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Query: UMLS:C0043167 (pertussis)
 19,595 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Muscarinic receptor-linked Ca2+ mobilization and changes in cyclic AMP were studied in SH-SY5Y and IMR 32 human neuroblastoma cell lines. Muscarinic agonists acetylcholine, carbachol, methacholine and muscarine induced an increase in cytosolic free Ca2+ in a pertussis toxin (100 ng/ml)-insensitive manner in both cell lines. The ED50 values in IMR 32 cells (8-98 microM) were one order of magnitude higher than in SH-SY5Y cells (0.3-1.6 microM). Oxotremorine and pilocarpine failed to mobilize Ca2+ in IMR 32 cells. Pirenzepine antagonized carbachol-induced Ca2+ mobilization in SH-SY5Y cells with a Ki value in the range of 150-189 nM whereas the corresponding values in IMR 32 cells were 24-28 nM. Atropine inhibited a carbachol-stimulated increase in cytosolic Ca2+ with an equal potency in both cell lines (Ki 2-3 nM). Carbachol stimulated cyclic AMP (cAMP) accumulation in SH-SY5Y cells in a pertussis toxin-insensitive manner. In IMR 32 cells carbachol inhibited prostaglandin E1-stimulated cAMP accumulation. Treatment of IMR 32 cells with pertussis toxin abolished the inhibition of stimulated cAMP accumulation. These results suggest that in SH-SY5Y cells the M3 muscarinic receptor couples to both Ca2+ mobilization and stimulation of cAMP accumulation. In IMR 32 cells the M1 receptor seems to couple to Ca2+ mobilization whereas the inhibition of stimulated cAMP accumulation is coupled to a non-M1 subtype by an inhibitory G-protein.
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PMID:Differential coupling of muscarinic receptors to Ca2+ mobilization and cyclic AMP in SH-SY5Y and IMR 32 neuroblastoma cells. 165 23

Carbachol produces both negative and positive inotropy in rat left atria. It is not clear whether these two effects are mediated by two separate cell surface muscarinic receptors or a single receptor interacting with two coupling proteins in the cell membrane. Pirenzepine, known to selectively block some biochemical muscarinic responses, was used in this study to block the biphasic response to carbachol in rat left atria. The negative inotropy to carbachol was blocked by pirenzepine, and Schild analysis indicated a -log dissociation constant (pKb) for the pirenzepine-receptor complex of 6.2. However, the Schild analysis may have been complicated by positive inotropy observed with pirenzepine. This positive inotropic effect was sensitive to blockade by other muscarinic antagonists. In atria from rats pretreated with pertussis toxin, carbachol produced a positive inotropic effect. Schild analysis with pirenzepine for antagonism of this response indicated a -log equilibrium dissociation constant for the pirenzepine-receptor complex of 6.7, significantly different from that for antagonism of negative inotropy. This ostensibly suggested a difference in the receptors mediating these responses. In view of the possible complicating effects of the positive inotropic effects of pirenzepine in this assay, an alternative method for the measurement of pirenzepine affinity was utilized. Resultant analysis was used to measure the pKb for pirenzepine antagonism of negative inotropy to carbachol. This method had the advantage of cancelling the positive inotropy to pirenzepine. Under these circumstances, pirenzepine had a pKb of 6.9, a value not significantly different from for antagonism of the positive inotropy to carbachol. The relevance of these findings is discussed in terms of a single promiscuous muscarinic receptor or heterogeneous receptors in this tissue. These data do not support the hypothesis that two separate receptors mediate these two effects.
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PMID:Promiscuous or heterogeneous muscarinic receptors in rat atria? II. Antagonism of responses to carbachol by pirenzepine. 209

The direct effect of muscarine on neurons of the rat dorsolateral septal nucleus (DLSN) was examined by using conventional microelectrode and voltage-clamp techniques. Muscarine (1-50 microM) caused a hyperpolarization accompanied by an increase of a voltage-independent potassium conductance. Pirenzepine competitively antagonized the muscarine-induced hyperpolarization with an apparent dissociation constant (Kd) value of 54 nM. Furthermore, intracellular loading with GTP gamma S, a non-hydrolyzable GTP analog, blocked irreversibly the muscarine-induced hyperpolarization. In addition, pretreatment of neurons with pertussis toxin (PTX) prevented the hyperpolarization produced by muscarine. These results suggest that muscarine hyperpolarizes DLSN neurons via a voltage-independent potassium conductance by acting at M4 subtype receptors which are coupled to a PTX-sensitive G-protein in DLSN neurons.
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PMID:Muscarine increases a voltage-independent potassium conductance through an M4 receptor in rat dorsolateral septal nucleus neurons. 762 35

