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A scanning electron microscope study of the in vitro development of dissociated hippocampal cells.

Cultures of dissociated hippocampal neurons from 18-day-old rat fetuses were examined by scanning electron microscopy after periods between one hour and 20 days following plating on a poly-L-lysine coated substrate. Cell attachment was virtually complete within one hour after plating, and at that stage many cells could be seen which had started to extend processes with broad growth cones. By four hours in culture, process formation was well advanced and some cells had already assumed a pyramidal configuration. After 16 hours in culture, numerous contacts were seen between neighboring growth cones, and this frequently led to fasciculation of the interacting fibers. During the next three weeks the cell bodies enlarged considerably and rounded-up, and two distinct types of processes became evident: large, rapidly tapering dendrite-like processes and finer, essentially uniform-diametered processes that resemble axons. In most of the older cultures a dense plexus of processes was formed, and many of the finer processes appeared to have "bouton-like" swellings as they traversed the upper surfaces of the neuronal perikarya. Non-neuronal elements, which comprised only about 5% of the cells initially plated, rapidly proliferated in our cultures and within three to six days formed a confluent monolayer beneath the neurons.

Animals↗

Early development of the circumferential axonal pathway in mouse and chick spinal cord.

The early development of the circumferential axonal pathway in the brachial and lumbar spinal cord of mouse and chick embryos was studied by scanning and transmission electron microscopy. The cellular processes which comprise this pathway grow in the transverse plane and along the lateral margin of the marginal zone (i.e., circumferentially oriented), as typified by the early embryonic commissural axons. The first formative event observed was in the ventrolateral margin of the primitive spinal cord ventricular zone. Cellular processes were found near the external limiting membrane that appeared to grow a variable distance either dorsally or ventrally. Later in development, presumptive motor column neurons migrated into the ventrolateral region, distal to these early circumferentially oriented processes. Concurrently, other circumferentially oriented perikarya and processes appeared along the dorsolateral margin. Due to their aligned sites of origin and parallel growth, the circumferential processes formed a more or less continuous line or pathway, which in about 10% of the scanned specimens could be followed along the entire lateral margin of the embryonic spinal cord. Several specimens later in development had two sets of aligned circumferential processes in the ventral region. Large numbers of circumferential axons were then found to follow the preformed pathway by fasciculation, after the primitive motor column had become established. Since the earliest circumferential processes appeared to differentiate into axons and were found nearly 24 hours prior to growth of most circumferential axons, their role in guidance as pioneering axons was suggested.

Animals↗

Axonal projections of mechanosensory neurons in the connectives and peripheral nerves of the leech, Haemopis marmorata.

The axonal projections of the seven mechanosensory neurons in the connectives and peripheral nerves were examined by horseradish peroxidase (HRP) injections. In the connective the closely functionally related mechanosensory neurons travel in two bilaterally symmetrical regions, which comprise less than 10% of the connective's cross-sectional area. This grouping may reflect the similar functional properties and synaptic connections of the cells. Serial sections through the neuropil-connective junction zone showed that the organization of the axons is independent of glial fasciculation. Fascicles are not rigid structures; they were found to part and fuse in a seemingly random manner. The glial cell seems therefore to have only a supportive structural function and not to play any role in axonal guidance or to define any specific nerve fascicles in the leech. The projections of the mechanosensory cells in the peripheral roots were not as stereotyped as in the connective. However, examination of the branching of the medial and lateral P cells in the posterior root showed that the roots are highly structured and that some axons may be confined to tracts which have specific positions related to the branching of the root. The possible role of the P cells in pioneering these tracts is discussed.

Animals↗

Projection patterns of lateral-line afferents in anurans: a comparative HRP study.

