Microglial pannexin-1 channel activation is a spinal determinant of joint pain

Mousseau, Michael; Burma, Nicole E.; Lee, Kwan Yeop; Leduc‐Pessah, Heather; Kwok, Charlie H.T.; Reid, Allison; O’Brien, Melissa; Sagalajev, Boriss; Stratton, Jo Anne; Patrick, Natalya; Stemkowski, Patrick L.; Biernaskie, Jeff; Zamponi, Gerald W.; Salo, Paul; McDougall, Jason J.; Prescott, Steven A.; Matyas, John R.

doi:10.1126/sciadv.aas9846

Cited by 76 publications

(70 citation statements)

References 47 publications

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“…Large-pore forming channels play important roles in cell to cell communication by responding to diverse stimuli and releasing signaling molecules like ATP and amino acids (Giaume et al, 2013;Ma et al, 2016;Okada et al, 2018;Osei-Owusu et al, 2018). Pannexins are a family of ubiquitously expressed large-pore forming channels which regulate nucleotide release during apoptosis (Chekeni et al, 2010), blood pressure (Billaud et al, 2011;Billaud et al, 2015), and neuropathic pain (Bravo et al, 2014;Weaver et al, 2017;Mousseau et al, 2018). While pannexins have limited sequence identity with innexins (~15% identity), they have virtually no sequence similarity to other large-pore forming channels (Panchin et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

The Cryo-EM structure of pannexin 1 reveals unique motifs for ion selection and inhibition

Michalski

Syrjänen

Henze

et al. 2020

eLife

121

150

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Pannexins are large-pore forming channels responsible for ATP release under a variety of physiological and pathological conditions. Although predicted to share similar membrane topology with other large-pore forming proteins such as connexins, innexins, and LRRC8, pannexins have minimal sequence similarity to these protein families. Here, we present the cryo-EM structure of a frog pannexin 1 (Panx1) channel at 3.0 Å. We find that Panx1 protomers harbor four transmembrane helices similar in arrangement to other large-pore forming proteins but assemble as a heptameric channel with a unique constriction formed by Trp74 in the first extracellular loop. Mutating Trp74 or the nearby Arg75 disrupt ion selectivity, whereas altering residues in the hydrophobic groove formed by the two extracellular loops abrogates channel inhibition by carbenoxolone. Our structural and functional study establishes the extracellular loops as important structural motifs for ion selectivity and channel inhibition in Panx1.

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Section: Introductionmentioning

confidence: 99%

The Cryo-EM structure of pannexin 1 reveals unique motifs for ion selection and inhibition

Michalski

Syrjänen

Henze

et al. 2020

eLife

121

150

View full text Add to dashboard Cite

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“…The data are presented as the mean ± SD inhibition of this channel in microglia cells in mouse models can reduce neuronal pain in arteritis. 36 In another study by Diezmos et al, the expression rate of this channel was reduced in the tissues of patients with inflammatory bowel disease. 37 However, the results obtained in this study demonstrate that the expression of PANX1 gene was not significantly different between patients and healthy individuals.…”

Section: Discussionmentioning

confidence: 93%

“…Most studies on this channel have been performed in neural cells and so far, the association of this channel with diseases such as Alzheimer's, diabetes and Crohn's has been investigated . Studies have shown that inhibition of this channel in microglia cells in mouse models can reduce neuronal pain in arteritis . In another study by Diezmos et al, the expression rate of this channel was reduced in the tissues of patients with inflammatory bowel disease .…”

Section: Discussionmentioning

confidence: 99%

P2 receptors mRNA expression profiles in macrophages from ankylosing spondylitis patients and healthy individuals

