Tyrosine Kinase Inhibitors Reduce NMDA NR1 Subunit Expression, Nuclear Translocation, and Behavioral Pain Measures in Experimental Arthritis
In the lumbar spinal cord dorsal horn, release of afferent nerve glutamate activates the neurons that relay information about injury pain. Here, we examined the effects of protein tyrosine kinase (PTK) inhibition on NMDA receptor NR1 subunit protein expression and subcellular localization in an acut...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-05-01
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| Series: | Frontiers in Physiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fphys.2020.00440/full |
| _version_ | 1818294645547859968 |
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| author | Karin N. Westlund Karin N. Westlund Karin N. Westlund Ying Lu Liping Zhang Todd C. Pappas Wen-Ru Zhang Giulio Taglialatela Giulio Taglialatela Sabrina L. McIlwrath Terry A. McNearney Terry A. McNearney Terry A. McNearney |
| author_facet | Karin N. Westlund Karin N. Westlund Karin N. Westlund Ying Lu Liping Zhang Todd C. Pappas Wen-Ru Zhang Giulio Taglialatela Giulio Taglialatela Sabrina L. McIlwrath Terry A. McNearney Terry A. McNearney Terry A. McNearney |
| author_sort | Karin N. Westlund |
| collection | DOAJ |
| description | In the lumbar spinal cord dorsal horn, release of afferent nerve glutamate activates the neurons that relay information about injury pain. Here, we examined the effects of protein tyrosine kinase (PTK) inhibition on NMDA receptor NR1 subunit protein expression and subcellular localization in an acute experimental arthritis model. PTK inhibitors genistein and lavendustin A reduced cellular histological translocation of NMDA NR1 in the spinal cord occurring after the inflammatory insult and the nociceptive behavioral responses to heat. The PTK inhibitors were administered into lumbar spinal cord by microdialysis, and secondary heat hyperalgesia was determined using the Hargreaves test. NMDA NR1 cellular protein expression and nuclear translocation were determined by immunocytochemical localization with light and electron microscopy, as well as with Western blot analysis utilizing both C- and N-terminal antibodies. Genistein and lavendustin A (but not inactive lavendustin B or diadzein) effectively reduced (i) pain related behavior, (ii) NMDA NR1 subunit expression increases in spinal cord, and (iii) the shift of NR1 from a cell membrane to a nuclear localization. Genistein pre-treatment reduced these events that occur in vivo within 4 h after inflammatory insult to the knee joint with kaolin and carrageenan (k/c). Cycloheximide reduced glutamate activated upregulation of NR1 content confirming synthesis of new protein in response to the inflammatory insult. In addition to this in vivo data, genistein or staurosporin inhibited upregulation of NMDA NR1 protein and nuclear translocation in vitro after treatment of human neuroblastoma clonal cell cultures (SH-SY5Y) with glutamate or NMDA (4 h). These studies provide evidence that inflammatory activation of peripheral nerves initiates increase in NMDA NR1 in the spinal cord coincident with development of pain related behaviors through glutamate non-receptor, PTK dependent cascades. |
| first_indexed | 2024-12-13T03:35:03Z |
| format | Article |
| id | doaj.art-fbc1653fd82641d5b376bdd486f292ed |
| institution | Directory Open Access Journal |
| issn | 1664-042X |
| language | English |
| last_indexed | 2024-12-13T03:35:03Z |
| publishDate | 2020-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Physiology |
| spelling | doaj.art-fbc1653fd82641d5b376bdd486f292ed2022-12-22T00:01:04ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2020-05-011110.3389/fphys.2020.00440521157Tyrosine Kinase Inhibitors Reduce NMDA NR1 Subunit Expression, Nuclear Translocation, and Behavioral Pain Measures in Experimental ArthritisKarin N. Westlund0Karin N. Westlund1Karin N. Westlund2Ying Lu3Liping Zhang4Todd C. Pappas5Wen-Ru Zhang6Giulio Taglialatela7Giulio Taglialatela8Sabrina L. McIlwrath9Terry A. McNearney10Terry A. McNearney11Terry A. McNearney12Research Division, New Mexico VA Health Care System, Albuquerque, NM, United StatesAnesthesiology, University of New Mexico Health Sciences Center, Albuquerque, NM, United StatesNeuroscience and Cell Biology, University of Texas Medical Branch at Galveston, Galveston, TX, United StatesNeuroscience and Cell Biology, University of Texas Medical Branch at Galveston, Galveston, TX, United StatesNeuroscience and Cell Biology, University of Texas Medical Branch at