Primary projections of the anterior and posterior lateral-line nerves were traced by means of transganglionic transport of horseradish peroxidase in species belonging to five of the six anuran superfamilies. Both anterior and posterior lateral-line nerve afferents each enter the brain via a single root which divides into two or more bundles. These bundles carry fibers from neuromasts only. No separate dorsal fascicle and no ampullary organs as in urodeles and gymnophions have been found. All bundles join in the neuropil of the nucleus intermedius to form ascending and descending fascicles. Two distinct fascicles are found in species showing little collateralization. No fasciculation is found in species with an elaborate telodendritic arborization. Afferents of the anterior lateral-line nerve run ventromedially and those of the posterior lateral-line nerve dorsolaterally within the ipsilateral nucleus intermedius neuropil. Rostrally they terminate in the vicinity of the eminentia granularis and caudally in the vicinity of the calamus scriptorius. The metamorphic changes in the alar plate do not support the hypothesis of Larsell ('34) as to a change in function of second-order lateral-line neurons into second-order auditory neurons. The rostral part of the nucleus intermedius shows numerous degenerating neurons at metamorphic climax whereas the caudal part becomes part of the nucleus caudalis of adult anurans. Besides the members of the Pipoidea, there is at least the genus Bombina which retains parts of the lateral-line system. The term "dorsal island," its relevance for any part of the anuran brain, and the possible relation between absence of electroreception and the development of the nucleus dorsolateralis are discussed.

Afferent Pathways↗

Postnatal changes in arborization patterns of murine retinocollicular axons.

The growth and arborization of murine retinocollicular axons have been studied by means of HRP axon filling during postnatal development. Transformations in arborization patterns have been correlated with changes in synaptic density in the superficial collicular neuropil and with the formation of synapses by HRP-filled axons. At all postnatal ages axons of the optic projection are fasciculated and most follow a rostrocaudally aligned path. On the day of birth the axons course through both stratum griseum superficiale (SGS) and stratum opticum (SO); during the following 4 days the axon trunks disappear from SGS and are subsequently found only in SO. From postnatal day (P) 0 to P3, the majority continue far caudally in the colliculus, giving rise to small ascending collaterals at multiple points along their course. Ultimately, usually by P3, one or two collaterals begin to branch profusely and by P5 the majority of axons give rise to a focal terminal ascending arborization. The general configuration of most arborizations at P3 approximates that of the mature axon. However, the richness of terminal branching increases from P3 through the first 2 postnatal weeks. Synaptic density is relatively low in the first postnatal week, and no synapses involving HRP-filled optic axons were identified in this interval. Subsequently, after elaboration of definitive arbors has begun, synaptogenesis in the surrounding neuropil accelerates. Synaptic density in the upper SGS approximates adult values early in the third postnatal week. By this time synaptic junctions involving the terminal arborizations of optic axons are abundant.

Aging↗

Map of retinal position onto the cross section of the optic pathway of goldfish.

The position of a retinal cell is defined by the two polar coordinates: r, the distance from the optic disc, and theta, the angular (or clock-face) position. Axons of similar theta value were labeled by the punctate application of horseradish peroxidase (HRP) to optic axons in the retina, and axons of similar r-value were labeled by the application of this same marker to a tectal fascicle. Labeled axons were traced in serial transverse sections of the optic pathway from the retina to the tectum to learn the map of the retinal surface onto the cross section of the pathway. Retinas were flat-mounted and treated for HRP to show the retinal origins of the labeled axons. Axons of similar r were clustered together, and the fraction of the pathway's cross-sectional area occupied by the cluster was about the same as the fraction of the retinal area occupied by the group of labeled somata. Axons of similar theta were also clustered, but the fraction of the cross-sectional area they occupied was larger than the fraction of retinal area occupied by their somata. The geometry of the clusters of labeled axons depended on the proximodistal location in the pathway. Near the retina both were strip-shaped, but the location and orientation of the strip varied. Both an r-strip and a theta-strip were labeled in some pathways by dual applications of HRP; the two strips were mutually orthogonal at all levels. Each of r and theta mapped onto a separate axis. The axons from most peripheral retina (largest r) were everywhere adjacent to the pia, and axons of progressively more central retina (smaller r) were progressively more separated from the pia (except in the nerve, where the secondary fasciculation complicates the geometry by wrapping old axons in new pia). The map of the circular variable, theta, onto a line, required a discontinuity, the location of which differed, depending on the proximodistal level. From the retina to the chiasm, the discontinuity was at the ventral retinal radius (i.e., the right retinal clock-face positions were ordered 6-9-12-3-6 o'clock across the line); just central to the chiasm, the fibers reordered to put the discontinuity at the nasal radius (clock-face positions ordered 3-6-9-12-3); at the brachial bifurcation, the 3-6-9 half turned dorsally, the 9-12-3 half, ventrally.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Axonal trajectories between mouse somatosensory thalamus and cortex.

