Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.49 (reverse transcriptase)
 31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

ATP-induced Ca2+ transients were examined in individual PC12 cells of a well defined clone, before and after treatment with nerve growth factor (NGF) to induce a neurone-like phenotype. Using reverse transcriptase PCR these cells were found to express mRNA for several P2 receptors. In undifferentiated cells the ATP-induced Ca2+ response was entirely dependent on Ca2+ influx, could not be mimicked by UTP, alpha,beta-methylene ATP or dibenzoyl ATP or be blocked by pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS). ATP had no significant effect on levels of cyclic AMP or inositol 1,4,5-trisphosphate (InsP3). These results suggest that in undifferentiated PC12 cells ATP mainly acts on a P2X receptor, possibly the P2X4 subtype. After treatment with NGF for 7 days the ATP response was increased and partially sensitive to PPADS. A component of the ATP-induced Ca2+ increase was due to mobilisation of intracellular Ca2+ stores and another to capacitative Ca2+ entry. UTP caused an increase in intracellular Ca2+, and InsP3 formation could be stimulated by ATP and UTP. ATP also caused a small increase in cyclic AMP, but this was abolished in the presence of indomethacin. Thus, after NGF treatment ATP acts partially via a P2Y receptor, possibly the P2Y2 subtype.
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PMID:P2Y receptors contribute to ATP-induced increases in intracellular calcium in differentiated but not undifferentiated PC12 cells. 1069 14

Extracellular ATP is a neurotransmitter and mediates a variety of responses. In the endocrine system, there are data suggesting a physiological role for ATP in Ca(2+) signalling and hormone secretion. However, the ATP receptor subtype involved has not been clearly elucidated in GH3 cells, a rat anterior pituitary cell line. BzATP- and ATP-induced [Ca(2+)](i) responses had EC(50) values of 18 and 651 microM, respectively. The maximal response to ATP was only 59+/-8% of that for BzATP. The BzATP-induced [Ca(2+)](i) increase was dependent upon the extracellular Ca(2+) concentration. Preincubation with oxidized ATP (oATP) nearly abolished the ATP- and BzATP-induced [Ca(2+)](i) increases. Both BzATP and ATP induced depolarization in GH3 cells, with EC(50) values of 31 microM and 1 mM, respectively. The maximal depolarization to BzATP and ATP were 152+/-21 and 146+/-16% of that elicited by 30 mM KCl. The rank order of agonist potency for [Ca(2+)](i) and depolarization responses was BzATP > > ATP >2-MeSATP and purine derivatives such as ADP, AMP, adenosine were ineffective. Neither UTP nor alpha, beta-methylene ATP showed any effect. In low-divalent conditions BzATP evoked non-desensitizing inward currents, which were reversed at approximately 0 mV. This nonselective cationic conductance was increased by repeated applications of BzATP and the cells became very permeable to NMDG. Longer applications (30 min) of BzATP stimulated ethidium bromide influx in low divalent conditions, suggesting increased permeability to larger molecules. We also identified the existence of P2X(7) mRNA on GH3 cells by using reverse transcriptase (RT)-polymerase chain reaction (PCR). These results suggest that the GH3 cells have an endogenous P2X(7) receptor and purinergic stimulation may play a potential role in neuroendocrine modulation on these cells.
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PMID:ATP-induced [Ca(2+)](i) changes and depolarization in GH3 cells. 1095 73

