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Article ; Online: Differential activation of mouse and human Panx1 channel variants

Antonio Cibelli / Preeti Dohare / David C. Spray / Eliana Scemes

PLoS ONE, Vol 18, Iss

2023 Volume 12

Keywords	Medicine ; R ; Science ; Q
Language	English
Publishing date	2023-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Differential activation of mouse and human Panx1 channel variants.

Cibelli, Antonio / Dohare, Preeti / Spray, David C / Scemes, Eliana

PloS one

2023 Volume 18, Issue 12, Page(s) e0295710

Abstract: Pannexins are ubiquitously expressed in human and mouse tissues. Pannexin 1 (Panx1), the most thoroughly characterized member of this family, forms plasmalemmal membrane channels permeable to relatively large molecules, such as ATP. Although human and ... ...

Abstract	Pannexins are ubiquitously expressed in human and mouse tissues. Pannexin 1 (Panx1), the most thoroughly characterized member of this family, forms plasmalemmal membrane channels permeable to relatively large molecules, such as ATP. Although human and mouse Panx1 amino acid sequences are conserved in the presently known regulatory sites involved in trafficking and modulation of the channel, differences are reported in the N- and C-termini of the protein, and the mechanisms of channel activation by different stimuli remain controversial. Here we used a neuroblastoma cell line to study the activation properties of endogenous mPanx1 and exogenously expressed hPanx1. Dye uptake and electrophysiological recordings revealed that in contrast to mouse Panx1, the human ortholog is insensitive to stimulation with high extracellular [K+] but responds similarly to activation of the purinergic P2X7 receptor. The two most frequent Panx1 polymorphisms found in the human population, Q5H (rs1138800) and E390D (rs74549886), exogenously expressed in Panx1-null N2a cells revealed that regarding P2X7 receptor mediated Panx1 activation, the Q5H mutant is a gain of function whereas the E390D mutant is a loss of function variant. Collectively, we demonstrate differences in the activation between human and mouse Panx1 orthologs and suggest that these differences may have translational implications for studies where Panx1 has been shown to have significant impact.
MeSH term(s)	Humans ; Adenosine Triphosphate/metabolism ; Cell Line ; Cell Membrane/metabolism ; Connexins/genetics ; Connexins/metabolism ; Neural Stem Cells/metabolism
Chemical Substances	Adenosine Triphosphate (8L70Q75FXE) ; Connexins ; Panx1 protein, mouse ; PANX1 protein, human
Language	English
Publishing date	2023-12-15
Publishing country	United States
Document type	Journal Article
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0295710
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Article ; Online: Activity and Stability of Panx1 Channels in Astrocytes and Neuroblastoma Cells Are Enhanced by Cholesterol Depletion.

Cibelli, Antonio / Scemes, Eliana / Spray, David C

Cells

2022 Volume 11, Issue 20

Abstract: Pannexin1 (Panx1) is expressed in both neurons and glia where it forms ATP-permeable channels that are activated under pathological conditions such as epilepsy, migraine, inflammation, and ischemia. Membrane lipid composition affects proper distribution ... ...

Abstract	Pannexin1 (Panx1) is expressed in both neurons and glia where it forms ATP-permeable channels that are activated under pathological conditions such as epilepsy, migraine, inflammation, and ischemia. Membrane lipid composition affects proper distribution and function of receptors and ion channels, and defects in cholesterol metabolism are associated with neurological diseases. In order to understand the impact of membrane cholesterol on the distribution and function of Panx1 in neural cells, we used fluorescence recovery after photobleaching (FRAP) to evaluate its mobility and electrophysiology and dye uptake to assess channel function. We observed that cholesterol extraction (using methyl-β-cyclodextrin) and inhibition of its synthesis (lovastatin) decreased the lateral diffusion of Panx1 in the plasma membrane. Panx1 channel activity (dye uptake, ATP release and ionic current) was enhanced in cholesterol-depleted Panx1 transfected cells and in wild-type astrocytes compared to non-depleted or Panx1 null cells. Manipulation of cholesterol levels may, therefore, offer a novel strategy by which Panx1 channel activation might modulate various pathological conditions.
MeSH term(s)	Humans ; Adenosine Triphosphate/metabolism ; Anticholesteremic Agents/pharmacology ; Astrocytes/metabolism ; Cholesterol/metabolism ; Connexins/metabolism ; Ion Channels/metabolism ; Lovastatin/pharmacology ; Membrane Lipids/metabolism ; Nerve Tissue Proteins/genetics ; Nerve Tissue Proteins/metabolism ; Neuroblastoma/metabolism ; Protein Stability
Chemical Substances	Adenosine Triphosphate (8L70Q75FXE) ; Anticholesteremic Agents ; Cholesterol (97C5T2UQ7J) ; Connexins ; Ion Channels ; Lovastatin (9LHU78OQFD) ; Membrane Lipids ; Nerve Tissue Proteins ; PANX1 protein, human
Language	English
Publishing date	2022-10-14
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells11203219
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Article ; Online: The Contribution of Astrocyte and Neuronal Panx1 to Seizures Is Model and Brain Region Dependent.