An in vitro brain slice preparation has been used to label fibers connecting the somatosensory thalamus and cortex of the mouse. In 400-800-micron brain slices, the pathway between the ventrobasal complex and somatosensory cortex was labeled under direct vision with horseradish peroxidase crystals (HRP), HRP-Nonidet P-40 (NP40) detergent chips, or a solution of HRP/dimethylsulfoxide. Thalamocortical and corticofugal fibers are organized into a plexiform system of bundles that appears to be fairly constant from animal to animal. Bundles of fibers projecting from the ventrobasal complex course between regularly spaced groups of thalamic neurons. Thalamocortical axons do not invariably leave the thalamus via the fiber bundle closest to the perikarya. Thus, nearest-neighbor relationships are abolished before these axons have even left the thalamus. The axon bundles traverse the thalamic reticular nucleus lateral to the complex. The axons then rotate about one another, analogous to the coiling of strands in rope about a central axis. This accounts for the well known 180 degrees rotation in the mediolateral direction between thalamic and cortical maps. Laterally, fiber bundles converge and diverge within the internal capsule so that nearest-neighbor relationships are lost. Individual thalamocortical axons do not bifurcate proximal to the subcortical white matter. After single bundles of fibers reach a point just below the subcortical white matter, their individual fibers diverge widely. Within the subcortical white matter most afferent fibers make a small dorsally concave loop prior to taking one of two possible courses: some of the fibers ascend directly into the overlying cortex usually angled towards the dorsal surface of the brain; other fibers run in the subcortical white matter for variable distances prior to ascending into cortex. Within somatosensory cortex, smooth axons branch near their terminals in layers IV and VI. Axonal terminal and branching patterns of these axons within somatosensory cortex are similar to those found in in vivo preparations. Most axons are smooth, but other axons are beaded. Some beaded axons project to layer I. Corticofugal fibers are labeled. Fibers leaving somatosensory cortex have an angle of descent opposite to the angle of ascent for afferent fibers, and are often fasciculated in the cortex and subcortical white matter. Within the subcortical white matter efferent fibers often loop in a direction opposite to that of afferent fibers. Corticofugal fibers occasionally give off a collateral corticostriatal branch within the internal capsule.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗

Factors guiding optic fibers in developing Xenopus retina.

We have characterized, by electron microscopy, the growth of pioneering axons from the retina into the visual pathway during early development of Xenopus laevis. The subsequent development of following fibers from the growing retinal margin as they accumulated in the ganglion cell fiber layer (GCFL) of the retina was also studied. Extracellular channels bordered by neuroepithelial cells appear in the developing retina in a dorsal to ventral gradient before any pioneering axons are seen. Pioneering axons are subsequently observed in these channels, usually surrounded by neuroepithelial cell processes. Ruthenium red treatment of embryonic retinas reveals extracellular matrix (ECM) within these retinal channels, while extracellular spaces in the proximal optic stalk, just beyond the optic disc, lack this material. ECM is also seen in optic tectum wherever ingrowing retinal and nonretinal axons are found. The channels and the ECM contained within them may provide guidance cues for pioneering retinal axons. The early association of pioneering retinal axons with neuroepithelial cell processes (putative glia) appears to be important in further development of the GCFL. The so-called following fibers of ganglion cells, arising later in development, fasciculate with pioneer axons in extracellular spaces and form fiber bundles of the GCFL on top of the layer of glial cell endfeet. It is not clear whether pioneering axons, glial cell surfaces, or both serve as guidance cues for following fiber migration.

