Purinergic receptors in human placenta: evidence for functionally active P2X₄, P2X₇, P2Y₂, and P2Y₆

Abstract

Appropriate regulation of ion transport by the human placental syncytiotrophoblast is important for fetal growth throughout pregnancy. In nonplacental tissues, ion transport can be modulated by extracellular nucleotides that raise intracellular calcium ([Ca2+]i) via activation of purinergic receptors. We tested the hypothesis that purinergic receptors are expressed by human placental cytotrophoblast cells and that their activation by extracellular nucleotides modulates ion (K+) efflux and [Ca2+]i. P2X/P2Y receptor agonists 5-bromouridine 5'-triphosphate (5-BrUTP), ADP, ATP, 2',3'-O-(4-benzoyl-benzoyl)adenosine 5'-triphosphate (BzATP), and UTP stimulated 86Rb (K+ tracer) efflux from cultured cytotrophoblast cells at early (mononuclear) or later (multinucleate syncytiotrophoblast-like) stages of differentiation, with ATP and UTP particularly potent. 2-Methylthioadenosine 5'-triphosphate (2-MeS-ATP), and UDP elevated 86Rb efflux only from multinucleated cells. All agonists caused a significant peak and plateau increase in [Ca2+]i, although the magnitude of responses was variable. The effect of BzATP, UTP, and UDP in multinucleated cells was unaffected, and that of ATP partially inhibited, by removal of extracellular Ca2+, implicating P2Y receptor activation. mRNA encoding P2X1, P2X2, P2X4, and P2X7 and P2Y1, P2Y2, P2Y4, P2Y6, and P2Y11 were identified in mono- and multinucleated cells, whereas P2X3 and P2X5 mRNA were absent from all samples. Western blot analysis revealed P2X4, P2X7, P2Y2, and P2Y6 protein in cytotrophoblast cells, but P2Y4 was not detected. On the basis of published agonist selectivity, the data indicate the presence of functionally active P2X4, P2X7, P2Y2, and P2Y6 receptors in cytotrophoblast cells. We propose that activation of these receptors, and subsequent elevation of [Ca2+]i, modulates syncytiotrophoblast homeostasis and/or maternofetal ion exchange in response to extracellular nucleotides.

Danger Signals From ATP and Adenosine in Pregnancy and Preeclampsia

“…Both subtypes of purinergic receptors have widespread tissue expression, including the placenta (see Tables S2 and S3). 5,24,25 …”

Purinergic signalling in the reproductive system in health and disease

“…In a study of P2 receptor mRNA and protein expression in human placental villus fragments at two stages of gestation, the first trimester and term [342], P2X4, P2X7 and P2Y 2 (but not P2Y 4 ) receptor subtypes were found and fluorescent calcium imaging experiments showed that ATP, UTP and 2′(3′)-O -(4-benzoylbenzoyl)adenosine 5′-triphosphate caused significant elevation of [Ca 2+ ] i in term compared to the first trimester. Systematic pharmacological studies of purinergic signalling in human placenta showed functionally active P2X4, P2X7, P2Y 2 and P2Y 6 receptors on cytotrophoblast cells and it was proposed that activation of these receptors and subsequent elevation of [Ca 2+ ] i modulates syncytiotrophoblast homeostasis and maternofoetal ion exchange [341]. P2Y 1 and P2Y 2 receptors are unevenly distributed along the human placental vascular tree; there is a 6-to 8-fold increase in these receptors from the cord to the chorionic or cotyledon vessels [69].…”

Purinergic signalling in endocrine organs

“…Since the human placenta is not innervated, autoc r i n e , p a r a c r i n e a n d e n d o c r i n e m o d u l a t i o n o f syncytiotrophoblast transport function is of pivotal importance. Functionally active P2X4, P2X7 , P2Y 2 and P2Y 6 receptors have been identified on human placental syncytiotrophoblast cells [446]. This group showed later that post-translational modifications of the syncytiotrophoblast P2X4 receptor are altered in preeclampsia [447].…”