Obot, Price / Velíšek, Libor / Velíšková, Jana / Scemes, Eliana

ASN neuro

2021 Volume 13, Page(s) 17590914211007273

Abstract: Pannexin1 (Panx1) is an ATP release channel expressed in neurons and astrocytes that plays important roles in CNS physiology and pathology. Evidence for the involvement of Panx1 in seizures includes the reduction of epileptiform activity and ictal ... ...

Abstract	Pannexin1 (Panx1) is an ATP release channel expressed in neurons and astrocytes that plays important roles in CNS physiology and pathology. Evidence for the involvement of Panx1 in seizures includes the reduction of epileptiform activity and ictal discharges following Panx1 channel blockade or deletion. However, very little is known about the relative contribution of astrocyte and neuronal Panx1 channels to hyperexcitability. To this end, mice with global and cell type specific deletion of Panx1 were used in one
MeSH term(s)	Animals ; Astrocytes/metabolism ; Brain/metabolism ; Brain/physiopathology ; Connexins/deficiency ; Connexins/genetics ; Disease Models, Animal ; Electroencephalography/methods ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Nerve Tissue Proteins/deficiency ; Nerve Tissue Proteins/genetics ; Neurons/metabolism ; Organ Culture Techniques ; Seizures/genetics ; Seizures/metabolism
Chemical Substances	Connexins ; Nerve Tissue Proteins ; Panx1 protein, mouse
Language	English
Publishing date	2021-08-30
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2485467-0
ISSN	1759-0914 ; 1759-0914
ISSN (online)	1759-0914
ISSN	1759-0914
DOI	10.1177/17590914211007273
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Article ; Online: Astrocyte and Neuronal Panx1 Support Long-Term Reference Memory in Mice.

Obot, Price / Subah, Galadu / Schonwald, Antonia / Pan, Jian / Velíšek, Libor / Velíšková, Jana / Stanton, Patric K / Scemes, Eliana

ASN neuro

2023 Volume 15, Page(s) 17590914231184712

Abstract: Pannexin 1 (Panx1) is an ubiquitously expressed protein that forms plasma membrane channels permeable to anions and moderate-sized signaling molecules (e.g., ATP, glutamate). In the nervous system, activation of Panx1 channels has been extensively shown ... ...

Abstract	Pannexin 1 (Panx1) is an ubiquitously expressed protein that forms plasma membrane channels permeable to anions and moderate-sized signaling molecules (e.g., ATP, glutamate). In the nervous system, activation of Panx1 channels has been extensively shown to contribute to distinct neurological disorders (epilepsy, chronic pain, migraine, neuroAIDS, etc.), but knowledge of the extent to which these channels have a physiological role remains restricted to three studies supporting their involvement in hippocampus dependent learning. Given that Panx1 channels may provide an important mechanism for activity-dependent neuron-glia interaction, we used Panx1 transgenic mice with global and cell-type specific deletions of Panx1 to interrogate their participation in working and reference memory. Using the eight-arm radial maze, we show that long-term spatial reference memory, but not spatial working memory, is deficient in Panx1-null mice and that both astrocyte and neuronal Panx1 contribute to the consolidation of long-term spatial memory. Field potential recordings in hippocampal slices of Panx1-null mice revealed an attenuation of both long-term potentiation (LTP) of synaptic strength and long-term depression (LTD) at Schaffer collateral-CA1 synapses without alterations of basal synaptic transmission or pre-synaptic paired-pulse facilitation. Our results implicate both neuronal and astrocyte Panx1 channels as critical players for the development and maintenance of long-term spatial reference memory in mice.
MeSH term(s)	Mice ; Animals ; Astrocytes/metabolism ; Neurons/metabolism ; Long-Term Potentiation/physiology ; Neuronal Plasticity/physiology ; Hippocampus/metabolism ; Synapses/metabolism ; Mice, Transgenic ; Mice, Knockout ; Nerve Tissue Proteins/genetics ; Nerve Tissue Proteins/metabolism ; Connexins/genetics ; Connexins/metabolism
Chemical Substances	Panx1 protein, mouse ; Nerve Tissue Proteins ; Connexins
Language	English
Publishing date	2023-07-10
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2485467-0
ISSN	1759-0914 ; 1759-0914
ISSN (online)	1759-0914
ISSN	1759-0914
DOI	10.1177/17590914231184712
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Article: Astrocyte and neuronal Panx1 support long-term reference memory in mice.

