Developmental changes in concentrations and distributions of neurotrophins in the monkey cerebellar cortex

Neurotrophins are involved in the survival, differentiation, migration and neurite outgrowth of various neuronal populations. Neurotrophins and their receptors are widely expressed in the developing cerebellum of various experimental animals. To gain some insight into the possible roles played by these molecules in monkey cerebellum, we examined the protein levels of BDNF, NT-4/5 and NT-3 and distributions of those neurotrophins and TrkC, a high affinity receptor for NT-3, in the cerebellum of developing macaque monkeys using ELISAs and immunohistochemical methods. We found that the level of BDNF increased during development, while the level of NT-3 was higher during embryonic stages and decreased toward adulthood. The level of NT-4/5 increased from embryonic stages to infant stages and gradually declined with age. Among the three neurotrophins, BDNF immunoreactivity was found in all kinds of cerebellar neurons, including all inhibitory interneurons, throughout the postnatal periods examined, indicating that BDNF may be an essential factor for the maintenance of cerebellar neural functions. The Bergmann glial fibers and the internal part of the external granule cell layer were strongly NT-3 immunopositive at the early postnatal stages, and more weakly immunoreactive toward adulthood. In addition, we found that the premigratory precursors of the granule cells were TrkC immunopositive at early postnatal stages. These findings suggest that NT-3 in Bergmann glial fibers may be involved in the migration of the premigratory granule cells.     

Morphological changes in the cerebellar cortex of aging Macaca nemestrina

The cerebellar cortices in 4, 10 and 20 year old Macaca nemestrina have been examined for the number of Purkinje (P) and granule cells and the deposition of lipofuscin in P cells in relation to aging. Lipofuscin distribution significantly increased with age in the P cells in these animals. The number of P cells was significantly reduced, while there were no changes in the number of granule cells. It appears from this and other studies that the Purkinje cells are more prone to aging changes than the granule cells of the cerebellum both in lipofuscin formation and cell loss. Although the precise functional significance of these changes in P cells is not clear, their vulnerability may be related to changes in motor function in old age.     

An HRP and autoradiographic study of cerebellar corticonuclear-nucleocortical reciprocity in the monkey

Summary Combined injections of ³H-leucine and HRP were made into the monkey cerebellar cortex in order to identify any reciprocal connections between the corticonuclear and the nucleocortical pathways. These combined intraaxonal labeling experiments have demonstrated a considerable overlap of orthogradely labeled Purkinje cell axons and terminals with retrogradely labeled HRP-positive neurons in the ventrolateral region of the dentate nucleus following combined injections into the lateral hemisphere, and in the dorsal area of the dentate following combined injections into medial cortical areas of the anterior lobe. There were also areas within the deep cerebellar nuclei where orthogradely labeled corticonuclear terminals did not overlap with retrogradely labeled nucleocortical neurons.     

Expression of TrkB subtypes in the adult monkey cerebellar cortex

BDNF and its specific receptor TrkB are concerned with synaptic plasticity as well as maintenance of the nervous system. TrkB has three subtypes: full-length TrkB (TK+), which has a tyrosine kinase containing intracellular domain, and two truncated TrkBs (TK-; T1 and T2), which lack tyrosine kinases. To understand the molecular interaction among these subtypes, we investigated the expression and distribution of BDNF, TK+, and T1 in the adult monkey cerebellum by single and double immunohistochemistry and Western blot analysis. We observed by single immunohistochemistry that BDNF, TK+, and T1 are distributed in almost all the somata and dendrites of Purkinje and granule cells. In the double-stained sections, three kinds of regions were observed: TK+ >T1; TK+ =T1; TK+ 相似文献   

Early postnatal corticosterone administration regulates neurotrophins and their receptors in septum and hippocampus of the rat

