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Radial glial cells represent a subpopulation of secondary neural precursor cells that differentiate from neuroepithelial progenitors and are transiently found in the developing CNS of mammals. There is ample evidence for a temporal and spatial arrangement of increasingly committed radial glial cells that is of critical importance for the organisation and specification of different brain regions. For the human ganglionic eminence, recent findings have shown an early molecular specification of this cell type by the CD15 carbohydrate epitope, beginning already at the end of the first trimester. Here we further characterise the CD15+ radial glia cells as bFGF/EGF responsive progenitors allowing its propagation in vitro. By magnet activated cell sorting, its trilineage differentiation potential can be shown by differentiation into (PSA-NCAM ß3 tubulin immunoreactive) neurones, GFAP expressing cells of astrocytic morphology, and O4 positive oligodendrocytes. Subcloning experiments under proliferation conditions reveal ongoing CD15 expression by dividing cells. Although the relative number of CD15+ progenitor cells is found to decrease in favour of CD15- precursor cells during continuous passaging, cell sorting experiments allow the repetitive purification of high numbers of positively selected precursor cells for up to 12 weeks. In conclusion, expression of the cell adhesion molecule CD15 by a subpopulation of proliferative cells from the lateral ganglionic eminence allows easy and reproducible purification of progenitor cells by cell sorting, enabling the generation of a compartment-specific cell pool as a prerequisite for a safe and standardised therapy of neurodegenerative basal forebrain diseases.

We have studied the distribution of the CD15 epitope in the developing brain of an Australian diprotodontid metatherian mammal, the tammar wallaby ( Macropus eugenii), using immunohistochemistry in conjunction with hematoxylin and eosin staining. At the time of birth (28 days after conception), CD15 immunoreactivity labeled somata in the primordial plexiform layer of the parietal cortex in a similar position to that seen in the early fetal eutherian brain. CD15 immunoreactivity in the brain of the developing pouch-young wallaby was found to be localized on the surface of radial glia at boundaries between developmentally significant forebrain compartments in a similar distribution to that seen in developing eutherian brain. These were best seen in the developing diencephalon, delineating epithalamus, ventral and dorsal thalamus and hypothalamic anlage, and in the striatum. Immunoreactivity for CD15 identified radial glia marking the lateral migratory stream at the striatopallial boundary, peaking in intensity at P19 to P25. From P37 to P54, CD15 immunoreactivity also demarcated patch compartments in the developing striatum. In contrast, CD15 immunoreactivity in hindbrain structures showed some differences from the temporospatial pattern seen in eutherian brain. These may reflect the relatively early brainstem maturation required for the newborn wallaby to be able to traverse the distance from the maternal genital tract to the pouch. The wallaby provides a convenient model for testing hypotheses concerning the role of CD15 in forebrain development because all events in which CD15 may play a critical role in forebrain morphogenesis occur during pouch life, when the young wallaby is accessible to experimental manipulation.

The distribution of the CD15 antigen (CD15, 3-fucosyl-N-acetyl-lactosamine, Lewis x) has been studied immunohistochemically in the fetal human thalamus. Its changing patterns could be related to three successive, but overlapping, periods primarily due to its association with radial glial cells, neuropil, and neural cell bodies, respectively. From 9 weeks of gestation (wg), a subset of CD15-positive radial glial cells distinguished the neuroepithelium of the ventral thalamus, a characteristic also seen in the developing mouse. Distal processes of the radial glial cells converged at the root of the forebrain choroid tenia, which was also CD15 positive. From 13 wg until approximately 20 wg, CD15-positive neuropil labeling marked the differentiation areas of prospective nuclei within the dorsal thalamus and progressively outlined their territories in a time sequence, which appeared specific for each nucleus. CD15 labeling of differentiating nuclei of the ventral, medial, anterior, and intralaminar thalamic divisions showed a transient topographic relationship with restricted areas of the ventricular wall. After 26 wg, CD15 immunoreactivity was observed in subpopulations of glial cells and neurons. Transient CD15 immunoreactivity was also found in delimited compartments within the subventricular region. The time of CD15 expression, its location, and cellular association suggest that CD15 is involved in segmentation of diencephalon, in the specification of differentiating nuclear areas and initial processes regarding the formation of intercellular contacts and cellular maturation., (Copyright 2001 Wiley-Liss, Inc.)

