Callosal projections of the striate cortex in the neonatal rabbit

Summary Callosal fiber projections of the striate cortex were studied in newborn rabbits using the methods of 1) retrograde transport of horseradish peroxidase (HRP) and 2) orthograde transport of tritiated leucine. Data collected from 6–8 day old animals revealed not only the existence of adult-like commissural fibers in the lateral striate cortex bordering the occipital area, but also an aberrant callosal projection from the medial striate cortex, an area not known to have any commissural fibers in the adult brain of those mammalian species studied.This research was supported by NIH grants NS 18512 and EY 00691 to K.L.C. and USPHS postdoctoral fellowship EY 5176 and grants NS 07012 to H.D.B.     

Reciprocal heterotopic callosal connections between the two striate areas in Tupaia

Summary WGA-HRP injections were placed into area 17 close to the border with area 18 of Tupaia belangeri in order to study the callosal connections of the striate area in this animal. Most callosal neurons were found in the striate cortex (57.6–86.9%), some in the extrastriate area 18 (10.6–28.1%), and a few in even more temporal regions (2.5–14.3%). Concerning only the area 17, reciprocal homotopic connections could be observed as a strip along the area 17/18 border. Additionally, heterotopic callosal connections could be seen in regions representing the binocular visual field, especially the lower part. The area 17 cells were mostly located in the supragranular layers II and III (94.1–97.2%). But neurons could also be found in the infragranular layers, especially layer VI (2.6–5.2%) and in layer IV (0.2–1.1%). Homotopic projections were mostly seen in layers IIIc and V. The majority of the supragranular and infragranular neurons are pyramidal cells. However, a newly defined subpopulation of neurons, most probably stellate cells, were discovered forming a band in sublayer IIIc, very close to the layer III/IV border.     

Psychosocial stress affects pineal function in the tree shrew (Tupaia belangeri)

Using a recently developed commercially available radioimmunoassay the concentration of the principal melatonin metabolite 6-sulfatoxymelatonin (aMT6s) in the morning urine of male tree shrews was determined. Chronic social confrontation elicited a drastic increase of aMT6s excretion in subordinate tree shrews, whereas there was a tendency to reduced excretion of the melatonin metabolite in dominant animals. These results substantiate the function of the pineal gland in transforming stimuli from the social environment to endocrine information and, therefore, are indicative for the relevant role the gland may play in the physiological reactions to chronic psychosocial stress.     

Architectonic Subdivisions of Neocortex in the Tree Shrew (Tupaia belangeri)

Tree shrews are small mammals that bear some semblance to squirrels, but are actually close relatives of primates. Thus, they have been extensively studied as a model for the early stages of primate evolution. In this study, subdivisions of cortex were reconstructed from brain sections cut in the coronal, sagittal, or horizontal planes, and processed for parvalbumin, SMI‐32‐immunopositive neurofilament protein epitopes, vesicle glutamate transporter 2 (VGluT2), free ionic zinc, myelin, cytochrome oxidase, and Nissl substance. These different procedures revealed similar boundaries between areas, suggesting the detection of functionally relevant borders and allowed a more precise demarcation of cortical areal boundaries. Primary cortical areas were most clearly revealed by the zinc stain, because of the poor staining of layer 4, as thalamocortical terminations lack free ionic zinc. Area 17 (V1) was especially prominent, as the broad layer 4 was nearly free of zinc stain. However, this feature was less pronounced in primary auditory and somatosensory cortex. In primary sensory areas, thalamocortical terminations in layer 4 densely express VGluT2. Auditory cortex consists of two architectonically distinct subdivisions, a primary core region (Ac), surrounded by a belt region (Ab) that had a slightly less developed koniocellular appearance. Primary motor cortex (M1) was identified by the absence of VGluT2 staining in the poorly developed granular layer 4 and the presence of SMI‐32‐labeled pyramidal cells in layers 3 and 5. The presence of well‐differentiated cortical areas in tree shrews indicates their usefulness in studies of cortical organization and function. Anat Rec, 2009. © 2009 Wiley‐Liss, Inc.     

