Rapid <it>S</it>-nitrosylation of actin by NO-generating donors and in inflammatory pain model mice
<p>Abstract</p> <p>Background</p> <p><it>S</it>-Nitrosylation, the reversible post-translational modification of reactive cysteine residues in proteins, has emerged as an important mechanism by which NO acts as a signaling molecule. We recently demonstrated...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2011-12-01
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| Series: | Molecular Pain |
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| Online Access: | http://www.molecularpain.com/content/7/1/101 |
| _version_ | 1828410209399209984 |
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| author | Lu Jingshan Katano Tayo Uta Daisuke Furue Hidemasa Ito Seiji |
| author_facet | Lu Jingshan Katano Tayo Uta Daisuke Furue Hidemasa Ito Seiji |
| author_sort | Lu Jingshan |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p><it>S</it>-Nitrosylation, the reversible post-translational modification of reactive cysteine residues in proteins, has emerged as an important mechanism by which NO acts as a signaling molecule. We recently demonstrated that actin is a major <it>S</it>-nitrosylated protein in the spinal cord and suggested that NO directly attenuates dopamine release from PC12 cells by causing the breakdown of F-actin. However, the occurrence of <it>S</it>-nitrosylation of actin remained unclarified in animal pain model. Kinetic analysis of <it>S</it>-nitrosylation of actin in the present study was made by using NO-generating donors. The biotin-switch assay and purification on streptavidin-agarose were employed for identification of <it>S</it>-nitrosylated actin.</p> <p>Results</p> <p>Dopamine release from PC12 cells was markedly attenuated by NOR1 (<it>t</it><sub>1/2 </sub>= 1.8 min) and much less by NOR3 (<it>t</it><sub>1/2 </sub>= 30 min), but not by <it>S</it>-nitroso-glutathione, an endogenous NO donor. A membrane-permeable cGMP analogue could not substitute for NOR1 as a suppressor nor could inhibitors of soluble guanylate cyclase and cGMP-dependent protein kinase attenuate the suppression. <it>S</it>-Nitrosylated actin was detected by the biotin-switch assay at 5 min after the addition of NOR1. Consistent with the kinetic analysis, actin in the spinal cord was rapidly and maximally <it>S</it>-nitrosylated in an inflammatory pain model at 5 min after the injection of 2% formalin into the hind paws. <it>In vivo </it>patch-clamp recordings of the spinal dorsal horn, NOR3 showed an inhibitory action on inhibitory synaptic transmission in interneurons of the substantia gelatinosa.</p> <p>Conclusions</p> <p>The present study demonstrates that rapid <it>S</it>-nitrosylation of actin occurred <it>in vitro </it>in the presence of exogenous NO-generating donors and <it>in vivo </it>in inflammatory pain model mice. Our data suggest that, in addition to the well-known cGMP-dependent protein kinase pathway, <it>S</it>-nitrosylation is involved in pain transmission via disinhibition of inhibitory neurons.</p> |
| first_indexed | 2024-12-10T12:08:15Z |
| format | Article |
| id | doaj.art-5d53261003c14a4db46c9e216395b3ab |
| institution | Directory Open Access Journal |
| issn | 1744-8069 |
| language | English |
| last_indexed | 2024-12-10T12:08:15Z |
| publishDate | 2011-12-01 |
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| series | Molecular Pain |
| spelling | doaj.art-5d53261003c14a4db46c9e216395b3ab2022-12-22T01:49:26ZengSAGE PublishingMolecular Pain1744-80692011-12-017110110.1186/1744-8069-7-101Rapid <it>S</it>-nitrosylation of actin by NO-generating donors and in inflammatory pain model miceLu JingshanKatano TayoUta DaisukeFurue HidemasaIto Seiji<p>Abstract</p> <p>Background</p> <p><it>S</it>-Nitrosylation, the reversible post-translational modification of reactive cysteine residues in proteins, has emerged as an important mechanism by which NO acts as a signaling molecule. We recently demonstrated that actin is a major <it>S</it>-nitrosylated protein in the spinal cord and suggested that NO directly attenuates dopamine release from PC12 cells by causing the breakdown of F-actin. However, the occurrence of <it>S</it>-nitrosylation of actin remained unclarified in animal pain model. Kinetic analysis of <it>S</it>-nitrosylation of actin in the present study was made by using NO-generating donors. The biotin-switch assay and purification on streptavidin-agarose were employed for identification of <it>S</it>-nitrosylated actin.</p> <p>Results</p> <p>Dopamine release from PC12 cells was markedly attenuated by NOR1 (<it>t</it><sub>1/2 </sub>= 1.8 min) and much less by NOR3 (<it>t</it><sub>1/2 </sub>= 30 min), but not by <it>S</it>-nitroso-glutathione, an endogenous NO donor. A membrane-permeable cGMP analogue could not substitute for NOR1 as a suppressor nor could inhibitors of soluble guanylate cyclase and cGMP-dependent protein kinase attenuate the suppression. <it>S</it>-Nitrosylated actin was detected by the biotin-switch assay at 5 min after the addition of NOR1. Consistent with the kinetic analysis, actin in the spinal cord was rapidly and maximally <it>S</it>-nitrosylated in an inflammatory pain model at 5 min after the injection of 2% formalin into the hind paws. <it>In vivo </it>patch-clamp recordings of the spinal dorsal horn, NOR3 showed an inhibitory action on inhibitory synaptic transmission in interneurons of the substantia gelatinosa.</p> <p>Conclusions</p> <p>The present study demonstrates that rapid <it>S</it>-nitrosylation of actin occurred <it>in vitro </it>in the presence of exogenous NO-generating donors and <it>in vivo </it>in inflammatory pain model mice. Our data suggest that, in addition to the well-known cGMP-dependent protein kinase pathway, <it>S</it>-nitrosylation is involved in pain transmission via disinhibition of inhibitory neurons.</p>http://www.molecularpain.com/content/7/1/101dopamine releaseF-actininflammatory painnitric oxidePC12 cell<it>S</it>-nitrosylationspinal cord<it>in vivo </it>patch-clamp recordings |
| spellingShingle | Lu Jingshan Katano Tayo Uta Daisuke Furue Hidemasa Ito Seiji Rapid <it>S</it>-nitrosylation of actin by NO-generating donors and in inflammatory pain model mice Molecular Pain dopamine release F-actin inflammatory pain nitric oxide PC12 cell <it>S</it>-nitrosylation spinal cord <it>in vivo </it>patch-clamp recordings |
| title | Rapid <it>S</it>-nitrosylation of actin by NO-generating donors and in inflammatory pain model mice |
| title_full | Rapid <it>S</it>-nitrosylation of actin by NO-generating donors and in inflammatory pain model mice |
| title_fullStr | Rapid <it>S</it>-nitrosylation of actin by NO-generating donors and in inflammatory pain model mice |
| title_full_unstemmed | Rapid <it>S</it>-nitrosylation of actin by NO-generating donors and in inflammatory pain model mice |
| title_short | Rapid <it>S</it>-nitrosylation of actin by NO-generating donors and in inflammatory pain model mice |
| title_sort | rapid it s it nitrosylation of actin by no generating donors and in inflammatory pain model mice |
| topic | dopamine release F-actin inflammatory pain nitric oxide PC12 cell <it>S</it>-nitrosylation spinal cord <it>in vivo </it>patch-clamp recordings |
| url | http://www.molecularpain.com/content/7/1/101 |
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