The expression of the terminal saccharide determinant CD15 (3[a1-3]-fucosyl-N-acetyl-lactosamine) was evaluated in the central auditory system of the human developing brain by using monoclonal antibodies against this epitope. CD15 immunoreactivity was first observed in the ventral cochlear nucleus at 10 weeks of gestation, whereas the dorsal cochlear nucleus became positive from 13 weeks of gestation. In both nuclei, the intensity of immunoreactivity increased until 16 weeks of gestation and lasted until 25 weeks of gestation. In the inferior colliculi, CD15 was poorly expressed in the central nucleus from 13 to 23 weeks of gestation and later with moderate levels until birth. Within the medial geniculate nucleus, a biphasic pattern of expression was observed with peaks around 14-17 and 21-24 weeks of gestation. Heterogeneous expression in the medial geniculate nucleus, which was associated either with neurons or the neuropil, allowed distinction of subnuclei. In many of the auditory pathway structures (e.g., ventral cochlear nucleus and central nucleus of the inferior colliculus), a heterogeneous pattern of CD15 expression in the form of repeating parallel bands, possibly related to tonotopic organization, became transiently apparent around 23 weeks of gestation, whereas in the magnocellular part of the medial geniculate nucleus, a striking modular or compartmental arrangement of immunoreactive structures (which could also be associated with tonotopic organization) was also noted at about 23 weeks of gestation. We propose that the initiation of CD15 expression in each nucleus heralds the appearance of functional contacts and that high levels of neuropil labeling are related to the formation of nonstabilized synaptic contacts. Thus, transient CD15 expression in the central auditory system is possibly correlated with phases of functional plasticity in this pathway.

The developmental expression pattern of the carbohydrate epitope CD15 (Lewis X, Le X) (alpha1-->3-fucosyl-N-acetyl-lactosamine) has been immunocytochemically evaluated in paraffin sections within the human basal ganglia from 10 weeks gestation to three years after birth. At 11 weeks of gestation, CD15 (Le X) positive radial glial cells were located in the anterior and dorsal parts of the lateral ganglionic eminence. Their processes ran from the subventricular zone radially in a highly ordered fashion to the dorsolateral margin of the caudate nucleus and further to the lateral rim of the putamen. At 12 weeks of gestation, strands of CD15 (Le X) material continued to the pial surface, forming a continuous CD15 (Le X) positive borderline separating the accumbens nucleus and olfactory tubercle from the piriform cortex. At 13 weeks of gestation the dorsal putamen was completely CD15 (Le X) immunoreactive along its perimeter and CD15 (Le X) patches, consisting of fine granular material, appeared at the dorsolateral margin of the putamen at this age; while the first CD15 (Le X) patches in the caudate nucleus were observed four weeks later. The matrix compartment of the caudate and dorsal putamen became gradually stained by granular CD15 (Le X) positive material into which CD15 (Le X) immunoreactive somata were embedded. The striking contrast in staining between patch and matrix compartments disappeared shortly after birth. The ventral striatum did not become immunoreactive until the last few weeks before birth. After the formation of CD15 (Le X) positive patches in the striatum (from 12 weeks of gestation), delicate CD15 (Le X) fibres, often accumulated in bundles and related to the striatal patches, became apparent coursing towards the external pallidal lamina and the globus pallidus. Immunoreactivity in the globus pallidus itself was transient, emerging from 16 weeks of gestation, reaching a peak at 21 weeks of gestation and disappearing by birth. Both processes, i.e. the occurrence of CD15 (Le X) striatopallidal fibres and the emerging immunoreactivity in their pallidal target, may be interrelated, so that ingrowing CD15 (Le X) positive axons from the striatum provoke CD15 (Le X) expression in the external and internal pallidum. The variable patterns and intensities of CD15 (Le X) expression are possibly related to periods of maturation of the striatum and the establishment of functional interactions within the basal ganglia. Differential staining of patch and matrix in the developing neostriatum suggests that a distinct phase of cellular adhesion or dishesion mediated by the CD15 (Le X) epitope occurs during establishment of the patch and matrix regions.

