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Intracellular recordings were made in a midbrain slice preparation of the rat brain containing the ventral tegmental area (VTA). Dopaminergic principal cells were identified by their electrophysiological properties and their hyperpolarizing responses to dopamine. Superfusion with dopamine (100 μM) caused hyperpolarization and a decrease of the apparent input resistance. By contrast, two structural analogues of ATP, 2-methylthio ATP (2-MeSATP; 10 μM) and α,β-methylene ATP (α,β-meATP; 30 μM) had no effect, when added to the superfusion medium. Pressure applied dopamine also hyperpolarized the membrane, while both 2-MeSATP and α,β-meATP were ineffective. Hence, dopaminergic principal neurons of the VTA do not possess somatic P2 purinoceptors present on peripheral and central noradrenergic neurons. 相似文献
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Abstract Although the concept of purinergic signalling arose from experiments designed to find the identity of the non-adrenergic, non-cholinergic (NANC) inhibitory neurotransmitter in the gut, it has taken many years for the more general importance of the various roles of ATP as a physiological messenger in the gut to be recognized. Firstly, vasoactive intestitial polypeptide (VIP) and later nitric oxide (NO) were considered the NANC transmitter and it was only later, after the concept of cotransmission was established, that ATP, NO and VIP were recognized as cotransmitters in NANC nerves, although the proportions vary in different gut regions. Recently, many purinoceptor subtypes have been identified on myenteric, submucosal motor, sensory and interneurons involved in synaptic neurotransmission and neuromodulation and reflex activity of several kinds, including ascending excitatory and descending inhibitory reflex pathways. Nucleotide receptors have been shown to be expressed on enteric glial cells and interstitial cells of Cajal. Purinergic mechanosensory transduction, involving release of ATP from mucosal epithelial cells during distension to stimulate subepithelial nerve endings of intrinsic and extrinsic sensory nerves to modulate peristalsis and initiate nociception respectively, is attracting current attention. Exciting new areas of interest about purinergic signalling in the gut include: involvement of purines in development, ageing and regeneration, including the role of stem cells; studies of the involvement of nucleotides in the activity of the gut of invertebrates and lower vertebrates; and the pathophysiology of enteric purinergic signalling in diseases including irritable bowel syndrome, postoperative ileus, oesophageal reflux, constipation, diarrhoea, diabetes, Chaga's and Hirschprung's disease. 相似文献
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目的研究不同方式给予α,β-MeATP对肠系膜动脉P2X1受体介导血管收缩反应的影响。方法制备大鼠离体肠系膜动脉环标本,采用非累积给药法和单浓度给药法两种方式给予α,β-MeATP,记录药物诱发的等长收缩反应。结果两种给药方式给予α,β-MeATP(10-7-10-4mol·L-1)均可使大鼠离体肠系膜动脉产生浓度依赖性收缩反应。以KCl最大收缩反应或以标本湿重标化α,β-MeATP诱发的收缩反应时,α,β-MeATP单浓度给药的收缩反应均大于非累积给药(P<0.01)。以标本湿重标化时,10-4mol·L-1浓度α,β-MeATP诱发的收缩反应分别是(0.73±0.10)g·mg-1(单浓度组)和(0.38±0.05)g·mg-1(非累积组);以KCl最大收缩反应标化时,分别是(53.17±6.0)%(单浓度组)和(36.78±5.71)%(非累积组)。在α,β-MeATP非累积给药组120mmol·L-1KCl诱发的动脉收缩反应小于单浓度给药组和NA对照组(P<0.01)。结论在大鼠肠系膜动脉,非累积给予α,β-MeATP降低P2X1受体介导的收缩反应以及高钾诱发的收缩反应,该给药方式可能导致错误的实验结论 。 相似文献
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- The ionotropic purinoceptors in isolated Deiters'' cells of guinea-pig cochlea were characterized by use of the whole-cell variant of the patch-clamp technique.
- Extracellular application of adenosine 5′-triphosphate (ATP) induced a dose-dependent inward current when the cells were voltage-clamped at −80 mV. The ATP-induced current showed desensitization and had a reversal potential around −4 mV.
- Increasing intracellular free Ca2+ by decreasing the concentration of EGTA in the pipette solution reduced the amplitude of the ATP-gated current.
- The order of agonist potency was: 2-methylthioATP (2-meSATP)>ATP>benzoylbenzoyl-ATP (BzATP)>α,β-methyleneATP (α,β,meATP>adenosine 5′-diphosphate (ADP)>uridine 5′-triphosphate (UTP)>adenosine 5′-monophosphate (AMP)=adenosine (Ad).
