Effect of protein kinase C blockade on phosphorylation of NR1 in dorsal horn and spinothalamic tract cells caused by intradermal capsaicin injection in rats.
We have previously reported that protein kinase A (PKA) is involved in the phosphorylation of NR1 subunits of N-methyl-d-aspartate (NMDA) receptors in dorsal horn neurons after intradermal injection of capsaicin (CAP). To see if protein kinase C (PKC) also participates in the phosphorylation of NR1, we used electron microscopic techniques to determine further where the phosphorylated NR1 subunits (pNR1) are expressed in the spinothalamic tract (STT) cells and immunohistochemistry to examine whether a PKC inhibitor, chelerythrine chloride, blocks the enhanced phosphorylation of NR1 on serine 896. The pNR1 subunits were in the soma and dendrites of STT cells and in presynaptic endings. Western blots showed that pretreatment with the PKC inhibitor caused a decrease in CAP-induced phosphorylation of NR1 protein. In immunofluorescence staining, the number of pNR1-like immunoreactive neurons was significantly decreased on the side ipsilateral to the injection when chelerythrine chloride was administered intrathecally before CAP injection. In addition, when STT cells were labeled by microinjection of the retrograde tracer, fluorogold (FG), into the thalamus, we found that the proportion of p-NR1-LI STT cells was markedly reduced after PKC inhibition. Combined with our previous findings, these results strongly suggest that NR1 subunits in spinal dorsal horn neurons are phosphorylated following CAP injection, and this phosphorylation is catalyzed by PKC, as well as by PKA.
Department of Anatomy and Neurosciences, Marine Biomedical Institute, The University of Texas Medical Branch, Galveston, TX 77555-1069, USA.
Xiaoju
X
Zou
Qing
Q
Lin
William D
WD
Willis
D000470:Alkaloids; D000818:Animals; D053119:Benzophenanthridines; D002211:Capsaicin; D004791:Enzyme Inhibitors / Q000008:administration & dosage; D005455:Fluorescent Antibody Technique; D007839:Functional Laterality / Q000502:physiology; D007271:Injections, Intradermal; D007278:Injections, Spinal; D008297:Male; D009619:Nociceptors / Q000187:drug effects / Q000378:metabolism*; D010146:Pain / Q000139:chemically induced / Q000201:enzymology*; D010617:Phenanthridines / Q000008:administration & dosage; D010766:Phosphorylation; D020671:Posterior Horn Cells / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000378:metabolism*; D013133:Spinothalamic Tracts / Q000187:drug effects / Q000201:enzymology* / Q000648:ultrastructure
D000470:Alkaloids; D053119:Benzophenanthridines; D004791:Enzyme Inhibitors; C409466:NR1 NMDA receptor; D010617:Phenanthridines; D016194:Receptors, N-Methyl-D-Aspartate; C016299:chelerythrine; D011493:Protein Kinase C; D002211:Capsaicin