The distribution of the expression of the CD15 epitope was characterized by immunohistochemistry in the developing mouse and human lung on embryonic days E9.5-E19 and gestational weeks GW7-GW25, respectively. In the earliest stages in the mouse, the tracheal epithelial cells expressed CD15 on their apical and lateral cell membranes and, in the more proximal regions, also showed a faint cytoplasmatic CD15 expression. Only very few epithelial cells in the bronchial bud regions expressed CD15 on their apical surfaces. In later stages (E12-E17), cells in the proximal parts of the bronchi and bronchioli expressed CD15 on their apical, but also on their lateral membranes, and increasing numbers of cells expressed CD15 cytoplasmatically. Cells in the distal, presumably proliferating, areas of the bud regions were CD15 negative. This distribution pattern of CD15 was consistent until the latest embryonic stages. These results are completely opposite to those found in human developing lung where up to GW20 bronchial and bronchiolar bud regions were CD15 positive, while in the proximal parts of the airways the vast majority of cells were CD15 negative. After GW20, CD15 immunoreactivity in the bud regions vanished and was completely absent on GW25. This difference between human and mouse adds further evidence to profound species differences in the expression of CD15 in various organs, e.g., in the cerebellum or the retina, and should be taken into account when considering functional roles of CD15 and also when relating results from a (transgenic) mouse model to the respective human organ system.

The formation of the nasal lining with its sensory and its nonsensitive respiratory epithelium requires a spatially ordered pattern of cellular differentiation. Aiming at identifying cell recognition molecules that may be involved in cellular differentiation steps, we applied a panel of antibodies to terminal carbohydrate sequences of the lactoseries on the developing chick olfactory epithelium. This approach is based on the idea that these terminal sugar residues may be involved in certain steps of maturation. Restricted expression of three epitopes NALA, HNK-1, and CD15 was observed in olfactory receptor neurons. The first immature olfactory receptor neurons were observed by day 3 of incubation, expressing the HNK-1 epitope, whereas a total epithelial staining was observed for NALA. By day 9 of incubation high numbers of HNK-1 positive immature olfactory receptor neurons were observed. At the same time mature olfactory receptor neurons showed immunoreactivity for CD15, whereas NALA was still expressed throughout the whole epithelial cell population. However, there was a pronounced staining in the population of mature olfactory receptor neurons. Around hatching only CD15 was detectable in (mature) olfactory receptor neurons, whereas HNK-1 and NALA immunoreactivity have switched to glandular and sustentacular cells respectively. The differentiation-dependent expression patterns of these three cell surface molecules suggest them as suitable markers to explore mechanisms that determine embryonic olfactory receptor neurogenesis.

A subpopulation of radial glial cells has been identified in the mouse prosencephalon during the second half of embryonic development. This subpopulation, specified by the putative cell adhesion molecule CD15 (LeX, FAL), is arranged in a segmented pattern within the telencephalon and diencephalon. Glial processes, spanning the prosencephalic wall, first appear at E10.5 and remain clearly visible until E19, when staining of discrete nuclei begins to appear. Registration of the correspondence between ventricular and pial surfaces, however, is still possible due to the persistence of individual CD15-positive fibres. These can be traced even when the initial simple linear (radial) orientation between ventricular and pial surfaces becomes complicated and distorted. After birth, CD15 immunoreactivity is distributed in a mosaic pattern in the forebrain. Because radial glial cells provide a scaffolding system for postmitotic neurones, the pattern of CD15-positive fibres in the embryonic prosencephalon may also demarcate future discrete regions of the postnatal brain.

The distribution of the 3-fucosyl-N-acetyl-lactosamine (FAL, CD15) epitope in the developing mouse cerebellum was examined with the aid of immunohistochemistry of paraffin sections. CD15 immunoreactivity first appeared at E15 as a discrete bundle of processes lying beneath, and slightly within, the deeper layers of the external granular layer. By E17, when the cerebellar anlagen had completed their midline fusion, these processes could be traced from the germinal trigone at the lateral limits of the cerebellar anlage around the posterior cerebellar midline to the opposite germinal trigone. By P2, this sling was no longer apparent and CD15 immunoreactivity was confined to astrocytes in the cerebellar white matter, surrounding the deep cerebellar nuclei. The CD15 immunoreactive processes pursue an unusual trajectory through the developing cerebellum, unlike any other previously described axonal or glial process bundle in the cerebellum. From its trajectory and association with the ventricular surface it seems that this structure, which we have named the transverse cerebellar sling, is composed of glial processes, although it was not immunoreactive for S-100 or glial fibrillary acidic protein. The transient appearance of this sling encircling the posterior cerebellum is suggestive of a role in prenatal cerebellar morphogenesis.

