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Functional and Molecular Characterization of Voltage-Gated Sodium Channels in Uteri from Nonpregnant Rats

AuthorsSeda, Marian; Pinto Pérez, Francisco M. ; Wray, Susan; Cintado, Cristina G.; Noheda Marín, Pedro ; Buschmann, Helmut; Candenas, M. Luz
KeywordsFemale reproductive tract
Signal transduction
Issue Date1-Aug-2007
PublisherSociety for the Study of Reproduction
CitationBiology of Reproduction 77(5): 855-863 (2007)
AbstractWe investigated the function and expression of voltage-gated Naþ channels (VGSC) in the uteri of nonpregnant rats using organ bath techniques, intracellular [Ca2þ] fluorescence measurements, and RT-PCR. In longitudinally arranged whole-tissue uterine strips, veratridine, a VGSC activator, caused the rapid appearance of phasic contractions of irregular frequency and amplitude. After 50–60 min in the continuous presence of veratridine, rhythmic contractions of very regular frequency and slightly increasing amplitude occurred and were sustained for up to 12 h. Both the early and late components of the contractile response to veratridine were inhibited in a concentration-dependent manner by tetrodotoxin (TTX). In small strips dissected from the uterine longitudinal smooth muscle layer and loaded with Fura-2, veratridine also caused rhythmic contractions, accompanied by transient increases in [Ca2þ]i, which were abolished by treatment with 0.1 lM TTX. Using end-point and real-time quantitative RTPCR, we detected the presence of the VGSC alpha subunits Scn2a1, Scn3a, Scn5a, and Scn8a in the cDNA from longitudinal muscle. The mRNAs of the auxiliary beta subunits Scbn1b, Scbn2b, Scbn4b, and traces of Scn3b were also present. These data show for the first time that Scn2a1, Scn3a, Scn5a, and Scn8a, as well as all VGSC beta subunits are expressed in the longitudinal smooth muscle layer of the rat myometrium. In addition, our data show that TTX-sensitive VGSC are able to mediate phasic contractions maintained over long periods of time in the uteri of nonpregnant rats.
Description9 páginas, 8 figuras, 2 tablas
Publisher version (URL)http://dx.doi.org/10.1095/biolreprod.107.063016
Appears in Collections:(IIQ) Artículos
(IQOG) Artículos
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