The principal glucocorticoid in rats, corticosterone, interacts with neurons in the limbic system and leads to morphological and behavioral changes. Putative corticosterone-triggered mediators are neurotrophins. In the present study we investigated the effects of early postnatal corticosterone treatment in rats on neurotrophic factors of the nerve growth factor (NGF) family and their receptors. Newborn rats were treated with corticosterone-containing polymers until postnatal day 12. The mRNA and protein levels of the neurotrophins of the NGF family (NGF, BDNF, NT-3 and NT-4/5) and their receptors (trkA, trkB, trkC and p75) were quantified in septum and hippocampus using RT-PCR. In the septal region, we found an unchanged mRNA expression after corticosterone treatment, whereas in the hippocampus there was a general increase in mRNA. Particularly, the gene expression of NGF, NT-3, and the high affinity receptors trkA, trkB and trkC increased significantly. Quantification of the neurotrophin protein levels using an ELISA revealed significant treatment effects for NGF and NT-4/5 in the hippocampus. The present study of corticosterone treatment in young rats demonstrates interactions of steroid hormones with neurotrophic factors and their receptors in the septo-hippocampal system during the first two postnatal weeks.     

Compensatory motor function of the somatosensory cortex for dysfunction of the motor cortex following cerebellar hemispherectomy in the monkey

Summary Electrical activities of the motor and somatosensory cortices preceding visually-initiated hand movements were recorded with electrodes chronically implanted on the surface and at 2.5–3.0 mm depth in the cortex of monkeys, and changes in field potentials in these cortices after cerebellar hemispherectomy were observed for many weeks. As previously reported, a unilateral cerebellar hemispherectomy including the lateral and interpositus nuclei eliminates the cerebellar-mediated superficial thalamo-cortical (T-C) responses recorded in the forelimb motor cortex contralateral to the hemispherectomy. These T-C responses normally precede the hand movement, and the operation results in the delay of movement initiation. The electrodes in the forelimb area of the contralateral primary somatosensory cortex showed an enhancement of superficial T-C responses of the somatosensory cortex for 30–40 days after the operation. The enhanced potentials preceded the delayed movement as do the cerebellar-mediated superficial T-C responses of the motor cortex in normal situations. Local cooling of the somatosensory cortex following the cerebellar hemispherectomy disturbed the reaction time movement for a few weeks after the operation. This effect was rarely encountered in normal monkeys. The present study suggests the compensatory motor function of the somatosensory cortex for the dysfunction of the motor cortex in early weeks after cerebellar hemispherectomy.Supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan     

Temporal changes in the level of neurotrophins in the spinal cord and associated precentral gyrus following spinal hemisection in adult Rhesus monkeys

Neurotrophins (NTs) appear to be crucial for the survival and potential regeneration of injured neurons. However, their temporal changes and remote regulations following spinal cord injury (SCI) have been only partially determined, especially in primates. In this study, ELISA was performed on the extracts of injured spinal cord and the associated precentral gyrus contralateral to the site of spinal cord hemisection to investigate the temporal changes in the levels of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and neurotrophin-4 (NT-4) in adult rhesus monkeys subjected to T8 spinal hemisection. Animals were allowed to survive 3, 7, 14, 30 and 90 days post-operation (dpo). In the spinal cord, the levels of NGF, BDNF and NT-3 sharply decreased between 3 and 7dpo. Thereafter, the levels of NGF and BDNF were transiently elevated while NT-3 level continuously increased and recovered to normal level at 30dpo. In the contralateral precentral gyrus (cPG), only the NT-3 level was altered and in fact elevated above the normal value. No obvious changes were observed in NT-4 level in any of the regions studied. Taken together, the present findings indicated that intrinsic NGF, BDNF and NT-3 may play a local role in the responses to the SCI in primates. Especially, the increase of NT-3 level occurred continuously in both the cPG and the spinal cord pointed to a possible transportation of NT-3 to the cord following SCI.     

Factoring neurotrophins into a neurite-based pathophysiological model of schizophrenia

Neurotrophins are growth factors that, through variations in concentration and changes in receptor expression, regulate the formation of axons and dendrites during development and throughout adult life. Here we review these growth factors, particularly in the context of schizophrenia, a psychiatric disorder characterized by neurodevelopmental abnormalities. We first discuss emerging information derived from physiologically relevant organotypic cultures and in vivo studies regarding the effects of neurotrophins on the neuronal structure including pruning and GABAergic neurons. We then review postmortem studies of neurotrophin levels and their receptors in brains of individuals with schizophrenia, and compare them with what is known about neurotrophin effects on neuronal structure. This comparison indicates that only some neuropathological defects encountered in patients with schizophrenia can be explained by the single action of neurotrophins on dendrites and axons. However, we propose that a number of inconsistent findings and apparently unrelated results in the schizophrenia field can be reconciled if neurons are considered structurally plastic cells capable of extending and retracting dendrites and axons throughout life.     

