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Biomedical subjects

A Dinopoulos

Publications and source records attributed to A Dinopoulos.

At least 19 recordsLinked to original sources

Chronic progressive autonomic dysfunction in a dog.

A 3.5-year-old intact male American Pit Bull was presented because of urinary incontinence and dysuria. Constipation, followed by diarrhoea, ocular disturbances and finally regurgitation developed over the next 4 years. Autonomic dysfunction was evidenced by clinical presentation, as well as positive ophthalmic pilocarpine test and subnormal Schirmer tear test. Diagnosis, however, was established through histopathological and immunohistochemical examination. Lesions attributable to inflammatory degenerative neuropathy of the autonomic ganglia, which represents one of the various types of human autonomic failure, were detected.

Animals↗

Mitochondrial dysfunction in a patient with Joubert syndrome.

Joubert syndrome is a genetically heterogeneous disorder. The diagnostic criteria include episodic hyperventilation, abnormal eye movements, psychomotor retardation, hypotonia, ataxia, and the characteristic neuro-imaging findings (molar-tooth sign). Many of these clinical features have been observed in new-borns with mitochondrial disorders as well. Congenital brain malformations, including cerebellar hypoplasia, have been described in pyruvate dehydrogenase deficiency. Malformations of the vermis and the cerebellar peduncles, with the lack of axonal decussations, however, are characteristic for Joubert syndrome but unique in patients with mitochondrial disorders. Here, we describe a child with Joubert syndrome presenting with primary lactic acidemia, decreased pyruvate oxidation rates, decreased ATP production, and a mildly decreased pyruvate dehydrogenase complex activity measured in a fresh muscle biopsy. Sequence analysis of the PDHc E1 alpha gene and the PDHX genes revealed no mutations. The patient received continuous feeding through a feeding tube for two years and showed a significant clinical improvement with a complete resolution of the chronic lactic acidemia. A second muscle biopsy revealed significantly decreased pyruvate oxidation rates and ATP production, but a normal pyruvate dehydrogenase complex activity. We suggest that the described mitochondrial dysfunction in our patient is secondary to an underlying mutation leading to Joubert syndrome.

Abnormalities, Multiple↗

Brain MRI and proton MRS findings in infants and children with respiratory chain defects.

OBJECTIVE: The aim of this study is to describe neuroimaging patterns in children with respiratory chain (RC) defects using magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy (MRS) and to assess their role in the diagnostic evaluation. METHOD: Neuroimaging studies of 49 children (newborn to 15 years old) with biochemical evidence of RC defect were reviewed. Patients were divided in 3 groups ("definite" = 24, "probable" = 14, "possible" = 11) according to Modified Adult Criteria for the diagnosis of RC defect. Eighty-one MRI studies were reviewed for deep gray and white matter changes, degree of myelination, cerebral and cerebellar atrophy, and 67 proton MRS studies were assessed for the presence or absence of lactate elevation, as well as NAA/Cr ratio. The findings were compared among the 3 groups with chi-square test. RESULTS: All patients with "pure" myopathy had normal imaging studies. In patients with CNS involvement, significant differences in the frequency of imaging abnormalities among groups were found for deep gray matter (43 %/8 %/0 %; p = 0.01) and for the presence of lactate elevation on proton MRS (81 %/31 %/0 %; p = 0.001). CONCLUSION: Brain MRI and proton MRS abnormalities were observed only in association with clinical CNS involvement. Deep gray matter signal abnormalities on structural imaging and lactate elevation on proton MRS were more frequently observed in the "definite" group and represent neuroimaging markers for RC mitochondriopathy.

Aspartic Acid↗

Postnatal development of the dopaminergic system of the striatum in the rat.

