Properties of presynaptic P2X7-like receptors at the neuromuscular junction

T S Moores, B Hasdemir, L Vega-Riveroll, J Deuchars, S H Parson

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Adenosine triphosphate is released into the synaptic cleft of the neuromuscular junction during normal synaptic transmission, and in much greater quantities following injury and ischaemia. There is much data to suggest roles for presynaptic P2 receptors but little to demonstrate which specific receptor subunits are present. Here we show P2X7 receptor subunits on presynaptic motor nerve terminals from birth, but no evidence for P2X1, P2X2, P2X3, P2X4, P2X5 or P2X6 receptor subunits. Further, P2X receptor subunits are present as multimeric, membrane-inserted receptors. A selective agonist, 2'-3'-O-(4-benzoylbenzoyl)-adenosine 5'-triphosphate (BzATP: 100 microM), triggers vesicle release from motor nerve terminals, which is blocked by P2X7RS-specific concentrations of periodate oxidised ATP (OxATP: 100 microM) and brilliant blue G (BBG: 1 microM), but not by suramin (100 microM). Vesicle release is enhanced in the absence of extracellular divalent cations and occurs through activation of the ion channel and not any associated large pore, as we failed to label nerve terminals with large membrane-impermeant molecules after addition of BzATP. We conclude that a P2X7-like receptor is present at mouse motor nerve terminals, and that their activation promotes vesicle release.

Original languageEnglish
Pages (from-to)40-50
Number of pages11
JournalBrain Research
Volume1034
Issue number1-2
DOIs
Publication statusPublished - 9 Feb 2005

Fingerprint

Purinergic P2X7 Receptors
Neuromuscular Junction
Adenosine Triphosphate
Presynaptic Receptors
Suramin
Membranes
Divalent Cations
Ion Channels
Synaptic Transmission
Ischemia
Parturition
Wounds and Injuries
3'-O-(4-benzoyl)benzoyladenosine 5'-triphosphate
coomassie Brilliant Blue
periodate-oxidized adenosine 5'-triphosphate

Keywords

  • Adenosine Triphosphate
  • Animals
  • Benzenesulfonates
  • Cations, Divalent
  • Ion Channels
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Electron, Transmission
  • Motor Neurons
  • Muscle, Skeletal
  • Neuromuscular Junction
  • Platelet Aggregation Inhibitors
  • Presynaptic Terminals
  • Protein Subunits
  • Purinergic P2 Receptor Agonists
  • Purinergic P2 Receptor Antagonists
  • Receptors, Purinergic P2
  • Receptors, Purinergic P2X7
  • Synaptic Membranes
  • Synaptic Transmission
  • Synaptic Vesicles
  • Time Factors

Cite this

Properties of presynaptic P2X7-like receptors at the neuromuscular junction. / Moores, T S; Hasdemir, B; Vega-Riveroll, L; Deuchars, J; Parson, S H.

In: Brain Research, Vol. 1034, No. 1-2, 09.02.2005, p. 40-50.

Research output: Contribution to journalArticle

Moores, T S ; Hasdemir, B ; Vega-Riveroll, L ; Deuchars, J ; Parson, S H. / Properties of presynaptic P2X7-like receptors at the neuromuscular junction. In: Brain Research. 2005 ; Vol. 1034, No. 1-2. pp. 40-50.
@article{a1560581ba704521a85da54bc1d1d4fb,
title = "Properties of presynaptic P2X7-like receptors at the neuromuscular junction",
abstract = "Adenosine triphosphate is released into the synaptic cleft of the neuromuscular junction during normal synaptic transmission, and in much greater quantities following injury and ischaemia. There is much data to suggest roles for presynaptic P2 receptors but little to demonstrate which specific receptor subunits are present. Here we show P2X7 receptor subunits on presynaptic motor nerve terminals from birth, but no evidence for P2X1, P2X2, P2X3, P2X4, P2X5 or P2X6 receptor subunits. Further, P2X receptor subunits are present as multimeric, membrane-inserted receptors. A selective agonist, 2'-3'-O-(4-benzoylbenzoyl)-adenosine 5'-triphosphate (BzATP: 100 microM), triggers vesicle release from motor nerve terminals, which is blocked by P2X7RS-specific concentrations of periodate oxidised ATP (OxATP: 100 microM) and brilliant blue G (BBG: 1 microM), but not by suramin (100 microM). Vesicle release is enhanced in the absence of extracellular divalent cations and occurs through activation of the ion channel and not any associated large pore, as we failed to label nerve terminals with large membrane-impermeant molecules after addition of BzATP. We conclude that a P2X7-like receptor is present at mouse motor nerve terminals, and that their activation promotes vesicle release.",
keywords = "Adenosine Triphosphate, Animals, Benzenesulfonates, Cations, Divalent, Ion Channels, Mice, Mice, Inbred C57BL, Microscopy, Electron, Transmission, Motor Neurons, Muscle, Skeletal, Neuromuscular Junction, Platelet Aggregation Inhibitors, Presynaptic Terminals, Protein Subunits, Purinergic P2 Receptor Agonists, Purinergic P2 Receptor Antagonists, Receptors, Purinergic P2, Receptors, Purinergic P2X7, Synaptic Membranes, Synaptic Transmission, Synaptic Vesicles, Time Factors",
author = "Moores, {T S} and B Hasdemir and L Vega-Riveroll and J Deuchars and Parson, {S H}",
year = "2005",
month = "2",
day = "9",
doi = "10.1016/j.brainres.2004.12.001",
language = "English",
volume = "1034",
pages = "40--50",
journal = "Brain Research",
issn = "0006-8993",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Properties of presynaptic P2X7-like receptors at the neuromuscular junction

