Regulation of acetylcholinesterase activity by nitric oxide in rat neuromuscular junction via N-methyl-d-aspartate receptor activation

Acetylcholinesterase (AChE) is an enzyme that hydrolyses the neurotransmitter acetylcholine, thereby limiting spillover and duration of action. This study demonstrates the existence of an endogenous mechanism for the regulation of synaptic AChE activity. At the rat extensor digitorum longus neuromuscular junction, activation of N-methyl-d-aspartate (NMDA) receptors by combined application of glutamate and glycine led to enhancement of nitric oxide (NO) production, resulting in partial AChE inhibition. Partial AChE inhibition was measured using increases in miniature endplate current amplitude. AChE inhibition by paraoxon, inactivation of NO synthase by Nω-nitro-l-arginine methyl ester, and NMDA receptor blockade by dl-2-amino-5-phosphopentanoic acid prevented the increase in miniature endplate current amplitude caused by amino acids. High-frequency (10 Hz) motor nerve stimulation in a glycine-containing bathing solution also resulted in an increase in the amplitude of miniature endplate currents recorded during the interstimulus intervals. Pretreatment with an NO synthase inhibitor and NMDA receptor blockade fully eliminated this effect. This suggests that endogenous glutamate, released into the synaptic cleft as a co-mediator of acetylcholine, is capable of triggering the NMDA receptor/NO synthase-mediated pathway that modulates synaptic AChE activity. Therefore, in addition to well-established modes of synaptic plasticity (e.g. changes in the effectiveness of neurotransmitter release and/or the sensitivity of the postsynaptic membrane), another mechanism exists based on the prompt regulation of AChE activity. NO molecules depress AChE activity in the neuromuscular junction thereby enhancing endplate current amplitude. Endogenous glutamate, released into the synaptic cleft as a co-mediator of acetylcholine, is capable of triggering the NMDA receptor-/NO synthase-mediated pathway that modulates synaptic AChE activity. In addition to well-established modes of synaptic plasticity another mechanism exists based on the prompt regulation of AChE activity. © 2012 Federation of European Neuroscience Societies and Blackwell Publishing Ltd

Enhancement of mouse diaphragm contractility in the presence of antagonists of GABA<inf>A</inf> and GABA<inf>B</inf> receptors

© 2019 The Authors. Experimental Physiology © 2019 The Physiological Society New Findings: What is the central question of this study? Do GABA receptors play any role at the neuromuscular junction? What is the main finding and its importance? In the presence of either ionotropic or metabotropic GABA receptor antagonists, diaphragm muscle force production elicited by stimulating the motor nerve at ≥50 Hz was increased. Our data indicate the presence of GABAergic signalling at the neuromuscular junction. Abstract: Despite the signalling role of GABA in the brain and spinal cord, the role of this molecule in the peripheral nervous system and, in particular, at the neuromuscular junction remains practically unexplored. In the present work, the force of mouse diaphragm contractions was measured in the presence of blockers of metabotropic GABAB receptors (CGP 55845) and ionotropic GABAA receptors (picrotoxin) with various patterns of indirect and direct stimulation of muscle by trains of 40 pulses delivered at 10, 20, 50 and 70 Hz. It was found that neither blocker affected the diaphragm contractility caused by indirect stimulation through the motor nerve at 10 and 20 Hz. However, when the stimulation frequency was increased to 50 or 70 Hz, the force of subsequent contractions in the train (when compared with the amplitude of contraction in response to the first pulse) was increased by both CGP 55845 and picrotoxin. With direct stimulation of the diaphragm, no significant changes in the contraction force were detected at any frequency used. The results obtained support the following conclusions: (i) pharmacological inhibition of GABA receptors increases the contractile activity of skeletal muscle; and (ii) frequency-dependent enhancement of GABA receptor activation takes place in the region of the neuromuscular junction

Enhancement of mouse diaphragm contractility in the presence of antagonists of GABA<inf>A</inf> and GABA<inf>B</inf> receptors

