Add time:09/26/2019         Source:sciencedirect.com

A set of well-defined antisera against neuronal and glial proteins were used to characterize patterns of protein expression in rat hippocampus following transection of the fimbira–fornix and perforant pathways or after administration of the selective neurotoxicant trimethyltin (8 mg/kg, i.p.). SNAP-25 (synaptosomal protein, mol. wt 25 000) is a neuron-specific, developmentally regulated presynaptic protein, stannin is a protein enriched in cells sensitive to trimethyltin, and GAP-43 (growth-associated protein, mol. wt 43 000) is associated with axonal growth and regeneration. Glial fibrillary acidic protein is an astrocyte-specific intermediate filament protein and a marker for reactive gliosis. SNAP-25 immunoreactivity was altered following both neurotoxicant and mechanical injury. Three days after fimbria–fornix/perforant path lesions, there was a loss of SNAP-25 immunoreactivity in hippocampal efferent pathways and in the lesioned entorhinal cortex. By day 12, there was evidence of reinnervation of hippocampal subfields by SNAP-25-immunopositive commissural afferent fibers. On day 3, immunoblots showed the appearance of SNAP-25a, a developmental isoform produced by alternative splicing of nine amino acids in exon 5, in lesioned tissues. This isoform declined by day 12 and was not found in contralateral control hippocampus or non-lesioned brain regions. Stannin immunoreactivity was unchanged, while GAP-43 was prominent on day 12 post-lesion. Glial fibrillary acidic protein immunoreactivity indicated gliosis near the site of pathway transection. In contrast, trimethyltin induced a marked loss of stannin immunoreactivity in hippocampal neurons seven days after injection. Trimethyltin increased glial fibrillary acidic protein staining in the hippocampus and other damaged regions. SNAP-25 immunoreactivity was markedly increased in mossy fibers and other hippocampal fields seven days following trimethyltin. Immunoblot analysis showed that only the adult SNAP-25b isoform was expressed after trimethyltin intoxication.These data suggest that SNAP-25 is a useful marker for presynaptic damage. Furthermore, re-expression of developmental isoforms of SNAP-25a may precede functional reinnervation when the postsynaptic target remains intact.

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