Highly sialylated N-CAM is expressed in adult mouse optic nerve and retina

Summary The localization of the neural cell adhesion molecule (N-CAM) and its highly sialylated form, which is prevalent in young tissues and has therefore been called embryonic neural cell adhesion molecule, was studied in the developing and adult mouse optic nerve and retina immunohistologically and immunochemically. At embryonic and early postnatal ages, neuroblasts and young postmitotic neurons, Müller cells and astrocytes in the retina, and retinal ganglion cell axons and all glial cells in the optic nerve express highly sialylated neural cell adhesion molecule. Beginning with the third postnatal week, highly sialylated neural cell adhesion molecule disappears from retinal ganglion cell axons in the optic nerve and from neuronal cell bodies and processes in the retina. In addition, it is not detectable on oligodendrocytes in 3-week-old animals. However, highly sialylated neural cell adhesion molecule continues to be expressed in the adult optic nerve and retina by astrocytes and Müller cells. On these cells it is only absent from cell membranes contacting basal lamina. Weakly sialylated neural cell adhesion molecule, in contrast, is expressed by all cell types of retina and optic nerve during development and in the adult. The loss of highly sialylated neural cell adhesion molecule from neurons and oligodendrocytes must therefore be considered as a cell type-specific conversion of the so-called embryonic to the adult form of neural cell adhesion molecule and does not simply reflect the disappearance of neural cell adhesion molecule from these cells. Weakly sialylated neural cell adhesion molecule, however, is absent from outer segments of photoreceptor cells and, as is the case for the highly sialylated form, from glial cell surfaces contacting basal lamina. Thus, the expression of highly sialylated neural cell adhesion molecule by pre- and postmitotic neurons and by oligodendrocytes is restricted mainly to the period of histogenetic events in retina and optic nerve, i.e. cell division, cell migration, dendritic and axonal growth and synaptogenesis. In addition to the observation that this form of neural cell adhesion molecule is less adhesive than the weakly sialylated, adult form, it is likely that highly sialylated neural cell adhesion molecule plays an important role during dynamic morphogenetic events. Furthermore, the expression of highly sialylated neural cell adhesion molecule by astrocytes and Müller cells in adult optic nerves and retinae suggests some histogenetically plastic functions for these cells in the adult mouse visual system.     

The development of astrocytes in the albino rabbit retina and their relationship to retinal vasculature

We have examined the development of astrocytes in the albino rabbit retina, using antibodies to glial fibrillary acidic protein (GFAP) and vimentin. Vimentin immunoreactive (vimentin⁺) astrocyte-like cells first appear at the 24th postconceptional day (24 PCD), in a pattern similar to that of the adult. GFAP immunoreactivity was first detected in astrocytes at the 29 PCD, in a similar pattern. Vessels enter the retina from 29 PCD. The presence of astrocytes in a mature distribution prior to the ingrowth of vessels indicates that astrocytes are not dependent on the vessels for their early positioning and differentiation. In contrast with the rat and cat, we found no evidence of migration of astrocytes into the rabbit retina from the optic nerve.     

The connective tissue and glial framework in the optic nerve head of the normal human eye: light and scanning electron microscopic studies

The arrangement of connective tissue components (i.e., collagen, reticular, and elastic fibers) and glial elements in the optic nerve head of the human eye was investigated by the combined use of light microscopy and scanning electron microscopy (SEM). Light-microscopically, the optic nerve head could be subdivided into four parts from the different arrangements of the connective tissue framework: a surface nerve fiber layer, and prelaminar, laminar, and postlaminar regions. The surface nerve fiber layer only possessed connective tissue elements around blood vessels. In the prelaminar region, collagen fibrils, together with delicate elastic fibers, formed thin interrupted sheaths for accommodating small nerve bundles. Immunohistochemistry for the glial fibrillary acidic protein (GFAP) showed that GFAP-positive cells formed columnar structures (i.e., glial columns), with round cell bodies piled up into layers. These glial columns were located in the fibrous sheaths of collagen fibrils and elastic fibers. In the laminar region, collagen fibrils and elastic fibers ran transversely to the optic nerve axis to form a thick membranous layer - the lamina cribrosa - which had numerous round openings for accommodating optic nerve fiber bundles. GFAP-positive cellular processes also ran transversely in association with collagen and elastin components. The postlaminar region had connective tissues which linked the lamina cribrosa with fibrous sheaths for accommodating nerve bundles in the extraocular optic nerve, where GFAP-positive cells acquired characteristics typical of fibrous astrocytes. These findings indicate that collagen fibrils, as a whole, form a continuous network which serves as a skeletal framework of the optic nerve head for protecting optic nerve fibers from mechanical stress as well as for sustaining blood vessels in the optic nerve. The lamina cribrosa containing elastic fibers are considered to be plastic against the mechanical force affected by elevation of the intraocular pressure. The present study has also indicated that glial cells with an astrocytic character play an important role in constructing the connective tissue framework characteristic of the optic nerve head.     

