József Takács

Metabotrop glutamate receptor type 1a expressing unipolar brush cells in the cerebellar cortex of different species: A comparative quantitative study

Morphology, distribution and number of unipolar brush cells (UBCs) was studied in the cerebellar vermal lobules I–X of the chicken, rat, guinea pig, cat, and monkey using monoclonal mGluR1a antibody as a marker to visualise these recently described nerve cells (Mugnaini and Floris [1994] J. Comp. Neurol. 339:174–180; Mugnaini et al. [1994] Synapse 16:284–311). The morphological appearance of mGluR1a immunopositive UBCs is similar in all species investigated: they are small cells, having a single, relatively short and thick dendrite, terminating in brush‐like dendrioles. Although this, probably excitatory, cell type can be found all over the cerebellar cortex, highest density of UBCs can be seen in the vermal cortex. The present study, therefore, was focused on the quantitative morphology and distribution of UBCs in the 10 lobules of the vermis. Calculating the number of UBCs/1 Purkinje cell (PC), we have found differences in this value (average in vermal lobules I–X) from 1.04 in rat, 1.10 in chicken, 1.16 in guinea pig, 2.27 in monkey, and up to 2.44 in cat. The highest density of UBCs was observed in lobules I, IX, and X, whereas the lowest number of UBCs/1 PC was found in lobules IV–VI (in the mammals) and in lobules VII–VIII (in the chicken). In mammals, particularly the monkey and cat, an increased presence of UBCs was observed in vermal sub‐lobules VIc–VIIb,c, a region defined as the oculomotor vermis because of its role in the control of saccadic eye movement. There is also a basic difference between chicken and mammals in the distribution of UBCs within the lobules: in mammals, the lowest density of these nerve cells was found in the peripheral portion of the lobules, near to the pia, while in the chicken, in contrast, the density of UBCs was the highest subpially with fewer UBCs located in the deepest curvature of the lobules. Finally, the functional significance of the differences in the density and in the distribution pattern of UBCs in the cerebellar vermis between the phylogenetically different species investigated is briefly discussed. J. Neurosci. Res. 55:733–748, 1999. 

Secretin and autism: a basic morphological study about the distribution of secretin in the nervous system.

For the first time, the relationship between secretin and autism has been demonstrated by one of us. Intravenous administration of secretin in autistic children caused a fivefold higher pancreaticobiliary fluid secretion than in healthy ones and, at least in some of the patients, better mental functions were reported after the secretin test. Because the precise localization of secretin in the brain is still not completely known, the abovementioned observation led us to map secretin immunoreactivity in the nervous system of several mammalian species. In the present work, the distribution of secretin immunoreactivity in cat and human nervous systems was compared with that of rats using an immunohistochemical approach. Secretin immunoreactivity was observed in the following brain structures of both humans and in colchicine-treated rats: (1) Purkinje cells in the cerebellar cortex; (2) central cerebellar nuclei; (3) pyramidal cells in the motor cortex; and (4) primary sensory neurons. Additionally, secretin immnoreactive cells were observed in the human hippocampus and amygdala and in third-order sensory neurons of the rat auditory system. In cats, secretin was only observed in the spinal ganglia. Our findings support the view that secretin is not only a gastrointestinal peptide but that it is also a neuropeptide. Its presence or the lack of its presence may have a role in the development of behavioral disorders.

Postnatal development of unipolar brush cells in the cerebellar cortex of cat

The postnatal developmental distribution pattern of metabotropic glutamate receptor (mGluR1a) immunoreactive unipolar brush cells (UBCs) was studied in the cerebellar cortex of kittens. On the day of birth (P0) UBCs are already present in the white matter in lobule X of the vermis, but only a few of these cell seemed to migrate to the deeper region of the internal granular layer. By the end of the first week (P8) UBCs were seen to invade the white matter + internal granular layer of lobules IX, VIII, I, and II of the vermis, and they spread further in the transitory area medio‐laterally from the vermis toward the cerebellar hemispheres. By P15, UBCs appeared in lobules III and VII of the vermis, as well as in corresponding lobules of the neocerebellum, with especially high numbers in lobule VII. By P22, UBCs migrated further after their medio‐lateral course in the neocerebellum, and began to invade lobules V and VI. At P62 the amount of UBCs in midsagittal planes of early developing vermal lobules (I, II, VII–X) resembled the P132 or adult pattern. The medio‐lateral migration and incorporation of UBCs into the late‐developing cerebellar lobules V and VI was completed only by P132, when the spatial distribution of UBCs in both the vermal and neocerebellar lobules was comparable to that seen in the 1 year old young adult cat. Although by P132 the postnatal migration of the vast majority of UBCs seemed to be completed, in the cerebellum of adult cats a few migrating UBCs could still be observed in the white matter of the cerebellar lobules, and beneath the ependyma of the fourth ventricle. It is concluded that during ontogenesis the migration course of UBCs follows essentially the developmental sequence of cerebellar lobules, although the incorporation of UBCs into the internal granular layer continues until 4 months postnatally, i.e., much beyond the apparent completion (about two months postnatally) of cytoarchitectonic built up of the cerebellar cortex of kittens. J. Neurosci. Res. 61:107–115, 2000.