Animals↗

Normal and regenerating optic fibers in goldfish tectum: HRP-EM evidence for rapid synaptogenesis and optic fiber-fiber affinity.

The distribution of normal and regenerating retinal fibers and synapses was studied on tectum in goldfish by light (LM) and electron microscopy (EM). Since labeling of the early regenerating fibers was previously reported to be difficult, a new 'cold-fill' HRP labeling protocol was developed, which labeled regenerating optic fibers and terminals on tectum as early as 14 days after nerve crush when they first arrive on tectum. In order to characterize the laminar distribution of optic afferents in normal fish and in fish regenerating for 14-240 days, EM photomontages of areas 14 microns wide by 160 microns deep through the HRP-labeled primary optic innervation layer (S-SO-SFGS) were constructed. The time points in regeneration that were examined spanned the period in which others have shown that an initially diffuse retinotopic map becomes spatially restricted. At the LM level regenerating optic fibers were restricted to the optic lamina. They reinnervated tectum in an anterior to posterior sequence as previously seen with autoradiography. In addition, at 14 days, some "pioneer" optic fascicles were found to have already grown to posterior tectum where they gave rise to branches with boutonlike terminations and growth-cone-like processes. Form the ultrastructural analysis it was clear that optic fibers and terminals observed strict laminar boundaries as they partitioned themselves in the optic laminae (S, SO and SFGS) in both normal and regenerating fish. The behavior of optic fibers was lamina specific with respect to synapse formation and the orientation of fiber outgrowth. As early as 14 days regeneration, optic fibers made synapses onto the four types of postsynaptic profiles observed in normal fish. Numerous optic terminals were labeled at 14 days, and there appeared to be no waiting period between fiber ingrowth to the SO and synapse formation in the S and SFGS. At 14-60 days, atypical synaptic contacts which appear to be nascent synapses were made by labeled optic fibers in fascicles and by growth-cone-like processes. By 21-30 days, the density of optic terminals was high and there were many more fasciculated optic fibers in the SFGS than normal as late as 350 days. These findings suggest that optic fiber lamination is highly constrained by tectal cues, that fibers rapidly regenerate many synaptic terminals before retinotopic map refinement is complete, and that fibers have a strong affinity for each other.

Animals↗

Implantation of cultured sensory neurons and Schwann cells into lesioned neonatal rat spinal cord. I. Methods for preparing implants from dissociated cells.

Our goal was to devise methods of implanting defined populations of the cellular constituents of peripheral nerve into regions of spinal cord injury. This objective derived from the knowledge that the cellular environment of peripheral nerve is known to be supportive of axon regeneration from both central and peripheral neurons. Two of the constituents of the peripheral nerve environment known to influence axonal growth are the Schwann cell and extracellular matrix (particularly basal lamina), both of which can be obtained in culture. We describe here large-scale methods of establishing purified populations of rat sensory neurons to which purified populations of Schwann cells were added. These essentially monolayer preparations were then scrolled and cut into lengths of proper shape and size to provide implants for sites of spinal cord injury in newborn rats. We also describe methods enabling the addition of leptomeningeal components to the implants; this addition contributes a proliferating population of vascular endothelial cells (identified by immunostaining) to the otherwise vasculature-free neuron/Schwann cell implant. Light and electron microscopic observations were made to characterize the implants. When the implant was ready for use, it contained Schwann cells that were differentiated, i.e., had begun to ensheathe axons and form basal lamina. The use of a medium containing human plasma to foster endothelial cell growth led to increased neurite fasciculation and Schwann cell migratory activity in the outgrowth, particularly when the neurons and Schwann cells were cultured on leptomeninges. The second paper in this series reports the deportment of these implants and their influence on corticospinal tract growth after placement into regions of dorsal column injury in neonatal rats (Kuhlengel et al., J. Comp. Neurol 293:74-91, 1990).