1. The effects of acetylcholine (ACh) on pyramidal neurons acutely dissociated from the rat cerebral cortex were studied in the whole-cell mode, by use of the nystatin-perforated patch recording configuration. 2. ACh induced a net inward current (IACh) accompanied by a membrane conductance decrease at a holding potential (VH) of -40 mV. IACh increased in a concentration-dependent manner with a half-maximum concentration (EC50) of 8.7 x 10(-7) M. 3. IACh mainly resulted from the suppression of the voltage- and time-dependent K+ current (M-current). 4. Muscarine and muscarinic agonists such as McN-A-343, oxotremorine and oxotremorine-M mimicked the ACh response. The potency was in the order of oxotremorine-M > McN-A-343 > or = muscarine > oxotremorine. 5. Pirenzepine shifted the concentration-response curve for ACh to the right and the corresponding Schild plot yielded a pA2 value of 7.81. Other muscarinic antagonists also reversibly blocked IACh in a concentration-dependent manner. The inhibitory potency was in the order of atropine > 4-DAMP > pirenzepine > AF-DX-116. 6. IACh could be induced normally even after pre-incubation of dissociated neurones in external solution with 200 ng ml-1 pertussis toxin (PTX) for 8 h, whereas the inhibitory effect of ACh on high-voltage-activated Ca2+ channels was completely abolished by the PTX treatment.
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PMID:Muscarinic acetylcholine response in pyramidal neurones of rat cerebral cortex. 795 77

1. The actions of muscarinic agonists and antagonists upon the Ca2+ current (ICa) in acutely dissociated magnocellular cholinergic basal forebrain neurones from 11 to 14-day-old postnatal rats were studied using the whole-cell patch-clamp technique. 2. In all cells studied, muscarinic agonists inhibited a transient component of high-voltage-activated (HVA) current, but had no effect upon the low-voltage-activated (LVA) current. The mean IC50 values for ACh and oxotremorine methiodide (oxo-M), obtained from non-cumulative dose-response curves, were 204 and 363 nM respectively. Superfusion with the K+ channel blocker, tetraethylammonium chloride (TEA; 30 mM) shifted the ACh dose-response curve to the right giving an IC50 value of 22:9 microM. 3. Pirenzepine (0.1-1 microM) and methoctramine (0.03-0.3 microM) produced parallel shifts to the right in the agonist dose-response curves. Schild analysis of the agonist dose ratios yielded pA2 (negative log of the apparent dissociation constant KB) values for pirenzepine and methoctramine of 6.8 and 8.2 respectively, indicating the involvement of an M2 receptor subtype. 4. In the presence of GTP-gamma-S (10-500 microM) in the patch pipette, the agonist-induced inhibition of ICa became irreversible. Dialysis with GDP-beta-S (1 mM) abolished all agonist-induced inhibition of the Ca2+ current. The agonist-induced inhibition of ICa was totally blocked by pretreatment with pertussis toxin (500 ng ml-1) but unaffected by preincubation with cholera toxin (500 ng ml-1). Superfusion with 8-bromo cAMP (0.5-1 mM) did not mimic or prevent the effect of agonist application. 5. Inhibition of the Ca2+ current by muscarinic agonists was only partially blocked by omega-conotoxin GVIA (omega-CgTX GVIA), with approximately 46% of the agonist-sensitive current being resistant to omega-CgTX GVIA. Both the agonist- and omega-CgTX GVIA-sensitive components of the current were abolished following maximal inhibition of ICa by GTP-gamma-S. 6. These results indicate that inhibition of the Ca2+ current by muscarinic agonists is mediated via an M2 muscarinic receptor coupled to a pertussis toxin-sensitive G-protein. The possible modulation of multiple HVA Ca2+ channels by muscarinic agonists and the role that these receptors may play in presynaptic modulation of neurotransmitter release are discussed.
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PMID:M2 muscarinic receptor-mediated inhibition of the Ca2+ current in rat magnocellular cholinergic basal forebrain neurones. 841 Jun 90