The expression of the 3-fucosyl-N-acetyl-lactosamine (FAL, CD15, Le(x)) epitope has been examined immunohistochemically in paraffin sections of the brainstem of the mouse. The initial appearance of labelling was at embryonic day (E)11, when immunoreactivity of radial glial fibres was noted in the medulla, pons and dorsal isthmus, while the spinal cord was immunoreactive from E12. Labelling remained faint until E14, when a distinctive radial pattern appeared in the medulla, pons and spinal cord. Immunoreactivity at E14 in both the spinal cord and medulla was strongest in a band in line with the sulcus limitans, passing ventrolaterally through the nucleus of the solitary tract in the case of the medulla. In both the spinal cord and the medulla, CD15 immunoreactivity divided the brainstem into radially arranged compartments, with an immunonegative paramedian region, strongly labelled ventrolateral segments, and moderately labelled lateral regions. Labelling of midline radial fibres was also apparent in the ventral mesencephalon at E14. After E18, labelling appeared in the paramedian region of the medulla, in particular around the inferior olivary nucleus, and gradually adopted a tufted appearance throughout the brainstem. Several regions of the developing brainstem showed specific labelling during fetal life. Distinct labelling of the developing red nucleus was visible from E15 to the time of birth, while some longitudinal bands of labelling were noted in the deeper layers of the superior colliculus from E17 until postnatal day (P)2. The adult pattern of immunolabelling was achieved by the end of the second postnatal week. The striking concentration of CD15 along the sulcus limitans of both the spinal cord and brainstem may serve to demarcate and separate dorsal (sensory) and ventral (motor) columns in a similar fashion to Pax gene expression. The precise timing of transient CD15 expression in the developing inferior olive and red nucleus is suggestive of a role for this epitope in developmental events of those structures.

The distribution of two glycoconjugates 3-fucosyl-N-acetyllactosamine (CD15) and HNK-1 epitope (CD57) in the inner ear of the NMRI mouse was analysed from the eighth day of gestation to the 16th day after birth. CD15 epitope distribution is developmentally regulated. The up- and down-regulation of expression, the change in the number of cells which are positive, the ingrowth of CD15-positive cells and their distribution, intracellular and/or cell-surface-associated expression, all assume a characteristic appearance at each developmental stage. Distribution of CD57 documented the nerve outgrowth and formation of the innervation of the vestibular apparatus and cochlear duct. Correlation between CD15 and CD57 expression patterns revealed differences in the interaction of the ingrowing fibres and epithelial tissue between the vestibular organ and the cochlea and differences in the development of the cristae and maculae.

The developmental expression of the 3-fucosyl-N-acetyl-lactosamine (CD15, Lex, SSEA-1) epitope has been examined immunohistochemically in paraffin sections of the mouse hippocampus. Labelling appeared initially at E10, with immunostaining of the ventricular surface of the hippocampal primordium and in the early hippocampal marginal zone. At E12 and E13, a group of cells in the marginal zone of the hippocampus was labelled. From E14 to E15, CD15 immunoreactivity delineated several boundaries in the developing hippocampal formation. A band of labelling was seen at these ages separating the fimbrioglial proliferative compartment from the primary dentate neuroepithelium. During late fetal and early postnatal life (E19-P5. 5), two strong bands of labelling were visible in the stratum lacunosum moleculare and stratum oriens of CA3, and in the marginal zone and deeper layers of the subiculum. The bands in CA3 corresponded in position to the earliest afferents to the hippocampus from the entorhinal cortex. In later postnatal life (from P5.5), astrocytic CD15 labelling predominated in a mosaic camouflage pattern, as seen in the adult.

