C. Batini

Autroradiographic demonstration with 2-[14C]deoxyglucose of frequency selectivity in the auditory system of cats under conditions of functional activity

2-[(14)C]deoxyglucose was injected into 3 cats, each of whom was presented with a tone of a different frequency under controlled auditory conditions. Clear selective labelling was observed in autoradiographs of the cochlear nucleus, parts of the superior olivary complex, the nuclei of the lateral lemniscus and the inferior colliculus. No selective labelling was observed in autoradiographs of the medial geniculate body and the auditory cortex. The labelling observed in the cochlear nucleus and the inferior colliculus was in agreement with previous studies of both the physiology and anatomy of these regions and for the first time provides an anatomical demonstration of the organization of frequency selectivity under conditions of functional activity.

Long term modification of cerebellar inhibition after inferior olive degeneration.

SummaryThe long term effects of inferior olive destruction on the activities of the Purkinje cells and their target neurones in the cerebellar nuclei were studied in the rat. Careful observations were also made of motor behaviour throughout the study. Albino rats were injected with 3-acetylpyridine to produce a neurotoxic destruction of the inferior olive and then were used for acute recording experiments at 1–2 days, 5–7 days, 12–18 days, 35–38 days, 75–97 days and 230–252 days. After degeneration of the inferior olive, there was an initial period lasting for a few days, characterized by a high firing frequency of Purkinje cells associated with a very low level of activity of the neurones in the cerebellar nuclei. During this period, there was a deep depression of motor activity. A period of adaptation follows during the first month, characterized by a slow recovery of the initial firing frequency of the cerebellar units and a gradual recovery of spontaneous locomotion; nevertheless the firing pattern and motor behaviour remain abnormal. From one month on the unit activities disturbances and the motor deficiencies stabilize. The hypothesis is advanced that Purkinje cell inhibition on their target neurones, which increases during the initial period, gradually diminishes during the adaptation time, and then stabilizes to a subnormal state.

Harmaline-induced tremor

SummaryChanges of local cerebral glucose consumption under the effect of tremogenic doses of harmaline were studied. To find the brain structures activated by the drug, the autoradiographic method using [14C]2-deoxyglucose was applied to young cats. After administration of harmaline, the animals were paralized with flaxedil. Results were compared to a group of control animals not injected with the drug, but submitted to the same experimental protocol.Increases of neuronal activity were observed in several structures. A)Among the relays of the olivo-cerebellofastigio (and vestibulo)-reticulo-spinal circuit that had been claimed to fire at the frequency of the tremor, labeling was found in:1.selected portions of the inferior olive including the medial accessory olive and the caudolateral part of the dorsal accessory olive;2.the molecular layers of the cerebellar cortex including vermian and paravermian zones. Labeling of the olivo-cerbellar system was therefore larger than the compartment controlling the fastigial nucleus and extended to that controlling the interpositus nucleus.B)Other structures not under the direct control of the olivo-cerebellar system displayed increased radioactivity under harmaline: lateral reticular nucleus, nucleus reticularis tegmenti pontis, red nucleus and basal ganglia. Part of nucleus ambiguus, intensely labeled in the control animals, showed decreased radioactivity under harmaline. The experiments were repeated with the same protocol in another group of animals with unilateral sections of the inferior cerebellar peduncle in order to distinguish between a direct pharmacological influence and a nervous one. Marking of the basal ganglia was not affected by pedunculotomy, suggesting a direct “pharmacological” action of the drug in this cases. On the other hand, marking of the other labeled structures was asymmetric or suppressed by pedunculotomy, and therefore could result from a “nervous” effect secondary to activation of the inferior olive.

The GABAergic neurones of the cerebellar nuclei in the rat: projections to the cerebellar cortex.

The presence of gamma-aminobutyric acid (GABA) in the neurones of the cerebellar nucleocortical pathway is here reported. The pathway was identified by retrograde tracer and the GABA content was revealed immunohistochemically. It was found that most of the neurones giving rise to the reciprocal, non-reciprocal and symmetrical projections are indeed GABA-immunoreactive. They were observed in all the subdivisions of the nucleus medialis, of the nucleus interpositus and of the nucleus lateralis sending axons respectively to the sagittal zones A, C1-3 and D of the cerebellar cortex. The nucleus vestibularis lateralis and the related sagittal zone B were devoid of such projections.

Glutamate, GABA, calbindin-D28k and parvalbumin immunoreactivity in the pulvinar-lateralis posterior complex of the cat : relation to the projection to the Clare-Bishop area

Neurons of the pulvinar-lateralis posterior complex (Pul-LP) containing glutamate (Glu) and GABA, as presumed neurotransmitters, and calbindin- D28k (calbindin) and parvalbumin (PV), as Ca-binding proteins, were identified in the cat by using immunohistochemical methods. In vibratome sections, neurons immunoreactive (IR) to each of the four antibodies were observed throughout the Pul-LP. In semithin sections, GABA-IR neurons were also PV-IR but not calbindin-IR and some of them also co-localized Glu. The Glu-IR neurons which were negative for GABA co-localized calbindin but not PV. The neurons of the Pul-LP projecting to the Clare-Bishop area (CB) in the suprasylvian gyrus were identified with a retrogradely transported tracer and the sections were then immunostained for Glu, GABA, calbindin and PV. Only Glu- and calbindin-IR neurons were retrogradely labeled. These results show that, if calbindin and PV have a Ca-binding role, the presumably excitatory Glu-IR neurons projecting to the CB are use calbindin whereas the presumably inhibitory GABA-IR neurons are intrinsic and use PV. This relationship implies that these proteins probably have other roles specifically related to the kind of agonist to be released at the neuron.

