ATP acts as fast neurotransmitter in rat habenula: neurochemical and enzymecytochemical evidence

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

The release of ATP and ADP, the putative central neurotransmitters, from the isolated habenula preparation was investigated in the rat, at rest and during electrical stimulation, using the luciferin-luciferase assay and the creatine phosphokinase assay. Electrical field stimulation (2 Hz, 360 pulses) released a considerable amount of ATP (2450 ± 280 pmol/g wet tissue) from the tissue; inhibition of the voltage Na+ entry by tetrodotoxin (1 μM) reduced significantly the evoked release (by 66.25 ± 6.65%), but not the resting release of ATP. Endogenous ADP also appeared in the effluent, but its amount differed during resting condition and after stimulation from that of ATP, suggesting that the majority of the released compound is ATP in response to stimulation. When ATP was added to the tissue, it readily decomposed to ADP and AMP ( Km = 811.6 ± 68.88 μM, vmax = 23.1±2.75nmol/min per prep., measured by high-performance liquid chromatography combined with ultraviolet detection), indicating that the habenula contains ectoATPases. In addition, the inactivation of extracellular ATP by the ectoATPase enzyme was also visualized by electron microscopic enzyme cytochemistry. The ectoATPase enzyme was present on the membranes of the dendrites and nerve terminals and in the synapses of the habenula. Taking into account the fact that ATP is ubiquitous in excitable cells (storage) and the findings published by Edwardset al. in 1992 ("ATP receptor-mediated synaptic currents in the central nervous system",Nature, Vol. 359, pp. 144-147), our data provides evidence for the release by axonal stimulation and extracellular decomposition of ATP, all needed for an endogenous substance qualified as a transmitter.

Original languageEnglish
Pages (from-to)915-920
Number of pages6
JournalNeuroscience
Volume66
Issue number4
DOIs
Publication statusPublished - 1995

Fingerprint

Habenula
Neurotransmitter Agents
Adenosine Triphosphate
Adenosine Diphosphate
Electric Stimulation
Enzymes
Purinergic P2 Receptors
Histocytochemistry
Tetrodotoxin
Adenosine Monophosphate
Creatine Kinase
Dendrites
Luciferases
Synapses
Central Nervous System
High Pressure Liquid Chromatography
Electrons

Keywords

  • bovine serum albumin
  • BSA
  • EDTA
  • ethylenediaminetetra-acetate
  • high-performance liquid chromatography with ultraviolet detection
  • HPLC-UV
  • tetrodotoxin
  • TTX

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

ATP acts as fast neurotransmitter in rat habenula : neurochemical and enzymecytochemical evidence. / Sperlágh, B.; Kittel, A.; Lajtha, A.; Vízi, E.

In: Neuroscience, Vol. 66, No. 4, 1995, p. 915-920.

Research output: Contribution to journalArticle

@article{418fcef1f1684c6ebf23f086a1507677,
title = "ATP acts as fast neurotransmitter in rat habenula: neurochemical and enzymecytochemical evidence",
abstract = "The release of ATP and ADP, the putative central neurotransmitters, from the isolated habenula preparation was investigated in the rat, at rest and during electrical stimulation, using the luciferin-luciferase assay and the creatine phosphokinase assay. Electrical field stimulation (2 Hz, 360 pulses) released a considerable amount of ATP (2450 ± 280 pmol/g wet tissue) from the tissue; inhibition of the voltage Na+ entry by tetrodotoxin (1 μM) reduced significantly the evoked release (by 66.25 ± 6.65{\%}), but not the resting release of ATP. Endogenous ADP also appeared in the effluent, but its amount differed during resting condition and after stimulation from that of ATP, suggesting that the majority of the released compound is ATP in response to stimulation. When ATP was added to the tissue, it readily decomposed to ADP and AMP ( Km = 811.6 ± 68.88 μM, vmax = 23.1±2.75nmol/min per prep., measured by high-performance liquid chromatography combined with ultraviolet detection), indicating that the habenula contains ectoATPases. In addition, the inactivation of extracellular ATP by the ectoATPase enzyme was also visualized by electron microscopic enzyme cytochemistry. The ectoATPase enzyme was present on the membranes of the dendrites and nerve terminals and in the synapses of the habenula. Taking into account the fact that ATP is ubiquitous in excitable cells (storage) and the findings published by Edwardset al. in 1992 ({"}ATP receptor-mediated synaptic currents in the central nervous system{"},Nature, Vol. 359, pp. 144-147), our data provides evidence for the release by axonal stimulation and extracellular decomposition of ATP, all needed for an endogenous substance qualified as a transmitter.",
keywords = "bovine serum albumin, BSA, EDTA, ethylenediaminetetra-acetate, high-performance liquid chromatography with ultraviolet detection, HPLC-UV, tetrodotoxin, TTX",
author = "B. Sperl{\'a}gh and A. Kittel and A. Lajtha and E. V{\'i}zi",
year = "1995",
doi = "10.1016/0306-4522(94)00588-V",
language = "English",
volume = "66",
pages = "915--920",
journal = "Neuroscience",
issn = "0306-4522",
publisher = "Elsevier Limited",
number = "4",

}

TY - JOUR

T1 - ATP acts as fast neurotransmitter in rat habenula

T2 - neurochemical and enzymecytochemical evidence

AU - Sperlágh, B.

