A critical role of SRC-suppressed C kinase substrate in rat astrocytes after chronic constriction injury.
Src-suppressed C kinase substrate (SSeCKS), is an in vivo and in vitro protein kinase C substrate that may have a role in both mitogenic regulation and cytoskeletal arrangement. In this study, we mainly investigated the mRNA and protein expression and cellular localization of SSeCKS during chronic constriction injury (CCI). Reverse transcriptase-mediated PCR and western blot analysis revealed that SSeCKS was present in normal whole spinal cord. It gradually increased, and reached a peak at 2 weeks for its mRNA level and 7 days for its protein level after CCI. The protein expression of SSeCKS was further analyzed by immunohistochemistry. The positively stained areas for SSeCKS changed with the similar pattern to that of protein expression detected by immunoblotting analysis. Double immunofluorescence staining showed SSeCKS immunoreactivity was mostly co-localized with neurons, partly with activated astrocytes and rarely with microglia in the superficial laminar of spinal dorsal horn. In cell culture, the expression of pro-inflammation cytokines, p-ERK, and SSeCKS was increased in the spinal astrocytes after stimulated by damaged sensory neurons. However, SSeCKS gene silencing by siRNA inhibited the up-regulation of p-ERK and the pro-inflammation cytokines. Taken together, activated astrocytes released cytokines and iNOS after neuropathic pain via SSeCKS-ERK signaling. SSeCKS might be critical for the activation of astrocytes in the neuropathic pain.
Xia
Yinyin
Y
Department of Microbiology and Immunology, Medical College, Nantong University, Nantong, Jiangsu, China.
Liu
Haiou
H
Shen
Aiguo
A
Liu
Yonghua
Y
Sun
Linlin
L
Tao
Tao
T
Ke
Qing
Q
Cheng
Chun
C
D054758:A Kinase Anchor Proteins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism* / Q000473:pathology*; D001522:Behavior, Animal; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism; D004789:Enzyme Activation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D020868:Gene Silencing; D008297:Male; D010147:Pain Measurement; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011984:Sensory Receptor Cells / Q000166:cytology / Q000378:metabolism / Q000473:pathology; D013116:Spinal Cord / Q000166:cytology / Q000378:metabolism / Q000473:pathology*
D054758:A Kinase Anchor Proteins; C102361:Akap12 protein, rat; D018797:Cell Cycle Proteins; D016207:Cytokines; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D048049:Extracellular Signal-Regulated MAP Kinases