Developmental changes of chick cerebellar cortex revealed by monoclonal antibodies

Immunohistochemistry studies of the embryonic and newly hatched chick cerebellum were performed with 13 monoclonal antibodies (MAbs) raised against the embryonic chick optic nerve and a MAb which binds to cell nuclei. Neural MAbs differentially stained Purkinje cells, the external granular layer, molecular layer, internal granular layer, climbing fibers, basket cell axons, Bergmann glia and Ramón y Cajal's ansiform fibers. At the different developmental stages each component responded to MAbs differently. For example, staining of Purkinje cells with MAbs 23C10, 82E10 and 94C2 appeared on day 11 of incubation and disappeared sequentially after day 18. These results reveal molecular heterogeneity not only in cerebellar neurons but also at various developmental stages.     

Two modes of cerebellar input to the parietal cortex in the cat

Summary The characteristics of cerebellar input to the parietal cortex through the ventroanterior-ventrolateral (VA-VL) complex of the thalamus were investigated in the adult cat by using combined electrophysiological and anatomical methods. Two distinct parietal regions were activated by stimulation of the cerebellar nuclei (CN). In the first region located in the depth of the bank of the ansate sulcus, stimulation of the CN induced early surface positive-deep negative potentials and late surface negative-deep positive potentials. In this cortical area, potentials of similar shape and time course were evoked at a shorter latency by stimulation of the ventrolateral part of the VA-VL complex where large negative field potentials were evoked by stimulation of the CN. After injection of the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) in this part of the VA-VL complex, axon terminals of thalamocortical (TC) fibers were found in layers I, III and IV in the depth of the bank of the ansate sulcus and layers I and III in the motor cortex. In the second region located in the suprasylvian gyrus, late surface negative-deep positive potentials were evoked by stimulation of the CN and similar potentials were evoked at a shorter latency from the dorsomedial part of the VA-VL complex where large cerebellar-evoked potentials could be recorded. PHA-L injection in this thalamic region stained TC fibers and their terminals in layer I of the suprasylvian gyrus, and in layers I and III of the motor cortex. The laminar distribution of TC axon terminals in two different regions of the parietal cortex could account for the depth profiles of the cerebellar- and the thalamic-evoked potentials in each region. These results show that cerebellar information is conveyed to two separate areas in the parietal cortex by two different TC pathways.     

Developmental changes in rhesus monkey placental villi and cell columns

Summary Developing rhesus monkey placentas were analyzed by scanning electron microscopy with special attention directed toward defining stages in the development of the villus branches. The initial phase in formation of villi was the conversion of reticulated trabeculae of syncytial trophoblast into chorionic villi by growth and proliferation of cell columns of cytotrophoblast. These villi were stout and unbranched. The second phase of development appeared to be the longitudinal splitting of the villi and cell columns to form groups of parallel branches but there was a common insertion of these into the basal plate. The third phase in formation of villi, which appeared to begin at about the same time as the longitudinal splitting occurred, was the outgrowth of largediameter side branches in a zone nearer the chorionic plate. At about 38–40 days of gestation the next stage in villus formation occurred, characterized by the emergence of numerous, small syncytial sprouts. Continued proliferation of villi at later stages of gestation resulted in a decreased diameter of the terminal villi and an increasing complexity in the course of fetal capillaries.     

Reacquisition of classical conditioning after removal of cerebellar cortex

Summary We trained rabbits with white noise and light conditioned stimuli and a face shock unconditioned stimulus for classical conditioning of the nictitating membrane response and then removed the ipsilateral cerebellar cortex by aspiration. Larsell's hemisphere lobule VI was completely removed in 5 rabbits; the pattern of degeneration in the inferior olive matches the projections to HVI reported in experiments with horseradish peroxidase. All rabbits showed an initial abolition of conditioned responses but then relearned within two days. This indicates that cerebellar cortical lobule HVI normally is involved but is not necessary for classical conditioning.     

