Pannexin1-mediated ATP release provides signal transmission between Neuro2A cells

Rodolfo M. Iglesias, David C. Spray

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Pannexin1 (Panx1), a protein related to the gap junction proteins of invertebrates, forms nonjunctional channels that open upon depolarization and in response to mechanical stretch and purinergic receptor stimulation. Importantly, ATP can be released through Panx1 channels, providing a possible role for these channels in non-vesicular signal transmission. In this study we expressed exogenous human and mouse Panx1 in the gap junction deficient Neuro2A neuroblastoma cell line and explored the contribution of Panx1 channels to cell-cell communication as sites of ATP release. Electrophysiological (patch clamp) recordings from Panx1 transfected Neuro2A cells revealed membrane conductance that increased beyond 0 mV when applying voltage ramps from -60 to +100 mV; threshold was correlated with extracellular K +, so that at 10 mM K +, channels began to open at -30 mV. Evaluation of cell-cell communication using dual whole cell recordings from cell pairs revealed that activation of Panx1 current in one cell of the pair induced an inward current in the second cell after a latency of 10-20 s. This paracrine response was amplified by an ATPase inhibitor (ARL67156, 100 μM) and was blocked by the ATP-degrading enzyme apyrase (6.7 U/ml), by the P2 receptor antagonist suramin (50 μM) and by the Panx1 channel blocker carbenoxolone. These results provide additional evidence that ATP release through Panx1 channels can mediate nonsynaptic bidirectional intercellular communication. Furthermore, current potentiation by elevated K + provides a mechanism for enhancement of ATP release under pathological conditions.

Original languageEnglish (US)
Pages (from-to)1355-1363
Number of pages9
JournalNeurochemical Research
Volume37
Issue number6
DOIs
StatePublished - Jun 2012

Fingerprint

Adenosine Triphosphate
Cell Communication
Communication
Carbenoxolone
Apyrase
Purinergic Receptors
Suramin
Architectural Accessibility
Mechanoreceptors
Connexins
Gap Junctions
Depolarization
Clamping devices
Patch-Clamp Techniques
Invertebrates
Cell membranes
Neuroblastoma
Adenosine Triphosphatases
Chemical activation
Cells

Keywords

  • Hemichannel
  • Intercellular signaling
  • Neuroblastoma
  • Panx1
  • Paracrine
  • Purinergic receptor

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Biochemistry

Cite this

Pannexin1-mediated ATP release provides signal transmission between Neuro2A cells. / Iglesias, Rodolfo M.; Spray, David C.

In: Neurochemical Research, Vol. 37, No. 6, 06.2012, p. 1355-1363.

Research output: Contribution to journalArticle

@article{61a96aa88a624c3981575d70c79c7cc7,
title = "Pannexin1-mediated ATP release provides signal transmission between Neuro2A cells",
abstract = "Pannexin1 (Panx1), a protein related to the gap junction proteins of invertebrates, forms nonjunctional channels that open upon depolarization and in response to mechanical stretch and purinergic receptor stimulation. Importantly, ATP can be released through Panx1 channels, providing a possible role for these channels in non-vesicular signal transmission. In this study we expressed exogenous human and mouse Panx1 in the gap junction deficient Neuro2A neuroblastoma cell line and explored the contribution of Panx1 channels to cell-cell communication as sites of ATP release. Electrophysiological (patch clamp) recordings from Panx1 transfected Neuro2A cells revealed membrane conductance that increased beyond 0 mV when applying voltage ramps from -60 to +100 mV; threshold was correlated with extracellular K +, so that at 10 mM K +, channels began to open at -30 mV. Evaluation of cell-cell communication using dual whole cell recordings from cell pairs revealed that activation of Panx1 current in one cell of the pair induced an inward current in the second cell after a latency of 10-20 s. This paracrine response was amplified by an ATPase inhibitor (ARL67156, 100 μM) and was blocked by the ATP-degrading enzyme apyrase (6.7 U/ml), by the P2 receptor antagonist suramin (50 μM) and by the Panx1 channel blocker carbenoxolone. These results provide additional evidence that ATP release through Panx1 channels can mediate nonsynaptic bidirectional intercellular communication. Furthermore, current potentiation by elevated K + provides a mechanism for enhancement of ATP release under pathological conditions.",
keywords = "Hemichannel, Intercellular signaling, Neuroblastoma, Panx1, Paracrine, Purinergic receptor",
author = "Iglesias, {Rodolfo M.} and Spray, {David C.}",
year = "2012",
month = "6",
doi = "10.1007/s11064-012-0720-6",
language = "English (US)",
volume = "37",
pages = "1355--1363",
journal = "Neurochemical Research",
issn = "0364-3190",
publisher = "Springer New York",
number = "6",

