Search

• P2X4 receptor on CD4 + T cells from PB and AT was increased in individuals with BMI ≥ 25 Kg/m2. • CD4 + T cells expressing P2X4 and P2X7 from PB and AT were augmented in individuals with BMI ≥ 25 Kg/m2. • CD4 + T cells with P2X4 and P2X7 showed a higher percentage in cells isolated from AT compared to PB. • ATP-induced shedding of CD62 L was higher in PBMC than in ATMC. • Influx of Ca2+ dependent of ATP-induced P2X4 activation in PBMC was higher than in ATMC. Obesity is characterized by immune cell infiltration and inflammation. Purinergic receptors such as P2X1, 4 and 7 are expressed on immune cells and their activation contributes with an inflammatory response. However, the simultaneous expression of P2X1, 4 and 7 during overweight or obesity have not been described. Therefore, the aim of this study was to determine single and simultaneously expression and function of the P2X1, 4 and 7 receptors in lymphocytes and CD4 + T cells from peripheral blood (PB) and adipose tissue (AT). Our results showed a higher expression of the P2X4 receptor on CD4 + T cells from PB regarding P2X7 and P2X1 receptor expression. In addition, P2X4 receptor expression on CD4 + T cells from PB and AT was increased in individuals with BMI ≥ 25 Kg/m2. Moreover, a higher simultaneous expression of the P2X4 and P2X7 receptors on CD4 + T cells from AT compared to CD4 + T cells expressing P2X1 and P2X7 receptors simultaneously. Besides, CD4 + T cells expressing P2X4 and P2X7 receptors from PB and AT were augmented in individuals with BMI ≥ 25 Kg/m2. In addition, the percentage of lymphocytes and also CD4 + T cells expressing P2X4 receptor were elevated both in PB and AT compared to cells expressing P2X7 or P2X1. However, CD4 + T cells expressing P2X4 and P2X7 were augmented in AT compared to PB. The function of the receptors showed a lower shedding of CD62 L in adipose tissue mononuclear cells (ATMC) compared with peripheral blood mononuclear cells (PBMC) and a greater participation of P2X4 in the mobilization of intracellular calcium. We concluded that it was possible to determine for the first time the simultaneous expression of purinergic receptors in ATMC, where the P2X4 receptor has a greater participation in the activation of CD4 + T cells possibly modulating the function of the other two receptors. [ABSTRACT FROM AUTHOR]

The pathologic hallmark of Parkinson's disease (PD) is the accumulation of alpha-synuclein (αsyn) in susceptible neurons in the form of Lewy bodies and Lewy neurites. The etiology of PD remains unclear. Because brain injury has been suggested to facilitate αsyn aggregation, we investigated whether cellular breakdown products from damaged cells can act on neighboring healthy cells and cause intracellular αsyn accumulation and/or aggregation. Using 2 neuronal cell models, we found that extracellular adenosine triphosphate (ATP) induced a significant increase in intracellular αsyn levels between 24 and 48 hours after treatment. Further investigation revealed that the observed αsyn accumulation is a result of lysosome dysfunction caused by extracellular ATP-induced elevation of lysosomal pH. Interestingly, P2X1 receptor appears to mediate the cells' response to extracellular ATP. Although Ca 2+ influx via P2X1 receptor is necessary for αsyn accumulation, Ca 2+ influx per se is not sufficient for increased αsyn accumulation. These findings provide new insight into our knowledge of the role of P2X receptors in PD pathogenesis and may be helpful in identifying new therapeutic targets for PD. [ABSTRACT FROM AUTHOR]

Abstract: Carboxylic acid derivatives of pyridoxal were developed as potent P2X1 and P2X3 receptor antagonists with modifications of a lead compound, pyridoxal-5′-phosphate-6-azophenyl-2′,5′-disulfonate (5b, iso-PPADS). The designing strategies included the modifications of aldehyde, phosphate or sulfonate groups of 5b, which may be interacted with lysine residues of the receptor binding pocket, to weak anionic carboxylic acid groups. The corresponding carboxylic acid analogs of pyridoxal-5′-phosphate (1), 13 and 14, showed parallel antagonistic potencies. Also, most of 6-azophenyl derivatives (24–28) of compound 13 or 14 showed potent antagonistic activities similar to that of 5b at human P2X3 receptors with 100 nM range of IC50 values in two-electrode voltage clamp (TEVC) assay system on the Xenopus oocyte. The results indicated that aldehyde and phosphoric or sulfonic acids in 5b could be changed to a carboxylic acid without affecting antagonistic potency at mouse P2X1 and human P2X3 receptors. [Copyright &y& Elsevier]