The retinotopic distribution of visual callosal projections in the suprasylvian visual areas compared to the classical visual areas (17, 18, 19) in the cat

Summary The distribution of the interhemispheric projection from area 17 and 18 was studied using the anterograde degeneration technique. Besides the classical visual areas (17, 18, 19), area 21 and several visual areas in the middle suprasylvian sulcus also received visual callosal input. In the four terminal areas of the middle suprasylvian sulcus the projection was found to be focused on representations of the vertical meridian including the area centralis, as in the classical visual areas. An increase of the width of visual field represented in the zone of callosal terminations can be seen from area 17 through area 18 to area 19 and possibly this trend continues in the suprasylvian visual areas.     

单胺类神经元在成年树鼩中枢神经系统中的分布

目的探讨酪氨酸羟化酶(TH)和色氨酸羟化酶2(Tph2)在成年树鼩中枢神经系统(CNS)中的分布特征。方法利用免疫组织化学技术观察TH和Tph2在成年树鼩CNS的分布特征。结果在成年树鼩CNS中,TH阳性信号主要分布于嗅球小球层、纹状体、第3脑室周围区域、未定带、黑质、腹侧被盖区、中央管周围灰质和蓝斑;而成年树鼩CNS中的Tph2阳性信号主要分布于中脑和脑桥中缝复合结构中。结论树鼩TH和Tph2阳性信号的分布模式与脊椎动物的总体分布模式大致相同;某些细节特征显示,树鼩在进化上更接近灵长动物。     

A cytoarchitectonic study of the hippocampal formation of the tree shrew (Tupaia belangeri)

Tree shrews constitute an interesting animal model to study the impact of stress or aging on the hippocampal formation, a brain structure known to be affected under such environmental or internal influences. To perform detailed investigations of the hippocampal formation, adequate knowledge of its anatomy should be present. Until now, the hippocampal formation of the tree shrew has not yet been studied extensively. The main objective of this study, therefore, was to describe the subfield boundaries in various levels of the dorsoventral hippocampal axis of the tree shrew (Tupaia belangeri) in detail. The secondary aim was to clarify whether a separate CA2 field can actually be distinguished in the tree shrew hippocampus, a fact that was denied in former reports. In addition, we aimed at investigating whether or not a CA4 subfield can be identified in the tree shrew's hippocampus. The immunocytochemical distribution of microtubule-associated protein 2 and the calcium-binding proteins, parvalbumin and calbindin, and the characteristics of Nissl staining in adjacent sections were compared. Because of the rather dorsoventral orientation of the long hippocampal axis in tree shrews, staining patterns were analyzed mainly in horizontal sections. The subiculum and the hippocampal CA1 and CA3 areas were easily identified. Moreover, we were able to demonstrate the existence of a distinct CA2 subfield in the tree shrew's Ammon's horn, contrary to previous reports. However, our results indicate that a CA4 field in the tree shrew hippocampal formation cannot be identified with the methods that we used. Therefore, supposed CA4 pyramidal neurons should be included into the CA3 field.     

Mapping of cytoskeletal components in the hippocampal formation of the tree shrew (Tupaia belangeri)

The distribution of the major cytoskeletal components in frontal cryosections of the hippocampal formation of adult male tree shrews (Tupaia belangeri) was immunohistochemically investigated by using commercially available antibodies. Actin-immunolabeling was evident in all layers of the dentate gyrus as well as in the regio superior (CA1) and the regio inferior (CA3). Neurofilament 160 was detected only in the molecular layer of the dentate gyrus and in the axons of the granule cells (mossy fibers). For beta-tubulin, the microtubule associated proteins (MAPs) MAP2AB, MAP2ABC and Tau, immunoreactivity was evident within the granule cells and within the somatodendritic compartment of pyramidal neurons. Granule cells and the somata of the pyramidal neurons were intensely labeled for kinesin. Our findings show the elaborate expression of cytoskeletal proteins in the hippocampal formation of the tree shrew, relatively similar to what is seen in other species but with also some important differences, such as the immunonegativity of the axonal compartment for Tau in the tree shrew, which is contrary to what we see in the mouse (unpublished data). These findings provide useful insights regarding the organization of the hippocampal formation of the tree shrew and are fundamental for further research in this field.     

