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Guanosine, a guanine-based nucleoside relaxed isolated corpus cavernosum from mice through cGMP accumulation

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Abstract

In corpus cavernosum (CC), guanosine triphosphate (GTP) is converted into cyclic guanosine monophosphate (cGMP) to induce erection. The action of cGMP is terminated by phosphodiesterases and efflux transporters, which pump cGMP out of the cell. The nucleotides, GTP, and cGMP were detected in the extracellular space, and their hydrolysis lead to the formation of intermediate products, among them guanosine. Therefore, our study aims to pharmacologically characterize the effect of guanosine in isolated CC from mice. The penis was isolated and functional and biochemical analyses were carried out. The guanine-based nucleotides GTP, guanosine diphosphate, guanosine monophosphate, and cGMP relaxed mice corpus cavernosum, but the relaxation (90.7 ± 12.5%) induced by guanosine (0.000001–1 mM) was greater than that of the nucleotides (~ 45%, P < 0.05). Guanosine-induced relaxation was not altered in the presence of adenosine type 2A and 2B receptor antagonists. No augment was observed in the intracellular levels of cyclic adenosine monophosphate in tissues stimulated with guanosine. Inhibitors of nitric oxide synthase (L-NAME, 100 μM) and soluble guanylate cyclase (ODQ, 10 μM) produced a significant reduction in guanosine-induced relaxation in all concentrations studied, while in the presence of tadalafil (300 nM), a significant increase was observed. Pre-incubation of guanosine (100 μM) produced a 6.6-leftward shift in tadalafil-induced relaxation. The intracellular levels of cGMP were greater when CC was stimulated with guanosine. Inhibitors of ecto-nucleotidases and xanthine oxidase did not interfere in the response induced by guanosine. In conclusion, our study shows that guanosine relaxes mice CC and opens the possibility to test its role in models of erectile dysfunction.

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Abbreviations

ACh:

Acetylcholine

ADP:

Adenosine diphosphate

ARL 67156:

6-N,N-Diethyl-β-γ-dibromomethylene-D-adenosine-5′-triphosphate

ATP:

Adenosine triphosphate

cAMP:

Cyclic adenosine monophosphate

CC:

Corpus cavernosum

CD39:

Ecto-NTPase

CD73:

Ecto-5′-nucleotidase

cGMP:

Cyclic guanosine monophosphate

DPCPX:

1,3-Dipropyl-8-cyclopentylxanthine

ENT 1/2:

Nucleoside transporters type 1 and 2

GDP:

Guanosine diphosphate

GMP:

Guanosine monophosphate

GTP:

Guanosine triphosphate

L-NAME:

Nω-Nitro-L-arginine methyl ester

MRP4/MRP5:

Multidrug resistance proteins type 4 and 5

MRS 1754:

8-[4-[((4-Cyanophenyl)carbamoylmethyl)oxy]phenyl]-1,3-di(n-propyl)xanthine

NBTI:

S-(4-Nitrobenzyl)-6-thioinosine

NECA:

5′-(N-Ethylcarboxamido)adenosine

NF-kB:

Kappa beta nuclear factor

NO:

Nitric oxide

NOS:

Nitric oxide synthase

ODQ:

1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one

OGD:

Oxygen/glucose deprivation

PDE5:

Phosphodiesterase 5

sGC:

Soluble guanylate cyclase

SNP:

Sodium nitroprusside

ZM 241385:

4-(-2-[7-amino-2-{2-furyl}{1,2,4}triazolo{2,3-a} {1,3,5}triazin-5-yl-amino]ethyl)phenol

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Funding

This work is supported by São Paulo State Research Support Foundation (FAPESP - 2017/15175-1, 2018/21880-2), Higher Education Personnel Improvement Coordination (CAPES-001), and the National Council for Scientific and Technological Development (CNPQ-167319/2018-3).

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Authors and Affiliations

  1. Department of Pharmacology, Faculty of Medical Sciences, University of Campinas (UNICAMP), Campinas, São Paulo, 13083-881, Brazil

    Aline de Souza Nicoletti, Gabriela Reolon Passos, Gabriela Maria Bertollotto, Caroline Honaiser Lescano, Mariana Gonçalves de Oliveira, Edson Antunes & Fabiola Zakia Mónica

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  1. Aline de Souza Nicoletti
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  2. Gabriela Reolon Passos
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  3. Gabriela Maria Bertollotto
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  4. Caroline Honaiser Lescano
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  5. Mariana Gonçalves de Oliveira
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  6. Edson Antunes
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  7. Fabiola Zakia Mónica
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All authors have participated in the research and/or article preparation.

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Correspondence to Fabiola Zakia Mónica.

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All experimental protocols were carried out according to the Ethical Principles in Animal Research adopted by the Brazilian College for Animal Experimentation and approved by the Institutional Committee for Ethics in Animal Research of the University of Campinas (CEUA/UNICAMP protocol number 4719-1).

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de Souza Nicoletti, A., Passos, G.R., Bertollotto, G.M. et al. Guanosine, a guanine-based nucleoside relaxed isolated corpus cavernosum from mice through cGMP accumulation. Purinergic Signalling 16, 241–249 (2020). https://doi.org/10.1007/s11302-020-09702-5

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