Animals↗

Serially homologous development of the peripheral nervous system in the mouthparts of the grasshopper.

Segmentally homologous neurogenesis and neuronal differentiation processes characterize the formation of the peripheral nervous system in the developing mouthparts of the grasshopper embryo. The peripherally located neurons derive from the ectoderm in each of the embryonic mouthparts. The major nerve branches serving the mandibles, maxillae, and labium are established by peripheral pioneer neurons, which project their axons into the central nervous system via a set of guidepost cells. The two secondary nerve branches in each appendage are established by fasciculation of peripheral afferent pioneers and central efferent pioneers or by efferent pioneers alone. Sensory cells differentiate and connect with the peripheral nerve branches after a basic peripheral nerve scaffold has been established. The serial homology of these developmental processes in the mouthparts and in the thoracic legs is striking and can be documented at the level of individual identified cells. Thus despite the enormous differences in gross structure and function among cephalic mouthparts and true thoracic legs, many aspects of neurogenesis and early neuronal differentiation are remarkably conserved in all of these appendages.

Aging↗

Phrenic motoneuron morphology in the neonatal rat.

The morphology of neonatal rat phrenic motoneurons was studied following retrograde labeling with horseradish peroxidase, which resulted in Golgi-like fills of phrenic motoneuron somata and dendrites. At birth, these neurons have well-developed dendritic trees with many characteristics described for phrenic motoneurons in the adult rat. The dendrites form tightly fasciculated bundles that emerge from the phrenic nucleus primarily along four axes: ventromedial, ventrolateral, dorsolateral, and rostral/caudal, with smaller and more variable projections directly lateral and ventral. Although sparse, some dendritic appendages were also present, and in a few animals, somata clustering was apparent. The most significant difference between adult and neonatal rat phrenic motoneurons is in the extent to which medially and laterally projecting dendrites extend beyond the borders of the ipsilateral gray matter. In the neonate, unlike the adult, these dendrites project extensively past the gray/white border to the edge of the hemicord. Ventromedial dendrites occasionally cross to the contralateral ventral horn in the ventral white commissure and laterally projecting dendrites could be seen reaching the edge of the cord, turning and traveling rostrally or caudally for up to 100 microns. Phrenic motoneurons are not unique in having long dendrites at birth. A brief comparative study showed that neonatal cervical, thoracic, and lumbar motoneurons also have long dendrites that project to the medial and lateral borders of the hemicord.

Afferent Pathways↗

Development of the olivocerebellar projection in the rat: I. Transient biochemical compartmentation of the inferior olive.

In the present study the early phases of the development of the inferior olive were examined by using immunocytochemical techniques. We observed that, from embryonic day 16 onward, antibodies against the calcium binding proteins parvalbumin and calbindin and the calcitonin gene related peptide stain partially overlapping territories of the inferior olive. This staining delimits a biochemical zonation of the inferior olive which is combinatory and transient. We have previously observed a biochemical parcellation of the cerebellar Purkinje cells which, like that of the inferior olive, is first observed at E16, involves the combined expression of marker proteins and is also transient. In order to know whether the biochemical compartmentations of the cerebellum and inferior olive arise independently, the time course of the development of the olivocerebellar projection was studied by anterograde and retrograde in vitro axonal tracing by using the fluorescent carbocyanine dye DiI. The olivocerebellar axons were found to reach the limit of the cerebellar plate at E16 and to enter it at E17. Even at this age the great majority of the climbing fibers are tightly fasciculated, which minimizes their interactions with the PC clusters. These observations indicate that the topographical heterogeneity of Purkinje cells and inferior olive neurons arise independently. The transient biochemical individualization of subgroups of neurons during development could contribute to recognition mechanisms.

Animals↗

Distinctive pattern of organisation in the retinofugal pathway of a marsupial: I. Retina and optic nerve.