Extracellular matrix and development of lamination in the dorsal lateral geniculate nucleus in the tree shrew (Tupaia belangeri)

In the tree shrew (Tupaia belangeri), the cytoarchitectonic lamination of the lateral geniculate nucleus cannot be detected at birth; it only appears during the early postnatal period. However, a laminated pattern was revealed with rapid Golgi staining and retinal afferents were segregated into the appropriate laminae well before cytoarchitectonic lamination could be seen. Both observations indicate that the extracellular matrix may play a role in the separation of lateral geniculate nucleus cells into laminae. In the present study, the organization of the extracellular matrix was investigated during development using immunohistochemical and in situ hybridization techniques. For immunohistochemistry, peanut agglutinin (PNA) lectin and antibodies against tenascin (TN) were chosen, while for in situ hybridization, mTN riboprobes were used, simultaneously, with antibodies against Vimentin (Vim) and microtubule associated protein (MAP-2). The results showed that the pattern of PNA-binding glycoproteins and that of tenascin were relatively similar, although tenascin appeared later and disappeared earlier. The first interlaminar spaces to be detected were those between layers innervated by opposite eyes. The TN specific mRNA was detected in the lateral geniculate nucleus at P0, but was no longer visible at P7. By comparing TN mRNA and Vim or MAP-2 stainings a correspondence could be observed. The extracellular matrix lamination therefore seems to precede cytoarchitectonic lamination, suggesting that the extracellular matrix may play a role in the development of laminated structures. The TN-producing cells seem to be developing astrocytes and neurons.     

c-Fos expression in the visual system of the tree shrew (Tupaia belangeri)

c-Fos is a nuclear phosphoprotein coded by the proto-oncogen c-fos which can be detected immunohistochemically after both physiological and pathological stimuli. This property is of great importance, because it offers a valuable tool for morphofunctional identification of activated neurons. We have studied the neuronal activity in the visual pathway of Tupaia belangeri within the following anatomical structures: retina, superior colliculus (SC), dorsal lateral geniculate nucleus (dLGN), pulvinar (Pu), parabigeminal (PBG) nucleus and primary visual cortex (V1) analyzing the c-Fos expression after exposing the tree shrews to different light stimuli (white light –control positive group–, green light, blue light and darkness conditions –control negative group–). Our findings suggest that in the retina, the ganglion cells and the cells of the inner nuclear layer respond better to blue and green light stimuli, when comparing the c-Fos expression between white, green, blue lights and darkness conditions. However, in the SC, dLGN, Pu, PBG nucleus and V1 another pattern of c-Fos expression is observed: a maximum expression for the control positive group, a minimum expression for the control negative group and intermediate expressions within the blue and green light groups. Conclusion: the expression levels of c-Fos protein are able to show significant differences between distinct light stimuli in all anatomical structures studied (retina, SC, dLGN, Pu, PBG and V1) of T. belangeri.     

The corticosterone receptive system in the brain of Tupaia belangeri visualized by in vivo autoradiography

Summary The present investigation deals with in vivo binding of ³H-corticosterone in the brains of tree shrews as visualized by autoradiography. Tree shrews were injected with ³H-corticosterone and brain sections were mounted on slides which were subsequently exposed on tritium sensitive film. The relative labeling of 20 different brain structures was determined densitometrically. The indusium griseum, which demonstrated the highest binding for corticosterone of all brain regions in the autoradiograms, was taken as reference and defined as 100% relative labeling (RL). As in other species, the hippocampal subdivisions of the tree shrew retained high amounts of the steroid (60 to 80% RL). In other parts of the limbic system, medium labeling intensities were observed with approximately 40% RL in the lateral septum. The amygdala was less intensely labeled revealing around 30% RL in the basal accessory, the cortical, central, and the lateral nuclei. Autoradiographic grey values in the ventral striatum and pallidum were comparable to those in the amygdala, but in the islands of Callejae they were approximately as high as in the lateral septum (44% RL). In contrast to previous reports dealing with other species, the tree shrew cerebellum also demonstrated a high binding capacity for corticosterone. The RL was nearly 60% in the cerebellar granular layer. This finding may indicate that the cerebellum also plays a role in mediating the effects of corticosterone in the central nervous system.Abbreviations Acb nucleus accumbens - ADX adrenalectomy - BAA nucleus basalis accessorius amygdalae - BLA nucleus basalis lateralis amygdalae - BMA nucleus basalis medialis amygdalae - bv blood vessel - CD nucleus caudatus - Ce nucleus centralis amygdalae - ChP plexus choroideus - Co nucleus corticalis amygdalae - DG fascia dentata; gran, granule cells - Hip hippocampus - I nucleus intercalatus amygdalae - ICj insulae Callejae - Ig indusium griseum - IO nucleus olivaris inferior - LA nucleus lateralis amygdalae - LS nucleus lateralis septi - OT optic tract - RL relative labeling - S subiculum - SFi septofimbrial nucleus - SI substantia innominata - SHy nucleus septohypothalamicus - Tu olfactory tubercle     