- Pretreatment with forskolin (10 μM), 8-bromoadenosine-3′,5′-cyclophosphate (8-Br-cyclic AMP, 1 mM), 3-isobutyl-1-methylxanthine (IBMX, 1 mM) or phorbol-12-myristate-13-acetate (PMA, 1 μM) reversibly reduced the ATP-induced peak current.
- The results are consistent with molecular biological data which indicate that P2X2 purinoceptors are present in Deiters'' cells. In addition, the reduction of the ATP-gated current by activators of protein kinase A and protein kinase C indicates that these P2X2 purinoceptors can be functionally modulated by receptor phosphorylation.
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We previously reported that slow depolarization of substantia gelatinosa neurons is evoked by repetitive stimulation of C-fibers of dorsal root in adult rat spinal cord transverse slices with the dorsal root attached, which was considered to be an event relevant to spinal nociception. In the present study, we investigated the effects of adenosine 5′-triphosphate (ATP) and its analogs on the slow depolarization. ATP (10–100 μM) significantly inhibited the amplitude of slow depolarization in a concentration-dependent manner. The inhibitory effect of ATP was not reversed by suramin, an antagonist for some P2-purinoceptors, and was mimicked by a P2Y selective agonist uridine 5′-triphosphate, but not a P2X selective agonist ,β-methylene ATP. These results suggest that ATP inhibits the slow depolarization of substantia gelatinosa neurons relevant to nociceptive transmission in the spinal dorsal horn, via suramin-insensitive P2Y purinoceptors. 相似文献
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Previous pharmacological experiments have indicated the existence of ATP P2X receptors in chick embryonic skeletal muscles. In this study we cloned a P2X4‐like cDNA encoding a protein of 385 amino acids, which shares 75% and 76% identity with rat and human P2X4 receptors, respectively. Functional studies of this cP2X4 receptor expressed in Xenopus oocytes showed that ATP induced a fast inward current, which was partially desensitized upon prolonged application of ATP. The ATP‐induced currents were concentration‐dependent, with an EC50 of 9.5 μM. Adenosine 5′‐O‐(thio)triphosphate and 2‐methylthioATP very weak agonists. α,β‐methyleneATP was almost inactive. In contrast to their potentiating effects on recombinant rat P2X4 receptors, both suramin and pyridoxalphosphate‐6‐azophenyl‐2′,4′‐disulfonic acid partially blocked ATP‐induced currents. TrinitrophenylATP was able to block ATP‐induced response completely, with an IC50 of 4.7 μM. Northern blot and RT‐PCR analysis showed that cP2X4 mRNAs were mainly expressed in skeletal muscle, brain, and gizzard of day 10 chick embryos. Lower levels of expression were also detected in liver, heart, and retina. Whole‐mount in situ hybridization showed that cP2X4 mRNAs were expressed in the brain, spinal cord, notochord, gizzard, and skeletal muscle. The physiological functions of cP2X4 receptors in embryonic skeletal muscle remain unclear at present. Drug Dev. Res. 53:22–28, 2001. © 2001 Wiley‐Liss, Inc. 相似文献
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Sean G. Brown Yong‐Chul Kim Soon‐Ai Kim Kenneth A. Jacobson Geoffrey Burnstock Brian F. King 《Drug development research》2001,53(4):281-291
Seven PPADS ( P yridoxal‐5′‐ P hosphate 6‐ A zophenyl 2′,4′‐ D i S ulfonate) analogs were investigated at Group 1 P2X receptors expressed in Xenopus oocytes. All seven analogs potently inhibited P2X1 (IC50 range, 5–32 nM) and P2X3 (IC50 range, 22–345 nM), the two Group I P2X receptor subtypes. Analogs showed greater inhibitory activity where the pyridoxal moiety of PPADS contained a 5′‐phosphonate group, rather than a 5′‐phosphate group. Analogs also showed greater potency where disulfonate groups were removed from, or substituted at, the azophenyl moiety. The most active analog was MRS 2257 (pyridoxal‐5′‐phosphonate 6‐azophenyl 3′,5′‐bismethylenephosphonate) at P2X1 (IC50, 5 nM) and P2X3 (IC50, 22 nM) receptors, being 14‐fold and 10‐fold more potent than PPADS itself. MRS 2257 produced a nonsurmountable inhibition when tested against a range of ATP concentrations, although blockade was reversed by about 85% after 20 minutes of washout. TNP‐ATP and Ip5I were equipotent with MRS 2257 at P2X1 receptors, whereas TNP‐ATP was 64‐fold more potent than MRS 2257 at P2X3 receptors. In conclusion, the PPADS template can be altered at the pyridoxal and phenyl moieties to produce P2X1 and P2X3 receptor antagonists showing higher potency and greater degree of reversibility than the parent compound at these Group I P2X receptors. Drug Dev. Res. 53:281–291, 2001. © 2001 Wiley‐Liss, Inc. 相似文献
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