et al. 2019

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Background Ankylosing spondylitis (AS) is a multifactorial rheumatic disease which mainly involves the axial skeleton. Macrophages and extracellular nucleotides have been shown to contribute to the inflammation process in autoimmune diseases. Membrane‐bound purinergic P2 receptors might be involved in the modulation of immune cells in AS. Therefore, we aimed to analyze the messenger RNA (mRNA) expression of P2 receptors in the macrophages of AS patients and healthy controls. Methods Twenty‐three AS patients and 23 age‐ and sex‐matched healthy individuals were included in our study. Whole blood‐separated monocytes of study participants were stimulated by macrophage colony‐stimulating factor for 7 days and differentiated to macrophages. Monocyte and macrophage markers were analyzed by flow cytometry. SYBR green real‐time polymerase chain reaction was used to measure the relative expression levels of P2RX1, P2RX2, P2RX3, P2RX4, P2RX5, P2RX6, P2RX7, P2RY1, P2RY2, P2RY4, P2RY6, P2RY11, P2RY12, P2RY13, P2RY14, and PANX1 genes. Results P2RY13 and P2RY6 genes had the highest expression levels in macrophages among P2RY genes. P2RY1 mRNA expression was significantly down‐regulated (−1.75 fold) and P2RY14 was up‐regulated (2.6 fold) in macrophages of AS patients compared to healthy individuals. P2RX4 gene had the highest expression in monocyte‐derived macrophages, followed by P2RX7 and P2RX1 genes. There was no significant difference in P2X receptor mRNA expression level between macrophages of AS patients and healthy individuals. Conclusions Our results indicate that AS patients show altered expression levels of P2 receptor genes. Moreover, these changes might be associated with disease activity and patients’ status.

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“…Recent studies indicate that myeloid Panx1 is critical for the progression of joint and nerve-injury pain, likely by attenuating microglial and macrophage activation at the spinal cord 41,42 . A major finding from our work is that myeloid Panx1 promotes acute infiltration of pro-inflammatory cells after brain trauma, which consequently impacts the neuroinflammatory response and outcomes after TBI.…”

Section: Discussionmentioning

confidence: 99%

Myeloid Pannexin-1 mediates acute leukocyte infiltration and leads to worse outcomes after brain trauma.

Seo

Dalal

Calderón

et al. 2020

Preprint

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Background: Neuroinflammation is a major component of secondary damage after traumatic brain injury (TBI). We recently reported that pharmacological inhibition of Pannexin-1 (Panx1) channels markedly reduced the inflammatory response after TBI. Panx1 channels have been shown to be important conduits for adenosine 5′-triphosphate (ATP) release and are associated with leukocyte infiltration and pyroptosis. Because Panx1 blockers significantly decrease ATP release and migration of activated microglia and other myeloid cells (such as monocyte derived macrophages and dendritic cells) in vitro , we hypothesized that myeloid Panx1 channels play a specific role in immune cell infiltration promoting tissue damage following TBI.Methods: The murine controlled cortical impact (CCI) model was used on myeloid-specific Panx1 conditional knockout ( Cx3cr1 - Cre :: Panx1 fl/fl ) mice to determine whether myeloid Panx1 mediates neuroinflammation and brain damage. Immune cell infiltration was measured using flow cytometry. Locomotor and memory functions were measured using the rotarod and Barnes maze test, respectively. The levels of biomarkers for tissue damage and blood brain barrier leakage were measured using western blot and magnetic resonance imaging. Panx1 channel activity was measured with ex vivo dye uptake assays, using flow cytometry and confocal microscopy.Results: CCI-injured Cx3cr1- Cre ::Panx1 fl/fl mice showed markedly reduced immune cell infiltration to the brain parenchyma compared with Panx1 fl/fl mice. As expected, Panx1 dependent activity, assessed by dye uptake, was markedly reduced only in myeloid cells from Cx3cr1- Cre ::Panx1 fl/fl mice. The expression of biomarkers of tissue damage was significantly reduced in the CCI-injured Cx3cr1- Cre ::Panx1 fl/fl mice compared to Panx1 fl/fl mice. In line with this, magnetic resonance imaging showed reduced blood brain barrier leakage in CCI-injured Cx3cr1- Cre ::Panx1 fl/fl mice. There was also a significant improvement in motor and memory function in Cx3cr1- Cre ::Panx1 fl/fl mice when compare to Panx1 fl/fl mice within a week post-CCI injury.Conclusion: Our data demonstrate that CCI-related outcomes correlate with Panx1 channel function in myeloid cells, indicating that activation of Panx1 channels in myeloid cells is a major contributor to acute brain inflammation following TBI. Importantly, our data indicate myeloid Panx1 channels could serve as an effective therapeutic target to improve outcome after TBI.

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Microglial pannexin-1 channel activation is a spinal determinant of joint pain

Abstract: A new therapeutic option for treating arthritis pain.

Cited by 76 publications

References 47 publications

The Cryo-EM structure of pannexin 1 reveals unique motifs for ion selection and inhibition

The Cryo-EM structure of pannexin 1 reveals unique motifs for ion selection and inhibition

P2 receptors mRNA expression profiles in macrophages from ankylosing spondylitis patients and healthy individuals

Myeloid Pannexin-1 mediates acute leukocyte infiltration and leads to worse outcomes after brain trauma.

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