The expression of the (alpha1-->3)-fucosyl-N-acetyl-lactosamine (CD15) epitope in the genital tract of the female rat during fetal and early postnatal ontogeny was investigated by means of immunohistochemistry. CD15 was exclusively associated with epithelial cells and was mainly located along the cell membrane. The CD15 expression was characterized, firstly, by considerable differences within the various structures and even substructures of the genital tract and secondly, by the high degree of time-related changes which accompanied the morphological development. In the Mullerian duct, CD15 was present from embryonic day (E) 14 until birth on the apical membranes throughout the epithelial cell layer. In the Wolffian duct, CD15 expression was present between E16 and E19. Along the longitudinal extent of the Wolffian duct, expression intensity differed, showing moderate to high levels in the epithelial cells of the cranial and caudal parts, but without recognizable CD15 expression in the intermediate part. In the urogenital sinus, CD15 was expressed from E15 until E21. In the cranial parts, all epithelial cells were positive, whereas in the caudal parts, CD15 was present only on their apical membranes. In the ovarian tube, uterine horn, and vagina, a moderate to high CD15 expression at birth gradually diminished to very low levels during postnatal days (P) 8 and 9. After P9, re-expression of CD15 occurred in the caudal part of the ovarian tube and in the uterus, increasing to a maximum at about P32. The findings provide (indirect) evidence for a correlation between the intensity of CD15 immunoreactivity and the serum concentrations of estrogens as well as of estrogen receptors in the urogenital tract. Since steroid hormone dependency can be regarded as a gauge of the differentiation of malignancies, it would be worthwhile correlating CD15 levels with those parameters.

The distribution of the glycoprotein CD15 (FAL, SSEA-1, Le(x)) in the developing cortex of the mouse has been examined by immunohistochemistry of paraffin sections. The pattern of CD15 immunoreactivity was observed to occur in 4 stages. In the first stage, the presumptive subplate region in the lateral and rostral parts of the developing isocortex was transiently labelled at E12. In the second stage, between E13 and E17, very faint label was found only in the ventricular germinal zone, possibly within radial glial fibres. The third stage began at E18, when transient labelling of several discrete cortical areas was apparent; the motor cortex (from E18 to P3), an area in the temporal lobe (presumptive primary auditory cortex; from E18 to P5.5) and the piriform and entorhinal cortices (from E18 to P3). The final stage began at P2, when a mosaic camouflage pattern of labelling started to appear throughout the developing cortex, particularly in layer I. The distribution of the transient label in the motor cortex during the perinatal period suggests that it may be associated with early events in corticospinal or other projection neuron maturation (apical dendrite growth and differentiation). The labelling in the presumptive auditory cortex (Te1) may also be involved in the differentiation of neurons in the upper cortical plate.

The effect of glycerol administration on the distribution pattern of the CD15 (3-fucosyl-N-acetyl-lactosamine) epitope has been studied immunohistochemically in hydropic and non-hydropic ears of adult guinea pigs from 80 min to 5 h after treatment with 2 g/kg glycerol. Glycerol administration characteristically modulated the immunoreactivity of the CD15 epitope in the endolymphatic sac and the tectorial membrane (TM) and revealed the CD15 epitope in the otoconia of the saccule, the stereocilia and the supporting cells of the ampullae. Our results suggest that glycerol interacts with glycoconjugates of the TM and otoconia, causing changes in their density and possibly the hydration state. As a consequence, the transduction process may be affected.

The distribution of the carbohydrate epitope CD15, a putative cell adhesion molecule, was studied in adult vertebrate retinas by light-microscopic immunohistochemistry. Except for Old World primates, in which no immunoreactivity was detectable, all other species expressed the epitope on retinal interneurones. Subpopulations of stratified amacrine cells were found in all species with the exception of bats and marmoset monkeys, and bipolar cells were immunoreactive in frogs and all amniotic animals. Ganglion cells were labelled in urodelian, in all sauromorphian, as well as in some mammalian species. In some species, the distribution of immunoreactive neurones was correlated to areas of retinal specialization such as the visual streak in frogs and the dorsotemporal field in birds. In these parts of the retina with enhanced visual acuity, more CD15 glycosylated bipolar cells were found than in other parts. Among mammals, labelled bipolar cells were found exclusively in species with cone-dominated retinas. This comparative study shows that CD15 expression is consistently membrane associated in morphologically defined subsets of amacrine, bipolar, and ganglion cells throughout the vertebrate phylum. Its distribution pattern was found to depend more on the visual behavior of a given species (cone-dominated or rod-dominated retina) than on phylogenetic proximity between species.

