A. Dinopoulos

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.

Somatostatinlike immunoreactive neurons in the hedgehog (Erinaceus europaeus) and the sheep (Ovis aries) central nervous system.

The morphology and distribution of somatostatinlike immunoreactive perikarya in the central nervous system of the hedgehog and sheep have been studied by means of the peroxidase‐antiperoxidase immunohistochemical method. Intracerebroventricular colchicine infusion not only enhanced the immunostaining but also revealed new immunoreactive cell bodies. In both hedgehog and sheep immunoreactive neurons of various forms, ranging from 12 to 28 μm in diameter, were observed in a number of homologous brain structures. However, some species‐related differences were noticed. Thus, somatostatinlike immunoreactive neurons were found only in the hedgehog anterior olfactory nucleus, olfactory tubercle, nucleus accumbens, medial parabrachial nucleus, raphe nuclei of the medulla, and spinal trigeminal nucleus, whereas some somatostatin‐positive neurons were observed in the locus coeruleus and the pontine reticular formation of the sheep only.

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.

Morphological characterization of cholinergic neurons in the horizontal limb of the diagonal band of Broca in the basal forebrain of the rat

SummaryA monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine synthesizing enzyme, was used to identify cholinergic neurons in the nucleus of the horizontal limb of the diagonal band of Broca at the light and electron microscopic levels. ChAT-labelled somata were fusiform, triangular or round in shape and varied considerably in size. Depending on the type of the cell, one to four dendrites emerged from the soma, but an axon could rarely be seen. The nuclei of most cells were round or oval, showed invaginations and displayed prominent nucleoli. The karyoplasm of the larger fusiform and triangular neurons contained abundant organelles including parallel arrays of granular endoplasmic reticulum. The synaptic input to labelled perikarya and proximal dendrites was sparse. It consisted chiefly of asymmetrical synaptic contacts, sometimes with postjunctional densities, but a few symmetrical synapses were also noted. ChAT-positive axon terminals were not identified which suggests that axon collaterals are rare within the nucleus of the horizontal limb of the diagonal band of Broca.

The development of basal forebrain projections to the rat visual cortex

SummaryThe development of the basal forebrain projections to the visual cortex of the rat were studied using retrograde tracer techniques. Injections of wheat germ agglutinin-horseradish peroxidase placed in the visual cortex of newborn animals resulted in labelling of neurons throughout the basal forebrain nuclei. Although at this time the overall distribution of retrogradely labelled cells within the basal forebrain appeared similar to that seen in the adult, cells were smaller and weakly stained. It was only at the end of the second postnatal week that the somata of stained neurons showed sizes and staining intensity comparable to the adult. This precedes or coincides with the reported significant increases in cortical and basal forebrain ChAT activity and the first detection of ChAT-labelled fibres in this cortical area. These data suggest an important developmental point around the end of the second postnatal week that may correspond to the time when a significant number of cholinergic axons first appear within the visual cortex. They also suggest that the cholinergic projections to the visual cortex develop late in comparison with the thalamic and other subcortical afferents in this cortical area.

Noradrenergic innervation of the developing and mature visual and motor cortex of the rat brain: A light and electron microscopic immunocytochemical analysis

The noradrenergic (NA) innervation of the developing and adult visual and motor cortex of the rat was examined with light and electron microscopic immunocytochemistry by using antibodies against dopamine‐β‐hydroxylase. At birth, NA fibers were present in both cortical areas, appearing as two tangential streams, one above and the other below the cortical plate. During the subsequent weeks, these two streams arborized gradually innervating all cortical layers. The adult pattern of distribution was attained by postnatal day 14, but the density of innervation, which was higher in the motor than in the visual cortex, appeared similar to the adult by the end of the third postnatal week. Electron microscopic analysis revealed that a low proportion of NA varicosities (the highest value was 12% in the adult motor cortex in single sections) were engaged in synaptic contact, throughout development, in both areas examined. The overwhelming majority of these synapses were symmetrical, involving predominantly small or medium dendrites. This evidence suggests that transmission by diffusion is the major mode of NA action in the developing and adult cerebral cortex. Noradrenaline released in the rare synaptic junctions may act mainly to reduce the activity of its cortical targets. The results altogether provide morphologic evidence for an involvement of noradrenaline in the development of the neocortex and, along with earlier data on the serotonergic system, indicate that the monoaminergic systems are endowed with a specific anatomic organization in various areas of the brain. J. Comp. Neurol. 445:145–158, 2002.

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.

NEUROCHEMICAL HETEROGENEITY AMONG CORTICOFUGAL AND CALLOSAL PROJECTIONS

Biochemical, physiological, and anatomical studies over the past 30 years have firmly established glutamate (Glu) as the major neurotransmitter of those cortical neurons which give rise to corticofugal pathways. In the present study we utilized immunohistochemistry, with an antibody directed against Glu, in conjunction with wheat germ agglutinin-horseradish peroxidase (WGA-HRP) histochemistry to examine the Glu-containing neurons which give rise to corticofugal and callosal projections of the rat. Injections of WGA-HRP into the pons labeled cells in layer V of both visual and somatosensory cortices. WGA-HRP-labeled cells which also stained for Glu were large pyramids and in the visual cortex constituted approximately 42% of the total number of neurons which had effectively transported WGA-HRP, while the percentage was 56% in the somatosensory cortex. Following caudate/putamen injections, WGA-HRP-labeled cells were confined to layer V of the somatosensory and motor cortices. Of these cells, 40% in the somatosensory cortex and 53% in the motor cortex were also stained for Glu. Finally, after WGA-HRP injections in the visual cortex numerous WGA-HRP-positive neurons were found throughout layers II-VI around the boundaries between area 17 and areas 18 and 18a of the contralateral hemisphere. Here, 38% of these cells were also labeled for Glu, but this percentage was higher (49%) when layers II-III were considered alone. These findings show that Glu is not the neurotransmitter used overwhelmingly in corticofugal and callosal projections and that different proportions of neurons are Glu-immunoreactive in the systems examined.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

Cortical and brain stem projections to the spinal cord of the hedgehog (Erinaceus europaeus)

SummaryCortical and brain stem neurons projecting to the spinal cord in the hedgehog were studied by means of the horseradish peroxidase (HRP) tracing method. HRP injections were placed in the first cervical segments, in the cervical enlargement (C5-T3) and in the lumbar enlargement. Following injections in the first cervical segments and in the cervical enlargement labelled neurons were observed in the somatic motor and somatic sensory cortices, the paraventricular and the dorsomedial hypothalamic nucleus, the lateral hypothalamic area, the nuclei of field H of Forel, the red nucleus, the mesencephalic reticular formation, the deep layers of the superior colliculus, the Edinger-Westphal nucleus, the periaqueductal grey, the mesencephalic trigeminal nucleus, the loci coeruleus and subcoeruleus, the nuclei raphe dorsalis, centralis superior, raphe magnus, raphe pallidus, and raphe obscurus, the rhombencephalic reticular formation, the lateral, medial and caudal vestibular nuclei, the nucleus ambiguus, the nucleus of the solitary tract and the gracile nucleus. After HRP injections in the lumbar enlargement, labelled neurons were not found in the cortex, the dorsomedial hypothalamic nucleus, the nuclei of field H of Forel, the superior colliculus and the mesencephalic trigeminal nucleus. These results show that cortical and brain stem projection to the spinal cord are comparable to those described in other species.