Human Placental Endothelium Research Articles

Insulin receptor isoforms: an integrated view focused on gestational diabetes mellitus.

The human insulin receptor (IR) exists in two isoforms that differ by the absence (IR-A) or the presence (IR-B) of a 12-amino acid segment encoded by exon 11. Both isoforms are functionally distinct regarding their binding affinities and intracellular signalling. However, the underlying mechanisms related to their cellular functions in several tissues are only partially understood. In this review, we summarize the current knowledge in this field regarding the alternative splicing of IR isoform, tissue-specific distribution and signalling both in physiology and disease, with an emphasis on the human placenta in gestational diabetes mellitus (GDM). Furthermore, we discuss the clinical relevance of IR isoforms highlighted by findings that show altered insulin signalling due to differential IR-A and IR-B expression in human placental endothelium in GDM pregnancies. Future research and clinical studies focused on the role of IR isoform signalling might provide novel therapeutic targets for treating GDM to improve the adverse maternal and neonatal outcomes.

Fc gamma receptor IIb participates in maternal IgG trafficking of human placental endothelial cells.

The human placental transfer of maternal IgG is crucial for fetal and newborn immunity. Low-affinity immunoglobulin gamma Fc region receptor IIb2 (FCGR2B2 or FcγRIIb2) is exclusively expressed in an IgG-containing, vesicle-like organelle (the FCGR2B2 compartment) in human placental endothelial cells; thus, we hypothesized that the FCGR2B2 compartment functions as an IgG transporter. In this study, to examine this hypothesis, we performed in vitro bio-imaging analysis of IgG trafficking by FCGR2B2 compartments using human umbilical vein endothelial cells transfected with a plasmid vector containing enhanced GFP-tagged FCGR2B2 (pFCGR2B2-EGFP). FCGR2B2-EGFP signals were detected as intracellular vesicular structures similar to FCGR2B2 compartments in vivo. The internalization and transcytosis of IgG was significantly higher in the pFCGR2B2-EGFP-transfected cells than in the mock-transfected cells, and the majority of the internalized IgG was co-localized with the FCGR2B2-EGFP signals. Furthermore, we isolated FCGR2B2 compartments from the human placenta and found that the Rab family of proteins [RAS-related protein Rab family (RABs)] were associated with FCGR2B2 compartments. Among the RABs, RAB3D was expressed predominantly in placental endothelial cells. The downregulation of RAB3D by small interfering RNA (siRNA) resulted in a marked reduction in the FCGR2B2-EGFP signals at the cell periphery. Taken together, these findings suggest that FCGR2B2 compartments participate in the transcytosis of maternal IgG across the human placental endothelium and that RAB3D plays a role in regulating the intracellular dynamics of FCGR2B2 compartments.

Role of equilibrative adenosine transporters and adenosine receptors as modulators of the human placental endothelium in gestational diabetes mellitus

Gestational diabetes mellitus (GDM) is a diseases that alters human placenta macro and microvascular reactivity as a result of endothelial dysfunction. The human placenta is a highly vascularized organ which lacks innervation, so blood flux is governed by locally released vasoactive molecules, including the endogenous nucleoside adenosine and the free radical nitric oxide (NO). Altered adenosine metabolism and uptake by the endothelium leads to increased NO synthesis which then turns-off the expression of genes coding for a family of nucleoside membrane transporters belonging to equilibrative nucleoside transporters, particularly isoforms 1 (hENT1) and 2 (hENT2). This mechanism leads to increased extracellular adenosine and, as a consequence, activation of adenosine receptors to further sustain a tonic activation of NO synthesis. This is a phenomenon that seems operative in the placental macro and microvascular endothelium in GDM. We here summarize the findings available in the literature regarding these mechanisms in the human feto-placental circulation. This phenomenon is altered in the feto-placental vasculature, which could be crucial for understanding GDM deleterious effects in fetal growth and development.

Gestational Diabetes Mellitus and the Role of Adenosine in the Human Placental Endothelium and Central Nervous System

Gestational Diabetes Mellitus and the Role of Adenosine in the Human Placental Endothelium and Central Nervous System

Gestational Diabetes Mellitus and the Role of Adenosine in the Human Placental Endothelium and Central Nervous System

Diabetes mellitus is a syndrome that alters macro and microvascular reactivity as a result of endothelial dysfunction. This phenomenon occurs in vascular beds from different regions in the adult human body, as well as the developing fetus including the placenta and the brain vasculature. The human placenta is highly vascularized lacking of innervation, so that blood flux is governed by locally released vasoactive moleculles, including the endogenous nucleoside adenosine and the gas nitric oxide (NO). Altered adenosine uptake by the endothelium leads to increased synthesis of NO resulting in repression of equilibrative nucleoside membrane transporters isoforms 1 (hENT1) and 2 (hENT2). The latter result in higher extracellular adenosine and activation of adenosine receptors (ARs) sustaining tonic activation of NO synthesis in gestational diabetes mellitus (GDM). Similar alterations are apparent in the fetus brain. Because GDM associates with serious alterations in the CNS functions leading to abnormal higher cognitive functions, among others complications, it is proposed that altered adenosine metabolism, uptake and dynamics of ARs activation in the brain is a factor leading to these pathologies. We here summarize the findings available in the literature regarding these mechanisms in the human fetus brain with reference to that in the feto-placental circulation. We emphasize the possibility that adenosine plays a key role in the functional link established by astrocytes connecting neurons and the brain vasculature. This phenomenon is altered in the brain and the feto-placental vasculature, which could be crucial for understanding GDM deleterous effects in fetal growth and development, as well as CNS abnormal function in adulthood.