Carbachol increased ventricular automaticity in a concentration-dependent fashion from a control rate of 72 +/- 5 (mean +/- SEM) to 86 +/- 4 beats per minute at 10(-4) M carbachol. Pirenzepine, an M1-selective antagonist, and AFDX 116, an M2-selective antagonist, both at 10(-7) M, did not block the carbachol-induced positive chronotropic response. In contrast, 10(-7) M HHSiD, an M3-selective antagonist, completely blocked the positive chronotropic effect of carbachol. Carbachol stimulated the accumulation of IP1 in a concentration-dependent manner at concentrations > or = 3 x 10(-6) M. AFDX 116 had no effect on carbachol-induced IP1 accumulation. HHSiD significantly inhibited IP1 accumulation at concentrations > or = 3 x 10(-8) M, while pirenzepine inhibited IP1 accumulation only at concentrations > or = 10(-5) M. McN A343 and methacholine, two muscarinic receptor agonists with minimal M2 activities, and carbachol did not alter basal cAMP concentration, but all three agonists significantly attenuated the increase in cAMP accumulation in response to isoproterenol. Carbachol inhibited isoproterenol-mediated cAMP accumulation at concentrations > or = 10(-7) M. AFDX 116, HHSiD, and pirenzepine blocked the carbachol-induced inhibition of isoproterenol-stimulated cAMP accumulation. At equimolar concentrations, the inhibitory effects of HHSiD and AFDX-116 were similar, while that of pirenzepine was much less. Pretreatment with pertussis toxin for 24 h did not prevent the carbachol-mediated positive chronotropic response or accumulation of IP1 but completely abolished the inhibition of isoproterenol-stimulated cAMP accumulation. These results indicate that (a) neonatal ventricular myocytes in culture have a heterogeneous population of muscarinic (M2 and M3) receptors, (b) the M3 receptor is coupled to pertussis toxin-sensitive and pertussis toxin-insensitive G proteins, (c) M3 receptor stimulation activates phosphoinositide hydrolysis and increases automaticity via a pertussis toxin-insensitive G protein-dependent pathway, and (d) both M2 and M3 receptors couple to pertussis toxin-sensitive G protein(s) to mediate the inhibition of intracellular cAMP accumulation in response to isoproterenol stimulation.
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PMID:Muscarinic receptor heterogeneity in neonatal rat ventricular myocytes in culture. 884 59

The coupling of muscarinic-cholinergic receptors (mAchR) with the phospholipase C (PLC) second messenger system has been demonstrated in central nervous system (CNS) tissue of many animal species. However, little information exists regarding this association in the developing human CNS. Due to the suggested role of acetylcholine in the regulation of development and differentiation of neural cells, the knowledge of these relationships during human fetal development acquires singular importance. Because of this, we examined the cholinergic stimulation of PLC in human fetal CNS organotypic tissue cultures. Agonist treatment of cultures, in the presence of lithium, resulted in a 4-6-fold increase in inositol phosphates formation. This increase was caused principally by the formation of inositol phosphate (IP). However, kinetic studies demonstrated that the levels of IP2, IP3 and IP4 also increased rapidly after stimulation reaching maximum levels before IP. These results support the hypothesis that muscarinic receptor activation results in an increase in the hydrolysis of PIP2. The inositol phosphate formation was dependent on agonist concentration. The obtained EC50 values were approximately 57 +/- 15 microM for carbachol, 8 +/- 2 microM for acetylcholine and 49 +/- 15 microM for oxotremorine. The agonist-dependent formation of inositol phosphates was inhibited by the muscarinic antagonists atropine and pirenzepine. Pirenzepine inhibited carbachol stimulation with high affinity (Ki = 2.90 +/- 1.15 nM), indicating that PLC activation is the result of activation of the m1 subtype of muscarinic receptors. Treatment of cultures with pertussis toxin did not result in inhibition of agonist-dependent activation of PLC. This result suggests that the m1 muscarinic receptor is coupled to PLC through Gq.
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PMID:Muscarinic receptor-dependent activation of phospholipase C in human fetal central nervous system organotypic tissue culture. 892 76

