Neonatal neurodevelopmental outcomes following tocolysis with glycerol trinitrate patches.

OBJECTIVE: The object of this study was to determine the effects of maternal tocolysis with glycerol trinitrate (GTN) patches on the neurodevelopment of infants. STUDY DESIGN: This was a randomized, multicenter, controlled trial comparing the efficacy of GTN patches with standard beta2 agonist as tocolytic therapy. The previously reported outcomes of this study indicated no difference in neonatal mortality or morbidity to hospital discharge. One hundred fifty-six surviving infants from 2 Australian centers were psychometrically assessed using the Griffiths Mental development Scales (revised) at 18 months of age. RESULTS: There was no difference in psychometric performance between those infants enrolled in either the GTN (81 infants) or beta2 agonist (75 infants) arm of the study. CONCLUSION: This randomized trial supports no significant difference between GTN patches in comparison with standard beta2 agonist for tocolytic therapy. The results underscore the association between premature labor and adverse infant outcomes.

Prostaglandins differentially modulate progesterone receptor-A and -B expression in human myometrial cells: evidence for prostaglandin-induced functional progesterone withdrawal.

We tested the hypothesis that prostaglandin (PGs), PGE2, and PGF2 alpha, stimulate labor and delivery in women, in part, by inducing functional progesterone withdrawal in myometrial cells by increasing the progesterone receptor (PR)-A/PR-B expression ration. PHM1-31 cells (an immortal pregnant human myometrial cell line) were exposed to PGE2, PGF2 alpha, cyclic-8-bromoadenosine monophosphate (8-Br-cAMP) and phorbol 12-myristate 13-acetate (PMA) at various concentrations for 24h. Effects on PR-A and PR-B expression were then assessed by quantitative RT-PCR. PGF2 alpha dose dependently increased PR-A mRNA and the PR-A/PR-B expression ration but did not effect PR-B mRNA. PGE2 dose-dependently increased mRNAs encoding PR-A and PR-B. The PGE2 dose-threshold for PR-A (0.01 nM) was lower than that for PR-B (0.1 nM), which resulted in an initial rise then a gradual fall in PR-A/PR-B expression ration to basal levels in response to PGE2. Activation of the protein kinase (PK)-A signaling pathway with 8-Br-cAMP coordinately increased expression of PR-A and PR-B and therefore did not alter the PR-A/PR-B expression ration. In contrast, activation of the PKC signaling pathway with PMA increased expression of PR-A without affecting PR-B and therefore significantly (P<0.05) increased the PR-A/PR-B expression ration. These data demonstrate differential control of myometrial PR-A and PR-B expression by PGE2 and PGF2 alpha and by specific intracellular signaling pathways. We conclude that PGs acting via the PKC pathway facilitate functional progesterone withdrawal by increasing the myometrial PR-A/PR-B expression ratio.

Intracrine control of estrogen action in human gestational tissues at parturition.

OBJECTIVE: We examined whether estrogen action in human parturition is regulated by an intracrine mechanism mediated by target tissue expression of specific 17beta-hydroxysteroid dehydrogenase (17betaHSD) isozymes that interconvert estrone (E1) and estradiol (E2), such that the onset of labor is associated with an increase in local E2 bioavailability. METHODS: The extent of 17betaHSD-1, -2, -3, -4, -5, and -7 expression (measured by quantitative reverse transcriptase polymerase chain reaction) and the capacity to interconvert E1 and E2 were compared in amnion, chorion, placenta, decidua, and myometrium obtained from women at term before (n = 6) and after (n = 6) the onset of labor. RESULTS: In chorion, abundance of 17betaHSD-1 (converts E1 to E2) mRNA decreased 2.7-fold (P <.05) in association with labor onset. In myometrium, 17betaHSD-1 and 17betaHSD-4 (converts E2 to E1) mRNAs increased two-fold and five-fold, respectively, with the onset of labor (P <.05 for each). No other statistically significant labor-associated change in 17betaHSD expression was observed. In chorion, 17betaHSD oxidative (E2 to E1) and reductive (E1 to E2) activities and the net E2 synthetic capacity increased with labor. In decidua, both activities decreased with the onset of labor, but there was no change in net E2 synthetic capacity. The capacity to interconvert E1 and E2 did not change in the other tissues. CONCLUSION: The increase in E2 synthetic capacity in the chorion might contribute to an increase in local estrogen bioactivity in association with the onset of labor. However, it cannot be explained by changes in 17betaHSD isozyme expression and is unlikely to account for the increased estrogen action at parturition. These data show that intracrine mechanisms based on 17betaHSD isozyme expression play a minor role, if any, in controlling estrogen action in gestational tissues during human parturition.

Recent advances in understanding the endocrinology of human birth.

The timing of human birth has a crucial impact upon the survival of the fetus. New knowledge on the regulation of human birth includes the role of endogenous retroviruses in the formation of the syncytiotrophoblast cells and consequently the secretion of corticotrophin releasing hormone, a hormone linked to gestational length determination. miRNAs have been identified that mediate progesterone withdrawal at labor by suppressing progesterone-induced transcription factors. Progress has also been made in understanding how the contractile machinery of the uterine myocytes is activated at labor and the role of small heat-shock proteins in this process. From this work, new therapeutic targets have been identified that may be used to regulate the onset of labor and improve neonatal mortality.

Nuclear progesterone receptor expression in the human fetal membranes and decidua at term before and after labor.

To explore how progesterone affects human pregnancy, we identified the progesterone target cells within the fetal membranes (amnion, chorion, and decidua) at term by assessing the extent of expression and localization of the nuclear progesterone receptors, progesterone receptor-A and progesterone receptor-B. Fetal membranes (separated into amnion and chorion-decidua) were obtained after term cesarean deliveries performed before (n = 7) and after (n = 7) labor onset. Nuclear progesterone receptor expression was determined by the abundance of nuclear progesterone receptor mRNAs (by quantitative reverse transcriptase-polymerase chain reaction) and proteins (by western blotting). Localization of nPRs was determined by immunohistochemistry. Progesterone receptor-A and progesterone receptor-B mRNA and protein levels were highest in the chorion-decidua and did not change in association with labor. Nuclear progesterone receptor mRNAs and proteins were barely detectable in amnion. Nuclear progesterone receptor immunostaining was detected only in the nucleus of decidual cells. These findings suggest that the decidua, and not the amnion and chorion, is a direct target for nuclear progesterone receptor-mediated progesterone actions during human pregnancy.

Progesterone receptor or cytoskeletal protein?

Immunoblotting is used to characterize the various nuclear progesterone receptor (nPR) isoforms present in tissues; however, the success of this technique is dependent on the specificity of the primary nPR antibody. The authors investigate the specificity of a frequently used nPR antibody, sc-538, in total protein from human myometrium and a myometrial cell line (PHM1-31). Using immunoblotting, 2 sc-538 immunoreactive bands at 100 and 55 kDa were detected. The bands were extracted and identified by 1-dimensional liquid chromatography mass spectrometry. The predominant protein in the 100-kDa band was alpha-actinin. The dominant proteins in the smaller band were vimentin (57 kDa) and desmin (53 kDa). Myometrial lysate was immunoprecipitated with sc-538, and immunoblotting of the immunoprecipitate with antibodies to alpha-actinin, desmin, and vimentin confirmed the presence of these proteins. The sc-538 nPR antibody therefore cross-reacts with cytoskeletal proteins that could be misinterpreted as nPR isoforms. Such misinterpretation has confused the progesterone response literature.