© 2019 The Authors. Experimental Physiology © 2019 The Physiological Society New Findings: What is the central question of this study? Do GABA receptors play any role at the neuromuscular junction? What is the main finding and its importance? In the presence of either ionotropic or metabotropic GABA receptor antagonists, diaphragm muscle force production elicited by stimulating the motor nerve at ≥50 Hz was increased. Our data indicate the presence of GABAergic signalling at the neuromuscular junction. Abstract: Despite the signalling role of GABA in the brain and spinal cord, the role of this molecule in the peripheral nervous system and, in particular, at the neuromuscular junction remains practically unexplored. In the present work, the force of mouse diaphragm contractions was measured in the presence of blockers of metabotropic GABAB receptors (CGP 55845) and ionotropic GABAA receptors (picrotoxin) with various patterns of indirect and direct stimulation of muscle by trains of 40 pulses delivered at 10, 20, 50 and 70 Hz. It was found that neither blocker affected the diaphragm contractility caused by indirect stimulation through the motor nerve at 10 and 20 Hz. However, when the stimulation frequency was increased to 50 or 70 Hz, the force of subsequent contractions in the train (when compared with the amplitude of contraction in response to the first pulse) was increased by both CGP 55845 and picrotoxin. With direct stimulation of the diaphragm, no significant changes in the contraction force were detected at any frequency used. The results obtained support the following conclusions: (i) pharmacological inhibition of GABA receptors increases the contractile activity of skeletal muscle; and (ii) frequency-dependent enhancement of GABA receptor activation takes place in the region of the neuromuscular junction

Regulation of acetylcholinesterase activity by nitric oxide in rat neuromuscular junction via N-methyl-d-aspartate receptor activation

Acetylcholinesterase (AChE) is an enzyme that hydrolyses the neurotransmitter acetylcholine, thereby limiting spillover and duration of action. This study demonstrates the existence of an endogenous mechanism for the regulation of synaptic AChE activity. At the rat extensor digitorum longus neuromuscular junction, activation of N-methyl-d-aspartate (NMDA) receptors by combined application of glutamate and glycine led to enhancement of nitric oxide (NO) production, resulting in partial AChE inhibition. Partial AChE inhibition was measured using increases in miniature endplate current amplitude. AChE inhibition by paraoxon, inactivation of NO synthase by Nω-nitro-l-arginine methyl ester, and NMDA receptor blockade by dl-2-amino-5-phosphopentanoic acid prevented the increase in miniature endplate current amplitude caused by amino acids. High-frequency (10 Hz) motor nerve stimulation in a glycine-containing bathing solution also resulted in an increase in the amplitude of miniature endplate currents recorded during the interstimulus intervals. Pretreatment with an NO synthase inhibitor and NMDA receptor blockade fully eliminated this effect. This suggests that endogenous glutamate, released into the synaptic cleft as a co-mediator of acetylcholine, is capable of triggering the NMDA receptor/NO synthase-mediated pathway that modulates synaptic AChE activity. Therefore, in addition to well-established modes of synaptic plasticity (e.g. changes in the effectiveness of neurotransmitter release and/or the sensitivity of the postsynaptic membrane), another mechanism exists based on the prompt regulation of AChE activity. NO molecules depress AChE activity in the neuromuscular junction thereby enhancing endplate current amplitude. Endogenous glutamate, released into the synaptic cleft as a co-mediator of acetylcholine, is capable of triggering the NMDA receptor-/NO synthase-mediated pathway that modulates synaptic AChE activity. In addition to well-established modes of synaptic plasticity another mechanism exists based on the prompt regulation of AChE activity. © 2012 Federation of European Neuroscience Societies and Blackwell Publishing Ltd

Regulation of acetylcholinesterase activity by nitric oxide in rat neuromuscular junction via N-methyl-d-aspartate receptor activation