1. The modulation of high-voltage-activated (HVA) Ca2+ channels by acetylcholine (ACh) was studied in the paratracheal ganglion cells acutely dissociated from 2-week-old Wistar rats by use of the nystatin perforated patch recording configuration under voltage-clamp conditions. 2. ACh inhibited the HVA Ca2+ currents in a concentration- and voltage-dependent manner. 3. The inhibition was mimicked by a muscarinic agonist, oxotremorine. Pirenzepine and methoctramine produced parallel shifts to the right in the ACh concentration-response curves. Schild analysis of the ACh concentration-ratios yield pA2 values for pirenzepine and methoctramine of 6.85 and 8.57, respectively, suggesting the involvement of an M2 receptor. 4. Nifedipine, omega-conotoxin-GVIA and omega-conotoxin-MVIIC reduced the HVA I(Ca) by 16.8, 59.2 and 6.3%, respectively. A current insensitive to all of these Ca2+ antagonists, namely 'R-type', was also observed. The results indicated the existence of L-, N-, P/Q-, and R-type Ca2+ channels. 5. The ACh-sensitive current component was markedly reduced in the presence of omega-conotoxin-GVIA, but not with both nifedipine and omega-conotoxin-MVIIC. ACh also inhibited the R-type HVA I(Ca) remaining in saturating concentrations of nifedipine, omega-conotoxin-GVIA and omega-conotoxin-MVIIC. 6. The inhibitory effect of ACh was prevented by pretreatment with pertussis toxin. 7. It was concluded that ACh selectively reduces both the N- and R-type Ca2+ channels, by activating pertussis toxin sensitive G-protein through the M2 muscarinic receptor in paratracheal ganglion cells.
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PMID:Acetylcholine modulation of high-voltage-activated calcium channels in the neurones acutely dissociated from rat paratracheal ganglia. 957 41

In isolated mouse left atria, acetylcholine (ACh) produced a biphasic inotropic response; a transient decrease in developed tension was followed by an increase. Both negative and positive responses were concentration dependent and were inhibited by atropine. The negative and positive inotropic responses were also observed with a nonselective muscarinic stimulant, oxotremorine-M, but not with an M1-receptor selective stimulant, McN-A343. Pirenzepine, an M1-receptor antagonist, inhibited both negative and positive inotropic responses at high concentrations. Gallamine, an M2-receptor antagonist, inhibited the negative response. Hexahydro-siladifenidol hydrochloride, p-fluoro analog (p-F-HHSiD), an M3-receptor antagonist, inhibited the positive response with no effect on the negative phase. In pertussis toxin (PTX) treated preparations, negative inotropic response to ACh was not observed. These results suggest that the negative and positive inotropic responses to acetylcholine in mouse atria are mediated by M2 and M3 receptors, respectively. The negative phase, but not the positive phase, was mediated by a PTX-sensitive G protein.
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PMID:Positive and negative inotropic effects of muscarinic receptor stimulation in mouse left atria. 1079 16

Acetylcholine has long been known to excite sympathetic neurons via M1 muscarinic receptors through an inhibition of M-currents. Nevertheless, it remained controversial whether activation of muscarinic receptors is also sufficient to trigger noradrenaline release from sympathetic neurons. In primary cultures of rat superior cervical ganglia, the muscarinic agonist oxotremorine M inhibited M-currents with half-maximal effects at 1 microM and induced the release of previously incorporated [3H]noradrenaline with half-maximal effects at 10 microM. This latter action was not affected by the nicotinic antagonist mecamylamine which, however, abolished currents through nicotinic receptors elicited by high oxotremorine M concentrations. Ablation of the signalling cascades linked to inhibitory G proteins by pertussis toxin potentiated the release stimulating effect of oxotremorine M, and the half-maximal concentration required to stimulate noradrenaline release was decreased to 3 microM. Pirenzepine antagonized the inhibition of M-currents and the induction of release by oxotremorine M with identical apparent affinity, and both effects were abolished by the muscarinic toxin 7. These results indicate that one muscarinic receptor subtype, namely M1, mediates these two effects. Retigabine, which enhances M-currents, abolished the release induced by oxotremorine M, but left electrically induced release unaltered. Moreover, retigabine shifted the voltage-dependent activation of M-currents by about 20 mV to more negative potentials and caused 20 mV hyperpolarisations of the membrane potential. In the absence of retigabine, oxotremorine M depolarised the neurons and elicited action potential discharges in 8 of 23 neurons; in its presence, oxotremorine M still caused equal depolarisations, but always failed to trigger action potentials. Action potential waveforms caused by current injection were not affected by retigabine. These results indicate that the inhibition of M-currents is the basis for the stimulation of transmitter release from sympathetic neurons via M1 muscarinic receptors.
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PMID:Activation of M1 muscarinic receptors triggers transmitter release from rat sympathetic neurons through an inhibition of M-type K+ channels. 1455 21


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