

• 

  FIG. 1 Immunohistochemical staining with antiserum raised in rabbit to heme oxygenase-2 in rat penile smooth muscle. (A) Negative staining with rabbit serum (primary antibody) and secondary antibody (CYTM3-conjugated AffiniPure donkey anti-rabbit IgG) (Control). (B) Positive staining with primary antibody (antiserum raised in rabbits to heme oxygenase-2) and secondary antibody (CYTM3-conjugated AffiniPure donkey anti-rabbit IgG). (C) Nerve fibers stained with primary antibody (antiserum raised in mouse to protein gene product 9.5) and secondary antibody (CYTM2-conjugated AffiniPure donkey anti-mouse IgG). (D) A composite of B and C.

• 

  FIG. 2 Effect of different carbon monoxide (CO) concentrations (1%, 2%, and 5%) on relaxation in Phe-precontracted corpus cavernosum smooth muscle before exposure to electrical field stimulation. HBSS: Hank's balanced salt solution.

• 

  FIG. 3 Effect of carbon monoxide (5%) on corpus cavernosum smooth muscle (CCSM) relaxation induced by electrical field stimulation (EFS). Rat CCSM was phenylephrine-precontracted and responded to EFS (0.5 to 32 Hz, 0.2 ms duration). HBSS: Hank's balanced salt solution.

• 

  FIG. 4 Frequency-dependent relaxation of phenylephrine-precontracted rat corpus cavernosum smooth muscle on different carbon monoxide concentrations (1%, 2%, and 5%). CTL: control, CO: carbon monoxide.

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  FIG. 5 Effect of nitric oxide synthase inhibitor, NG-nitro-L-arginine, on rat corpus cavernosum smooth muscle relaxation induced by electrical field stimulation in the presence of carbon monoxide (5%). CO: carbon monoxide, HBSS: Hank's balanced salt solution, L-NOARG: NG-nitro-L-arginine, EFS: electrical field stimulation.

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  FIG. 6 Inhibition of electrical field stimulation-induced relaxation of rat corpus cavernosum smooth muscle by antagonists (NG-nitro-L-arginine, zinc protoporphyrin-IX) in the presence of carbon monoxide (5%). CTL: control, CO: carbon monoxide, L-NOARG: NG-nitro-L-arginine, ZnPP: zinc protoporphyrin.

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