Concentration of astrocytic filaments at the retinal optic nerve junction is coincident with the absence of intra-retinal myelination: comparative and developmental evidence

The structure of the lamina cribrosa (LC) and astrocytic density were examined in various species with and without intra-retinal myelination. Sections of optic nerve from various species were stained with Milligan's trichrome or antibodies to glial fibrillary acidic protein, myelin basic protein (MBP) and antibody O4. Marmoset, flying fox, cat, and sheep, which lack intraretinal myelination, were shown to possess a well-developed LC as well as a marked concentration of astrocytic filaments distal to the LC. Rat and mouse, which lack intraretinal myelination, lacked a well-developed LC but exhibited a marked concentration of astrocytic filaments in this region. Rabbit and chicken, which exhibit intraretinal myelination, lacked both a well-developed LC and a concentration of astrocytes at the retinal optic nerve junction (ROJ). A marked concentration of astrocytes at the ROJ of human fetuses was also apparent at 13 weeks of gestation, prior to myelination of the optic nerve; in contrast, the LC was not fully developed even at birth. This concentration of astrocytes was located distal to O4 and MBP immunoreactivity in human optic nerve, and coincided with the site of initial myelination of ganglion cell axons in marmoset and rat. Myelination proceeded from the chiasm towards the retinal end of the human optic nerve. Moreover, the outer limit of oligodendrocyte precursor cells (OPC) migration into the rabbit retina was restricted by the outer limit of astrocyte spread. These observations indicate that a concentration of astrocytic filaments at the ROJ is coincident with the absence of intraretinal myelination. Differential expression of tenascin-C by astrocytes at the ROJ appears to contribute to the molecular barrier to OPC migration (see Bartsch et al., 1994), while expression of the homedomain protein Vax 1 by glial cells at the optic nerve head appears to inhibit migration of retinal pigment epithelial cells into the optic nerve (see Bertuzzi et al., 1999). These observations combined with our present comparative and developmental data lead us to suggest that the astrocytes at the ROJ serve to regulate cellular traffic into and out of the retina.     

The peripapillary glia of the optic nerve head in the chicken retina

Abstract The eye of reptiles and birds is characterized by an avascular retina and a vascular convolute called conus papillaris in reptiles and pecten oculi in birds which arises from the papilla nervi optici (PNO) or optic nerve head into the vitreous. At least in birds, this central part of the retina is the site of a heterogeneous population of glial cells. Müller cells reside in the retina, astrocytes in the optic nerve, and pecteneal glial cells in the pecten. The latter are developmentally related to the pigment epithelial cells. In addition to these established types of cells, there is a population of glial cells lining the base of the pecten oculi. In the present study, we investigated both the morphology and the development of these glial cells of the PNO in a series of chicken embryos. These cells were called peripapillary glial cells. They were characterized by their morphology and by their spatiotemporal expression of antigens typical of glial cells (intermediate filaments and glutamine synthetase). They reside at the border between the retina and the optic nerve and at the innermost border of the ventricular cleft representing transitional forms among Müller cells, astrocytes, and pigment epithelial cells. The developmental data suggest a migration of the perikarya of the peripapillary glia in vitread direction, which may coincide with that of the pecteneal glia. Whereas the pecteneal glial cells differentiate morphologically from E16 on, the peripapillary glia retain characteristics of radial glia by spanning the distance from the vitreous to the ventricular cleft. Blood vessels only occurred in the optic nerve head and the pecten oculi. No capillaries were found in the retinal tissue, beyond the peripapillary glia, leading us to suggest that these cells may play a role in demarcating the outer limit of vascularization. The functional properties of these cells are unknown but were discussed to include prevention of vessel growth into the avascular retina and/or axonal guidance during development.     