Acetylcholinesterase (AChE) is an enzyme that hydrolyses the neurotransmitter acetylcholine, thereby limiting spillover and duration of action. This study demonstrates the existence of an endogenous mechanism for the regulation of synaptic AChE activity. At the rat extensor digitorum longus neuromuscular junction, activation of N-methyl-d-aspartate (NMDA) receptors by combined application of glutamate and glycine led to enhancement of nitric oxide (NO) production, resulting in partial AChE inhibition. Partial AChE inhibition was measured using increases in miniature endplate current amplitude. AChE inhibition by paraoxon, inactivation of NO synthase by Nω-nitro-l-arginine methyl ester, and NMDA receptor blockade by dl-2-amino-5-phosphopentanoic acid prevented the increase in miniature endplate current amplitude caused by amino acids. High-frequency (10 Hz) motor nerve stimulation in a glycine-containing bathing solution also resulted in an increase in the amplitude of miniature endplate currents recorded during the interstimulus intervals. Pretreatment with an NO synthase inhibitor and NMDA receptor blockade fully eliminated this effect. This suggests that endogenous glutamate, released into the synaptic cleft as a co-mediator of acetylcholine, is capable of triggering the NMDA receptor/NO synthase-mediated pathway that modulates synaptic AChE activity. Therefore, in addition to well-established modes of synaptic plasticity (e.g. changes in the effectiveness of neurotransmitter release and/or the sensitivity of the postsynaptic membrane), another mechanism exists based on the prompt regulation of AChE activity. NO molecules depress AChE activity in the neuromuscular junction thereby enhancing endplate current amplitude. Endogenous glutamate, released into the synaptic cleft as a co-mediator of acetylcholine, is capable of triggering the NMDA receptor-/NO synthase-mediated pathway that modulates synaptic AChE activity. In addition to well-established modes of synaptic plasticity another mechanism exists based on the prompt regulation of AChE activity. © 2012 Federation of European Neuroscience Societies and Blackwell Publishing Ltd

Regulation of acetylcholinesterase activity by nitric oxide in rat neuromuscular junction via N-methyl-d-aspartate receptor activation

Acetylcholinesterase (AChE) is an enzyme that hydrolyses the neurotransmitter acetylcholine, thereby limiting spillover and duration of action. This study demonstrates the existence of an endogenous mechanism for the regulation of synaptic AChE activity. At the rat extensor digitorum longus neuromuscular junction, activation of N-methyl-d-aspartate (NMDA) receptors by combined application of glutamate and glycine led to enhancement of nitric oxide (NO) production, resulting in partial AChE inhibition. Partial AChE inhibition was measured using increases in miniature endplate current amplitude. AChE inhibition by paraoxon, inactivation of NO synthase by Nω-nitro-l-arginine methyl ester, and NMDA receptor blockade by dl-2-amino-5-phosphopentanoic acid prevented the increase in miniature endplate current amplitude caused by amino acids. High-frequency (10 Hz) motor nerve stimulation in a glycine-containing bathing solution also resulted in an increase in the amplitude of miniature endplate currents recorded during the interstimulus intervals. Pretreatment with an NO synthase inhibitor and NMDA receptor blockade fully eliminated this effect. This suggests that endogenous glutamate, released into the synaptic cleft as a co-mediator of acetylcholine, is capable of triggering the NMDA receptor/NO synthase-mediated pathway that modulates synaptic AChE activity. Therefore, in addition to well-established modes of synaptic plasticity (e.g. changes in the effectiveness of neurotransmitter release and/or the sensitivity of the postsynaptic membrane), another mechanism exists based on the prompt regulation of AChE activity. NO molecules depress AChE activity in the neuromuscular junction thereby enhancing endplate current amplitude. Endogenous glutamate, released into the synaptic cleft as a co-mediator of acetylcholine, is capable of triggering the NMDA receptor-/NO synthase-mediated pathway that modulates synaptic AChE activity. In addition to well-established modes of synaptic plasticity another mechanism exists based on the prompt regulation of AChE activity. © 2012 Federation of European Neuroscience Societies and Blackwell Publishing Ltd