Calcitriol affects hCG gene transcription in cultured human syncytiotrophoblasts.
BACKGROUND
In pregnancy, maternal serum concentrations of calcitriol significantly rise as a result of increased renal and placental contribution in order to assure calcium supply for the developing fetus. Considering that placenta is a site for vitamin D activation, and the versatility and potency of calcitriol, it is feasible that this hormone participates in fetal/placental development and physiology. In the present work we studied calcitriol actions upon human chorionic gonadotropin (hCG) secretion and expression in cultured trophoblasts, as well as vitamin D receptor (VDR) and CYP27B1 immunolocalization in placental villi.


METHODS
Quantification of hCG in culture media was performed by immunoassay. Expression studies were carried out by real time PCR. Analysis of CYP27B1 and VDR localization in placental slides were performed by immunohistochemistry. Statistical significance was established by one way ANOVA using Tukey test for comparisons.


RESULTS
Calcitriol regulated hCG in a time-dependent manner: at 6 h the secosteroid stimulated hCG, whereas longer incubations (24 h) showed opposite effects. Interestingly, calcitriol stimulatory effects on hCG were accompanied by an increase in intracellular cAMP content and were abolished by pre-incubation of the cells with a selective protein kinase A inhibitor. Immunohistochemical techniques showed differential VDR localization in the syncytiotrophoblast layer or in the vascular smooth muscle cells depending on the epitope to which the antibodies were raised (specific for the carboxy- or amino-terminal regions, respectively). CYP27B1 was immunolocalized in the syncytiotrophoblast layer of placental villi.


CONCLUSIONS
The presence and location of the vitamin D activating enzyme CYP27B1 as well as the specific receptor for vitamin D were shown in placental sections. The latter, together with findings demonstrating specific effects of calcitriol acting through the VDR and the cAMP/PKA signaling pathway upon hCG expression and secretion, indicate that there is a functional vitamin D endocrine system in the placenta, and recognize calcitriol as an autocrine regulator of hCG.
Barrera
David
D
Departamento de Biología de la Reproducción, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Vasco de Quiroga No, 15, Tlalpan 14000, México, DF, México. barrera1912@gmail.com
Avila
Euclides
E
Hernández
Guillermo
G
Méndez
Isabel
I
González
Leticia
L
Halhali
Ali
A
Larrea
Fernando
F
Morales
Angélica
A
Díaz
Lorenza
L
D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase / Q000032:analysis; D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D002117:Calcitriol / Q000494:pharmacology*; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D018167:Receptors, Calcitriol / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic / Q000187:drug effects*; D014327:Trophoblasts / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*
D006063:Chorionic Gonadotropin; D012333:RNA, Messenger; D018167:Receptors, Calcitriol; D015124:8-Bromo Cyclic Adenosine Monophosphate; D000242:Cyclic AMP; D015090:25-Hydroxyvitamin D3 1-alpha-Hydroxylase; D002117:Calcitriol