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[Ultrastructural changes in cerebral cortex and cerebellar cortex of rats under simulated weightlessness].

OBJECTIVE: To study the ultrastructural changes in the cerebral cortex and cerebellar cortex of rats under simulated weightlessness and the possible mechanism. METHOD: The tail-suspended rats model (-30 degrees head down tilt) was adopted to simulate weightlessness in the experiment. The rats were suspended for 7 d, 14 d, 21 d, and 28 d, and then were perfused through the hearts. The specimens were drawn from the rats' cerebral cortex and cerebellar cortex for electron microscopy. RESULT: The results showed that under simulated weightlessness, the main changes in the neuron can be described as follows: swelling of mitochondria, endoplasmic reticulum and Golgi complex, even formation of big empty vesicles; reduction of number of synaptic vesicles in IV layer; increase corrugation of capillary lumen and thickening of basement membrane. Degranulation of rough endoplasmic reticulum in Purkinje's cells of the cerebellar cortex occurred obviously. On the 14th and the 21st day of suspension, the changes were most significant and tended to return to normal on the 28th day. CONCLUSION: The experimental results demonstrated that simulated weightlessness led to ultrastructural changes in the cerebral cortex and cerebella cortex of rats. The ultrastructure changed with the course of simulated weightlessness and tended to return to normal. It showed an adaption to the simulated weightlessness.

Adaptation, Physiological↗

Peripheral stimuli excite coronal beams of Golgi cells in rat cerebellar cortex.

Cerebellar granule cells constitute the largest neurone population of the brain. Their axons run as parallel fibres along the coronal axis, and the one-dimensional spread of excitation that is expected to result from this arrangement is a key assumption of theories of cerebellar function. In many studies using various techniques, however, it was not possible to evoke such a beam-like propagation of excitation with natural stimuli. We recorded, in Crus I and II of anaesthetised rats, pairs of Golgi cells aligned along the parallel fibre axis and synchronising spontaneously. Each pair was subjected to two stimulation protocols: punctate and semi-continuous. Local punctate facial stimulation evoked distinct fast and late responses of variable strength and latency (fast: 4.0-10.2 ms; late: 13.6-22.7 ms). Semi-continuous stimulation with a brush increased the firing rate, and modified the precision and phase of synchronisation. Differences between a pair in response strength and phase to brush stimulation correlated strongly with the difference in latency to punctate stimulation. These observations were reproduced in a model of the granular layer. The stimulus activated a central patch of mossy fibres, and Golgi cells received short- and long-range excitation from mossy and parallel fibres, respectively. The strength and latency of the punctate response of a model Golgi cell were found to vary with its position, reflecting a systematic change in the contribution of mossy and parallel fibres to its excitation with distance from the activated patch. During brush stimulation, model Golgi cells inside the patch fired more precisely synchronised, whereas the other Golgi cells responded with a lag proportional to their distance from the patch, thereby reproducing the experimentally observed changes in synchronisation. Taken together with the previously reported large receptive fields of Golgi cells and with their spontaneous synchronisation, the variable, position-dependent latency of evoked Golgi cell responses indicates a beam-like spread of excitation along the parallel fibres in rat cerebellar cortex.

Animals↗

The significance of foliation and fissuration of cerebellar cortex. The cerebellar folium as a fundamental unit of sensorimotor integration.

I propose the general hypothesis that each individual folium in the cerebellum is an integrative module that is involved in unique sets of sensorimotor transactions. Although the basic types of operations carried out by cerebellar cortex may be similar in all folia, the mosaic of afferent sources, intrinsic organization and efferent destinations appear to be unique for each folium. I believe that this conception is supported by: 1) comparative data which illustrate species-typical folial patterns, 2) neuroanatomical data which reveal not only different structural features of folial crowns and fundi, but differential afferent and efferent connectivity of different folia as well, 3) physiological data, which demonstrate unique patterns of afferent activity in different folia, and especially by 4) ontogenetic data which establish that each folial crown expands and differentiates into an architecturally distinct cortical entity. Taken together, all these lines of evidence suggest that the numbers and patterns of folia exhibited by the cerebellar cortex of different mammals are morphological indicators of differential organization of sensorimotor control functions in each animal. Even intraspecific individual variations in folial number, size and pattern may signify structural-functional determinants of some individual differences in sensorimotor transactions. Since so little research has addressed the many testable ideas embodied in these general hypotheses, it seems to me that neuroscientists have a long way to go to clarify how the many different folia and lobules of cerebellar cortex actually function in the common, everyday, orderly, dynamic and ongoing reflex, postural, learned and deliberate behavioral sequences that characterize the normal behavioral repertoires of different animals. The enormous advances in understanding brought forth by the extensive research and writings of Professor Brodal and his colleagues have expanded our horizons to avail us of an enormous range of new vistas into cerebellar functional morphology. It is now the task of neurobiologists to explore these diverse new domains in ever greater depth and detail.