AU - Moores, T S

AU - Hasdemir, B

AU - Vega-Riveroll, L

AU - Deuchars, J

AU - Parson, S H

PY - 2005/2/9

Y1 - 2005/2/9

N2 - Adenosine triphosphate is released into the synaptic cleft of the neuromuscular junction during normal synaptic transmission, and in much greater quantities following injury and ischaemia. There is much data to suggest roles for presynaptic P2 receptors but little to demonstrate which specific receptor subunits are present. Here we show P2X7 receptor subunits on presynaptic motor nerve terminals from birth, but no evidence for P2X1, P2X2, P2X3, P2X4, P2X5 or P2X6 receptor subunits. Further, P2X receptor subunits are present as multimeric, membrane-inserted receptors. A selective agonist, 2'-3'-O-(4-benzoylbenzoyl)-adenosine 5'-triphosphate (BzATP: 100 microM), triggers vesicle release from motor nerve terminals, which is blocked by P2X7RS-specific concentrations of periodate oxidised ATP (OxATP: 100 microM) and brilliant blue G (BBG: 1 microM), but not by suramin (100 microM). Vesicle release is enhanced in the absence of extracellular divalent cations and occurs through activation of the ion channel and not any associated large pore, as we failed to label nerve terminals with large membrane-impermeant molecules after addition of BzATP. We conclude that a P2X7-like receptor is present at mouse motor nerve terminals, and that their activation promotes vesicle release.

AB - Adenosine triphosphate is released into the synaptic cleft of the neuromuscular junction during normal synaptic transmission, and in much greater quantities following injury and ischaemia. There is much data to suggest roles for presynaptic P2 receptors but little to demonstrate which specific receptor subunits are present. Here we show P2X7 receptor subunits on presynaptic motor nerve terminals from birth, but no evidence for P2X1, P2X2, P2X3, P2X4, P2X5 or P2X6 receptor subunits. Further, P2X receptor subunits are present as multimeric, membrane-inserted receptors. A selective agonist, 2'-3'-O-(4-benzoylbenzoyl)-adenosine 5'-triphosphate (BzATP: 100 microM), triggers vesicle release from motor nerve terminals, which is blocked by P2X7RS-specific concentrations of periodate oxidised ATP (OxATP: 100 microM) and brilliant blue G (BBG: 1 microM), but not by suramin (100 microM). Vesicle release is enhanced in the absence of extracellular divalent cations and occurs through activation of the ion channel and not any associated large pore, as we failed to label nerve terminals with large membrane-impermeant molecules after addition of BzATP. We conclude that a P2X7-like receptor is present at mouse motor nerve terminals, and that their activation promotes vesicle release.

KW - Adenosine Triphosphate

KW - Animals

KW - Benzenesulfonates

KW - Cations, Divalent

KW - Ion Channels

KW - Mice

KW - Mice, Inbred C57BL

KW - Microscopy, Electron, Transmission

KW - Motor Neurons

KW - Muscle, Skeletal

KW - Neuromuscular Junction

KW - Platelet Aggregation Inhibitors

KW - Presynaptic Terminals

KW - Protein Subunits

KW - Purinergic P2 Receptor Agonists

KW - Purinergic P2 Receptor Antagonists

KW - Receptors, Purinergic P2

KW - Receptors, Purinergic P2X7

KW - Synaptic Membranes

KW - Synaptic Transmission

KW - Synaptic Vesicles

KW - Time Factors

U2 - 10.1016/j.brainres.2004.12.001

DO - 10.1016/j.brainres.2004.12.001

M3 - Article

VL - 1034

SP - 40

EP - 50

JO - Brain Research

JF - Brain Research

SN - 0006-8993

IS - 1-2

ER -