Protective effect of the antiepileptic drug candidate talampanel against AMPA-induced striatal neurotoxicity in neonatal rats.

2,3-Benzodiazepines represent a family of specific, noncompetitive AMPA receptor antagonists with anticonvulsant and neuroprotective properties. In this study, the antiexcitotoxic potency of the clinical antiepileptic drug candidate, talampanel (4 x 2 mg/kg), and that of two related 2,3-benzodiazepines, 5-(4-aminophenyl)-8-methyl-9H-1,3-dioxolo[4,5-h][2,3]-benzodiazepine (GYKI 52466) (4 x 10 mg/kg) and GYKI 53784 (4 x 2 mg/kg), was investigated in 7-day-old rats. The AMPA antagonists were applied in four consecutive i.p. injections at 1-h intervals, the first dosage was given shortly after the intrastriatal injection of (S)-alpha-amino-3-hydroxy-5,7-methylisoxazole-4-propionic acid (AMPA) (2.5 nmol). All tested compounds protected animals from brain damage induced by AMPA as assessed 5 days later by using a tissue volume determination method based on computer-aided serial section reconstruction. GYKI 53784 (56.1 +/- 5.0% protection) and talampanel (42.5 +/- 5.3% protection) were more potent neuroprotective agents than GYKI 52466 (21.8 +/- 2.8% protection). Furthermore, the three compounds attenuated the unilateral AMPA injection-induced turning behavior and seizure-like events.Our present findings are in agreement with those of other investigators who found talampanel neuroprotective in various in vivo experimental models. These data indicate that besides being a promising antiepileptic drug candidate talampanel may have a value in the pharmacotherapy of acute and chronic neurodegenerative diseases, including perinatal ischemia/hypoxia-induced brain injuries, as well.

Calretinin-immunoreactive unipolar brush cells in the developing human cerebellum

We have studied the temporal and spatial characteristics of the development of unipolar brush cells (UBCs) in the human cerebellar vermis. Consistently with previous studies in rodents and cat, we have found that unipolar brush cells appear at a relatively late phase of cerebellar development and their development continues up to and beyond the first postnatal year. A series of 23 normal human brains, including 5 adult and 18 fetal or infant brains (between the 24th gestational week and the 11th postnatal month) were used. In order to visualize unipolar brush cells, calretinin‐immunocytochemistry was performed on formaldehyde‐fixed, paraffin‐embedded blocks of the cerebellar vermis. Our results show that calretinin‐immunoreactive unipolar brush cells are not yet present in the cerebellar vermis at the 28th gestational week. At birth, they are present in a relatively small number, mostly in the vestibular lobules. At the 3rd, 5th, 8.5th and 11th postnatal months the number of calretinin‐immunoreactive unipolar brush cells gradually increase, first appearing in the vestibular lobules, followed by the invasion of the later developing vermal lobules, spreading in a rostro‐caudal and proximo‐distal direction. Although at the 11th postnatal month unipolar brush cells exhibited adult‐like morphological and distributional features, their number appeared to be lower than in the adult cerebellum. The late maturation of unipolar brush cells implies that the cytoarchitectonical development of the human cerebellum is not completed by the end of the first postnatal year.

Distribution of mGluR1α and SMI 311 immunoreactive Lugaro cells in the kitten cerebellum