The dopaminergic innervation of the developing caudate-putamen (patches and matrix) and nucleus accumbens (shell and core) of the rat was examined with light and electron microscope immunocytochemistry, using antibodies against dopamine. Light microscopic analysis showed, in accordance with previous studies, that early in life, dopaminergic fibers were relatively thick and present throughout the striatum. Their distribution was heterogeneous, showing dense aggregations, the so-called dopamine islands. The pattern of innervation became more uniform during the third postnatal week with most of the dopamine islands no longer detectable. For electron microscopic analysis, parts of the caudate-putamen containing dopamine islands or matrix, and of the nucleus accumbens, from the shell and the core of the nucleus, were selected. This analysis revealed that symmetrical synapses between immunoreactive profiles and unlabeled dendritic shafts predominated throughout development but, at the late stages, symmetrical axospinous synapses also became a prominent feature. These findings indicate that: (1) although the caudate-putamen and the nucleus accumbens have different connections and functions, they exhibit similar types of dopaminergic synapses, and (2) the relatively late detection of dopaminergic axospinous synapses suggests that the development of the dopaminergic system in the striatum is an active process, which parallels the morphological changes of striatal neurons and may contribute to their maturation.

Aging↗

Variations of ferritin levels over a period of 15 years as a compliance chelation index in thalassemic patients.

We studied the changing pattern of the distribution of ferritin levels in 430 regularly-transfused patients with thalassemia in an attempt to evaluate compliance of chelation with deferoxamine. The study covered 15 years and was divided in three periods: 1981-1985, 1986-1990, and 1991-1995. The patients were stratified in age-groups. The mean ferritin levels of each period were calculated for each patient individually. The study showed that: (i) When all the patients were compared as a group, there was a significant decrease in mean ferritin between 1981-1985 and 1991-1995, despite a significant change in the patients' mean age; (ii) When patients of same age were compared between periods, there was a decrease in mean ferritin between 1981-1985 and 1991-1995, as well as a decrease in the proportion of patients with ferritin >4000 microg/L, with a parallel increase in the proportion of patients who had ferritin <2000 microg/L; (iii) When the same patients were followed longitudinally, they showed a decrease in their ferritin levels in all age groups with the exception of the late adolescence period. The decrease in iron overload observed in patients on close follow up implies that compliance with chelation therapy has improved with time and therefore, a favourable influence in survival could be expected.

Adolescent↗

The development of the synaptic organization of the serotonergic system differs in brain areas with different functions.

The serotonergic innervation of the developing superior colliculus and ventrolateral nucleus of the thalamus of the rat were studied with light and electron microscope immunocytochemistry. We compared the pattern of innervation and synaptic organization of the serotonin (5-HT) system in the superficial and deep layers of the superior colliculus. We also compared the developmental pattern of synaptic incidence of 5-HT varicosities in the superior colliculus with that in the ventrolateral nucleus. Serotonin fibers were present in the superior colliculus at birth, concentrated mainly in the deep layers, whereas the superficial layers were only sparsely innervated. By the end of the first postnatal week the overall density of 5-HT fibers increased, but was still higher in the deep than in the superficial layers. The distribution pattern, density, and morphology of serotonergic axons acquired mature features by the end of the third postnatal week. In the adult, these axons were thin, varicose, forming a complex network which was denser in the lower part of the superficial layers and the upper part of the deep layers. Electron microscopical analysis revealed that the vast majority of 5-HT varicosities established symmetrical synapses with dendritic shafts in all layers of the superior colliculus throughout development. In the superficial layers, known to be involved in visual functions, the proportion of varicosities forming synapses increased gradually from birth to reach a peak at the end of the first postnatal week, then declined markedly in the subsequent 2 weeks before rising again at later stages. In contrast, in the deep layers and in the ventrolateral nucleus of the thalamus, areas involved in motor functions, the proportion of 5-HT varicosities engaged in synaptic contacts showed a continuous increase from birth until adulthood. Considering these results together with data from our previous studies, we speculate that the regional heterogeneity in the synaptic organization of the serotonergic system may reflect a differential role of 5-HT in the development of brain areas with different functions.

Aging↗

Distribution and synaptology of dopaminergic fibers in the mature and developing lateral septum of the rat.

The dopamine innervation of the adult and developing lateral septum of the rat was investigated with light and electron microscope immunocytochemistry using anti-dopamine antibodies. Light microscopic analysis showed that the pattern of innervation of the lateral septum exhibited a marked reorganization during the first 2 postnatal weeks, when it acquired features comparable to the adult. Ultrastructural analysis suggested that there may be two different dopamine inputs in the lateral septum. The first develops earlier in life and, through symmetrical axodendritic synapses, affects remote parts of neurons and may cause inhibition. The second develops later and, through asymmetrical axosomatic synapses, affects neuronal somata and may cause excitation. These findings may explain the reported contradictory results concerning the physiological role of dopamine in neurons of the lateral septum.