Galveston, Galveston, TX, United StatesNeuroscience and Cell Biology, University of Texas Medical Branch at Galveston, Galveston, TX, United StatesNeuroscience and Cell Biology, University of Texas Medical Branch at Galveston, Galveston, TX, United StatesNeuroscience and Cell Biology, University of Texas Medical Branch at Galveston, Galveston, TX, United StatesNeurology, University of Texas Medical Branch at Galveston, Galveston, TX, United StatesResearch Division, New Mexico VA Health Care System, Albuquerque, NM, United StatesNeuroscience and Cell Biology, University of Texas Medical Branch at Galveston, Galveston, TX, United StatesMicrobiology and Immunology, University of Texas Medical Branch at Galveston, Galveston, TX, United StatesInternal Medicine, University of Texas Medical Branch at Galveston, Galveston, TX, United StatesIn the lumbar spinal cord dorsal horn, release of afferent nerve glutamate activates the neurons that relay information about injury pain. Here, we examined the effects of protein tyrosine kinase (PTK) inhibition on NMDA receptor NR1 subunit protein expression and subcellular localization in an acute experimental arthritis model. PTK inhibitors genistein and lavendustin A reduced cellular histological translocation of NMDA NR1 in the spinal cord occurring after the inflammatory insult and the nociceptive behavioral responses to heat. The PTK inhibitors were administered into lumbar spinal cord by microdialysis, and secondary heat hyperalgesia was determined using the Hargreaves test. NMDA NR1 cellular protein expression and nuclear translocation were determined by immunocytochemical localization with light and electron microscopy, as well as with Western blot analysis utilizing both C- and N-terminal antibodies. Genistein and lavendustin A (but not inactive lavendustin B or diadzein) effectively reduced (i) pain related behavior, (ii) NMDA NR1 subunit expression increases in spinal cord, and (iii) the shift of NR1 from a cell membrane to a nuclear localization. Genistein pre-treatment reduced these events that occur in vivo within 4 h after inflammatory insult to the knee joint with kaolin and carrageenan (k/c). Cycloheximide reduced glutamate activated upregulation of NR1 content confirming synthesis of new protein in response to the inflammatory insult. In addition to this in vivo data, genistein or staurosporin inhibited upregulation of NMDA NR1 protein and nuclear translocation in vitro after treatment of human neuroblastoma clonal cell cultures (SH-SY5Y) with glutamate or NMDA (4 h). These studies provide evidence that inflammatory activation of peripheral nerves initiates increase in NMDA NR1 in the spinal cord coincident with development of pain related behaviors through glutamate non-receptor, PTK dependent cascades.https://www.frontiersin.org/article/10.3389/fphys.2020.00440/fullpainmembrane traffickinginflammationglutamatecentral sensitizationgenistein |
| spellingShingle | Karin N. Westlund Karin N. Westlund Karin N. Westlund Ying Lu Liping Zhang Todd C. Pappas Wen-Ru Zhang Giulio Taglialatela Giulio Taglialatela Sabrina L. McIlwrath Terry A. McNearney Terry A. McNearney Terry A. McNearney Tyrosine Kinase Inhibitors Reduce NMDA NR1 Subunit Expression, Nuclear Translocation, and Behavioral Pain Measures in Experimental Arthritis Frontiers in Physiology pain membrane trafficking inflammation glutamate central sensitization genistein |
| title | Tyrosine Kinase Inhibitors Reduce NMDA NR1 Subunit Expression, Nuclear Translocation, and Behavioral Pain Measures in Experimental Arthritis |
| title_full | Tyrosine Kinase Inhibitors Reduce NMDA NR1 Subunit Expression, Nuclear Translocation, and Behavioral Pain Measures in Experimental Arthritis |
| title_fullStr | Tyrosine Kinase Inhibitors Reduce NMDA NR1 Subunit Expression, Nuclear Translocation, and Behavioral Pain Measures in Experimental Arthritis |
| title_full_unstemmed | Tyrosine Kinase Inhibitors Reduce NMDA NR1 Subunit Expression, Nuclear Translocation, and Behavioral Pain Measures in Experimental Arthritis |
| title_short | Tyrosine Kinase Inhibitors Reduce NMDA NR1 Subunit Expression, Nuclear Translocation, and Behavioral Pain Measures in Experimental Arthritis |
| title_sort | tyrosine kinase inhibitors reduce nmda nr1 subunit expression nuclear translocation and behavioral pain measures in experimental arthritis |
| topic | pain membrane trafficking inflammation glutamate central sensitization genistein |
| url | https://www.frontiersin.org/article/10.3389/fphys.2020.00440/full |
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