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Volume 17, Number 10, Issue of May 15, 1997 pp. 3710-3726
Copyright ©1997 Society for Neuroscience

Purkinje Cell Survival and Axonal Regeneration Are Age Dependent: An In Vitro Study

Received Dec. 23, 1996; revised Feb. 24, 1997; accepted March 3, 1997.

Isabelle Dusart¹, Matti S. Airaksinen², and Constantino Sotelo¹

¹ Institut National de la Santé et de la Recherche Médicale U106, Hôpital de la Salpêtrière, 75651 Paris Cedex 13, France, and ² Institute of Biotechnology, University of Helsinki, 00014 Helsinki, Finland

Purkinje cells are among the most resistant neurons to axotomy and the most refractory to axonal regeneration. By using organotypic cultures, we have studied age- and environment-related factors implicated in Purkinje cell survival and axonal regeneration. Most Purkinje cells taken from 1- to 5-d-old rats, the period in which these neurons are engaged in intense synaptogenesis and dendritic remodeling, die 1 week after plating, whereas if cultured before or after this period, Purkinje cells survive, even in the absence of deep nuclear neurons, their postsynaptic targets. Cerebellar slices taken from 10-d-old rats and kept in vitro for 1 week acquire a cellular composition resembling mature cerebellum. Their Purkinje cells are resistant to axotomy, but even when confronted with permissive environments (sciatic nerves or fetal cerebellar slices), their axons do not regenerate. In contrast, fetal rat and mouse Purkinje cells are able to regenerate their axons on mature cerebellar slices. This regeneration is massive, and the regrowing axons invade all cerebellar regions of the apposed mature slices, including white matter. These results show that Purkinje cell survival and axonal regeneration are age-related and independent from environmental constraints. Moreover, our observations suggest strongly that the onset of synaptogenesis of Purkinje cell axons could provide a signal to turn off their growth program and that, thereafter, permissive microenvironment alone is unable to reestablish such a program.

Key words: axonal regeneration; neuronal survival; cerebellum; Purkinje cell maturation; cerebellar organotypic cultures; axonal growth

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