Brain chimeras in birds: application to the study of a genetic form of reflex epilepsy

A strain of chicken, called here FEpi (for Fayoumi epileptic), bearing an autosomal recessive mutation, exhibits a form of reflex epilepsy with EEG interictal paroxysmal manifestations and generalized seizures in response to either light or sound stimulations. By using the brain chimera technology, we demonstrate here that the epileptic phenotype can be partially or totally transferred from an FEpi to a normal chick by grafting specific regions of the embryonic brain. The mesencephalon contains the generator of all epileptic manifestations whether they involve visual or auditory neuronal circuits, with the exception of the abnormal EEG which is transmitted exclusively by telencephalic grafts. This analysis supports the hypothesis that certain forms of human and mammalian epilepsies have a brainstem origin.

Pattern of electroencephalographic activity during light induced seizures in genetic epileptic chicken and brain chimeras

Genetic epilepsy was studied in Fayoumi epileptic (F.Epi) chickens and in neural chimeras obtained by selective substitution of embryonic brain vesicles of F.Epi donors in normal recipient chickens. Typical motor seizures accompanied by convulsions were evoked by intermittent light stimulation in F.Epi and in chimeras having embryonic substitution of the prosencephalon and the mesencephalon. The motor seizure was less severe in chimeras receiving only the prosencephalon. In the F.Epi, as well as in all the chimeras, the EEG during seizures was characterized by a desynchronized (or a flattening) pattern of activity. F.Epi and chimeras had a lower threshold to Metrazol induced seizures than control chickens. The experimental animals show that, in this model, large prosencephalic and mesencephalic areas are involved in the epileptic disease. The epileptic character of this genetic dysfunction is discussed.

Development of epileptic activity in embryos and newly hatched chicks of the Fayoumi mutant chicken

Summary: The homozygous Fayoumi strain of epileptic chickens (Fepi) is affected by generalized convulsions consistently induced by intermittent light stimulation (ILS) and by intense sound. Although interictal EEG recordings show continuous spikes and spike and wave activity, desynchronization and flattening (DF) of the EEG are observed during seizures. We have studied development of the epileptic phenotype in embryonic (E) and posthatching (P) Fepi. As compared with those of chicken embryos of a normal strain, no differences were observed in the EEG before embryonic day (E) 16. Clearly differentiated spikes and spike and waves appeared at E17 in Fepi. Metrazol‐induced EEG seizures were observed at E16 in normal embryos and at E17 in Fepi. The Fepi showed some characteristics: Spontaneous EEG seizure‐like discharges also appeared at E17 but decreased to‐ward hatching; visual or acoustic hyperexcitability developed at E20 together with evoked responses in normal chickens; desynchronization of the EEG, typical of the epileptic seizure of the adult, could be induced by ILS at E20, but ILS‐ or sound‐induced generalized motor seizures appeared at P1, a few hours after hatching. Results show that Fepi phenotype reaches full expression at P1, but the electric paroxysms are expressed earlier, paralleling synaptic maturation.

Avian photogenic epilepsy and embryonic brain chimeras: neuronal activity of the adult prosencephalon and mesencephalon

Photogenic genetic epilepsy was studied in an avian model, using either the Fayoumi epileptic chicken (Fepi) or neural chimeras obtained by replacement of em bryonic brain vesicles in normal chickens with those of Fepi embryos. In these two kinds of animals motor seizures accompanied by electroencephalographic (EEG) desynchronization and flattening (DF) were evoked by intermittent light stimulation (ILS). In chimeras with on ly the prosencephalon grafted, motor seizures were less severe but DF remained. ILS-induced DF persisted un der paralysis by gallamine triethiodide (Flaxedil). Extra cellular recordings were made in the prosencephalon (wulst) and in the mesencephalon (optic tectum) of paralysed animals. Units recorded in the prosencephalon of Fepi and chimeras showed abnormal interictal burst ing activity, distinctly different from the non-epileptic Fayoumi heterozygotes (Fhtz) and normal chickens. The mesencephalic units of Fepi and chimeras having both prosencephalon and mesencephalon grafted showed two types of abnormal activities during ILS-induced DF, which were distinct from the non-epileptic chickens: type I neurons displaying early, high sensitivity to ILS fol lowed by a prolonged suppression of activity; type II neurons displaying an early and prolonged suppression of activity. The results are discussed with respect to the brain structures generating ictal and interictal EEG ac tivities and motor seizures.

The immunocytochemical distribution of calbindin-D28k and parvalbumin in identified neurons of the pulvinar-lateralis posterior complex of the cat

The calbindin-D28k and parvalbumin immunoreactivities of the neurons of the pulvinar-lateral posterior complex (Pul-LP) were studied in the cat. The neurons of the Pul-LP projecting to the cerebral cortex were identified by a retrogradely transported tracer injected in the suprasylvian gyrus. Two populations of cells were found, a calbindin-D28k-immunoreactive, large-diameter population and a parvalbumin-immunoreactive, small-diameter group. The two kinds of cells are closely intermingled. The former includes the neurons retrogradely marked, and therefore projecting to the suprasylvian gyrus. The latter includes neurons which were not retrogradely marked, and therefore presumably intrinsic elements.