TY - JOUR
T1 - Tyrosine Kinase Inhibitors Reduce NMDA NR1 Subunit Expression, Nuclear Translocation, and Behavioral Pain Measures in Experimental Arthritis
AU - Westlund, Karin N.
AU - Lu, Ying
AU - Zhang, Liping
AU - Pappas, Todd C.
AU - Zhang, Wen Ru
AU - Taglialatela, Giulio
AU - McIlwrath, Sabrina L.
AU - McNearney, Terry A.
N1 - Funding Information:
Funding. These studies were funded by the Merit Review Award BX002695, Office of Research and Development, Medical Research Service, United States Department of Veterans Affairs. This communication does not necessarily reflect the views of the Department of Veterans Affairs or the U.S. government. These studies were also supported by grants from the Sealy Endowment; NIH NS32778 to KW; NIH P01NS11255 to KW, TM, and GT. NIH R21 AR48371 to KW and TM; the Dana Foundation; and a UTMB endowment award to KW and TM.
Funding Information:
These studies were funded by the Merit Review Award BX002695, Office of Research and Development, Medical Research Service, United States Department of Veterans Affairs. This communication does not necessarily reflect the views of the Department of Veterans Affairs or the U.S. government. These studies were also supported by grants from the Sealy Endowment; NIH NS32778 to KW; NIH P01NS11255 to KW, TM, and GT. NIH R21 AR48371 to KW and TM; the Dana Foundation; and a UTMB endowment award to KW and TM.
Publisher Copyright:
© Copyright © 2020 Westlund, Lu, Zhang, Pappas, Zhang, Taglialatela, McIlwrath and McNearney.
PY - 2020/5/27
Y1 - 2020/5/27
N2 - 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.
AB - 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.
KW - central sensitization
KW - genistein
KW - glutamate
KW - inflammation
KW - membrane trafficking
KW - pain
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UR - http://www.scopus.com/inward/citedby.url?scp=85086230078&partnerID=8YFLogxK
U2 - 10.3389/fphys.2020.00440
DO - 10.3389/fphys.2020.00440
M3 - Article
AN - SCOPUS:85086230078
SN - 1664-042X
VL - 11
JO - Frontiers in Physiology
JF - Frontiers in Physiology
M1 - 440
ER -