Comparative studies on the pre-and postterminal blood vessels in the cerebellar cortex of rhesus monkey, cat, and rat

Summary In the rhesus monkey, cat and rat, pial arteries give off branches which run vertically through all three layers of the cerebellar cortex. The large cortical arteries are surrounded by a perivascular space in the molecular layer. Their wall consists of several layers of smooth-muscle cells and the luminal endothelium. As the arteries reach the deeper layers of the cerebellar cortex, the number of smooth-muscle cells is reduced. In the rat, sometimes no smooth-muscle cells are detectable in the preterminal arterial vessels. If these deep arteries branch off by dichotomy of terminal vessels there occurs a gradual or complete loss of myocytes in all three species. In the cat, where cortical arteries give off branches at rightangles, there is a sphincter-like accumulation of smooth-muscle cells at the opening to the smaller branch.The postterminal vessels and veins in all species exhibit the smae mural structure found in capillaries. The wall consists only of an endothelium and occasional pericytes embedded in the basal lamina. Even the large veins which run to the pial veins show this simple mural structure.Dedicated to Prof. Dr. Dr. A. Dabelow in honour of his 80th birthdaySupported by the Deutsche Forschungsgemeinschaft (La 184/5)     

Lipofuscin in the cerebellar cortex of albino rats: An electron microscopic study

Summary The ultrastructure of autofluorescent, PAS-positive lipofuscin in Purkinje, granule, Golgi epithelial, basket and stellate, microglial and perivascular cells in the cerebellar cortex of senescent rats is described. The membrane-bounded pigment is composed of three elements: 1) electron-lucent homogeneous droplets, 2) a granular matrix and 3) intensely osmiophilic patches. The proportions of these three components vary between cell types and one can grossly differentiate a neuronal and a glial lipofuscin. The lipofuscin granules of stellate and perivscular cells are different from lipofuscin of other cerebellar neurons and glia. It can be concluded from these morphological observations that each cerebellar cell type has its distinct lipofuscin.Supported by the Deutsche Forschungsgemeinschaft La 184/5I would like to thank Mrs. v. Bronewski and Mr. H. Boffin for their technical assistance     

A computer model of mammalian cerebellar cortex

A computer-simulated model of mammalian cerebellar cortex is described, in which the individual units correspond to 300 μm square regions of the cerebellar cortical sheet. Local properties of these units as well as their pattern of connectivity were derived from anatomical and physiological data. Selection of an appropriate set of parameters produced predicted neuronal firing patterns and excitability changes similar to those obtained from anesthetized cats.The role of Purkinje recurrent collaterals was investigated by varying the strength of Purkinje cell inhibitory coupling to Golgi and basket cells. It is shown that the recurrent collaterals may exert important effects upon the time course of Purkinje cell responses to parallel fiber inputs. For spatially-distributed mossy fiber inputs, a focusing effect of long time course is demonstrated.     

Development of full-length Trk B-immunoreactive structures in the prefrontal and visual cortices of the macaque monkey

Distribution and morphological changes of cells containing the signal transducing neurotrophin receptor, full-length Trk B (fl-Trk B), were investigated in the prefrontal cortex (area FD) and the primary visual cortex (area OC) of the macaque monkey between embryonic day 140 and the adult stage. In area FD at the adult stage, fl-Trk B immunoreactivity was mainly observed in the pyramidal cells in layers II/III, V and VI. Small numbers of granule cells in layer IV were immunopositive. Bipolar and multipolar cells in layer II were rarely immunoreactive. At embryonic day 140, the number of fl-Trk B immunoreactive pyramidal cell was high, and gradually decreased until the adult stage. In layer IV, the number of fl-Trk B-ir cells was also high at embryonic day 140, and decreased remarkably from postnatal day 7 to the adult stage. On the other hand, in area OC at the adult stage, cells in layers II/III, IV, V and VI were fl-Trk B immunopositive. From embryonic day 140 until adulthood, the cells in layer IVc were fl-Trk B immunoreactive. The strongest fl-Trk B immunoreactivity in areas FD and OC occurred at postnatal month 6, coinciding with the time of the synapse overproduction. These findings suggest that ligands of fl-Trk B, such as BDNF and NT4/5 may be involved in the development and maintenance of the monkey cerebral cortices. Accepted: 26 July 1999     