Gestational Diabetes Mellitus and the Role of Adenosine in the Human Placental Endothelium and Central Nervous System

Gestational Diabetes Mellitus and the Role of Adenosine in the Human Placental Endothelium and Central Nervous System

IgG is transported across the mouse yolk sac independently of FcγRIIb

While generally accepted that FcRn of the human syncytiotrophoblast and the mouse yolk sac endoderm is the major IgG transporter, the finding of a different Fc receptor FcgammaRIIb (RIIb) in the human placental endothelium has suggested the existence of an additional IgG transporter. Testing our hypothesis in mouse, we found that while RIIb is expressed in the yolk sac vasculature, IgG concentrations in fetuses of wild-type mice (RIIb(+/+)) and mice with a null mutation in the gene encoding RIIb (RIIb(-/-) mice) are not different, and we thus reject our hypothesis that yolk sac RIIb transports IgG in utero in the mouse. However, the capillary bed in the mouse yolk sac is structurally more complex than in human placenta, consisting of three types of cells: an RIIb-negative endothelium, a unique RIIb-bearing cell that also expresses 2 out of 4 macrophage markers but not endothelial cell or pericyte markers, and pericytes. As in the human placenta the b2 isoform of RIIb predominates in the mouse yolk sac. Remarkably only a single capillary channel rather than 2 channels with a loop is found in each yolk sac villus, which, along with intracapillary erythrocytes, suggests that blood flow is peristaltic, mediated by pericytes. It is not clear whether RIIb in the human placental villus might mediate an IgG transport function in light of the mouse yolk sac equivalent failing to do so.

Insulin Action on the Human Placental Endothelium in Normal and Diabetic Pregnancy

The placental endothelium is unique among the entire human vasculature. The blood enriched in oxygen and nutrients is transported in the veins, whereas the arteries contain deoxygenated blood coming from the fetus. The placental vasculature has to develop rapidly to ensure adequate supply of the fetus. Therefore, factors present in the fetal circulation will stimulate placental angiogenesis. In the third trimester of pregnancy the placental endothelium is richly endowed with insulin receptors. In a pregnancy complicated by maternal diabetes, fetal hyperinsulinemia resulting from maternal and, hence, fetal hyperglycaemia induces changes in the placental vasculature such as increased growth and angiogenesis. This review will discuss general effects of insulin on endothelial cells and further focus on insulin effects on the placental endothelium. Isolation and culture of placental endothelial cells has allowed the identification of insulin effects in vitro. These include metabolic effects of insulin i.e. stimulation of glycogen synthesis, and modulation of angiogenesis on the placental arterial endothelium i.e. regulation of ephrin-B2 expression, an arterial specific signalling molecule implicated in sprouting. The effect of insulin on ephrin-B2 in placental arterial endothelial cells as well as their particularly high expression levels of insulin receptors and receptors for vascular endothelial growth factors indicate that placental angiogenesis is likely to emanate from the arterial compartment and is stimulated by insulin.

Structure and permeability of human placental microvasculature.

Endothelial paracellular junctions are important structures for the regulation of vascular permeability, junctional organisation being systematically related to the functional properties of the endothelium. Electron microscopic studies, immunocytochemistry, and single-passage permeability measurements have established that the placental microvessels resemble the fairly tight continuous microvessels of skeletal muscle both in structure and permeability. The endothelial paracellular clefts of these microvessels contain two distinct junctional entities which may influence permeability: the tight junction and the adherens junction. These clefts impose a substantial restriction to molecules above RMM 1000 Da and large haemproteins cannot cross the clefts. The 18 nm-wide zones of the clefts possess the transmembrane adhesion molecules PECAM-1 and VE-cadherin, which have been implicated in junctional assembly and permeability. Inflammatory mediators such as histamine and tumour necrosis factor cause a redistribution of these adhesion molecules to non-junctional regions, and histamine (100 microM) causes a rapid and sustained rise in extraction of radio-labeled tracers. Electron microscopy has also revealed possible first indications of tight junctional disassembly. Both the endothelia of larger placental vessels and isolated placental microvascular endothelial cells express cadherins and PECAM-1 and contain an extensive F-actin cytoskeleton, which is implicated in changes of cell shape and junctional assembly/disassembly. Thus, the human placental endothelium, using perfusion techniques and in vitro experiments, offers a valuable model for vascular permeability studies.

Concordant expression of tissue factor and class II MHC antigens in human placental endothelium

Villitis of unestablished etiology is a placental lesion frequently associated with high risk pregnancies: it is also found in placentae from normal term pregnancies. The etiology of the lesion is unknown. Vasculitis and thrombosis have been described in villitis areas of placentae from normal and high risk pregnancies. We asked if fetal stem vessel endothelium in villitis lesions expresses MHC class II antigens, and if this is associated with a thrombogenic activity of these vessels. We found that endothelium of fetal stem vessels in villitis areas was usually MHC class II (HLA-DR, DP and DQ) reactive. Reactivity of fetal stem vessel endothelium for MHC class II antigens was associated with the presence of tissue factor reactivity and the absence of thrombomodulin reactivity. These changes on endothelial plasma membranes can promote intravascular coagulation, ischemic necrosis, vasculitis and other histological changes characteristic of villitis.