The Lugaro cell is a feedback interneuron of the cerebellar cortex, recognizable by its characteristic morphology. Postnatal neuronal migration to the cortex has been described for several cerebellar interneurons. Since in our previous studies we observed Lugaro-like cells (LCs) in the white matter (WM) and internal granular layer (IGL) of the cerebellum of young cats, we assumed that a proportion of these cells migrate also postnatally to their destination. In the present study using and immunostaining for the metabotropic glutamate receptor mGluR1α and neurofilament protein SMI 311 the number and spatial distribution of LCs at different postnatal days were investigated.We found that the number and distribution of both mGluR1a-immunoreactive (ir) and of SMI 311-ir LCs changed with age in the developing cerebellar cortex of kittens: developing LCs express mGluR1α already in the newborn, while expression of SMI 311-ir in LCs appears only about a week later. At postnatal day 1 (P1) relatively few mGluR1-ir LCs were detected in the WM and at the border of WM and IGL. Later, their number increased sharply until P15 (6–7 fold) and decreased continuously between P15 and P135. SMI 311-ir LCs were not present at P1 and even at P8 only a few were observed in the WM or in infraganglionic positions. Their number increased gradually (12–14 fold) until adulthood when their number was stabilized at 8.000–10.000/cerebellum. At the same time the number of probably ectopic SMI 311-ir LCs decreased with age: at P22 about one third of them was found in “ectopic” position, whereas in the adult cat only about 10–12% of LCss was either in the WM or scattered in the whole depth of the granular layer.These results suggest that: (1) most LCs appear in the cerebellar cortex postnatally; and (2) postnatal migration and incorporation of LCs to the cortex is a much longer process than previously expected, occurring even after the cytoarchitectonic built-up (about P65–P70 in cat) of the cerebellum.

Quantitative analysis of the postnatal development of Purkinje neurons in the cerebellum of the cat

We have studied the postnatal quantitative changes of cortical Purkinje neurons in the cerebellum of the cat at the following postnatal groups of age: P0, P42, P72 and adults. An unbiased counting method, the optical fractionator was used for the estimation of Purkinje cell numbers. A significant increase of Purkinje cell number was found between P0 (1.097 × 106) and P42/P72 (1.805 × 106 and 1.895 × 106) declining to 1.429 × 106 in the adult, still 30% higher than in the newborn. It was also observed that during the first few postnatal weeks large “gaps” were present in the Purkinje monolayer as revealed by Nissl staining and metabotropic glutamate receptor 1α immunocytochemistry. These Purkinje cell gaps were observed most frequently in well‐definable areas, especially in the intermediate zone of the neocerebellum. Simultaneously with the numerical increase of Purkinje neurons between the P0 and P72 age groups, these gaps disappeared after the third postnatal week resulting in the completion of the Purkinje monolayer in the whole cerebellum.

An in vitro electrophysiological and Co2+-uptake study on the effect of infraorbital nerve transection on the cortical and thalamic neuronal activity

Changes of neuronal membrane characteristics in somatosensory barrel cortex and barreloid thalamus were investigated in rats following unilateral transection of the infraorbital nerve. Kainate induced Co2+-uptake method and image analysis were used to assess the Ca2+ permeability of non-NMDA (N-methyl-D-aspartate) glutamate receptors. Changes in some biophysical parameters of the affected cortical neurons were also investigated by intracellular recording in slice experiments. The altered neuronal activity was measured on days 1, 5 and 14 after surgery. Kainate induced Co2+ uptake increased markedly reflecting enhanced Ca2+ permeability of alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionate/kainate (AMPA/KAIN)-type receptors. Changes were more pronounced in the cortex than in the thalamus and peaked on the first day following nerve transection. After that, parameters gradually returned to the normal level. However, a small enhancement was still detectable in the cortex at the end of the 2-week-long observation period. In parallel with the increased Co2+-uptake, moderate membrane potential changes, stronger spiking activity and enhanced excitability were characteristic for cortical neurons. The observed alterations in neuronal characteristics underlie the reorganization and regeneration processes following injuries or surgeries. We can conclude that immediate change of the receptive field in the barrel cortex following unilateral nerve transection is based on changes in biophysical parameters of the neurons. Altered peripheral activation evokes changes in the neuronal activity, thus providing opportunity for a quick synaptic rearrangement. AMPA/KAIN-type glutamate receptors have a decisive role in the regulation of these processes. This kind of synaptic plasticity is more significant in the cortex than in the thalamus.

Retardation in somatosensory cortex development induced by postnatal BrdU treatment in mice

Cerebral dysgeneses are in the background of several neurological and mental disturbances. The aim of the present study was to investigate structural and activity changes following disturbed postnatal neuronal development in mice. Newborn C57Bl6 mice were exposed to 5‐bromo‐2′‐deoxyuridine (BrdU: daily 50 μg/g body weight) during a period between postnatal days P0–P5 or P0–P11, respectively, and neuronal malformation and malfunctioning of somatosensory (barrel field) cortex was analyzed in adolescent animals. Alterations in histological architecture of interneuronal and glial elements were studied and correlated with electrophysiological modifications. Between P30 and P35 days litters underwent ex vivo electrophysiological experiments to examine the changes in basic excitability and in synaptic efficacy. Parallel immunohistochemistry was performed to detect BrdU, GABA and GFAP.