小鼠放射状胶质细胞和小脑皮质的发育

目的探讨小鼠小脑皮质发育过程中放射状胶质细胞的分化。方法应用免疫荧光及5-溴脱氧尿嘧啶核苷(BrdU)检测技术,标记小鼠胚胎8d至生后180d小脑(57例,分为19组,每组3只)的神经干细胞、放射状胶质细胞、普肯耶细胞及颗粒细胞。结果放射状胶质细胞于胚胎13d的神经上皮出现,尔后该细胞分化为各种神经元和贝格曼胶质细胞,并在小脑皮质层状结构的形成中起着重要作用。结论放射状胶质细胞来源于神经上皮细胞,是神经细胞和神经胶质细胞的前体细胞。在小脑皮质的发育过程中,放射状胶质细胞能分化为普肯耶细胞和颗粒细胞,并为神经细胞的迁移提供路径和支架。     

The feline oculomotor nucleus: morphological subdivisions and projection to the cerebellar cortex and nuclei

Summary The cytoarchitecture of the feline oculomotor nucleus was examined in sections stained with thionin and neutral red. Five different subdivisions (caudal central, paramedian, ventral, dorsomedial and dorsolateral divisions) can be identified on each side of the midline. This observation is discussed, and our findings are compared to previous studies of the cytoarchitecture or central muscular representation of the oculomotor nucleus in which different subgroups have been distinguished. Implants or injections of the wheat germ agglutinin-horseradish peroxidase complex have revealed that all five subdivisions project to different parts of the cerebellar cortex and nuclei. Retrogradely labelled cells were found in the oculomotor nucleus in 18 cases following deposition of tracer in the fastigial and interposed nuclei and certain regions of the anterior, posterior and flocculonodular lobes. The projection is bilateral and appears to have its main termination in flocculus. It originates from small neurons, especially from those located along the dorsal border of the oculomotor nucleus.     

Projection on the motor cortex of thalamic neurons with pallidal input in the monkey

Summary The cortical projection areas of thalamic neurons with basal ganglia and/or cerebellar inputs were studied electrophysiologically in unanesthetized monkeys. Thalamic neurons which receive inhibition from the pallidum were found to project to the motor cortex (area 4) as well as to premotor cortex. The neurons with pallidal input and motor cortical projection were located mainly in VLo. This result indicates that the basal ganglia innervate the motor cortex through the thalamus. Thus the basal ganglia can modify the cortical output for controlling movements directly through this pathway as compared with its influence through the prefrontal and premotor cortices.     

Expression of some neurotrophins in the spinal motoneurons after cord hemisection in adult rats

There are numerous studies reporting on the crucial roles of neurotrophins (NTFs) in neuronal survival and sprouting after spinal cord injury (SCI). But studies on endogenous changes of neurotrophins after SCI are few. In this study we explored by means of immunohistochemistry the localization of NGF, BDNF and NT-3 in the normal adult spinal cord (SC) and the changes in the expression of these chemicals in the ventral horn after right cord hemisection at T9-10. The results showed an obvious increase in the numbers of NGF, BDNF and NT-3-immunoreactive neurons in the ventral horn and also an increase in their intracellular optical density (O.D.) at 3, 7 and 21 days after cord hemisection, when compared with sham-operated rats. The expression of NGF peaked at 7 days postoperation (dpo), while BDNF and NT-3 expressions peaked at 3 dpo. Evaluation of hindlimb functions by Basso Beattie Bresnahan (BBB) scoring showed that the hindlimb support and stepping function improved very quickly at 7 dpo. This study indicated that NGF, BDNF and NT-3 could play important but different roles in the mechanisms of spinal neuroplasticity at different times after SCI.