Ciliogenesis in photoreceptor cells of the tree shrew retina

 Transmission electron microscopy of the retinal cones from several prenatal, young postnatal and adult tree shrews (Tupaia belangeri) reveals that the centrioles, from which the ciliary precursors of the outer segments grow out, are not transported into a pre-existing inner segment, but are positioned under the apical plasma membrane of cone precursor cells all through the inner segment formation. Ciliogenesis starts before or on embryonic day 20 and thus precedes initial formation of the inner segment by 20 days, which is half the gestation period. Thus, the maturation of the outer segment covers a considerably longer period than has been previously described. Published observations from other mammals can be interpreted as conforming with the situation in Tupaia. In other vertebrates, compared to mammals, marked heterochronies do occur. In Tupaia, the centrioles and the cilium are located close to the central longitudinal axis of the photoreceptor precursor cell from the 20-day-old embryo to the 5-day-old juvenile. In this position the microtubule apparatus originating from the centrioles should be most effective in transporting the mitochondria into the inner segment. In the 12-day-old tree shrew, when transport of the mitochondria into the inner segment has been completed, centrioles and cilium have shifted into an eccentric position and the light-collecting megamitochondria have approached the disks of the outer segment. This eccentric position is maintained in all later developmental stages. In certain of the retinal areas of the adult Tupaia, the connecting cilia of neighbouring cones are always positioned on the same side of the inner segments. Accepted: 18 March 1997     

The distribution of crossed and uncrossed optic fibers in the different layers of the lateral geniculate nucleus in the tree shrew (Tupaia glis)

Summary The laminar distribution of crossed and uncrossed optic fibers was studied in the lateral geniculate nucleus (LGN) in the tree shrew (Tupaia glis) following unilateral enucleation. For the investigation of the termination of optic fibers the transneuronal degeneration method and experimental EM were employed. By using formvar film-coated slot grids, all six layers of the LGN could be studied in a single ultrathin section. Degeneration of crossed optic fibers was observed in layers 1, 3, 4 and 5 of the contralateral LGN. The uncrossed retinofugal fibers supply layers 2 and 6 of the LGN. The degeneration in layer 4 was less pronounced than that in the other layers. Ipsilateral and contralateral optic fibers were well separated. Filamentous as well as dark types of degeneration were found in the LGN after enucleation. The optic terminals (RL boutons) were seen only in the synaptic glomeruli.     

Postnatal development of 3H-rauwolscine binding sites in the dorsal lateral geniculate nucleus and the striate cortex of the tree shrew (Tupaia belangeri)

Summary Noradrenaline has been shown to play an important role within the visual system of the brain. To analyze the postnatal development of alpha₂-noradrenergic receptors in the visual system of tree shrews, we localized and quantified binding sites for the antagonist [³H]-rauwolscine by in vitro-autoradiography in the dorsal lateral geniculate nucleus and the striate cortex at different postnatal ages. At birth, the dorsal lateral geniculate nucleus is only slightly labeled by [³H]-rauwolscine. During the postnatal period, the number of binding sites increases to reach a maximum around postnatal day 20. Since the young tree shrews open their eyes at approximately day 19, it appears that this high concentration of alpha₂-adrenoceptors is related to eye opening. In the adult animal, [³H]-rauwolscine labeling shows a laminated pattern in the dorsal lateral geniculate nucleus. Laminae 1, 2, and 3 are more strongly labeled than laminae 4, 5, and 6. In the striate cortex, the pattern of [³H]-rauwolscine-binding sites changes dramatically during the early postnatal period. Immediately after birth, there is only one layer, located within the subplate zone, which is labeled. From postnatal day 5 onwards, all cortical layers which can be distinguished on histologically stained sections reveal [³H]-rauwolscine-binding sites, but in layer IV, which is known to receive major inputs from the dorsal lateral geniculate nucleus, there is very little labeling during the first two postnatal weeks. In this layer, a large number of [³H]-rauwolscine-binding sites occurs between postnatal day 15 and 20, that is slightly before and around the time of eye opening. From this time onwards, the pattern of [³H]-rauwolscine binding in the striate cortex is very similar to that in the adult, where all layers are labeled although to different degrees. Since around postnatal day 20, maximal numbers of [³H]-rauwolscine binding sites are present in the dorsal lateral geniculate nucleus and a large number of these binding sites also emerges in layer IV of the striate cortex, alpha₂-noradrenergic receptors are probably important for processes related to the opening of the eye and/or for the visual system to function.     

