BPTU, an allosteric antagonist of P2Y1 receptor, blocks nerve mediated inhibitory neuromuscular responses in the gastrointestinal tract of rodents

Mañé, N.; Jiménez-Sábado, Verónica; Jiménez, Marcel

doi:10.1016/j.neuropharm.2016.07.033

Cited by 11 publications

(14 citation statements)

References 58 publications

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“…However, a lower concentration of BPTU was needed to reduce the IJPf, both in mice (EC 50 = 0.06 μmol L −1 ) and as we previously reported, rats (EC 50 = 0.3 μmol L −1 ). 13 We also found that under the same experimental conditions BPTU blocked the purinergic relaxation, but the concentration needed to reduce the response by 50% was higher in pig tissue compared to the values previously reported in rodents (see Table 1). 13 Due to these differences, we repeated the experiments with mouse tissue and we obtained similar results to those previously reported.…”

Section: Discussionsupporting

confidence: 73%

“…In previous experiments, we showed that the IJPf was reduced by BPTU in a concentration-dependent manner in colonic tissues. 13 Accordingly, we determined the effect of BPTU on the IJPf in pig small intestine. Slow waves (sw) had an amplitude of 8.0 ± 1.5 mV and a frequency of 9.8 ± 0.3 contractions per minute and the rmp was −62.1 ± 1.3 mV.…”

Section: Effect Of Bptu On Ijpfmentioning

confidence: 99%

“…In 2016, we reported that BPTU blocked the IJPf and the mechanical purinergic relaxation in colonic muscle strips of rodents. 13 Recently, we demonstrated that both MRS2500 and BPTU blocked the hyperpolarization and relaxation induced by diadenosine tetraphosphate (Ap4A) in the rat colon. 14 Accordingly, the aim of this work was to determine the effects of BPTU first in a large animal model and then in human tissue.…”

Section: Introductionmentioning

confidence: 99%

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Different responses of the blockade of the P2Y1 receptor with BPTU in human and porcine intestinal tissues and in cell cultures

Traserra

Barber²,

Maclnnes

et al. 2021

Neurogastroenterology Motil

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Background: Gastrointestinal smooth muscle relaxation is accomplished by activation of P2Y 1 receptors, therefore this receptor plays an important role in regulation of gut motility. Recently, BPTU was developed as a negative allosteric modulator of the P2Y 1 receptor. Accordingly, the aim of this study was to assess the effect of BPTU on purinergic neurotransmission in pig and human gastrointestinal tissues.Methods: Ca 2+ imaging in tSA201 cells that express the human P2Y 1 receptor, organ bath and microelectrodes in tissues were used to evaluate the effects of BPTU on purinergic responses. Key results: BPTU concentration dependently (0.1 and 1 µmol L −1 ) inhibited the rise in intracellular Ca 2+ evoked by ADP in tSA201 cells. In the pig small intestine, 30 µmol L −1 BPTU reduced the fast inhibitory junction potential by 80%. Smooth muscle relaxations induced by electrical field stimulation were reduced both in pig ileum (EC 50 = 6 µmol L −1 ) and colon (EC 50 = 35 µmol L −1 ), but high concentrations of BPTU (up to 100 µmol L −1 ) had no effect on human colonic muscle. MRS2500(1 µmol L −1 ) abolished all responses. Finally, 10 µmol L −1 ADPβS inhibited spontaneous motility and this was partially reversed by 30 µmol L −1 BPTU in pig, but not human colonic tissue and abolished by MRS2500 (1 µmol L −1 ). Conclusions & inferences:BPTU blocks purinergic responses elicited via P2Y 1 receptors in cell cultures and in pig gastrointestinal tissue. However, the concentrations needed are higher in pig tissue compared to cell cultures and BPTU was ineffective in human colonic tissue.

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Section: Discussionsupporting

confidence: 73%

Section: Effect Of Bptu On Ijpfmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Different responses of the blockade of the P2Y1 receptor with BPTU in human and porcine intestinal tissues and in cell cultures

Traserra

Barber²,

Maclnnes

et al. 2021

Neurogastroenterology Motil

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“…In view of these results, we next checked whether P2Y1 pharmacological inhibition was enough to reduce ankyrinG levels. For this purpose, we used the non-nucleotide allosteric and none ADP-competitive P2Y1 antagonist, BPTU (1-(2-(2-(tert-butyl)phenoxy)pyridin-3-yl)-3-(4-(trifluoromethoxy)phenyl)urea; Mañé et al, 2016). First, we determined the most efficient dose in cultured hippocampal neurons.…”

Section: Resultsmentioning

confidence: 99%

P2Y1 Purinergic Receptor Modulate Axon Initial Segment Initial Development

Zhang

Bonadiman

Ciorraga

et al. 2019

Front. Cell. Neurosci.

