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Phorbol ester-induced neuritic alterations in the rat neocortex

Structural and immunocytochemical studies

  • Original Articles
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Molecular and Chemical Neuropathology

Abstract

In order to explore the effect of aberrant sprouting in the CNS, phorbol 12-myristate 13-acetate (PMA) was administered into the neocortex of adult rats. PMA is a growth-promoting agent that activates and eventually downregulates protein kinase C (PKC), and induces in the rat the expression of several genes, including amyloid precursor protein (APP). We found that multiple injections of 100 nM PMA into the rat neocortex promote, in the first week postinjection, a widespread vacuolization of the neuropil with a subsequent disruption of the synapses in the injection site, followed, at d 15, by the formation of abnormally distended clusters of neurites that resembled aberrant, sprouting axons. At d 30, fewer aberrant sprouts were observed, and many degenerating neurites were found. At the ultrastructural level, the PMA-induced abnormal neurites at d 7–15 resembled growth cones, whereas the dystrophic neurites at d 30 contained abundant dense and laminated bodies. Immunohistochemical analysis indicated that the abnormal neurites in the areas of denervation and PMA administration were positive with antisynaptophysin and antigrowth-associated protein 43 (GAP-43), with an increased APP immunoreactivity surrounding them. APP immunoreactivity around the injection site was mostly associated with pyramidal neurons and glial cells. Control experiments, where saline alone or 4α-phorbol 12, 13-didecanoate (PDD, an inactive phorbol derivative) was injected, failed to show aberrant sprouting neurites. Further immunohistochemical analysis showed that the PMA-treated animals presented increased amyloidΒ immunoreactivity in the pyramidal cells at the site of injection, when compared with control injections. These findings suggest that aberrant sprouting induced by overstimulation could be followed by neurodegeneration. Alternatively, PKC downregulation could directly induce the neurodegeneration, with a secondary sprouting response.

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Authors and Affiliations

  1. Department of Neurosciences, University of California, San Diego, School of Medicine, 92093-0624, La Jolla, CA

    Eliezer Masliah, Margaret Mallory, Nianfeng Ge & Tsunao Saitoh

  2. Department of Pharmacology, University of California, San Diego, School of Medicine, 92093-0624, La Jolla, CA

    Catherine Godson

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  1. Eliezer Masliah
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  2. Margaret Mallory
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  3. Nianfeng Ge
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  4. Catherine Godson
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  5. Tsunao Saitoh
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Masliah, E., Mallory, M., Ge, N. et al. Phorbol ester-induced neuritic alterations in the rat neocortex. Molecular and Chemical Neuropathology 20, 125–145 (1993). https://doi.org/10.1007/BF02815367

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