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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 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 N-acetyl-lactosamine (NALA), a cell-surface carbohydrate epitope of the lactoseries, has been studied in the retina of representative species of all vertebrate classes by light microscope immunohistochemistry. In only some species of different classes (fish, amphibia and mammals) was NALA expression detected, and in these animals the distribution showed profound interspecies variability. In fishes and amphibia in which NALA was present, patterns ranged from single immunopositive cells to homogeneous labelling of cell layers. In mammals, NALA was found only in retinas that are cone dominated (tree squirrel and primates). In the tree squirrel, there was a dense cellular staining of the photoreceptor cell layer; whereas in primates, the carbohydrate epitope occurred only on some photoreceptor cells. From these receptor cells, positive axons could be traced to the inner plexiform layer. In spite of the profound interspecies differences, NALA is not randomly expressed, as its exclusive expression in mammals with cone-dominated vision indicates. The suggestion of a functional relevance for NALA glycosylation of retinal cells is supported by the labelling pattern for HNK-1 in these species, which was different from the pattern found in rod-dominated mammalian retinas.

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 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 distribution of the carbohydrate epitope 3-fucosyl-N-acetyl-lactosamine (CD15) has been immunocytochemically evaluated in coronal paraffin sections through the magnocellular basal forebrain system--the nucleus basalis of Meynert, the nucleus of the diagonal band of Broca and the medial septal nucleus--of 202 human brains. The brains derived from differently aged controls (n = 54) and from patients suffering from organic brain diseases (n = 129) or psychiatric disorders (n = 19). In 30 cases dementia was clinically diagnosed. CD15 first appeared around birth when it became localized on singular astrocytes. The astrocyte number and process density steadily increased, and at approximately 12 years the typical adult-type pattern was acquired. Considerable variations in the expression patterns were noted with regard to the astrocyte number, the intensity in immunostaining and the process relations of CD15-positive astrocytes with the magnocellular neurons. In the light of these variations, and of conflicting additional changes in other areas of most diseased brains, it was difficult to correlate different intensities and patterns to specific diseases. The results, however, provide evidence for an increase in CD15 expression and in process network density of astrocytes in the lateral part of the nucleus basalis of Meynert in cases of Huntington's disease.

The age-related distribution of the trisaccharide epitope 3-fucosyl-N-acetyl-lactosamine (CD15) was evaluated in the human lateral geniculate nucleus (LGN). Coronal paraffin sections from individuals between the 12th week of gestation to 99 years of age were processed for immunohistochemistry using monoclonal antibodies against the CD15 epitope. CD15 immunoreactivity was present in the neuropil from the 14th week of gestation with a graded pattern along the anteroposterior and mediolateral axes of the LGN. Immunoreactivity then became preferentially located within the future cell layers, shortly before cellular segregation was visible in Cresyl Violet stained sections. Maximal CD15 expression occurred from the 22nd week of gestation until the beginning of visual experience (second week of postnatal life). During the subsequent period the spatial pattern of CD15 expression changed. Whereas immunoreactivity in the cell layers gradually disappeared, CD15 positive astrocytes became transiently concentrated in the intercellular layers. The staining within the interlaminar region was best developed at about one year of postnatal life. The adult pattern was found at around 10 years of age, when the LGN appeared almost unstained. Two stages of CD15 expression can thus be separated. The first is characterized by neuropil staining and is synchronized with the time profile of neuronal maturation and of formation of non-stabilized contacts. CD15 is at this time possibly correlated with structural instability and increased vulnerability but at the same time with a high degree of plasticity. The second, peri- and postnatal stage is characterized by CD15 positive astrocytes. These appeared when CD15 in the neuropil disappeared. This loss of CD15 expression in the neuropil occurs during the phase of experience-dependent establishment of the mature interconnectivity and probably heralds loss in plasticity. The time-related expression pattern of CD15 is therefore compatible with the idea that CD15 levels reflect different degrees of developmental determination of retino-geniculate interaction.

This study reports a specific cross-reactivity of the three anti-human-hematopoietic-cell monoclonal antibodies, anti-Leu-7 (HNK-1), anti-Leu-11a (NKP-15), and anti-Leu-M1 (MMA), with different epitopes in the brain of the adult rat. The distribution of these epitopes in rat brain is determined by means of immunohistochemistry in paraffin-embedded frontal serial sections. The reaction pattern of anti-Leu-11a monoclonal antibody is very similar to that of polyclonal antibodies against the myelin basic protein. Both antisera give a specific reaction with myelinated fibers. Immunoreaction products with the anti-Leu-7 monoclonal antibody are found as diffuse, mostly punctiform material in the neuropil and even more evident as small granules coating the cell surface of many neurons. In the white matter anti-Leu-7 reveals a moderate reactivity, which occurs predominantly as spots and fine-stranded material within the myelinated fiber tracts. Anti-Leu-M1 immunoreactivity is present between myelinated fiber bundles of the white matter, where it has a reticulate appearance, and as fine-granulated material within the grey matter of the cortex and the nuclei. The characteristic feature in the grey matter is that of irregularly shaped immunopositive plaques, which are often located around small blood vessels. The cytoplasm of glial and neuronal cells appeared negative with this MAB. The exact topographical distribution of the Leu-7 and Leu-M1 epitopes throughout the rat brain is described. The present hypotheses concerning the nature of this shared antigenicity between hematopoietic cells and nervous tissue are discussed.