Ryanodine receptor type 3 does not contribute to contractions in the mouse myometrium regardless of pregnancy.

Abstract

Ryanodine receptor type 3 (RyR3) is expressed in myometrial smooth muscle cells (MSMCs). The short isoform of RyR3 is a dominant negative variant (DN-RyR3) and negatively regulates the functions of RyR2 and full-length (FL)-RyR3. DN-RyR3 has been suggested to function as a major RyR3 isoform in non-pregnant (NP) mouse MSMCs, and FL-RyR3 may also be upregulated during pregnancy (P). This increase in the FL-RyR3/DN-RyR3 ratio may contribute to the strong contractions by MSMCs for parturition. In the present study, spontaneous contractions by the myometrium in NP and P mice were highly susceptible to nifedipine but were not affected by ryanodine. Ca2+ image analyses under a voltage clamp revealed that the influx of Ca2+ through voltage-dependent Ca2+ channels did not cause the release of Ca2+ from the sarcoplasmic reticulum (SR). Cytosolic Ca2+ concentrations ([Ca2+]cyt) in MSMCs were not affected by caffeine. Despite the abundant expression of large conductance Ca2+-activated K+ channels in MSMCs, spontaneous transient outward currents were not observed in the resting state because of the substantive lack of Ca2+ sparks. Quantitative PCR and Western blot analyses indicated that DN-RyR3 was strongly expressed in the NP myometrium, while the expression of FL-RyR3 and DN-RyR3 was markedly reduced in the P myometrium. The messenger RNA (mRNA) expression of RyR2 and RyR1 was negligible in the NP and P myometria. Moreover, RyR3 knockout mice may become pregnant and deliver normally. Thus, we concluded that none of the RyR subtypes, including RyR3, play a significant role in the regulation of [Ca2+]cyt in or contractions by mouse MSMCs regardless of pregnancy.