Extracellular nucleotides were used to characterise the contractile P2 receptors in the rat basilar artery. The isometric tension was recorded in vitro and receptor mRNA expression was examined by reverse transcriptase polymerase chain reaction (RT-PCR) after endothelium-denudation. Transient vasoconstriction was evoked by alphabeta-methylene-adenosine triphosphate (alphabeta-MeATP), indicating the presence of P2X(1) receptors. The P2Y receptors were analysed after P2X receptor desensitisation with 10 microM alphabeta-MeATP. Uridine diphosphate (UDP) and uridine triphosphate (UTP) induced sustained contractions of similar magnitude. The stable nucleotide analogue, uridine 5'-O-thiodiphosphate (UDPbetaS), was clearly more potent than uridine 5'-O-3-thiotriphosphate (UTPgammaS), suggesting prominent contractile effects of P2Y(6) receptors. P2Y(2) and P2Y(4) receptors might also be involved in nucleotide responses, since UTPgammaS and adenosine 5'-O-3-thiotriphosphate (ATPgammaS) were of similar potency. The P2Y(1) selective agonists, adenosine 5'-O-thiodiphosphate (ADPbetaS) and 2-methylthioadenosine diphosphate (2-MeSADP) did not induce contractions. RT-PCR analysis demonstrated P2X(1), P2Y(1), P2Y(2) and P2Y(6) receptor mRNA expression, while the P2Y(4) band was weak. In conclusion, extracellular nucleotides induce contractions of cerebral arteries primarily by activation of P2Y(6) receptors on smooth muscle cells, with a lesser contribution of P2Y(2) and P2X(1) receptors. Although mRNA for the P2Y(1) receptor was detected by RT-PCR, it does not mediate contraction.
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PMID:The stable pyrimidines UDPbetaS and UTPgammaS discriminate between contractile cerebrovascular P2 receptors. 1250 87

In the present study, we developed a novel method to analyze the calcium (Ca2+) signal in living slices of mouse caput epididymides by applying calcium imaging on Fura-2-loaded vibratome slices. The data revealed that in epithelial cells of mouse caput epididymides, ATP induces a rapid Ca2+ signal that is sustained after 60 sec. Preincubating the sections in Ca2+-free medium in the presence of EGTA did not affect the amplitude of the ATP-induced Ca2+ signal, indicating the presence of P2Y type purinergic receptors and phospholipase C activity. Furthermore, ATP induced a similar Ca2+ signal in the different subregions of caput epididymides. The P2X type ion-gated purinergic receptors could also be responsible for the ATP-induced Ca2+ signal because immunohistochemical and reverse transcriptase-polymerase chain reaction analyses showed that P2X1, P2X2, P2X4, P2X7, P2Y1, and P2Y2 receptors were expressed in the epididymis. We propose that P2X and P2Y receptor expression is vital for the normal function of epididymal epithelium and sperm maturation. Furthermore, the method we developed allows us to analyze the activity of various G protein-coupled receptors in intact epithelial cells of mouse epididymides, and other reproductive tissues as well.
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PMID:Adenosine triphosphate induces Ca2+ signal in epithelial cells of the mouse caput epididymis through activation of P2X and P2Y purinergic receptors. 1260 41

In this study, we have used reverse transcriptase polymerase chain reaction and immunocytochemistry to show that P2X1, P2X2, P2X4, P2X5, P2X6 and P2X7 receptor messenger RNA and protein are expressed in the rat choroid plexus taken from the fourth and lateral ventricles. In some epithelial cells, the apical surfaces were stained more intensely than the basal surfaces. Some of the epithelial cells showed strong staining, others weak or no staining. Double immunostaining with P2X and cytokeratin antibodies or isolectin B4 binding confirmed that P2X receptors were localized in subpopulations of the choroid plexus epithelial cells, rather than the endothelial cells of capillaries. The results suggest that P2X receptors might be involved in the regulation of cerebrospinal fluid composition.
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PMID:Expression of P2X receptors in rat choroid plexus. 1593 Oct 59