Obot, Price / Subah, Galadu / Schonwald, Antonia / Pan, Jian / Velíšek, Libor / Velíšková, Jana / Stanton, Patric K / Scemes, Eliana

bioRxiv : the preprint server for biology

2023

Abstract: Pannexin 1 (Panx1) are ubiquitously expressed proteins that form plasma membrane channels permeable to anions and moderate sized signaling molecules (e.g., ATP, glutamate). In the nervous system, activation of Panx1 channels have been extensively shown ... ...

Abstract	Pannexin 1 (Panx1) are ubiquitously expressed proteins that form plasma membrane channels permeable to anions and moderate sized signaling molecules (e.g., ATP, glutamate). In the nervous system, activation of Panx1 channels have been extensively shown to contribute to distinct neurological disorders (epilepsy, chronic pain, migraine, neuroAIDS, etc.) but knowledge of extent to which these channels have a physiological role remains restricted to three studies supporting their involvement in hippocampus dependent learning. Given that Panx1 channels may provide an important mechanism for activity-dependent neuron-glia interaction, we used Panx1 transgenic mice with global and cell-type specific deletions of Panx1 to interrogate their participation in working and reference memory. Using the 8-arm radial maze, we show that long-term spatial reference memory, but not spatial working memory, is deficient in Panx1-null mice and that both astrocyte and neuronal Panx1 contribute to the consolidation of long-term spatial memory. Field potential recordings in hippocampal slices of Panx1-null mice revealed an attenuation of both long-term potentiation (LTP) of synaptic strength and long-term depression (LTD) at Schaffer collateral - CA1 synapses without alterations basal synaptic transmission or pre-synaptic paired-pulse facilitation. Our results implicate both neuronal and astrocyte Panx1 channels as critical players for the development and maintenance of long-term spatial reference memory in mice.
Language	English
Publishing date	2023-01-18
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.01.16.524236
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Article: Nature of plasmalemmal functional "hemichannels".

Scemes, Eliana

Biochimica et biophysica acta

2011 Volume 1818, Issue 8, Page(s) 1880–1883

Abstract: The molecular identity of the protein forming "hemichannels" at non-junctional membranes is disputed. The family of gap junction proteins, innexins, connexins, and pannexins share several common features, including permeability characteristics and ... ...