AU - Kittel, A.

AU - Lajtha, A.

AU - Vízi, E.

PY - 1995

Y1 - 1995

N2 - The release of ATP and ADP, the putative central neurotransmitters, from the isolated habenula preparation was investigated in the rat, at rest and during electrical stimulation, using the luciferin-luciferase assay and the creatine phosphokinase assay. Electrical field stimulation (2 Hz, 360 pulses) released a considerable amount of ATP (2450 ± 280 pmol/g wet tissue) from the tissue; inhibition of the voltage Na+ entry by tetrodotoxin (1 μM) reduced significantly the evoked release (by 66.25 ± 6.65%), but not the resting release of ATP. Endogenous ADP also appeared in the effluent, but its amount differed during resting condition and after stimulation from that of ATP, suggesting that the majority of the released compound is ATP in response to stimulation. When ATP was added to the tissue, it readily decomposed to ADP and AMP ( Km = 811.6 ± 68.88 μM, vmax = 23.1±2.75nmol/min per prep., measured by high-performance liquid chromatography combined with ultraviolet detection), indicating that the habenula contains ectoATPases. In addition, the inactivation of extracellular ATP by the ectoATPase enzyme was also visualized by electron microscopic enzyme cytochemistry. The ectoATPase enzyme was present on the membranes of the dendrites and nerve terminals and in the synapses of the habenula. Taking into account the fact that ATP is ubiquitous in excitable cells (storage) and the findings published by Edwardset al. in 1992 ("ATP receptor-mediated synaptic currents in the central nervous system",Nature, Vol. 359, pp. 144-147), our data provides evidence for the release by axonal stimulation and extracellular decomposition of ATP, all needed for an endogenous substance qualified as a transmitter.

AB - The release of ATP and ADP, the putative central neurotransmitters, from the isolated habenula preparation was investigated in the rat, at rest and during electrical stimulation, using the luciferin-luciferase assay and the creatine phosphokinase assay. Electrical field stimulation (2 Hz, 360 pulses) released a considerable amount of ATP (2450 ± 280 pmol/g wet tissue) from the tissue; inhibition of the voltage Na+ entry by tetrodotoxin (1 μM) reduced significantly the evoked release (by 66.25 ± 6.65%), but not the resting release of ATP. Endogenous ADP also appeared in the effluent, but its amount differed during resting condition and after stimulation from that of ATP, suggesting that the majority of the released compound is ATP in response to stimulation. When ATP was added to the tissue, it readily decomposed to ADP and AMP ( Km = 811.6 ± 68.88 μM, vmax = 23.1±2.75nmol/min per prep., measured by high-performance liquid chromatography combined with ultraviolet detection), indicating that the habenula contains ectoATPases. In addition, the inactivation of extracellular ATP by the ectoATPase enzyme was also visualized by electron microscopic enzyme cytochemistry. The ectoATPase enzyme was present on the membranes of the dendrites and nerve terminals and in the synapses of the habenula. Taking into account the fact that ATP is ubiquitous in excitable cells (storage) and the findings published by Edwardset al. in 1992 ("ATP receptor-mediated synaptic currents in the central nervous system",Nature, Vol. 359, pp. 144-147), our data provides evidence for the release by axonal stimulation and extracellular decomposition of ATP, all needed for an endogenous substance qualified as a transmitter.

KW - bovine serum albumin

KW - BSA

KW - EDTA

KW - ethylenediaminetetra-acetate

KW - high-performance liquid chromatography with ultraviolet detection

KW - HPLC-UV

KW - tetrodotoxin

KW - TTX

UR - http://www.scopus.com/inward/record.url?scp=0029010019&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029010019&partnerID=8YFLogxK

U2 - 10.1016/0306-4522(94)00588-V

DO - 10.1016/0306-4522(94)00588-V

M3 - Article

C2 - 7651618

AN - SCOPUS:0029010019

VL - 66

SP - 915

EP - 920

JO - Neuroscience

JF - Neuroscience

SN - 0306-4522

IS - 4

ER -