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

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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.

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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

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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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Authors and Affiliations

  1. Department of Orthopedics, The Second Affiliated Hospital of Nantong University, Nantong, 226001, People’s Republic of China

    Dawei Xu, Gang Pan, Xinhui Zhu, Wei Liu, Gang Cai & Zhiming Cui

  2. Department of Neurosurgery, The Second Affiliated Hospital of Nantong University, Nantong, 226001, People’s Republic of China

    Wei Zhao, Ming Qian & Gang Cai

Authors
  1. Dawei Xu
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  2. Wei Zhao
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  3. Gang Pan
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  7. Gang Cai
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  8. Zhiming Cui
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Correspondence to Zhiming Cui.

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Dawei Xu and Wei Zhao contributed equally to this work.

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Xu, D., Zhao, W., Pan, G. et al. Expression of Nemo-like kinase after spinal cord injury in rats. J Mol Neurosci 52, 410–418 (2014). https://doi.org/10.1007/s12031-013-0191-5

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