Animals↗

Transient features of tachykinin peptide innervation of the dorsal lateral geniculate nucleus of the rabbit during postnatal development.

The tachykinin peptide innervation of the developing dorsal lateral geniculate nucleus of the rabbit was studied with immunocytochemical techniques at the light and electron microscopic levels by using an antibody directed to the C-terminal of the mammalian tachykinin peptides. At birth, the rabbit's lateral geniculate nucleus was densely innervated by a large number of labeled fibers. These relatively unbranched axons were dispersed throughout the nucleus but showed a higher density of fiber packing in the external layer of the alpha sector. Over the next three weeks this pattern of distribution changed dramatically. Immunoreactive fibers were gradually eliminated from most of the nucleus, and, by the end of the third postnatal week, they appeared as a narrow plexus deep to the optic tract. At the same time, these axons showed major morphological alterations as they gradually became thicker and developed terminal arborizations characterized by spherical or elongated enlargements. Overall, by the end of the third postnatal week, the pattern of distribution and appearance of tachykinin-labelled fibers in the dorsal lateral geniculate nucleus were similar to those described in the adult rabbit (Brecha et al., 1987). Ultrastructural analysis has shown that during the first week, tachykinin-immunoreactive profiles appeared as round or elongated varicosities making asymmetrical synapses with dendritic shafts throughout the lateral geniculate nucleus. Thereafter, as they were progressively restricted to the external layer of the alpha sector of the nucleus, they began to form multiple synaptic contacts with neuronal processes in complex glomerular neuropil. On the basis of the present and previous findings, we suggest that tachykinin peptides not only play a role as putative neurotransmitters in the retinogeniculate pathway, but they may also play a role in the development of the lateral geniculate nucleus and of the retinogeniculate projection system in the rabbit.

Animals↗

The serotonin innervation of the basal forebrain shows a transient phase during development.

The serotonergic innervation of the adult and developing basal forebrain nuclei of the rat was studied with immunocytochemical techniques at the light and electron microscopic levels. A substantial number of relatively thick serotonergic fibers with few varicosities and random orientation were observed at the time of birth. During the subsequent weeks, the serotonergic fibers increased in number and became thinner with many varicosities. They were also re-oriented, and around the end of the third postnatal week they exhibited the pattern of distribution and density seen in the adult. Electron microscopic analysis revealed that serotonin varicosities formed symmetrical or asymmetrical synapses mainly with dendritic shafts throughout postnatal life. Stereological extrapolation from single sections to the whole volume of varicosities showed that the percentage of serotonin varicosities engaged in synaptic junctions varied according to age. The proportion of labelled varicosities forming synapses increased from birth (21.3%) to the end of the second postnatal week (42.5%), then declined markedly in the following week (17.1%) before increasing again to an adult value of 46%. These findings suggest that the formation of synaptic connections by serotonin axons in the basal forebrain shows two distinct phases in postnatal development: exuberant synapses present in the first two weeks of life may be related to the involvement of serotonin in the maturation of this area, whereas synapses formed later in development may affect the functional state of basal forebrain projections to the neocortex and hippocampus. Thus, at these late stages of development and in the adult, serotonin may influence the activity of these forebrain structures both directly and indirectly.

Animals↗

Regional differences in the ontogeny of the serotonergic projection to the cerebral cortex.