Abstract: P2X1 receptors and adrenoceptors are mainly responsible for vasoconstriction in a variety of blood vessels. However, previous studies have shown that α,β-methylene adenosine 5′-triphosphate (α,β-MeATP), a stable analogue of ATP, can induce both pressor and depressor responses in laboratory animals. In this study, the effects of increasing intravenous doses of α,β-MeATP and noradrenaline (NA) (0–30nmol/kg) administered at 20min intervals on systolic (SBP), diastolic (DBP) and mean (MBP) blood pressure in groups of anesthetized mice (n=6) were compared. Both α,β-MeATP and NA caused transient, dose-dependent increases in SBP, DBP and MBP but the effect of α,β-MeATP was more rapid and significantly larger at doses of 10 and 30nmol/kg (P<0.01). At the dose of 30nmol/kg, α,β-MeATP increased SBP, DBP and MBP by 65.8±7.0, 65.7±5.0 and 65.7±5.5mmHg, respectively, compared to increases of 36.8±4.6, 33.3±4.9 and 34.5±4.7mmHg, respectively, produced by NA. These results indicate P2X1 receptors play an important role in BP regulation although purinergic vasoconstriction alone may not explain the more potent pressor response to α,β-MeATP in the anesthetized mouse. [Copyright &y& Elsevier]

Objective: To investigate the expression of P2X1 receptors and connexin 43 in gap junctions between smooth muscle cells. Contraction mediated by P2X receptors is known to occur in the bladder and male reproductive tract, and cell–cell coupling of smooth muscle via gap junctions is essential for synchronized rhythmic activity of these tissues. Design: We selected for this study rat myometrial smooth muscle during pregnancy and at postpartum day l. Setting: University medical school. Animal(s): Laboratory rats. Intervention(s): Rats were mated and became pregnant. Main Outcome Measure(s): Immunostaining and fluorescence and confocal microscopy. Result(s): The level of P2X1 receptor expression remained low throughout pregnancy (days 4 to 20) but was greatly up-regulated at day 22 (postpartum day 1). Connexin 43 expression showed a pattern of up-regulation, with progression through pregnancy and peaking near labor, but exhibited a rapid down-regulation after parturition. Conclusion(s): The functional significance of the changes in connexin 43 and P2X1 receptor expression that have been observed is discussed in relation to triggering and modulation of uterine contractility during and after pregnancy. [Copyright &y& Elsevier]

Background: It has been suggested that gap-junctional conductance between cardiomyocytes is regulated through a specific ligand-receptor interaction between ATP and connexins. In this study we examined the localization of P2X1 ionotropic receptors and their relation to connexin43 in gap junctions in human left ventricles. Methods and results: Using immunohistochemistry, we detected P2X1 expression predominantly in the intercalated discs. Labelling of the P2X1 receptor and the gap junction protein connexin43 showed close association in some gap junctions, while in others the two proteins often appeared to be spatially discrete. Western blotting detected four major bands at 45, 60, 95 and 120 kDa in the protein extracts from human left ventricles corresponding to equivalent bands from rat vas deferens. The most prominent band in human left ventricles was at 95 kDa, possibly a dimer of the native P2X1 receptor, whereas in rat vas deferens it was at 60 kDa. After preincubation of the antibody with its epitope peptide, the 45 and 60 kDa bands almost disappeared and the 95 and 120 kDa bands were significantly attenuated. Conclusions: P2X1 receptors in human myocardium are densely localized in gap junctions at intercalated discs between muscle cells. Close association of P2X1 receptors and connexin 43 occurred in some regions of some gap junctions, but in others they were spatially separate. Little difference in the pattern of distribution of P2X1 receptors was found in failing left ventricles of patients with dilated cardiomyopathy, although Western blots showed an enhancement of P2X1 receptor protein. [Copyright &y& Elsevier]