The spatiotemporal distribution pattern of the carbohydrate epitope CD15 was examined in the developing chick retina. CD15 expression appeared for the first time at E13 in the INL and GCL. The developmental profile of the INL, from E13 to E16, showed increasing numbers of stratified amacrine cells, whereas diffuse amacrine subtypes appeared later, beginning at E15. Smaller populations of bipolar cells were seen at E17. Three types of CD15-positive ganglion cells could be differentiated by E15. A gradient in the appearance of identified immunoreactive amacrine cells extended from the dorsotemporal to the ventral and nasal retina. The adult-like pattern of CD15 expression did not become established until E19. Adult-like densities of immunoreactive cells were reached toward the end of the embryonic period between E18 in the dorsotemporal and ventral retina, and E19 in the dorsonasal retina. In the adult-like retina, labelled cells became particularly numerous at its greatest circumference, and were most densely packed in the dorsotemporal retinal quadrant. From E16 to P5, three membrane-bound, CD15-positive glycoproteins of 20, 32 and 34 kDa were identified by Western blots. The time course in the appearance of the membrane-associated CD15 recognition molecule on differentiating amacrine, bipolar and ganglion cells is correlated to the establishment of synaptic contacts.

The expression of the 3-fucosyl-N-acetyl-lactosamine (CD15) epitope during the embryonic development of the mouse inner ear has been immunohistochemically studied on paraffin sections from day 8.5 to day 19 of gestation. Consecutive steps of morphological development are accompanied by up- and down-regulation of CD15 expression and changing cell patterns. Expression levels are reduced once structural maturation has been accomplished. Since CD15 has been described as a determinant that is expressed during critical developmental steps, it is speculated that the changes in the epitope expression in the inner ear reflect relevant stages of differentiation. This system may thus serve as a model for understanding inner ear development.

The present study examined the effect of glycerol administration on 3-fucosyl-N-acetyl-lactosamine (CD15) epitope expression in the endolymphatic sac (ES) of the guinea pig's inner ear. Adult guinea pigs were injected intravenously with glycerol (2 g/kg body wt.). CD15 expression was studied at 80 min up to 5 h after treatment. In untreated animals single cells and cell groups in the ES expressing CD15 epitope intra- and intercellularly were identified by immunohistochemistry to be mainly in the epithelial layer of the rugosal and distal part of the sac. Glycerol administration modulated the expression of CD15 epitope. In the epithelial layer, expression decreased and was nearly depleted after 3 h. After 4 h of glycerol administration, CD15 expression reappeared and reached the comparable level of controls. The numbers of CD15-positive cells in the lumen of the ES increased steadily and arrived at their the highest levels in 2-h specimens. The localization of CD15-epitope expression and its modulation after glycerol administration within the ES implies that this molecule may play a role in re-establishing the sac's normal function. In addition, we speculate that CD15 may be associated with processes of an immune response in the inner ear.

The expression of the CD 15 (3-fucosyl-N-acetyl-lactosamine) epitope was immunohistochemically studied on paraffin sections of adult guinea pig inner ears. Two regions of the inner ear expressed the epitope for CD 15: the tectorial membrane of the cochlea and the endolymphatic sac. The upper part of the main body of the tectorial membrane was deeply stained. In the rugosal and distal part of the endolymphatic sac several unevenly distributed cells showed strong intra- and extracellular localization of the CD15 epitope. The CD15 epitope is associated with a transduction structure (tectorial membrane) and with a "volume regulating" compartment (endolymphatic sac) and may be involved in the maintenance of the structural integrity of both.

The profile of the CD15 epitope-expression was studied in the lateral geniculate nucleus (LGN) of a monkey species (Cercopithecus aethiops aethiops L.) during the peri- and postnatal development. Intense labeling of the neuropil and of astroglial cells was detected. Morphological examination of the spatio-temporal appearance of the CD15-expression showed different consecutive patterns of expression: expression of the CD15 epitope was high one week prenatally and during the first postnatal weeks. During this perinatal period it was mainly expressed in the interlaminar zone of the magnocellular portion of the LGN. This was followed by a time period when immunoreactivity gradually decreased to become almost absent by approximately 10 weeks of age. Toward adulthood a different pattern of immunoreactivity occurred, revealing a pattern of lamination that was attributable to CD15 positive astrocytes, most prominent in the cellular layers 1 and 2. They were later on seen spread out through the entire LGN such that the LGN of adult animals was entirely filled with positive material. From the temporal correlation of CD15-expression and LGN histogenesis it is concluded that the high level of CD15, that is observed in the neuropil during the perinatal period, matches with high synaptic plasticity within the visual system; in contrast, low levels of CD15-expression correlate with synaptic reorganization processes.