We used light and electron microscope immunocytochemistry to compare the development of the pattern of innervation and synaptic organization of the serotonin (5-HT) afferent system in sensory (visual and somatosensory) and motor areas of the rat cerebral cortex. Serotonin-labeled fibers were present in all cortical areas at birth appearing as two tangential streams, one above and one below the cortical plate. These fibers gradually arborized sending branches into all cortical layers in an "inside-out" sequence that broadly paralleled the gradient of neurogenesis and differentiation in the cortex. A striking feature at the early stages of postnatal development of the visual and somatosensory cortex was the transient presence of a dense accumulation of 5-HT fibers in layer IV. In agreement with earlier reports, transient aggregations of serotonergic axons characterized sensory but not motor areas of the cortex. The innervation pattern characteristic of the adult cortex was attained by the end of the 3rd postnatal week. Electron microscopic analysis of the developing visual cortex showed that 5-HT-containing axonal varicosities formed synaptic contacts, predominantly of the asymmetrical variety, throughout postnatal life. The proportion of varicosities forming synapses increased gradually from birth to reach a peak at the end of the 2nd week, then declined markedly in the subsequent week before rising again at later stages. It appears that the formation of exuberant synapses by 5-HT axons coincides with the period of maturation of the neuronal circuitry of this cortical area. When quantitative analysis was restricted only to layer IV, the proportion of varicosities forming synapses reached a peak at the end of the 1st week of life, when transient innervation was most prominent in this layer. These results suggest that the effects of this dense band of serotonergic axons may be mediated through axodendritic synapses. The types of postsynaptic elements involved in the formation of synapses varied according to age, suggesting progressive morphological differentiation of cortical target neurons or, alternatively, a continuous process of removal and establishment of new connections by 5-HT axons in the visual cortex. In contrast, analysis in the motor cortex revealed a continuous increase in the proportion of 5-HT varicosities engaged in synaptic contacts from birth to the late sages of development. The results presented here provide anatomical evidence for a differential role of 5-HT in the maturation of the cerebral cortex.

Age Factors↗

Serotonergic innervation of the lateral geniculate nucleus of the rat during postnatal development: a light and electron microscopic immunocytochemical analysis.

The serotonergic innervation of the developing lateral geniculate nucleus of the rat was studied with immunocytochemical techniques at the light and electron microscope levels. A relatively small number of thick serotonergic fibers were observed at the time of birth, distributed more densely in the ventral portion of the nucleus and in the intergeniculate leaflet than in the dorsal lateral geniculate nucleus. By the end of the first postnatal week, this distribution pattern was more clearly established, but the number of immunoreactive fibers was increased. Thereafter, and until the adult pattern was established at the end of the third postnatal week, serotonergic fibers increased further in number and changed morphologically (e.g., they became finer and more ramified with closely spaced varicosities), but their pattern of distribution remained unchanged. Electron microscopical analysis of the dorsal lateral geniculate nucleus revealed that the vast majority of serotonin varicosities formed asymmetrical synapses with dendritic shafts; axosomatic synapses were a feature of the nucleus only at the time of birth. The proportion of serotonin varicosities forming synapses increased gradually from birth to reach a peak at the end of the second postnatal week, then declined markedly in the following week before increasing again at a later stage. It may be speculated that synapses formed during the first two weeks of life may be related to the involvement of serotonin in the morphogenesis of the lateral geniculate nucleus, whereas those formed later in development may be involved in the mediation of neurotransmitter effects.

Aging↗

The development of the serotonergic fiber network of the lateral ventricles of the rat brain: a light and electron microscopic immunocytochemical analysis.

The development of the serotonergic innervation of the lateral ventricles of the rat brain during the first five postnatal weeks was studied with immunocytochemical techniques at the light and electron microscopic levels. In the lateral ventricles of newborn rats serotonergic fibers are only rarely seen. During the first postnatal week the number of serotonergic fibers increases but they are straight and thick, bearing only a few varicosities. By the end of the second postnatal week, however, they become finer, exhibit a large number of varicosities, and form a dense supraependymal network. During the following weeks this network becomes slightly denser but the morphology of fibers as well as their distribution pattern remain unchanged. Examination of sagittal vibratome sections revealed that a group of serotonergic fibers leaves the medial forebrain bundle and turning dorsocaudally between the corpus callosum and the caudate/putamen enter the lateral ventricle from its rostral pole. They then spread to form the supraependymal network of the lateral ventricles and probably of the rest of the ventricular system. Ultrastructural analysis showed that serotonin varicosities are located close to the ventricular surface of the ependymal lining but never make synapses with the ependymal cells. Examination of a large number of labeled fibers and varicosities showed that they are never located between the ependymal cells or in the subependymal layer. This finding was confirmed by examining series of semithin sections. On the basis of these and previous findings we suggest that serotonergic fibers arising in the midbrain raphe nuclei enter the lateral ventricle from its rostral pole, form a dense network within the ventricles, and release their content into the cerebrospinal fluid. This system, as judged with morphological criteria, matures by the end of the second postnatal week.

Aging↗

Reciprocal connections of the motor neocortical area with the contralateral thalamus in the hedgehog (Erinaceus europaeus) brain.