Animals↗

[Hippocampus-cerebellar cortex-cerebellar nuclei projection in the rat: electrophysiological and HRP studies].

The spatial distribution of the hippocampus-cerebellar projection and the connection between the projection area of hippocampus in the cerebellum and cerebellar nuclei were studied by means of electrophysiological and HRP labelling techniques. Polysynaptic responses of Purkinje cell's simple and complex spike in the lobule VI could be evoked by stimulation of the area CA1/CA3 of the dorsal hippocampus. These results suggest that there are polysynaptic projections from CA1 and CA3 areas of dorsal hippocampus to the lobule VI of cerebellar cortex, and the last relay fibers for these projections are mossy fiber and/or climbing fiber. It is also shown that this hippocampus-cerebellar projection predominantly terminates at the lobule VI of cerebellar cortex, 0.8-1.4 mm lateral to the midline. The projection from CA1 area is mainly contralateral, whereas the projection from CA3 area is mainly ipsilateral. The results with HRP labelling indicate that the projection area of hippocampus is an interpositus zone of the lobule VI. Reciprocal connections may exist between this cortical area and the interpositus nucleus.

Animals↗

Posture-correlated responses to vestibular polarization in vermal versus intermediate posterior cerebellar cortex.

Cerebellar lesion experiments have led to the concept that the medial longitudinal zone controls postural tone while the intermediate zone controls discrete movement. This study is a test of the hypothesis that, of the two zones, the medial zone is more closely linked to the resting discharge of vestibular afferent fibers, a prime source of neural tonus underlying the tonus of posture. Unilateral polarizations of the vestibular apparatus via the round window in awake, unrestrained guinea pigs caused step changes of postural attitude, the direction of which was polarity dependent. In anesthetized animals, these currents caused nonadapting step changes, or posture-correlated responses in the level of resting discharge in vestibular primary afferent fibers. In the medial and the intermediate cerebellar cortices of the posterior lobe, the proportion of step-like responses was similar, in contradiction to the hypothesis. This suggests that the cerebellar computations for controlling both postural tonus and discrete movements require information about vestibular tonus in terms of simple spike activity.

Action Potentials↗

The content of amino acids in the developing cerebellar cortex and deep cerebellar nuclei of granule cell deficient mutant mice.

Glutamic acid is the only free amino acid to be clearly reduced in mature granule cell deficient cerebellum. The correlation between concentration of glutamic acid and extent of granule cell loss suggests that it may serve as a neurotransmitter. Curiously, in two neurological mouse mutants, glutamic acid is also decreased in the deep cerebellar nuclei where there are no granule cells. We have now examined the amino acid content of cerebellar cortex and deep cerebellar nuclei of the granule cell deficient mutants, weaver and staggerer, during the postnatal period in which granule cell development takes place. We have found: (1) an early and transient deficit in taurine in weaver cerebellar cortex during the period of granule cell migration, (2) deficits during the second postnatal week in taurine, aspartic and glutamic acids in both weaver and staggerer cerebellar cortex, (3) that aspartic and glutamic acid deficits result from failure to increase concentrations at the normal rate after birth rather than from a fall from normal levels, (4) decreased concentrations of glutamic acid but not of taurine and aspartic acids apparent in the deep nuclei of both weaver and staggerer at about the same time as in cerebellar cortex, (5) amino acid changes in weaver heterozygote cerebellum which result in values intermediate in magnitude between normal and homozygous weaver animals and (6) an early and persistent reduction in staggerer deep nuclei of gamma-aminobutyric acid (GABA), the Purkinje cell transmitter, indicating early denervation or lack of full innervation of deep nuclei by Purkinje cells.