}

TY - JOUR

T1 - Pannexin1-mediated ATP release provides signal transmission between Neuro2A cells

AU - Iglesias, Rodolfo M.

AU - Spray, David C.

PY - 2012/6

Y1 - 2012/6

N2 - Pannexin1 (Panx1), a protein related to the gap junction proteins of invertebrates, forms nonjunctional channels that open upon depolarization and in response to mechanical stretch and purinergic receptor stimulation. Importantly, ATP can be released through Panx1 channels, providing a possible role for these channels in non-vesicular signal transmission. In this study we expressed exogenous human and mouse Panx1 in the gap junction deficient Neuro2A neuroblastoma cell line and explored the contribution of Panx1 channels to cell-cell communication as sites of ATP release. Electrophysiological (patch clamp) recordings from Panx1 transfected Neuro2A cells revealed membrane conductance that increased beyond 0 mV when applying voltage ramps from -60 to +100 mV; threshold was correlated with extracellular K +, so that at 10 mM K +, channels began to open at -30 mV. Evaluation of cell-cell communication using dual whole cell recordings from cell pairs revealed that activation of Panx1 current in one cell of the pair induced an inward current in the second cell after a latency of 10-20 s. This paracrine response was amplified by an ATPase inhibitor (ARL67156, 100 μM) and was blocked by the ATP-degrading enzyme apyrase (6.7 U/ml), by the P2 receptor antagonist suramin (50 μM) and by the Panx1 channel blocker carbenoxolone. These results provide additional evidence that ATP release through Panx1 channels can mediate nonsynaptic bidirectional intercellular communication. Furthermore, current potentiation by elevated K + provides a mechanism for enhancement of ATP release under pathological conditions.

AB - Pannexin1 (Panx1), a protein related to the gap junction proteins of invertebrates, forms nonjunctional channels that open upon depolarization and in response to mechanical stretch and purinergic receptor stimulation. Importantly, ATP can be released through Panx1 channels, providing a possible role for these channels in non-vesicular signal transmission. In this study we expressed exogenous human and mouse Panx1 in the gap junction deficient Neuro2A neuroblastoma cell line and explored the contribution of Panx1 channels to cell-cell communication as sites of ATP release. Electrophysiological (patch clamp) recordings from Panx1 transfected Neuro2A cells revealed membrane conductance that increased beyond 0 mV when applying voltage ramps from -60 to +100 mV; threshold was correlated with extracellular K +, so that at 10 mM K +, channels began to open at -30 mV. Evaluation of cell-cell communication using dual whole cell recordings from cell pairs revealed that activation of Panx1 current in one cell of the pair induced an inward current in the second cell after a latency of 10-20 s. This paracrine response was amplified by an ATPase inhibitor (ARL67156, 100 μM) and was blocked by the ATP-degrading enzyme apyrase (6.7 U/ml), by the P2 receptor antagonist suramin (50 μM) and by the Panx1 channel blocker carbenoxolone. These results provide additional evidence that ATP release through Panx1 channels can mediate nonsynaptic bidirectional intercellular communication. Furthermore, current potentiation by elevated K + provides a mechanism for enhancement of ATP release under pathological conditions.

KW - Hemichannel

KW - Intercellular signaling

KW - Neuroblastoma

KW - Panx1

KW - Paracrine

KW - Purinergic receptor

UR - http://www.scopus.com/inward/record.url?scp=84862861555&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84862861555&partnerID=8YFLogxK

U2 - 10.1007/s11064-012-0720-6

DO - 10.1007/s11064-012-0720-6

M3 - Article

C2 - 22359052

AN - SCOPUS:84862861555

VL - 37

SP - 1355

EP - 1363

JO - Neurochemical Research

JF - Neurochemical Research

SN - 0364-3190

IS - 6

ER -