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Journal of Molecular Neuroscience

, Volume 52, Issue 3, pp 410–418 | Cite as

Expression of Nemo-like kinase after spinal cord injury in rats

  • Dawei Xu
  • Wei Zhao
  • Gang Pan
  • Ming Qian
  • Xinhui Zhu
  • Wei Liu
  • Gang Cai
  • Zhiming Cui
Article

Abstract

Wnt can induce signal transduction via the canonical pathway, which was involved in many processes in the nervous system. Nemo-like kinase (NLK) acts as a negative regulator of β-catenin/T-cell factor/lymphoid enhancer factor (LEF) and functions downstream of transforming growth factor β-activated kinase-1 in the Wnt signaling pathway. In this study, we performed a spinal cord injury (SCI) test in adult Sprague–Dawley rats and investigated the dynamic changes and role of NLK expression in the spinal cord. Western blot analysis revealed that NLK expression was low in normal spinal cord. It then increased markedly, peaked at 3 days, and declined to basal levels from 5 days after injury. Immunohistochemistry confirmed that NLK immunoactivity was expressed at low levels in gray and white matter under normal conditions and increased prominently in gray matter after the SCI test. Double immunofluorescent staining for NLK, caspase-3, β-catenin, and NeuN (neuronal nuclei) revealed that NLK and β-catenin were markedly increased and colocalized in apoptotic neurons. Coimmunoprecipitation data demonstrated that overexpression of NLK protein reduced β-catenin binding to LEF-1. Our results suggested that NLK was associated with neuronal apoptosis through attenuating the Wnt/β-catenin signaling pathway after SCIs.

Keywords

Nemo-like kinase Apoptosis Signaling pathway Spinal cord injury 

Abbreviations

BSA

Bovine serum albumin

CDKs

Cyclin-dependent protein kinases

CNS

Central nervous system

DAB

Diaminobenzidin

ERKs/MAPKs

Extracellular-signal-regulated kinases/mitogen-associated protein kinases

ECL

Enhanced chemiluminescence system

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

LEF

Lymphoid enhancer factor

NeuN

Neuronal nuclei

NLK

Nemo-like kinase

PAGE

Polyacrylamide gel electrophoresis

TAK-1

Transforming growth factor β-activated kinase-1

TCF

T-cell factor

SCI

Spinal cord injury

Notes

Acknowledgments

This work was supported by the National Natural Scientific Foundation of China grant (no. 81300955), projects of Nantong (no. BK2011013 and no. BK2012075).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Dawei Xu
    • 1
  • Wei Zhao
    • 2
  • Gang Pan
    • 1
  • Ming Qian
    • 2
  • Xinhui Zhu
    • 1
  • Wei Liu
    • 1
  • Gang Cai
    • 1
    • 2
  • Zhiming Cui
    • 1
  1. 1.Department of OrthopedicsThe Second Affiliated Hospital of Nantong UniversityNantongPeople’s Republic of China
  2. 2.Department of NeurosurgeryThe Second Affiliated Hospital of Nantong UniversityNantongPeople’s Republic of China

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