The CD15-epitope in the cerebellar cortex is expressed by Bergmann glial fibers which are organized in symmetrical iterative stripes. Their arrangement appears complementary to the pattern described for the organization of compartments demonstrated by any other marker used so far. HNK-1 however, reveals stripes that are not complementary to the general pattern for the organization of compartments.

We have investigated the distribution of CD15 immunoreactivity (IR) in chick sensory neurons during the embryonic development. IR first appeared around embryonic day 6 (E6) preferentially in the lateroventral part of the dorsal root ganglion (drg). Here it was located in the cytoplasm of the ganglion cells. Starting at E11, CD15 positive cells have been detected also in the dorsomedial part of the drug. Towards the end of embryonic development, these immunoreactive (ir) cells were evenly distributed within the drg. Around E 16 nerve fibres could also be found, expressing the CD15 epitope. These fibres run both in centripetal and centrifugal direction, and could be followed throughout peripheral nerves. IR was confined to peripheral nerves of the skin, where after its subepidermal arborisation fine nerve terminals entered the basal stratum of the epidermis or the dermal bulbous part of feathers. Until hatching no staining in visceral structures was detected. Our results suggest that CD15 is restricted to a subset of drg neurons innervating somatosensory targets. A role for the CD15 epitope for myelination is discussed.

The distribution of the 3-fucosyl-N-acetyl-lactosamine (FAL) epitope within the adult mouse brain was studied by immunohistochemistry using the monoclonal antibody Leu-M1. Leu-M1-positive elements comprised astrocytes and neurons. FAL-positive astrocytes were particularly abundant in barrier structures of the brain, but were also prominent at the periphery of most medullated fiber tracts. Their intracerebral distribution led to a distinct pattern of organization, which in some locations, including the cerebral cortex, could be used for an extended regional architectonic description. Since only some FAL-positive astrocytes were also positive for glial fibrillary acid protein (GFAP), the emerging topography of the FAL-positive astrocytes often differed from the GFAP-distribution. In the cerebellum, Bergmann glia cells expressed the FAL epitope and, in the vermis, their arrangement had a band-like appearance. Positive oligodendrocytes could not be identified. The common ependymal cells were negative, whereas tanycytes were highly immunoreactive. The Leu-M1 antibody also stained some neurons. These occurred in selected neocortical regions, within the dorsal and ventral striatum, in the globus pallidus, the nucleus basalis of Meynert, the nucleus diagonalis and some hypothalamic areas. In some instances, their morphology and location indicated an association with neurochemically specified cell groups.

The distribution of the carbohydrate epitope 3-fucosyl-N-acetyl-lactosamine was investigated on paraffin sections of the normal adult human central nervous system by means of immunohistochemistry, using the mouse monoclonal antibody anti-Leu-M1. This antibody is one among many others recognizing this epitope, which is also known as stage specific embryonic antigen I or X-hapten. We found this epitope predominantly localized on astrocytes especially along their numerous cell processes. There was a striking association of immunoreactive astrocytes with intracortical blood vessels and with distinct brain areas. Leu-M1-positive oligodendrocytes were present in the white matter with highest densities in the centrum semiovale and lateral pontine regions. Most cells of the ependymal lining, including tanycytes, showed Leu-M1 immunoreactivity but there was some topographical variability. Double-labelling experiments revealed that Leu-M1-positive glial cells may or may not be immunoreactive to glial fibrillary acidic protein or S-100 protein. The Leu-M1 antibody also stained a large number of different sets of neurons from cerebral cortex to spinal cord. Groups of immunostained cells were found in the hypothalamus, dorsal thalamus, ventral and mesial mesencephalic tegmentum, and several lower brainstem regions such as the lateral reticular formation, the raphe nuclei and the pons. Leu-M1 immunoreactivity appeared associated to neurochemically specified neuronal cell groups such as the mesencephalic dopaminergic system and the hypothalamic neurophysin system. In summary, our study demonstrates a distinct topographical distribution of the Leu-M1 epitope in the adult human central nervous system. Its exact functions, however, have not yet been elucidated, but there is evidence for the involvement of this epitope in cell-cell-adhesion and -interaction processes.