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Glutamatergic Neurotransmission Expression Profiling in the Mouse Hippocampus After Perforant-Path Transection

Received March 30, 2005; revised April 26, 2005; accepted June 21, 2005. From the Center for Dementia Research, Nathan Kline Institute, Department of Psychiatry, and Department of Physiology and Neuroscience, New York University School of Medicine, Orangeburg, NY 10962 Send correspondence and reprint requests to Stephen D. Ginsberg, Ph.D., Center for Dementia Research, Nathan Kline Institute, New York University School of Medicine, 140 Old Orangeburg Road, Orangeburg, NY 10962. e-mail: ginsberg{at}nki.rfmh.org
© American Association for Geriatric Psychiatry

Objective: The goal is to determine cellular and molecular mechanisms that regulate regenerative and neurodegenerative responses within the adult mouse dentate gyrus after axotomy of the principal glutamatergic input, the perforant path (PP). Methods: A "molecular fingerprint" of the dentate gyrus was generated to provide an extensive, concurrent representation of genes, with an emphasis on glutamate receptor subunits and related markers of glutamatergic neurotransmission. Reorganization of the hippocampal formation was evaluated by regional microdissection of the dentate gyrus, followed by terminal continuation RNA amplification and custom-designed cDNA microarray analysis after unilateral PP transections at five time-points (0, 1, 10, 14, and 30 days post-lesion). Gene-expression profiles garnered from the ipsilateral side of PP transected hippocampal formation (including the dentate gyrus) were compared and contrasted with those of naive subjects, sham surgical subjects, and mice subjected to unilateral occipital cortex lesions. Specific gene array observations were validated by immunoblot analysis. Results: Results indicated selective regulation of specific transcripts, including AMPA and NMDA glutamate-receptor subunits, excitatory amino acid transporters, glutamate receptor interacting protein genes, and glial-associated markers across the time-course of the lesion study. Conclusion: The goal was to identify messenger RNAs from specific classes of relevant transcripts that change over time in relationship to the synaptic and cellular alterations to help understand mechanisms that underlie lesion-induced synaptic plasticity.

Key Words: Excitotoxicity • Functional Genomics • Molecular Biology • Dentate Gyrus • Microarray

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