Although recent studies have suggested that purinergic receptors are expressed in the anterior pituitary gland, their involvement in the regulation of pituitary hormone gene expression is not completely understood. In the present study, we examined the expression of purinergic receptors and the effects of purinergic receptor ligands on pro-opiomelanocortin (POMC) gene expression, in AtT20 mouse corticotroph cells. We identified the expression of most of the purinergic receptor subtypes (A1, A2, P2X1, 3-7, P2Y1, 2, 4) mRNAs, analysed by the reverse transcriptase-polymerase chain reaction. We also found that adenosine and ATP, two representative and endogenous agonists of A1-3 and P2X/P2Y receptors, respectively, stimulated the 5'-promoter activity of the POMC gene in a dose- and time-related manner. When these ligands were simultaneously used with corticotrophin-releasing hormone (CRH), effects that were more than additive were observed, suggesting an enhancing role of these compounds in CRH-mediated adrenocorticotrophic hormone (ACTH) synthesis. These ligands also stimulated the expression of transcription factors involved in the regulation of the POMC gene, but did not enhance ACTH secretion. Finally, the positive effect of adenosine as well as CRH was completely inhibited by the protein kinase A inhibitor H89, whereas that of ATP was not influenced, indicating that different intracellular signalling pathways mediate these effects. Altogether, our results suggest a stimulatory role for these purinergic receptor ligands in the regulation of POMC gene expression in corticotroph cells. Because adenosine and ATP are known to be produced within the pituitary gland, it is possible they may be acting in an autocrine/paracrine fashion.
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PMID:Purinergic receptor ligands stimulate pro-opiomelanocortin gene expression in AtT-20 pituitary corticotroph cells. 1650 22

In this study, the P2X(5) receptor was found to be distributed widely in the rat hypothalamus using single and double labeling immunofluorescence and reverse transcriptase-polymerase chain reaction (RT-PCR) methods. The regions of the hypothalamus with the highest expression of P2X(5) receptors in neurons are the paraventricular and supraoptic nuclei. The intensity of P2X(5) immunofluorescence in neurons of the ventromedial nucleus was low. 70-90% of the neurons in the paraventricular nucleus and 46-58% of neurons in the supraoptic and accessory neurosecretory nuclei show colocalization of P2X(5) receptors and arginine vasopressin (AVP). None of the neurons expressing P2X(5) receptors shows colocalization with AVP in the suprachiasmatic and ventromedial nuclei. 87-90% of the neurons in the lateral and ventral paraventricular nucleus and 42-56% of the neurons in the accessory neurosecretory, supraoptic and ventromedial nuclei show colocalization of P2X(5) receptors with neuronal nitric oxide synthase (nNOS). None of the neurons expressing P2X(5) receptors in the suprachiasmatic nucleus shows colocalization with nNOS. These findings provide a morphological basis for possible functional interactions between the purinergic and nitrergic or vasopressinergic neurotransmitter systems.
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PMID:P2X5 receptors are expressed on neurons containing arginine vasopressin and nitric oxide synthase in the rat hypothalamus. 1676 18

The expression of the purinergic receptor subtype P2X(7)R, a nonselective cationic channel activated by high levels of adenosine triphosphate (ATP), has been studied in adult microglia obtained from Alzheimer disease (AD) and nondemented (ND) brains, in fetal human microglia exposed to Abeta(1-42) peptide and in vivo in Abeta(1-42)-injected rat hippocampus. Semiquantitative reverse transcriptase-polymerase chain reaction showed enhanced expression (increase of 70%) of P2X(7)R in AD microglia compared with ND cells (analysis of 6 AD and 8 ND cases). Immunohistochemical analysis showed prominent P2X(7)R expression in association with Abeta plaques and localized to HLA-DR-immunoreactive microglia. In cultured fetal human microglia, cells exposed to Abeta(1-42) (5 microM for 18 hours) had significantly elevated levels of P2X(7)R (by 106%) compared with untreated cells. Amplitudes of Ca(2+) responses in these cells, induced by the selective P2X(7)R agonist BzATP, were increased by 145% with Abeta(1-42) pretreatment relative to control (no peptide pretreatment) and were largely blocked if the P2X(7)R inhibitor-oxidized ATP (oxATP) was added with peptide in pretreatment solution. In vivo, double immunostaining analysis showed considerable P2X(7)R colocalized with microglia after injection of Abeta(1-42) (1 nmol) into rat hippocampus. The overall results suggest roles of P2X(7)R in mediating microglial purinergic inflammatory responses in AD brain.
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PMID:Upregulated expression of purinergic P2X(7) receptor in Alzheimer disease and amyloid-beta peptide-treated microglia and in peptide-injected rat hippocampus. 1708 6

