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Upregulation of CRM1 Relates to Neuronal Apoptosis after Traumatic Brain Injury in Adult Rats

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Abstract

Traumatic brain injury (TBI) initiates a complex series of neurochemical and signaling changes that leads to neuronal dysfunction and over-reactive astrocytes. There is increasing evidence that CRM1 mediated P27Kip1, which is a potent inhibitor of G1 cyclin-dependent kinases complexes, nuclear export-dependent or -independent Jab1/CSN5, and cytoplasmic degradation in cells. Up to now, the function of CRM1 in central nervous system (CNS) is still with limited acquaintance. In our study, to investigate whether CRM1 is involved in CNS lesion, we performed a TBI model in adult rats. Western blot and RT-PCR analysis revealed that the level of protein and mRNA of CRM1 increased in ipsilateral brain cortex in comparison to the contralateral. Immunohistochemistry and immunofluorescence double labeling indicated that CRM1 was shutting into nucleus around the wound, and increased CRM1 co-localized with P27Kip1. Terminal deoxynucleotidyl transferase deoxy-UTP-nick end labeling (TUNEL) staining suggested that CRM1 was involved in neuronal apoptosis after brain injury. We also investigated co-localization of CRM1 and active-caspase-3 in the ipsilateral brain cortex. In addition, the expression patterns of Bax and active-caspase-3 were parallel with that of CRM1. Based on our data, we suggested that CRM1 might play an important role in neuronal apoptosis following TBI, and might provide a basis for the further study on its role in regulating the expression of P27Kip1 and cell cycle re-entry in TBI.

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Abbreviations

TBI:

Traumatic brain injury

CRM1:

Chromosomal region maintenance 1

P-P27Ser10:

Phosphorylation of P27Kip1 on serine 10

PCNA:

Proliferating cell nuclear antigen

TUNEL:

Terminal deoxynucleotidyl transferase-mediated biotinylated-dUTP nick end labeling

PVDF:

Polyvinylidene difluoride filter

CNS:

Central nervous system

NeuN:

Neuronal nuclei

GFAP:

Glial fibrillary acidic protein

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

DAPI:

4′,6-Diamidino-2-phenylindole

PAGE:

Polyacrylamide gel electrophoresis

CDK:

Cyclin-dependent kinase

SCF:

Skp1/Cul-1/F box protein

ERK:

Extracellular signal-regulated kinase

KPC:

Kip1 ubiquitination-promoting complex

LMB:

Leptomycin B

SDS:

Sodium dodecyl sulfate

BSA:

Bovine serum albumin

DAB:

Diaminobenzidine

PBS:

Phosphate buffer solution

GILZ:

Glucocorticoid-induced Leucine Zipper

JNK:

c-Jun NH2-terminal kinase

VEEV:

Venezuelan equine encephalitis virus

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No.31071288) and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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Authors and Affiliations

  1. Department of Neurology, Affiliated Hospital of Nantong University, Nantong, Jiangsu, 226001, People’s Republic of China

    Aihong Li, Feihui Zou, Hongran Fu, Gang Cui & Yaohua Yan

  2. Jiangsu Key Lab of Neuroregeneration, Nantong, People’s Republic of China

    Qiyun Wu & Xingxing Gu

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  1. Aihong Li
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  2. Feihui Zou
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  3. Hongran Fu
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  4. Gang Cui
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  5. Yaohua Yan
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  6. Qiyun Wu
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  7. Xingxing Gu
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Correspondence to Qiyun Wu or Xingxing Gu.

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Aihong Li and Feihui Zou contributed equally to this work.

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Li, A., Zou, F., Fu, H. et al. Upregulation of CRM1 Relates to Neuronal Apoptosis after Traumatic Brain Injury in Adult Rats. J Mol Neurosci 51, 208–218 (2013). https://doi.org/10.1007/s12031-013-9994-7

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  • DOI: https://doi.org/10.1007/s12031-013-9994-7

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