Horseradish peroxidase unilateral injections in various neocortical areas (prefrontal, somatosensory, auditory, visual) of the hedgehog (Erinaceus europaeus) brain resulted in labelling of nuclei in the ipsilateral thalamus known from studies in other species and in the hedgehog to project to these areas. However, injections in the motor area resulted in retrograde and anterograde labelling of nuclei in both the ipsilateral and contralateral thalamus. These nuclei included the ventral lateral nucleus (VL), the intralaminar nuclei (ILN), the mediodorsal nucleus (MD) and midline nuclei. Large unilateral injections located mainly laterally in the thalamus labelled cells, contralaterally, in the ventral lateral geniculate nucleus, the intergeniculate leaflet and the reticular nucleus of the thalamus, but never in VL, ILN and MD. The present results confirm previously described bilateral thalamocortical projections from the VL to the somatosensorimotor area in this species (Regidor and Divac, Brain Behav. Evol., 39, 265-269, 1992) and in addition demonstrate that (i) bilateral thalamocortical projections are established preferentially with the motor area, (ii) several nuclei are involved in such connections, (iii) these connections are reciprocal and topographically organized, and (iv) labelling in the contralateral thalamus observed in the present study is not a result of transneuronal transport of the tracer through thalamothalamic connections. This organization is unique among mammals and supports previous anatomical and electrophysiological findings, on the basis of which it has been suggested that the hedgehog retains a primitive character in neocortical and thalamic evolution.

Animals↗

Serotonergic innervation of the mature and developing lateral septum of the rat: a light and electron microscopic immunocytochemical analysis.

The serotonergic innervation of the adult and developing lateral septum of the rat was studied with immunocytochemical techniques at the light and electron microscopic levels. A few, relatively thick serotonergic fibres are found in the lateral septum at the time of birth, but they are restricted to its medial part. They subsequently extend towards the lateral ventricle, increase in number and attain their final distribution pattern by the end of the first postnatal week. Thereafter they become finer, with regularly spaced varicosities, show a higher density, and generally exhibit features, density, and pattern of innervation comparable to the adult at the end of the third postnatal week. In the dorsal portion of the lateral septum, serotonergic fibres form characteristic pericellular basket-like arrangements around cell somata and their primary dendrites. These baskets are first observed at P7, and subsequently increase both in number and in terms of the number of serotonergic terminals which they comprise. The present findings suggest that the development of serotonergic innervation of the lateral septum parallels the neuronal differentiation in this area. Ultrastructural analysis has shown that the vast majority (congruent to 95%) of serotonin varicosities make symmetrical synapses with somata, dendritic shafts and spines. These varicosities in new-born animals are in close association with neuronal elements, without any intervening neuroglial processes, but towards the end of the first postnatal week they exhibit well-defined synaptic specializations. The mean diameter of serotonergic varicosities making synapses does not change substantially with age. Serotonin-receptive neurons have several morphological features in common with the type I cells described in a previous Golgi study of the lateral septum [Alonso and Frotscher (1989) J. comp. Neurol. 286, 472-487]. Some speculations on the chemical identity of the serotonin-receptive cells have been put forward in the present study but double-labelling studies will certainly shed more light on the organization of the serotonergic innervation of the lateral septum.

Animals↗

Proportion of glutamate- and aspartate-immunoreactive neurons in the efferent pathways of the rat visual cortex varies according to the target.

Immunohistochemistry, with antisera directed against glutamate (Glu) or aspartate (Asp), was combined with wheat germ agglutinin-horseradish peroxidase (WGA-HRP) histochemistry to examine the distribution, morphology, and proportions of Glu- and Asp-containing neurons that give rise to corticofugal and callosal projections of the rat visual cortex. WGA-HRP injections in the dorsal lateral geniculate nucleus resulted in retrograde labelling of small and medium-sized cells throughout layer VI of the visual cortex. Of these cells, 60% were also Glu-immunoreactive and 61% Asp-positive. WGA-HRP injections in the superior colliculus labelled large and medium-sized neurons in the upper portion of layer V of the visual cortex. Of these cells, 46% were also stained for Glu and 66% for Asp. Injections in the pontine nuclei resulted in retrograde labelling of cells in the deeper part of cortical layer V. Retrogradely labelled cells, which were also immunoreactive for Glu or Asp, were large pyramidal cells. Corticopontine neurons, which were also Glu-positive, accounted for 42% of the total number of WGA-HRP labelled cells, whilst for Asp-positive neurons this percentage was 51%. Finally, after injections in the visual cortex, retrogradely labelled small and medium-sized cells were found throughout layers II-VI in the contralateral visual cortex. Of these neurons, 38% were also labelled for Glu while 49% were also Asp-immunoreactive. The present results demonstrate that substantial proportions of projection neurons in the rat visual cortex are immunoreactive for Glu or Asp, suggesting that these excitatory amino acids are the major transmitters used by the cortical efferent systems examined. Furthermore, the proportions of these immunoreactive neurons in the efferent pathways vary according to the target.

