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Expression of G-protein-coupled receptor kinase 6 (GRK6) after acute spinal cord injury in adult rat

Journal of Molecular Histology volume 44, pages 259–270 (2013)Cite this article

Abstract

Spinal cord injury frequently results in permanent loss of neurological function. It includes many complex molecular and biochemical mechanisms. G-protein-coupled receptor kinase 6 (GRK6) is an intracellular kinase that regulates the sensitivity of certain G-protein-coupled receptors. Some studies reported GRK2 and GRK5 modulate the NFκB pathway in macrophages. Additionally, GRK2 is referred to as regulating activation of spinal cord microglia and GRK6 expression is significantly elevated in most brain regions in the MPTP-lesioned parkinsonian monkeys. However, the expression and function of GRK6 in nervous system lesion and repair are not well understood. In this study, we performed an acute spinal cord injury (SCI) model in adult rats. Western blot analysis showed the expression of GRK6 was upregulated significantly at protein level in spinal cord after SCI. Immunohistochemistry and immunofluorescence revealed wide expression of GRK6 in the normal spinal cord. After injury, GRK6 expression was increased predominantly in microglia, which expressed F4/80 (marker of macrophages and activated microglia) strongly. To understand whether GRK6 played a role in microglia activation, we applied lipopolysaccharide (LPS) to induce microglia activation in vitro. Western blot analysis demonstrated up-regulation in GRK6 protein expression after LPS stimulation was time- and dose-dependent and that up-regulation in F4/80 expression was concomitant with GRK6. These data suggested that GRK6 might be involved in the pathophysiology of SCI.

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Abbreviations

BSA:

Bovine serum albumin

DAB:

Diaminobenzidine

ECL:

Enhanced chemiluminescence system

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GFAP:

Glial fibrillary acidic protein

GPCRS:

G protein-coupled receptors

GRK:

G-protein-coupled receptor kinase

HAPI:

Highly aggressively proliferating immortalized

IκBα:

Inhibitor of κBα

LPS:

Lipopolysaccharide

LTB4:

Leukotriene B4

NF-κB:

Nuclear factor κB

PAGE:

Polyacrylamide gel electrophoresis

RT:

Room temperature

SCI:

Spinal cord injury

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 81070992); A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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Affiliations

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

    Binbin Sun, Yilu Gao, Dong Lou, Xiujie Wu, Haixiang Wei, Hai Wen, Xiaolong Deng & Feng Zhang

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  1. Binbin Sun
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  2. Yilu Gao
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  7. Xiaolong Deng
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  8. Feng Zhang
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Correspondence to Feng Zhang.

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Sun, B., Gao, Y., Lou, D. et al. Expression of G-protein-coupled receptor kinase 6 (GRK6) after acute spinal cord injury in adult rat. J Mol Hist 44, 259–270 (2013). https://doi.org/10.1007/s10735-013-9486-7

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  • DOI: https://doi.org/10.1007/s10735-013-9486-7

Keywords

  • SCI
  • GRK6
  • Microglia
  • Activation
  • Rats