Increased potency of α1-adrenergic receptors to induce inositol phosphates production correlates with the up-regulation of α1d/Ghα/phospholipase Cδ1 signaling pathway in term rat myometrium

Abstract

In the present study, we studied the potential regulation by rat myometrial α1-adrenergic receptors (α1-AR) of the newly identified Ghα protein/phospholipase Cδ1 (PLCδ1) signaling pathway and compared myometrial inositol phosphates (InsP) production and activity of the uterine circular muscle in response to α1-AR activation between mid-pregnancy and term. For this, we quantified the level of rat myometrial α1-AR coupling to Ghα protein by photoaffinity-labeling, the cytosolic amount of PLCδ1 enzyme by immunoblotting, and the expression level of α1-AR subtypes by RT-PCR. The results showed an increased level of α1-AR/Ghα protein coupling and the amount of PLCδ1 at term (+147 and +65% respectively, versus mid-pregnancy). This was correlated with an up-regulation of α1d-AR subtype (+70% versus mid-pregnancy). Incubation of myometrial strips with phenylephrine (Phe), a global α1-agonist, increased InsP production in a dose-dependent manner at both mid-pregnancy and term, but with an enhanced potency (tenfold decrease in EC50value) at term. Phe also dose-dependently induced contraction of the circular muscle at both mid-pregnancy and term. However, unlike InsP response, no amelioration of potency was observed at term. Similar results were obtained with the endogenous agonist norepinephrine. Our results show, for the first time, that rat myometrial α1d-AR/Ghα/PLCδ1 signaling pathway is up-regulated at term. This is associated with an increased potency of α1-AR to elicit InsP production but not uterine contraction at this period. It is thus hypothesized that α1-AR, through activation of Ghα/PLCδ1 system, are not primarily involved in the initiation of labor but may rather regulate responses such as myometrial cell proliferation or hypertrophy.