Abstract	The molecular identity of the protein forming "hemichannels" at non-junctional membranes is disputed. The family of gap junction proteins, innexins, connexins, and pannexins share several common features, including permeability characteristics and sensitivity to blocking agents. Such overlap in properties renders the identification of which of these protein species actually establishes the non-junctional membrane conductance and permeability quite complicated, especially because in vertebrates pannexins and connexins have largely overlapping distributions in tissues. Recently, attempts to establish criteria to identify events that are "hemichannel" mediated and those to allow the distinction between connexin- from pannexin-mediated events have been proposed. Here, I present an update on that topic and discuss the most recent findings related to the nature of functional "hemichannels" focusing on connexin43 and pannexin1. This article is part of a Special Issue entitled: The Communicating junctions, composition, structure and characteristics.
MeSH term(s)	Animals ; Biophysics/methods ; Carrier Proteins/chemistry ; Carrier Proteins/metabolism ; Cell Communication ; Cell Membrane/metabolism ; Connexin 43/chemistry ; Connexins/chemistry ; Gap Junctions ; HeLa Cells ; Humans ; Membrane Proteins/chemistry ; Membrane Proteins/metabolism ; Models, Biological ; Nerve Tissue Proteins/chemistry ; Permeability ; Protein Binding ; Proteins/chemistry ; Signal Transduction
Chemical Substances	Carrier Proteins ; Connexin 43 ; Connexins ; Membrane Proteins ; Nerve Tissue Proteins ; PANX1 protein, human ; PLVAP protein, human ; Proteins
Language	English
Publishing date	2011-06-16
Publishing country	Netherlands
Document type	Journal Article ; Review
ZDB-ID	60-7
ISSN	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
ISSN (online)	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650
ISSN	0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
DOI	10.1016/j.bbamem.2011.06.005
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Article ; Online: Aquaporin-4 and transient receptor potential vanilloid 4 balance in early postnatal neurodevelopment.

Cibelli, Antonio / Mola, Maria Grazia / Saracino, Emanuela / Barile, Barbara / Abbrescia, Pasqua / Mogni, Guido / Spray, David C / Scemes, Eliana / Rossi, Andrea / Spennato, Diletta / Svelto, Maria / Frigeri, Antonio / Benfenati, Valentina / Nicchia, Grazia Paola

Glia

2024 Volume 72, Issue 5, Page(s) 938–959

Abstract: In the adult brain, the water channel aquaporin-4 (AQP4) is expressed in astrocyte endfoot, in supramolecular assemblies, called "Orthogonal Arrays of Particles" (OAPs) together with the transient receptor potential vanilloid 4 (TRPV4), finely regulating ...

Abstract	In the adult brain, the water channel aquaporin-4 (AQP4) is expressed in astrocyte endfoot, in supramolecular assemblies, called "Orthogonal Arrays of Particles" (OAPs) together with the transient receptor potential vanilloid 4 (TRPV4), finely regulating the cell volume. The present study aimed at investigating the contribution of AQP4 and TRPV4 to CNS early postnatal development using WT and AQP4 KO brain and retina and neuronal stem cells (NSCs), as an in vitro model of astrocyte differentiation. Western blot analysis showed that, differently from AQP4 and the glial cell markers, TRPV4 was downregulated during CNS development and NSC differentiation. Blue native/SDS-PAGE revealed that AQP4 progressively organized into OAPs throughout the entire differentiation process. Fluorescence quenching assay indicated that the speed of cell volume changes was time-related to NSC differentiation and functional to their migratory ability. Calcium imaging showed that the amplitude of TRPV4 Ca
MeSH term(s)	Astrocytes/metabolism ; TRPV Cation Channels/metabolism ; Aquaporin 4/metabolism ; Neuroglia/metabolism ; Brain/metabolism
Chemical Substances	TRPV Cation Channels ; Aquaporin 4
Language	English
Publishing date	2024-02-16
Publishing country	United States
Document type	Journal Article
ZDB-ID	639414-0
ISSN	1098-1136 ; 0894-1491
ISSN (online)	1098-1136
ISSN	0894-1491
DOI	10.1002/glia.24512
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Article ; Online: Exciting and not so exciting roles of pannexins.

Scemes, Eliana / Velíšková, Jana

Neuroscience letters

2017 Volume 695, Page(s) 25–31

Abstract: It is the current view that purinergic signaling regulates many physiological functions. Pannexin1 (Panx1), a member of the gap junction family of proteins is an ATP releasing channel that plays important physio-pathological roles in various tissues, ... ...

