Temporal-spatial expressions of p27kip1 and its phosphorylation on Serine-10 after acute spinal cord injury in adult rat: Implications for post-traumatic glial proliferation.
P27kip1, as a member of Cip/Kip family of cyclin-dependent kinase inhibitors, plays important roles in cell cycle regulation and neurogenesis in the developing central nervous system. Serine-10 is the major phosphorylation site of p27kip1, and post-translational regulation of p27kip1 by different phosphorylation events is critical for its function. To elucidate the expressions and possible functions of p27kip1 and its phosphorylation in central nervous system lesion and repair, we performed an acute spinal cord contusion injury model in adult rats. Our work studied the temporal-spatial expression patterns of p27kip1 and Serine-10 phosphorylated p27kip1 (p-p27s10). Western blot analysis showed p27kip1 level significantly decreased at day 3 after damage, while p-p27s10 was detected at a high-level at the same time reaching the uninjured level. Moreover, immunofluorescence double labeling suggested these changes were striking in microglia and astrocytes, which were largely proliferated. Immunohistochemical analysis revealed subcellular localization changes of p27kip1 and p-p27s10 staining between nucleus and cytoplasm after injury in about 20% of total positive cells including neurons and glial cells. We also investigated the increased interactions of p27kip1 and p-p27s10 with CRM1 3 days after injury by co-immunoprecipitation studies. Taken together, we hypothesized spinal cord injury stimulated mitogenic signals to induce a serine-threonine kinase KIS (kinase interacting stathmin) to phosphorylate p27kip1 on Serine-10, so that p27kip1 could bind to CRM1 and be exported from nuclei for degradation. Such an event facilitated cell cycle progression of glial cells, especially microglia and astrocytes which had a prevalent proliferation.
Shen
Aiguo
A
The Jiangsu Province Key Laboratory of Neuroregeneration, Nantong University, Nantong 226001, China.
Liu
Yonghua
Y
Zhao
Jian
J
Qin
Jing
J
Shi
Shuxian
S
Chen
Mengling
M
Gao
Shangfeng
S
Xiao
Feng
F
Lu
Qiuyan
Q
Cheng
Chun
C
D021581:Active Transport, Cell Nucleus / Q000502:physiology; D000208:Acute Disease; D000595:Amino Acid Sequence / Q000502:physiology; D000818:Animals; D001253:Astrocytes / Q000378:metabolism; D049109:Cell Proliferation; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000737:chemistry / Q000378:metabolism*; D018450:Disease Progression; D015536:Down-Regulation / Q000502:physiology; D005455:Fluorescent Antibody Technique; D005911:Gliosis / Q000209:etiology / Q000378:metabolism* / Q000503:physiopathology; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D028884:Karyopherins / Q000378:metabolism; D008297:Male; D017628:Microglia / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D012694:Serine / Q000378:metabolism*; D013116:Spinal Cord / Q000378:metabolism* / Q000503:physiopathology; D013119:Spinal Cord Injuries / Q000150:complications / Q000378:metabolism* / Q000503:physiopathology; D013997:Time Factors; D015854:Up-Regulation / Q000502:physiology; D000097606:Exportin 1 Protein
D050760:Cyclin-Dependent Kinase Inhibitor p27; D047908:Intracellular Signaling Peptides and Proteins; D028884:Karyopherins; D017346:Protein Serine-Threonine Kinases; D018160:Receptors, Cytoplasmic and Nuclear; D012694:Serine; D000097606:Exportin 1 Protein; C493962:Cdkn1b protein, rat