Search

We have investigated the long-term structural changes which affect Purkinje cells exposed to a single dose of kainic acid. Following intraparenchymal injection of the excitotoxin in the cerebellar cortex (1 microliter of a 1 mg/ml solution), Purkinje cells which survived within the lesioned area or close to its edges showed remarkable axonal abnormalities, involving the formation of torpedoes, hypertrophy of recurrent collaterals and atrophy of the corticofugal portion of the axon. In addition, their dendritic trees were often affected by conspicuous regressive alterations. The climbing fibres contacting these Purkinje cells were characterized by thick perisomatic plexuses, whereas their peridendritic branches were atrophic. The dendrites innervated by such atrophic olivary arbours were studded with huge numbers of newly formed spines. These alterations were already present a few days after kainic acid administration and persisted for the total period of observation of 6 months after the lesion. The remarkable similarity between the abnormalities of Purkinje cells exposed to kainic acid and those observed after axotomy indicates that in these two conditions common mechanisms determine analogous long-lasting modifications in the affected neurons. It is proposed that kainic acid-induced intracellular calcium overload disrupts cytoskeletal components and impairs axonal transport, thus depriving the affected Purkinje cells of retrograde trophic influences from their target neurons. As a consequence the affected neurons undergo long-lasting regressive modifications and compensatory remodelling phenomena.

By grafting solid pieces of cerebellar anlage onto the surface of the adult rat cerebellum, we have investigated the problem of the interactions between embryonic and adult neurons in an intact brain. A few days after grafting, embryonic astrocytic processes crossed the graft--host interface and radiated into the recipient molecular layer. Several grafted Purkinje cells also migrated into the host brain along such processes as well as adult Bergmann glia. Adult climbing fibres, labelled by means of Phaseolus vulgaris leucoagglutinin (PHA-L), sprouted new collateral branches which terminated on embryonic Purkinje cells at both extra- and intraparenchymal levels. No sign of activation of host astroglia or microglia was evident in the host cerebellum in relation to these processes. Embryonic Purkinje cells which migrated into the host cerebellum developed an adult-like morphology. Intraparenchymal grafts of neocortical embryonic tissue induced conspicuous growth of host olivary axons, characterized by a pattern which was different from that observed following cerebellar grafts. By contrast, when neocortical tissue was placed onto the surface of the recipient cerebellum, graft--host interactions were limited and climbing fibre sprouting was rarely seen. These results show that (i) supernumerary Purkinje cells can penetrate and settle in the adult intact cerebellar cortex, (ii) adult climbing fibres are able to innervate these new targets in the absence of any injury or activation of non-neuronal cells of the adult brain, and (iii) in the absence of damage to the adult brain, the plasticity of adult olivary axons is specifically elicited and controlled by embryonic Purkinje cells.

The abnormal location of large numbers of neurones is characteristic of genetic mutations which impair the migratory processes of developing nerve cells. Nevertheless, the presence of small amounts of ectopic neurones is a fairly common finding even in normal adult animals. The first aim of this study was to investigate a series of features of ectopic Purkinje cells in normal adult rats and particularly to assess whether these cells are still capable of interacting with their normal afferents. Several displaced Purkinje cells, identified by anti-D28k calbindin immunolabelling as well as by typical morphological features, were present in the brainstem and cerebellum of all the examined animals. Two distinct morphological types of such cells could be recognized: 1) noncortical Purkinje cells, located in several areas of the dorsal brainstem and cerebellum, characterised by poorly developed and randomly oriented dendrites; and 2) cortical Purkinje cells, exclusively located in the dorsal cochlear nucleus, characterised by large dendritic trees oriented along parasagittal planes. Tracing experiments, in which Phaseolus vulgaris leucoagglutinin (PHA-L) was injected into the inferior olive, revealed that several ectopic Purkinje cells, belonging to both types, were contacted by the terminal arbours of olivary axons, structurally similar to cerebellar climbing fibres. On the basis of the observation that ectopic Purkinje cells were more frequent in the dorsal cochlear nucleus than in any other of the examined regions, we tested the hypothesis that this nucleus might represent a particularly favourable environment for the survival and development of Purkinje cells. By grafting embryonic cerebellar tissue in the fourth ventricle, only minimal migration of Purkinje cells into the recipient parenchyma was observed when the transplant was placed on the brainstem surface caudal to the cochlear nucleus. By contrast, when the graft was apposed to the latter nucleus, large numbers of Purkinje cells migrated and developed in its superficial layers. These Purkinje cells passed through all the different phases which characterise the normal ontogenesis of this neuronal population and finally developed mature structural features similar to those displayed by cortical ectopic Purkinje cells. This study demonstrates that at least some of the ectopic Purkinje cells receive their physiological olivary input. This fact indicates that Purkinje cells are able to attract olivary axons and establish specific connections, even if they are displaced in an abnormal environment. In addition, we show that the dorsal cochlear nucleus represents a particularly favourable environment for the survival and the development of Purkinje cells.

The presence of an injury or the selective degeneration of specific neuronal populations is commonly assumed to be a necessary prerequisite for the survival and the integration of grafted neurons in the recipient brain. In the present study we have placed solid grafts of cerebellar anlage in the fourth ventricle of adult rats, in close contact with the host cerebellar cortex, to assess the capacity of embryonic Purkinje cells to interact with adult neurons and integrate in the unlesioned cerebellar cortex. Numerous grafted Purkinje cells are indeed able to leave the implant and migrate into the host molecular layer, where they develop adult structural features. In addition, such cells are able to elicit the growth of host climbing fibre sprouts which end in newly formed arborizations impinging upon their dendritic trees. Climbing fibre collateral branches also penetrate the implant to innervate Purkinje cells which have not migrated in the host cerebellum. These results show that embryonic Purkinje cells are able to survive and integrate in an adult unlesioned cerebellar cortex. In addition, adult olivary axons respond to the increased size of the target population by expanding their terminal domain to innervate grafted Purkinje cells.