The nasotemporal division in the retina and the pattern of crossed and uncrossed axons in the optic nerve were determined in an Australian marsupial, a wallaby, Setonix brachyurus (the quokka), following unilateral horseradish peroxidase injections into primary visual centres. The gross morphology of the nerve was also examined. Ipsilaterally projecting ganglion cells were restricted to the temporal retina, whereas those that project contralaterally were located in all retinal regions. The morphological study of the nerve showed that fasciculation patterns, evident along much of the length of the nerve, became indistinct centrally and were replaced in the prechiasmatic region by dorsoventrally oriented fissures. In this prechiasmatic region, axons were oriented in two directions. Whereas the majority were aligned centroperipherally with the long axis of the nerve, a proportion were aligned dorsoventrally in the fissures. Labelling with HRP revealed that uncrossed axons were restricted to the lateral region of the optic nerve and possibly to discrete fascicles, whereas those destined to cross at the chiasm occupied all regions of the nerve but were less dense on the lateral side. This spatial distribution of crossed and uncrossed projections did not change along the length of the nerve. These results demonstrate that fibre organisation in the marsupial optic nerve is different than that found in eutherian mammals.

Animals↗

Distinctive pattern of organisation in the retinofugal pathway of a marsupial: II. Optic chiasm.

In the mammalian optic chiasm retinal axons from each eye divide into two populations, those that decussate and those that remain uncrossed. In eutherian (placental) mammals, the separation of these pathways is not reflected in the structure of the chiasm. The two populations from each eye are mixed through each hemichiasm, segregating only at the midline, where the uncrossed projection turns back. In this study the optic chiasm of a marsupial, the wallaby, Setonix brachyurus (quokka) has been investigated with staining and neuronal tracing techniques. The chiasm of this mammal is quite different from that of eutherian mammals. In coronal section it can be morphologically subdivided into three regions, a central body in which fasciculated groups of axons from each eye interdigitate across the midline, and two distinct lateral regions, one on each side, which contain the uncrossed retinal projections. In the rostral chiasm the lateral regions are separated from the main body of the chiasm by vertically oriented fibre-free regions. Caudally, the lateral regions increase in size and become less distinct as increasing numbers of contralaterally projecting axons that have crossed the midline project into them. However, the two populations remain predominantly segregated in this region. As the lateral regions develop, the central body of the chiasm becomes thinner and finally detaches at the midline to form the two optic tracts. The routes taken by retinal axons through the eutherian and marsupial chiasm appear to be fundamentally different. Therefore, the developmental factors that determine the laterality of retinal projections are likely to show significant differences in the two mammalian groups.

Animals↗

A nonrandom interneuronal pattern in the developing frog spinal cord.

In the developing spinal cord of the frog, Xenopus laevis, a population of interneurons assumes a pattern that represents a previously undescribed level of organization. Glyoxylic acid treatment and immunocytochemistry show that the neurons contain catecholamines and their synthetic enzyme, tyrosine hydroxylase. Cells are located within the ependymal layer of the floor plate region of the larval spinal cord. The cells have several processes including a long one that projects toward the brain without fasciculating with other labeled processes. In addition, the cytoplasm of the catecholaminergic cells extends into the central canal, showing that they are a population of cerebrospinal fluid-contacting neurons. The spatial domain of catecholaminergic neurons starts abruptly at the boundary between the hindbrain and spinal cord and continues to the tip of the tail. The neurons occupy two longitudinal columns within the sheet of floor plate cells, which includes cells that do not exhibit the catecholaminergic phenotype. Unlabeled cells are intercalated between catecholaminergic cells in each column, giving the labeled cells the appearance of being spaced along the length of the spinal cord. This general arrangement is evident at the time of hatching. Spatial analysis showed that the position of cells along a column is not random. The nonrandom behavior is due to cells being excluded from the area immediately surrounding other catecholaminergic cells. Further analysis showed that the cellular pattern lacks segmental or other periodic repeats. Ultimately, the location of a cell within a column depends upon the position of its closest catecholaminergic neighbor.