Extracellular matrix and development of lamination in the dorsal lateral geniculate nucleus in the tree shrew (Tupaia belangeri).

 In the tree shrew (Tupaia belangeri), the cytoarchitectonic lamination of the lateral geniculate nucleus cannot be detected at birth; it only appears during the early postnatal period. However, a laminated pattern was revealed with rapid Golgi staining and retinal afferents were segregated into the appropriate laminae well before cytoarchitectonic lamination could be seen. Both observations indicate that the extracellular matrix may play a role in the separation of lateral geniculate nucleus cells into laminae. In the present study, the organization of the extracellular matrix was investigated during development using immunohistochemical and in situ hybridization techniques. For immunohistochemistry, peanut agglutinin (PNA) lectin and antibodies against tenascin (TN) were chosen, while for in situ hybridization, mTN riboprobes were used, simultaneously, with antibodies against Vimentin (Vim) and microtubule associated protein (MAP-2). The results showed that the pattern of PNA-binding glycoproteins and that of tenascin were relatively similar, although tenascin appeared later and disappeared earlier. The first interlaminar spaces to be detected were those between layers innervated by opposite eyes. The TN specific mRNA was detected in the lateral geniculate nucleus at P0, but was no longer visible at P7. By comparing TN mRNA and Vim or MAP-2 stainings a correspondence could be observed. The extracellular matrix lamination therefore seems to precede cytoarchitectonic lamination, suggesting that the extracellular matrix may play a role in the development of laminated structures. The TN-producing cells seem to be developing astrocytes and neurons. Accepted: 23 November 1998     

The patterns of cell death and of macrophages in the developing forebrain of the tree shrew Tupaia belangeri

The patterns of cell death and of macrophages were investigated in the forebrain and eyes of the tree shrew Tupaia belangeri during five phases of optic cup formation. Seventeen embryos were studied. Three- dimensional reconstructions were made from one embryo of each phase. In phase 1 (V-shaped optic evagination) a midline band of cell death passes through the closing anterior neuroporus. From phases 2 (optic vesicle) to 5 (far-advanced invagination) the midline band of cell death extends in the dorsal wall of the forebrain to its rostral pole and, further, into its ventral wall. At the approximate future position of the optic chiasm this ventral pycnotic area, predicted but so far unidentified by others, is connected to a previously described second band of cell death passing through the optic anlagen. Recently, evidence has been presented that chicken embryos develop holoprosencephaly and cyclopia when ventral forebrain structures are lost secondary to experimentally induced apoptosis. Our findings in Tupaia suggest that, in cases of spontaneous malformations of this kind, such an atypical pycnotic area in the ventral telencephalon might result from the defective regulation of cell death processes during optic cup formation. In the forebrain and eyes of Tupaia, the occurrence of bands of cell death precedes the appearance of the earliest intraepithelial macrophages. From phase 3 (onset of invagination) onwards almost all of them are concentrated along the band of cell death. Accepted: 28 September 1999     

Neurons with callosal projections in visual areas of newborn kittens: an analysis of their dendritic phenotype with respect to the fate of the callosal axon and of its target

Summary Combined retrograde transport of Rhodamine-labeled latex beads and intracellular injection of Lucifer Yellow in aldehyde-fixed slices of areas 17 and 18 in kittens indicate that neurons with similar dendritic morphology send axons into the corpus callosum from the 17/18 border and from parts of area 17 destined to become acallosal. At both sites callosally projecting neurons (callosal neurons) include pyramids, spiny stellate cells and star-pyramids; two types of pyramidal neurons can be distinguished on the basis of the complexity of their apical dendrites. At both sites, the dendritic morphology of callosal neurons appears basically unaffected by the ablation at the beginning of the second postnatal week of the contralateral areas 17 and 18 to which they have sent their axon. Thus the dendritic morphology of this type of cortical neuron seems independent of retrograde signals coming from their contralateral target and may instead depend on programs intrinsic to the neurons and/or conditions acting locally on their cell bodies, dendrites or initial axon collaterals.