Monitoring real-time release of ATP from the molluscan central nervous system

Gruenhagen, Jason; Lovell, Peter V.; Moroz, Leonid L.; Yeung, Edward S.

doi:10.1016/j.jneumeth.2004.03.008

Cited by 32 publications

(22 citation statements)

References 36 publications

(34 reference statements)

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“…The presence of Lym P2X mRNA at a similar level in the majority of neurones in all CNS ganglia implies a general role of the Lym P2X channel in CNS function rather than a specific role in a particular neuronal pathway. Indeed, this widespread localisation of Lym P2X is consistent with the similarly widespread release of ATP from all Lymnaea ganglia upon addition of neuroexcitatory compounds [29] suggesting that purinergic synaptic transmission could be widely utilised in Lymnaea CNS.…”

Section: Discussionsupporting

confidence: 71%

“…Furthermore, the neuronal pathways underlying complex physiological processes such as feeding and respiration have been elucidated in this organism [26]–[28], making it an attractive model for investigating neural networks. The demonstration that ATP is released from Lymnaea stagnalis CNS ganglia [29] suggests the presence of a purinergic signalling system and it is therefore possible that Lymnaea could potentially be developed as a simple model system to study P2X receptor function in the CNS.…”

Section: Introductionmentioning

confidence: 99%

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Cloning and Characterization of a P2X Receptor Expressed in the Central Nervous System of Lymnaea stagnalis

et al. 2012

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P2X receptors are membrane ion channels gated by extracellular ATP. Mammals possess seven distinct P2X subtypes (P2X1-7) that have important functions in a wide array of physiological processes including roles in the central nervous system (CNS) where they have been linked to modulation of neurotransmitter release. We report here the cloning and functional characterization of a P2X receptor from the mollusc Lymnaea stagnalis. This model organism has a relatively simple CNS consisting of large readily identifiable neurones, a feature which together with a well characterized neuronal circuitry for important physiological processes such as feeding and respiration makes it an attractive potential model to examine P2X function. Using CODEHOP PCR we identified a single P2X receptor (LymP2X) in Lymnaea CNS which was subsequently cloned by RT-PCR. When heterologously expressed in Xenopus oocytes, LymP2X exhibited ATP evoked inward currents (EC50 6.2 µM) which decayed during the continued presence of agonist. UTP and ADP did not activate the receptor whereas αβmeATP was a weak agonist. BzATP was a partial agonist with an EC50 of 2.4 µM and a maximal response 33% smaller than that of ATP. The general P2 receptor antagonists PPADS and suramin both inhibited LymP2X currents with IC50 values of 8.1 and 27.4 µM respectively. LymP2X is inhibited by acidic pH whereas Zn2+ and Cu2+ ions exhibited a biphasic effect, potentiating currents up to 100 µM and inhibiting at higher concentrations. Quantitative RT-PCR and in situ hybridization detected expression of LymP2X mRNA in neurones of all CNS ganglia suggesting this ion channel may have widespread roles in Lymnaea CNS function.

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Section: Discussionsupporting

confidence: 71%

Section: Introductionmentioning

confidence: 99%

Cloning and Characterization of a P2X Receptor Expressed in the Central Nervous System of Lymnaea stagnalis

et al. 2012

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“…Br − , I − , NO 3 − , etc.) [24] and high salt concentrations in general [25]. The anion transport inhibitor DIDS [26], the cation channel blocker gadolinium [27] or P2 receptor antagonist suramin [28] inhibits the luciferase reaction.…”

Section: The Luciferin-luciferase Techniquementioning

confidence: 99%

“…The luciferin/luciferase technique has been used successfully to visualise ATP release [23,25,26,35,36]. In this approach, the cells (or tissue) are bathed with the ATP-specific luminescence indicator and the ATP is imaged directly as it is being released after a given stimulus.…”

Section: Imaging Of Atp Releasementioning

confidence: 99%

ATP release from non-excitable cells

Praetorius

Leipziger

2009

Purinergic Signalling

207

229

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All cells release nucleotides and are in one way or another involved in local autocrine and paracrine regulation of organ function via stimulation of purinergic receptors. Significant technical advances have been made in recent years to quantify more precisely resting and stimulated adenosine triphosphate (ATP) concentrations in close proximity to the plasma membrane. These technical advances are reviewed here. However, the mechanisms by which cells release ATP continue to be enigmatic. The current state of knowledge on different suggested mechanisms is also reviewed. Current evidence suggests that two separate regulated modes of ATP release co-exist in nonexcitable cells: (1) a conductive pore which in several systems has been found to be the channel pannexin 1 and (2) vesicular release. Modes of stimulation of ATP release are reviewed and indicate that both subtle mechanical stimulation and agonist-triggered release play pivotal roles. The mechano-sensor for ATP release is not yet defined.

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“…Its relatively simple CNS contains <22,000 neurons making it amenable to study processes of associative memory and taste. ATP release in molluscan CNS has been studied in real-time (Gruenhagen et al, 2004) highlighting a potential for purinergic receptors in invertebrate CNS function. A full-length P2X receptor orthologue has been cloned from L. stagnalis CNS ( Lym P2X).…”

Section: Cloned Non-vertebrate and Primitive P2x Receptorsmentioning

confidence: 99%

Primitive ATP-activated P2X receptors: discovery, function and pharmacology

Fountain

2013

Front. Cell. Neurosci.

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Adenosine 5-triphosphate (ATP) is omnipresent in biology. It is therefore no surprise that organisms have evolved multifaceted roles for ATP, exploiting its abundance and restriction of passive diffusion across biological membranes. A striking role is the emergence of ATP as a bona fide transmitter molecule, whereby the movement of ATP across membranes serves as a chemical message through a direct ligand-receptor interaction. P2X receptors are ligand-gated ion channels that mediate fast responses to the transmitter ATP in mammalian cells including central and sensory neurons, vascular smooth muscle, endothelium, and leukocytes. Molecular cloning of P2X receptors and our understanding of structure-function relationships has provided sequence information with which to query an exponentially expanding wealth of genome sequence information including protist, early animal and human pathogen genomes. P2X receptors have now been cloned and characterized from a number of simple organisms. Such work has led to surprising new cellular roles for the P2X receptors family and an unusual phylogeny, with organisms such as Drosophila and C. elegans notably lacking P2X receptors despite retaining ionotropic receptors for other common transmitters that are present in mammals. This review will summarize current work on the evolutionary biology of P2X receptors and ATP as a signaling molecule, discuss what can be drawn from such studies when considering the action of ATP in higher animals and plants, and outline how simple organisms may be exploited experimentally to inform P2X receptor function in a wider context.

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Monitoring real-time release of ATP from the molluscan central nervous system

Cited by 32 publications

References 36 publications

Cloning and Characterization of a P2X Receptor Expressed in the Central Nervous System of Lymnaea stagnalis

Cloning and Characterization of a P2X Receptor Expressed in the Central Nervous System of Lymnaea stagnalis

ATP release from non-excitable cells

Primitive ATP-activated P2X receptors: discovery, function and pharmacology

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