Amino Acids↗

Somatotopically organized transient projections from the primary somatosensory cortex to the cerebellar cortex.

The organization of transient projections from the primary somatosensory cortex (S-I) to the cerebellar cortex in neonatal kittens was examined using orthograde intraaxonal labeling techniques. Tritiated amino acid injections into face, forelimb and hindlimb areas of representation in S-I labeled mossy fiber-like terminals of cerebrocerebellar axons in different areas of the cerebellar cortex bilaterally. The hindlimb area of S-I projected to lobules I-IV in the anterior lobe and to ventral folia of the paramedian lobule (PML). Injections into forelimb areas of S-I labeled terminals in lobules IV and V and in intermediate and dorsal folia of the PML. The face area of S-I projected to the lobules V and VI, to medial folia in the ansiform and simplex lobules and to dorsal PML folia. Labeled terminals were more numerous in the cerebellar cortex contralateral to the S-I injections, except in lobules I and II and the ventral PML where the density of hindlimb input was approximately the same on both sides. These observations were supplemented by findings that small wheat germ agglutinin-horseradish peroxidase (WGA-HRP) injections into the dorsal or ventral PML resulted in retrogradely labeled layer V pyramidal neurons in lateral (face and forelimb) and medial (hindlimb) areas of S-I respectively. The somatotopic organization of transient S-I cerebrocerebellar projections is very similar to the topography of cerebellar somatosensory afferent pathways in adult cats.

Afferent Pathways↗

Differential roles of cerebellar cortex and deep cerebellar nuclei in learning and retention of a spatial task: studies in intact and cerebellectomized lurcher mutant mice.

Lurcher mutant mice (+/Lc) exhibit a massive loss of neurons in the cerebellar cortex and the inferior olivary nucleus, while deep cerebellar nuclei are essentially intact. To discriminate the relative participation of the cerebellar cortex and deep structures in learning and memory, 3 to 6-month-old +/Lc mice were subjected to a spatial learning task derived from the Morris water escape. They were able to learn to escape as well as their strain-matched controls (+/+). Seven days later, their scores showed that they had memorized the spatial environment but not as accurately as +/+ mice. Cerebellectomy before training did not significantly alter the escape learning capabilities of either group, whereas cerebellectomy performed after learning completely abolished retention in +/+, as well as in +/Lc, mice. These results suggest that the cerebellum, although not necessary for learning a spatial task, plays a crucial role in its retention, and that the storing structure of spatial information differs in +/+ and +/Lc mice.

Analysis of Variance↗

Delayed spontaneous alternation in intact and cerebellectomized control and lurcher mutant mice: differential role of cerebellar cortex and deep cerebellar nuclei.

Lurcher mutant (+/Lc) mice exhibit a massive loss of neurons in the cerebellar cortex and in the inferior olivary nucleus while deep cerebellar nuclei are essentially intact. To discriminate the respective participation of the cerebellar cortex and deep structures in learning and memory, the authors subjected 3- to 6-month-old +/Lc mice to a delayed spontaneous alternation task to test their working and long-term spatial memories. Results show that wild type (+/+) mice alternated above chance even after a 1-hr delay between the forced and choice trials, whereas in +/Lc mice, long-term memory was impaired. Cerebellectomized +/+ mice behave as +/Lc mice (working memory was preserved but long-term memory was not), whereas in the cerebellectomized +/Lc mice, both working and long-term memories were altered. These results are discussed in terms of relationships between the cerebellum and the hippocampus.

Animals↗

Dopaminergic and non-dopaminergic neurons in the ventral tegmental area of the rat project, respectively, to the cerebellar cortex and deep cerebellar nuclei.

