P2Y2 receptor expression is altered in rats after spinal cord injury

Abstract

Spinal cord injury increases inhibitory factors that may restrict neurite outgrowth after trauma. The expression of repulsive molecules in reactive astrocytes and the formation of the glial scar at the injury site produce the non-permissive environment for axonal regeneration. However, the mechanism that triggers this astrogliotic response is unknown. The release of nucleotides has been linked to this hypertrophic state.Our goal is to investigate the temporal profile of P2Y2 nucleotide receptor after spinal cord injury in adult female Sprague-Dawley rats. Molecular biology, immunofluorescence studies, and Western Blots were used to evaluate the temporal profile (2, 4, 7, 14, and 28 days post-injury) of this receptor in rats injured at the T-10 level using the NYU impactor device.Real time RT-PCR showed a significant increase of P2Y2 mRNA after 2 days post-injury that continues throughout 28 days post-injury. Double labeling studies localized P2Y2 immunoreactivity in neuronal cell bodies, axons, macrophages, oligodendrocytes and reactive astrocytes. Immunofluorescence studies also demonstrated a low level of P2Y2 receptor in sham samples, which increased after injury in glial fibrillary acidic protein positive cells. Western Blot performed with contused spinal cord protein samples revealed an upregulation in the P2Y2 42kDa protein band expression after 4 days post-injury that continues until 28 days post-injury. However, a downregulation of the 62kDa receptor protein band after 2 days post-injury that continues up to 28 days post-injury was observed.Therefore, the spatio-temporal pattern of P2Y2 gene expression after spinal cord injury suggests a role in the pathophysiology response generated after trauma. © 2010.