Sciatic nerve regeneration in KLF7-transfected acellular nerve allografts

Neurol Res. 2016 Mar;38(3):242-54. doi: 10.1080/01616412.2015.1105584. Epub 2016 Apr 19.

Abstract

Objective: Krüppel-like Factor 7 (KLF7) is a transcription factor that promotes axon regeneration in the central nervous system. Here, we assessed whether KLF7 stimulates regeneration after peripheral nerve injury.

Methods: C57BL/6 mice received an acellular nerve allograft (ANA) injected with either adeno-associated virus 2 (AAV2) vector or AAV2-KLF7 for sciatic nerve gap repair. After 4 weeks, KLF7 was detected by RT-PCR, western blot and immunohistochemistry in regenerated nerves. Axonal regeneration and functional recovery were examined by immunohistochemistry, Fluorogold (FG) and cholera toxin B (CTB) retrograde neural tracing, sciatic function index (SFI), angle of ankle, Hargreaves test and electrophysiological analysis.

Results: With AAV2-KLF7 injection, KLF7 expression increased in regenerated nerves, and amplitude, score of SFI, angle of ankle and FG-labelled spinal cord neurons were increased. We observed elevated CTB-labelled neurons in dorsal root ganglia (DRG), neurofilaments, P0 (peripheral myelin) and S100 and decreased latency period and withdrawal latencies in the Hargreaves test. The SFI was significantly correlated with amplitude and regenerated axon number. Tyrosine kinase A (TrkA) and B (TrkB) receptors were also increased in the DRG.

Conclusions: Our findings suggest that KLF7 promoted peripheral nerve axonal regeneration, further supporting a role for KLF7 as a growth-promoting transcription factor in the injured nervous system.

Keywords: Acellular nerve allograft; Axonal regeneration; Fluorogold (FG); KLF7; Krüppel-like Factor 7 (KLF7); Peripheral nerve injury; acellular nerve allograft (ANA); adeno-associated virus 2 (AAV2); central nervous system (CNS); cholera toxin B (CTB); day (d); dorsal root ganglia (DRG); electroneurogram (ENG); glial fibrillary acidic proteins (GFAP); neurofilaments (NF); peripheral nervous system (PNS); sciatic function index (SFI); small proline-rich repeat protein 1A (SPRR1A); tyrosine kinase A (TrkA).

MeSH terms

  • Allografts / metabolism*
  • Allografts / ultrastructure
  • Animals
  • Cholera Toxin / metabolism
  • Dependovirus / genetics
  • Disease Models, Animal
  • Female
  • Ganglia, Spinal / metabolism
  • Ganglia, Spinal / pathology
  • Gene Expression Regulation / genetics*
  • Glial Fibrillary Acidic Protein / metabolism
  • Intermediate Filaments / metabolism
  • Kruppel-Like Transcription Factors / genetics
  • Kruppel-Like Transcription Factors / metabolism*
  • Kruppel-Like Transcription Factors / therapeutic use
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred ICR
  • Myelin P0 Protein / metabolism
  • Nerve Regeneration / physiology*
  • S100 Calcium Binding Protein beta Subunit / metabolism
  • Sciatic Neuropathy / pathology
  • Sciatic Neuropathy / surgery*
  • Spinal Cord / pathology
  • Time Factors
  • Transduction, Genetic

Substances

  • Glial Fibrillary Acidic Protein
  • Klf7 protein, mouse
  • Kruppel-Like Transcription Factors
  • Myelin P0 Protein
  • S100 Calcium Binding Protein beta Subunit
  • Cholera Toxin