Abstract	It is the current view that purinergic signaling regulates many physiological functions. Pannexin1 (Panx1), a member of the gap junction family of proteins is an ATP releasing channel that plays important physio-pathological roles in various tissues, including the CNS. Upon binding to purinergic receptors expressed in neural cells, ATP triggers cellular responses including increased cell proliferation, cell morphology changes, release of cytokines, and regulation of neuronal excitability via release of glutamate, GABA and ATP itself. Under pathological conditions such as ischemia, trauma, inflammation, and epilepsy, extracellular ATP concentrations increases drastically but the consequences of this surge is still difficult to characterize due to its rapid metabolism in ADP and adenosine, the latter having inhibitory action on neuronal activity. For seizures, for instance, the excitatory effect of ATP on neuronal activity is mainly related to its action of P2X receptors, while the inhibitory effects are related to activation of P1, adenosine receptors. Here we provide a mini review on the properties of pannexins with a main focus on Panx1 and its involvement in seizure activity. Although there are only few studies implicating Panx1 in seizures, they are illustrative of the dual role that Panx1 has on neuronal excitability.
MeSH term(s)	Adenosine Triphosphate/metabolism ; Animals ; Connexins/chemistry ; Connexins/genetics ; Connexins/metabolism ; Connexins/physiology ; Gap Junctions/metabolism ; Gap Junctions/physiology ; Humans ; Nerve Tissue Proteins/chemistry ; Nerve Tissue Proteins/genetics ; Nerve Tissue Proteins/metabolism ; Nerve Tissue Proteins/physiology ; Neurons/metabolism ; Neurons/physiology ; Signal Transduction
Chemical Substances	Connexins ; Nerve Tissue Proteins ; Adenosine Triphosphate (8L70Q75FXE)
Language	English
Publishing date	2017-03-08
Publishing country	Ireland
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	194929-9
ISSN	1872-7972 ; 0304-3940
ISSN (online)	1872-7972
ISSN	0304-3940
DOI	10.1016/j.neulet.2017.03.010
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Article ; Online: Astrocyte and Neuronal Pannexin1 Contribute Distinctly to Seizures.

Scemes, Eliana / Velíšek, Libor / Velíšková, Jana

ASN neuro

2019 Volume 11, Page(s) 1759091419833502

Abstract: ATP- and adenosine-mediated signaling are prominent types of glia-glia and glia-neuron interaction, with an imbalance of ATP/adenosine ratio leading to altered states of excitability, as seen in epileptic seizures. Pannexin1 (Panx1), a member of the gap ... ...

Abstract	ATP- and adenosine-mediated signaling are prominent types of glia-glia and glia-neuron interaction, with an imbalance of ATP/adenosine ratio leading to altered states of excitability, as seen in epileptic seizures. Pannexin1 (Panx1), a member of the gap junction family, is an ATP release channel that is expressed in astrocytes and neurons. Previous studies provided evidence supporting a role for purinergic-mediated signaling via Panx1 channels in seizures; using mice with global deletion of Panx1, it was shown that these channels contribute in maintenance of seizures by releasing ATP. However, nothing is known about the extent to which astrocyte and neuronal Panx1 might differently contribute to seizures. We here show that targeted deletion of Panx1 in astrocytes or neurons has opposing effects on acute seizures induced by kainic acid. The absence of Panx1 in astrocytes potentiates while the absence of Panx1 in neurons attenuates seizure manifestation. Immunohistochemical analysis performed in brains of these mice, revealed that adenosine kinase (ADK), an enzyme that regulates extracellular levels of adenosine, was increased only in seized GFAP-Cre:Panx1
MeSH term(s)	Adenosine Kinase/antagonists & inhibitors ; Adenosine Kinase/metabolism ; Adenosine Triphosphate/metabolism ; Animals ; Astrocytes/drug effects ; Astrocytes/metabolism ; Brain/drug effects ; Brain/metabolism ; Connexins/genetics ; Connexins/metabolism ; Disease Models, Animal ; Epilepsy/drug therapy ; Epilepsy/metabolism ; Kainic Acid ; Mice, Inbred C57BL ; Mice, Transgenic ; Nerve Tissue Proteins/genetics ; Nerve Tissue Proteins/metabolism ; Neurons/drug effects ; Neurons/metabolism ; Seizures/drug therapy ; Seizures/metabolism
Chemical Substances	Connexins ; Nerve Tissue Proteins ; Panx1 protein, mouse ; Adenosine Triphosphate (8L70Q75FXE) ; Adenosine Kinase (EC 2.7.1.20) ; Kainic Acid (SIV03811UC)
Language	English
Publishing date	2019-04-10
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2485467-0
ISSN	1759-0914 ; 1759-0914
ISSN (online)	1759-0914
ISSN	1759-0914
DOI	10.1177/1759091419833502
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