Basic Study
Copyright ©The Author(s) 2018. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Feb 28, 2018; 24(8): 882-893
Published online Feb 28, 2018. doi: 10.3748/wjg.v24.i8.882
Relaxin influences ileal muscular activity through a dual signaling pathway in mice
Eglantina Idrizaj, Rachele Garella, Fabio Francini, Roberta Squecco, Maria Caterina Baccari
Eglantina Idrizaj, Rachele Garella, Fabio Francini, Roberta Squecco, Maria Caterina Baccari, Department of Experimental and Clinical Medicine, Section of Physiological Sciences, University of Florence, Florence 50134, Italy
Author contributions: Idrizaj E and Garella R contributed equally to this work. Idrizaj E and Garella R performed the electrophysiological and the functional experiments, respectively; Baccari MC, Squecco R, Idrizaj E and Garella R designed the research study and analyzed the data; Francini F contributed to design the research study and analyzed the data; Baccari MC and Squecco R wrote the paper; Idrizaj E, Garella R, Francini F, Squecco R and Baccari MC critically revised the manuscript.
Supported by University of Florence (ex 60%) ROBERTASQUECCORICATEN15 (to Squecco R).
Institutional animal care and use committee statement: The experimental protocol was designed in compliance with the guidelines of the European Communities Council Directive 2010/63/UE and the recommendations for the care and use of laboratory animals approved by the Animal Care Committee of the University of Florence, Italy, with authorization from the Italian Ministry of Health nr. 787/2016-PR.
Conflict-of-interest statement: No conflicts of interest, financial or otherwise, are declared by the authors.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Roberta Squecco, PhD, Research Scientist, Department of Experimental and Clinical Medicine, Section of Physiological Sciences, University of Florence, Viale G.B. Morgagni 63, Florence 50134, Italy. roberta.squecco@unifi.it
Telephone: +39-55-2751600 Fax: +39-55-4379506
Received: October 19, 2017
Peer-review started: October 23, 2017
First decision: November 8, 2017
Revised: November 14, 2017
Accepted: November 28, 2017
Article in press: November 28, 2017
Published online: February 28, 2018
Processing time: 130 Days and 16 Hours
Abstract
AIM

To investigate the signaling pathways involved in the relaxin (RLX) effects on ileal preparations from mice through mechanical and electrophysiological experiments.

METHODS

For mechanical experiments, ileal preparations from female mice were mounted in organ baths containing Krebs-Henseleit solution. The mechanical activity was recorded via force-displacement transducers, which were coupled to a polygraph for continuous recording of isometric tension. Electrophysiological measurements were performed in current- and voltage-clamp conditions by a microelectrode inserted in a single smooth muscle cell (SMC) of the ileal longitudinal layer. Both the membrane passive properties and inward voltage-dependent L-type Ca2+ currents were recorded using suitable solutions and voltage stimulation protocols.

RESULTS

Mechanical experiments showed that RLX induced a decay of the basal tension and a reduction in amplitude of the spontaneous contractions. The effects of RLX were partially reduced by 1H-[1,2,4]oxadiazolo[4,3-a]-quinoxalin-1-one (ODQ) or 9-cyclopentyladenine mesylate (9CPA), inhibitors of guanylate cyclase (GC) and adenylate cyclase (AC), respectively, and were abolished in the concomitant presence of both drugs. Electrophysiological experiments demonstrated that RLX directly influenced the biophysical properties of ileal SMCs, decreasing the membrane conductance, hyperpolarizing the resting membrane potential, reducing the L-type calcium current amplitude and affecting its kinetics. The voltage dependence of the current activation and inactivation time constant was significantly speeded by RLX. Each electrophysiological effect of RLX was reduced by ODQ or 9CPA, and abolished in the concomitant presence of both drugs as observed in mechanical experiments.

CONCLUSION

Our new findings demonstrate that RLX influences ileal muscle through a dual mechanism involving both GC and AC.

Keywords: Relaxin; Gastrointestinal motility; Smooth muscle; AC/cAMP; GC/cGMP

Core tip: Up to now relaxin (RLX) was described to act only through the NO/guanylate cyclase (GC)/cGMP/PKG pathway in different gastrointestinal tracts. The results of the present study, achieved on mice ileal preparations and carried out by a combined mechanical and electrophysiological approach, demonstrate for the first time that both GC and adenylate cyclase are involved in the effects of RLX in this intestinal region. The activation of this dual signaling pathway by RLX might represent a reinforcing (redundant) myorelaxant mechanism in the ileum, underlying the physiological importance of the hormone and leading to speculate translational perspectives in the treatment of intestinal dysmotilities.