I. Dori

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.

The cholinergic innervation of the rat cerebral cortex shows two distinct phases in development

SummaryThe cholinergic innervation of the rat cerebral cortex was examined in pre- and postnatal life using immunohistochemistry with a monoclonal antibody directed against choline acetyltransferase (ChAT). Our observations show that there are two separate phases in the development of the cholinergic innervation of the rat neocortex. The first, a transient phase, occurs in the late stages of gestation and in the perinatal period. During this time, ChAT-labelled cells (neuroblasts, as well as immature pyramidal and non-pyramidal neurons) are present throughout the entire rostro-caudal extent of the primordial cortex. The fate of these cells, which are not visible shortly after birth, is unknwon as is their functional role in the developing cortex. The second phase in the development of the cholinergic innervation begins in the middle of the second postnatal week. At this stage only a few faintly stained neurons and fibres appear in the cortex. Their numbers and staining intensity increase gradually until the fifth postnatal week when ChAT-labelled neurons and axonal arbours appear indistinguishable from their adult counterparts. The pattern of development observed in the second phase parallels closely that shown in a recent analysis of cortical ChAT activity during postnatal life.

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.

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.

Noradrenergic innervation of the developing and mature septal area of the rat

ABSTRACT The noradrenergic innervation of the developing and mature septal area of the rat was examined with light and electron microscopic immunocytochemistry using an antibody against dopamine‐β‐hydroxylase. At birth, a small number of relatively thick noradrenergic fibers were found to innervate the lateral septum (mainly its intermediate part) and the nuclei of the vertical and horizontal limbs of the diagonal band of Broca. By postnatal day 7, a substantial increase in their density was observed. At this age some labeled fibers left the medial forebrain bundle and invaded the nucleus of the horizontal limb of the diagonal band. These fibers then ran in a ventrodorsal direction and innervated the nucleus of the vertical limb before entering the medial septum. Immunoreactive fibers were finer and more varicose than at birth. In the subsequent 2 weeks, the density of labeled fibers in the septal area was further increased. By postnatal day 21, the distribution pattern and density of the noradrenergic innervation appeared similar to the adult. In the adult, noradrenergic fibers exhibited more varicosities than in younger rats. Electron microscopic analysis revealed a low proportion (peaked at P7) of noradrenergic varicosities engaged in synaptic contacts throughout development. The overwhelming majority of these synapses were symmetrical, predominantly with small or medium‐sized dendrites. The present findings provide the morphological basis for the functional interactions between noradrenergic afferents and neuronal elements in the septal area. The low proportion of synaptic contacts found in this study suggests that noradrenaline may exert its action in the septal area mainly through transmission by diffusion (volume transmission), as has been suggested for other areas of the developing and adult brain. J. Comp. Neurol. 476:80–90, 2004.

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.