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Morphological and functional polarization of neurons depends on the generation and maintenance of the axon initial segment (AIS). This axonal domain maintains axonal properties but is also the place where the action potential (AP) is generated. All these functions require the AIS, a complex structure that is not fully understood. An integrated structure of voltage-gated ion channels, specific cytoskeleton architecture, as well as, scaffold proteins contributes to these functions. Among them, ankyrinG plays a crucial role to maintain ion channels and membrane proteins. However, it is still elusive how the AIS performs its complex structural and functional regulation. Recent studies reveal that AIS is dynamically regulated in molecular composition, length and location in response to neuronal activity. Some mechanisms acting on AIS plasticity have been uncovered recently, including Ca 2+ , calpain or calmodulin-mediated modulation, as well as post-translational modifications of cytoskeleton proteins and actin-associated proteins. Neurons are able to respond to different kind of physiological and pathological stimuli from development to maturity by adapting their AIS composition, position and length. This raises the question of which are the neuronal receptors that contribute to the modulation of AIS plasticity. Previous studies have shown that purinergic receptor P2X7 activation is detrimental to AIS maintenance. During initial axonal elongation, P2X7 is coordinated with P2Y1, another purinergic receptor that is essential for proper axon elongation. In this study, we focus on the role of P2Y1 receptor on AIS development and maintenance. Our results show that P2Y1 receptor activity and expression are necessary during AIS initial development, while has no role once AIS maturity is achieved. P2Y1 inhibition or suppression results in a decrease in ankyrinG, βIV-spectrin and voltage-gated sodium channels accumulation that can be rescued by actin stabilization or the modulation of actin-binding proteins at the AIS. Moreover, P2X7 or calpain inhibition also rescues ankyrinG decrease. Hence, a dynamic balance of P2Y1 and P2X7 receptors expression and function during AIS assembly and maturation may represent a fine regulatory mechanism in response to physiological or pathological extracellular purines concentration.

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“…While purinergic neuromuscular transmission has been shown to cause a transient mechanical relaxation, the long‐lasting relaxation can only be accomplished with activation of the nitrergic pathway. Furthermore, there is evidence to suggest that P2Y 1 receptors participate in neuromuscular transmission since: 1‐ P2Y 1 receptor KO animals lack IJPf, IJPf is concentration dependently reduced by competitive (orthosteric) P2Y 1 receptor blockers such as MRS2179, MRS2279, and MRS2500 and non‐competitive (allosteric) blockers such as BPTU . Spontaneous IJPf (MRS2500 sensitive), not associated with electrical or chemical stimulation, are often present in electrophysiological recordings and additionally, ongoing release of nitric oxide (NO) by inhibitory neurons causes a hyperpolarized membrane potential that often reduces contractility.…”

Section: Introductionmentioning

confidence: 99%

The asymmetric innervation of the circular and longitudinal muscle of the mouse colon differently modulates myogenic slow phasic contractions

Traserra

Villarte

Traini

et al. 2019

Neurogastroenterology Motil

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Background Neuromuscular transmission has been extensively studied in the circular layer of the mouse colon where a co‐transmission of purines acting on P2Y1 receptors and NO has been previously described. However, the corresponding mechanisms in the longitudinal layer are less known. Methods Electrophysiological and myography techniques were used to evaluate spontaneous phasic contractions (SPC) and neural‐mediated responses in the proximal, mid, and distal colon devoid of CD1 mice. Immunohistochemistry against c‐kit and PDGFRα was performed in each colonic segment. Key Results SPC were recorded in both muscle layers at a similar frequency being about four contractions per minute (c.p.m.) in the proximal and distal colon compared to the mid colon (2 c.p.m.). In non‐adrenergic, non‐cholinergic conditions, L‐NNA (1 mmol/L) increased contractility in the circular but not in the longitudinal layer. In the longitudinal muscle, both electrophysiological and mechanical neural‐mediated inhibitory responses were L‐NNA and ODQ (10 µmol/L) sensitive. NaNP (1 µmol/L) caused cessation of SPC and the response was blocked by ODQ. Neither ADPßS (10 µmol/L) nor CYPPA (10 µmol/L), which both targeted the purinergic pathway, altered longitudinal contractions. PDGFRα + cells were located in both muscle layers and were more numerous compared with cKit + cells, which both formed a heterologous cellular network. A decreasing gradient of the PDGFRα labeling was observed along the colon. Conclusion An inhibitory neural tone was absent in the longitudinal layer and neuronal inhibitory responses were mainly nitrergic. Despite the presence of PDGFRα + cells, purinergic responses were absent. Post‐junctional pathways located in different cell types might be responsible for neurotransmitter transduction.

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BPTU, an allosteric antagonist of P2Y1 receptor, blocks nerve mediated inhibitory neuromuscular responses in the gastrointestinal tract of rodents

Cited by 11 publications

References 58 publications

Different responses of the blockade of the P2Y1 receptor with BPTU in human and porcine intestinal tissues and in cell cultures

Different responses of the blockade of the P2Y1 receptor with BPTU in human and porcine intestinal tissues and in cell cultures

P2Y1 Purinergic Receptor Modulate Axon Initial Segment Initial Development

The asymmetric innervation of the circular and longitudinal muscle of the mouse colon differently modulates myogenic slow phasic contractions

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