It has been suggested recently that dopamine in the cerebellum not only acts as a precursor for noradrenaline in afferent fibers supplied by locus coeruleus neurons, but also subserves an independent transmitter role in a separate neural system. The present study was initiated to investigate the possible sources for dopaminergic innervation of the cerebellum. Employing anterograde and retrograde axonal tracing with cholera toxin and a combination of fluorescent retrograde axonal tracing with Fluoro-Gold and tyrosine hydroxylase immunofluorescence histochemistry, we found in the rat that the ventral tegmental area, containing the A10 dopaminergic cell group, sends projection fibers to the cerebellum bilaterally with a slight contralateral predominance. The projections from the ventral tegmental area to the cerebellum were segregated into the dopaminergic one to the cerebellar cortex and the non-dopaminergic one to the deep cerebellar nuclei. Dopaminergic fibers projecting from the ventral tegmental area to the cerebellar cortex terminated mainly in the granular layer, additionally in the Purkinje cell layer, but not at all in the molecular layer. They were distributed predominantly in the crus I ansiform lobule and paraflocculus, and to a lesser extent in the crus II ansiform lobule. On the other hand, non-dopaminergic fibers projecting from the ventral tegmental area to the deep cerebellar nuclei were seen to terminate mainly in the lateral nucleus, to a lesser extent in the interpositus nucleus, but not at all in the medial nucleus. The ventral tegmental area was also observed to receive projection fibers from the lateral and interpositus cerebellar nuclei bilaterally with a contralateral predominance. The projections from the ventral tegmental area to the cerebellum revealed in the present study might exert limbic influences upon the cerebro-cerebellar loops subserving the execution and co-ordination of voluntary movements.

Animals↗

Differential roles of cerebellar cortex and deep cerebellar nuclei in the learning of the equilibrium behavior: studies in intact and cerebellectomized lurcher mutant mice.

Three- to 6-month-old lurcher mutant mice (+/lc), which exhibit a massive loss of neurons in the cerebellar cortex and in the inferior olivary nucleus but whose deep cerebellar nuclei are essentially intact, were trained daily, for 9 days, to maintain their equilibrium upon a rota rod rotating at 20 or 30 revolutions per minute (rpm). Their scores were measured and their behavior upon the rotating rod quantified in comparison to those of matched control (+/+) mice. Lurcher mice were able to learn to maintain their equilibrium efficiently when rotated at 20 rpm but were not when rotated at 30 rpm. After cerebellectomy, the equilibrium capabilities of the animals were much altered, especially in +/lc. These results show that the deep cerebellar nuclei are sufficient for motor learning, provided the task is not too difficult (20 rpm), but that the cerebellar cortex is required when the task is more difficult (30 rpm). Therefore, it can be concluded that the adaptive motor capabilities of lurcher mice are less developed than those of control animals.

Animals↗

Molecular identification of the Lugaro cell in the cat cerebellar cortex.

The cerebellar cortex contains five major classes of neurons that can be differentiated from one another on the basis of their location, size, shape, and, in some cases, molecular characteristics. The cerebellar cortex also contains other, less numerous neuronal types, including the Lugaro cell, which has been described on only a few occasions. The Lugaro cell is a relative rare cell type and is characterized by a fusiform cell body with thick, horizontally oriented dendrites. It is located in or slightly below the Purkinje cell layer. Because the Lugaro cell shares some morphological characteristics with the other large granular layer neurons, it often has been classified as a Golgi cell. In the present study we have taken advantage of differences in the molecular properties of neurons and have used monoclonal antibodies to identify and classify the Lugaro cell. Three large neuronal types in the cerebellar cortex were examined with cell-type-specific antibodies: Cat-301 and Cat-304 for Lugaro cells; Rat-303 for Golgi cells; and anti-calbindin for Purkinje cells. Double label immunocytochemistry on sections of the cat cerebellum was performed with subclass- or species-specific secondary antibodies. Each of the three antibodies was selective for one of the three large neuron classes. Cat-301 and Cat-304 recognized Lugaro cells but not Golgi or Purkinje cells. Our results demonstrate that the Lugaro cells are molecularly, as well as morphologically, distinct from Purkinje and Golgi cells and thus constitute a distinct cell type in the cerebellar cortex.

Animals↗

Pattern formation in the cerebellar cortex.

The cerebellar cortex is subdivided rostrocaudally and mediolaterally into a reproducible array of zones and stripes. This makes the cerebellum a valuable model for studying pattern formation in the vertebrate central nervous system. The structure of the adult mouse cerebellar cortex and the series of embryological events that generate the topography are reviewed.

Adult↗