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The molecular cloning of glial fibrillary acidic protein in Gekko japonicus and its expression changes after spinal cord transection

Abstract

The glial fibrillary acidic protein (GFAP) is an astrocyte-specific member of the class III intermediate filament proteins. It is generally used as a specific marker of astrocytes in the central nervous system (CNS). We isolated a GFAP cDNA from the brain and spinal cord cDNA library of Gekko japonicus, and prepared polyclonal antibodies against gecko GFAP to provide useful tools for further immunochemistry studies. Both the real-time quantitative PCR and western blot results revealed that the expression of GFAP in the spinal cord after transection increased, reaching its maximum level after 3 days, and then gradually decreased over the rest of the 2 weeks of the experiment. Immunohistochemical analyses demonstrated that the increase in GFAP-positive labeling was restricted to the white matter rather than the gray matter. In particular, a slight increase in the number of GFAP positive star-shaped astrocytes was detected in the ventral and lateral regions of the white matter. Our results indicate that reactive astrogliosis in the gecko spinal cord took place primarily in the white matter during a short time interval, suggesting that the specific astrogliosis evaluated by GFAP expression might be advantageous in spinal cord regeneration.

Abbreviations

GFAP:

glial fibrillaryacidic protein

IPTG:

isopropyl-β-D-thiogalactopyranoside

RACE:

rapid amplification of cDNA ends

SDS-PAGE:

sodium dodecyl sulfate polyacrylamide gel electrophoresis

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Correspondence to Xiaosong Gu or Zhengli Li.

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Key words

  • Glial fibrillary acidic protein
  • Gecko
  • Regeneration
  • Spinal cord
  • Real-time PCR