Adenosine triphosphate acts as a fast excitatory neurotransmitter by binding to and activating seven structurally related subtypes of purinergic P2X receptors, which act as ligand-gated ion channels. Besides its role in neurotransmission, ATP also has trophic functions during development of the neuronal system. P2X receptor expression, mainly of P2X(4) and P2X(6) subtypes, has been detected in adult brain and also during neuronal development. We have used the mouse teratocarcinoma P19 cell line as an in vitro model to study P2X(6) receptor expression during early neuronal differentiation. We have detected a full-length and an alternatively spliced form of the mouse P2X(6) receptor gene in P19 cells using reverse transcriptase-polymerase chain reaction. The alternatively spliced form was already present at the stage of pluripotent undifferentiated P19 cells, and was predominant compared to the full-length form during the whole course of neuronal differentiation of P19 cells. Alternative splicing of P2X(6) receptor subunits was also confirmed during postnatal development of mouse brain. During postnatal development, however, the full-length form was predominant compared to the spliced form. Alternative splicing is suggested to regulate P2X(6) receptor function during neuronal differentiation.
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PMID:Alternative splicing of P2X6 receptors in developing mouse brain and during in vitro neuronal differentiation. 1725 1

Pharmacological studies have indirectly shown the possible presence of cannabinoid receptors in the urinary bladder and their potential role in reducing bladder inflammatory pain. However, the localization of cannabinoid receptors in the urinary bladder remains unknown and there are no published data on the effects of cannabinoids on the sensory system of the bladder. The present study was performed to evaluate the expression of the cannabinoid CB(1) receptors in the mouse urinary bladder and to assess their co-localization with the purinergic P2X(3) receptor, a major player in the transduction of sensory events in the bladder. Also, the effect of intravesical administration of a cannabinoid agonist on the electrical activity of bladder afferent fibers was studied. The expression of mRNA coding for CB(1) receptor was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR). Immunofluorescence experiments were performed to study CB(1) and P2X(3) protein expression in the bladder. The electrical activity of bladder afferent fibers was recorded using an ex vivo bladder-nerve preparation. Mechanical stimulation of the bladder was performed by a controlled slow inflation with an external pump. A bolus of a cannabinoid agonist (AZ12646915) was administered intravesically prior to a second inflation. Afferent activity was measured before and after administration of the cannabinoid compound or its vehicle. The effects of CB(1) receptor antagonist (AM251) on the AZ12646915 response were also analyzed. Cannabinoid receptor CB(1) mRNA was detected in the urinary bladder of the mouse. The protein was found in the urothelium, as well as in nerve fibers. CB(1) and P2X(3) receptors were found to be co-expressed in urothelial cells and in some nerve fibers. In addition, intravesical administration of a cannabinoid receptor agonist reduced the mechanically-evoked activity of bladder afferents in the pelvic nerve. This effect was abolished by the previous administration of the CB(1) antagonist AM251. These data demonstrate the presence of cannabinoid CB(1) receptor mRNA and the protein in the mouse urinary bladder. CB(1) and P2X(3) protein co-localization supports the hypothesis of an interaction between the cannabinoid and the purinergic systems in the transduction of sensory information in the urinary bladder. Finally, the reduction of nerve activity induced by cannabinoid-receptor activation implicates CB(1) receptors in the peripheral modulation of bladder afferent information.
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PMID:Cannabinoid CB1 receptors are expressed in the mouse urinary bladder and their activation modulates afferent bladder activity. 1935 96


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