Animals↗

Claustrum in the hedgehog (Erinaceus europaeus) brain: cytoarchitecture and connections with cortical and subcortical structures.

The cytoarchitecture of the claustrum in the hedgehog (Erinaceus europaeus) brain, the morphology of its neurons, and the efferent connections with cortical and subcortical structures were studied with the Nissl and Klüver-Barrera, the Golgi, and the horseradish peroxidase methods. It was found that the claustrum is a well developed nucleus in the hedgehog telencephalon and, as in other mammals, is divided into dorsal and ventral parts. In Golgi-stained sections, spiny multipolar cells are the predominant neurons of both the dorsal and the ventral claustrum and are projection neurons. Aspiny multipolar neurons with fewer, often beaded, dendrites constitute a minority in both divisions and are interneurons. Injections of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) in the prefrontal, motor, somatosensory, auditory and visual areas, and HRP or WGA-HRP injections in the thalamus showed that: (1) the claustroneocortical projections originate in the dorsal claustrum and are distributed to the entire neocortex; these projections are mainly ipsilateral but some also originate contralaterally; (2) the claustroneocortical projections show a rough topographic organization; there exists a substantial degree of overlap; and (3) the claustrothalamic projection, arising throughout the dorsal claustrum, is strictly ipsilateral. No evidence of a thalamoclaustral projection was found. The present results suggest that, although the hedgehog has been referred to as a "paleocortical mammal" owing to the great development of its rhinencephalic structures in comparison with its small neocortex, the dorsal claustrum is well developed and is connected with all neocortical areas as well as with the thalamus, establishing it as a key structure in the hedgehog forebrain.

Animals↗

The development of neurons in the nuclei of the horizontal and vertical limb of the diagonal band of Broca of the rat: a qualitative and quantitative analysis of Golgi preparations.

We have studied the morphological alterations of neurons in the nuclei of the horizontal (NHL) and vertical (NVL) limbs of the diagonal band of Broca of rats from late embryonic life to maturity using the Golgi-Stensaas and Golgi-Cox methods. During late embryonic life and in the first postnatal days, the two nuclei of the diagonal band of Broca were found to be located near the ventral surface of the brain. Shortly thereafter, neurons in the NHL and NVL gradually take up the positions which they normally occupy in adulthood. At this stage neurons were small with round or elongated somata and 1-3 primary dendrites that only occasionally bore spines and very seldom showed varicosities, features commonly shown by neurons at later postnatal ages. At birth, cells showing varying soma shapes and dendritic morphology were present, and by postnatal day 4 (P4) the three forms of neurons previously described in adult rats (Dinopoulos et al., J. Comp. Neurol., 272 (1988) 461-474) were readily distinguished. During the second postnatal week, the size of cell somata as well as the number, size and extent of dendritic branching underwent considerable increases in both nuclei and at P14 neurons showed features typical of their adult counterparts. In addition they showed a dramatic increase in the number of spines which was followed during the next 10 days by a substantial decrease. Overall, the dendritic geometry of neurons in the NHL and NVL did not change significantly after P14, although their cell bodies continued to increase in size until the middle of the fourth and fifth postnatal weeks respectively. These findings suggest that neurons in the nuclei of the diagonal band of Broca show continuous growth from embryonic life to the end of the second postnatal week when they acquire morphological features comparable to the adult. Thereafter they exhibit only minor morphological alterations with the exception of extensive spine elimination which is pronounced during the third postnatal week and continues until adulthood.

Animals↗