Animals↗

NILE/L1 and NCAM-polysialic acid expression on growing axons of isolated neurons.

The neuron adhesion molecules NILE/L1 and NCAM may be involved in axonal guidance and cell recognition. To investigate all exposed membrane domains of single neurons, something which has not previously been done for any adhesion molecule, we used digitally processed scanning electron microscopy with a high-energy backscatter electron detector. This allowed a quantitative analysis of immunogold staining densities on all surfaces of isolated rat hippocampal neurons in culture to study NILE/L1 and NCAM expression independent of potentially inductive innervation. During early stages of neuritic extension, all growth cones showed similar NILE/L1 expression, but as soon as a single process extended farther than the others (by 20 hours), this putative axon and its growth cone generally showed a stronger level of NILE/L1 immunogold labeling than the other neurites. This is the earliest evidence of plasma membrane differentiation between axons and dendrites. With further neuritic growth, the relative NILE/L1 expression on axons and their growth cones continued to increase. In contrast to some earlier reports, NILE/L1 was expressed on axonal growth cones growing on both polylysine-coated glass and astrocyte substrates. Strong immunostaining for NCAM-related polysialic acid (PSA) was found on axonal growth cones and filopodia, suggesting that the homophilic adhesive action of NCAM may be reduced during axonal growth. PSA showed greater labeling on distal axons than on other areas of the neuron, indicating a variable NCAM-mediated adhesion on different regions of the same cell. Neither NILE/L1, NCAM, nor PSA appeared to show regional differences in axons fasciculating or defasciculating on themselves. A strong intercellular heterogeneity of NILE/L1, NCAM, and PSA expression levels on neurons in the same culture dish was found, suggesting that subsets of cells from the hippocampus may express biologically relevant differences in adhesion molecules compared to neighboring neurons. In light of the growing body of evidence pointing to the multifaceted array of homophilic and heterophilic binding interactions that NILE/L1 and NCAM may exhibit, and the functional importance of molecular densities, the quantitative data here support the hypothesis that sufficient cellular and subcellular heterogeneity exists for these molecules to be involved in some aspects of axonal guidance.

Animals↗

Differential expression of N- and R-cadherin in functional neuronal systems and other structures of the developing chicken brain.

Cadherins are a family of cell surface molecules mediating calcium-dependent cell-cell adhesion in a variety of tissues. More than a dozen cadherins are expressed in the vertebrate brain. To obtain insight into the biological significance of this diversity in cadherin expression, we mapped the expression of N- and R-cadherin in the brain of the developing chicken embryo (days 2-19 of incubation) by immunohistochemical and in situ hybridization techniques. Whereas the expression of N- and R-cadherin is relatively uniform or weak in early (about 2-5 days of incubation) and late development (15 days of incubation to hatching stage), these two molecules are differentially expressed in specific nuclei and fiber tracts between days 6-11 of incubation. For example, in the mes- and diencephalon, one of the tectofugal pathways and its target nuclei, here called the tecto-pretecto-rotundal system, express N-cadherin. R-cadherin is expressed by a different tectofugal system, the tectoisthmic pathway. The other tectofugal systems express neither N- nor R-cadherin. In addition, a small number of other mes- and diencephalic nuclei express N- or R-cadherin. On the basis of these results and experimental evidence from other studies, we speculate that the two cadherins are involved in the formation and segregation of particular functional systems within the vertebrate central nervous system (CNS) by regulating the formation of nuclei, and the pathfinding and/or the selective fasciculation of neurites. Apart from neuronal elements, a variety of vascular and ependymal structures also express N-cadherin or R-cadherin, e.g., the parenchymal blood vessels, the choroid plexus, the floor and roof plates, and the ventricular lining. These findings suggest that the two cadherins play a variety of roles during the development of neuronal and nonneuronal epithelial structures throughout CNS development.

Animals↗