Two populations of glial cells from fish optic nerve/tract with distinct electrophysiological properties

Summary The electrophysiological properties of the two major glial cell types in cultures from the regenerating goldfish optic nerve/tract were studied with patch-clamp techniques. Spindle-shaped cells express myelin proteins. These oligodendrocyte-like cells possess outwardly rectifying currents, do not show glutamate activated currents and are rarely electrically coupled to neighboring cells. Cells of epitheloid morphology probably represent astrocytes. They are GFAP-positive and do not exhibit myelin proteins. These cells have glutamate activated currents, display a linear current to voltage relationship and are extensively electrically coupled thus displaying properties similar to mammalian astrocytes.     

BMP-4在视网膜与视神经中的分布及对少突胶质前体细胞分化的影响

目的:研究骨形态发生蛋白4(bone morphogenetic protein4,BMP-4)在视网膜与视神经上的表达情况及其对少突胶质前体细胞(oligodendrocyte precursor cells,OPCs)分化的影响,进一步探讨视神经乳头处无髓鞘形成的原因。方法:取生后7d SD大鼠视神经,用免疫组织化学方法研究BMP-4的表达情况。采用恒温振荡法和差速贴壁法分离纯化新生SD大鼠OPCs。用BMP-4诱导OPCs分化,通过免疫细胞化学方法研究OPCs的分化方向。通过Western Blot方法研究BMP-4浓度与OPCs细胞体系内Olig2蛋白表达水平之间的相互关系。结果:(1)在发育过程中,BMP-4仅选择性表达在视神经乳头处;(2)成功纯化的OPCs在PDGF和bFGF撤除后自发分化为少突胶质细胞;(3)OPCs经BMP-4诱导后向II型星形胶质细胞分化;(4)OPCs内Olig2蛋白的表达量随着BMP-4浓度的逐渐增高而逐渐降低。结论:在视神经乳头表达的BMP-4蛋白有可能作用于迁移至此处的OPCs,通过激活相关信号途径以下调OPCs内Olig2蛋白的表达水平,抑制OPCs向少突胶质细胞方向分化,从而抑制视神经乳头髓鞘形成。     

Astrocyte invasion and vasculogenesis in the developing ferret retina

We studied the time course of astrocyte invasion and blood vessel formation in the developing ferret retina using glial fibrillary acidic protein (GFAP)-immunohistochemistry for astrocytes and isolectin B4 histochemistry for blood vessels. As in other mammals, strongly GFAP positive astrocytes invade the ferret retina from the optic nerve. At birth, strongly GFAP positive astrocytes have reached about 22% of the distance between optic disc and outer retinal edge whereas weakly GFAP positive processes already extend to the edge of the retina. At postnatal days P30–P37 about 82% of the distance between optic disc and outer retinal edge and in the adult 88% of this distance is covered with strongly labelled astrocytes. Superficial blood vessels form from the optic disc. They reach up to about 24% of the retinal radius at birth and grow radially across the retina during further development. At P30–P37, the whole retina is covered with superficial blood vessels. The deep vascular layer forms later (around P30) through sprouting from superficial vessels. The radial pattern of astrocyte and vessel growth from the optic disc is not affected by the formation of the area centralis and visual streak.     

Development of astrocytes in the lamina cribrosa sclerae of the mouse optic nerve,with special reference to myelin formation

In the mouse optic nerve, the optic nerve fiber layer in the retina, the optic papilla and the lamina cribrosa sclerae (LCS) just after penetrating the eyeball failed to generate myelin, whereas the optic nerve proper in the orbit was occupied by myelinated nerve fibers. The present study investigated development of the architecture of LCS, where the axons develop from unmyelinated to myelinated type, to elucidate how the initial part of axons was unmyelinated. At the LCS of the adult optic nerve, well developed astrocytes densely formed a cytoplasmic mesh-like frame through which unmyelinated fibers passed. The astrocytes here contained numerous and densely packed intermediate glial filaments and cell organelles. This framework formed by astrocytes appeared to be completed between 7 and 14 postnatal days before oligodendrocyte progenitors, migrated from the chiasm side, reached the proximal end of LCS, and began myelin formation. Thus the failure in myelin formation at the intraocular part and LCS possibly depended upon unsuccessful migration of oligodendrocytes beyond LCS constructed by specialized astrocytes, although other inhibitory factors for myelin formation, such as adhesion molecules distributed around LCS, may be unsolved.     

The origin and development of retinal astrocytes in the mouse

Summary Astrocytes, a class of glia which appear in the mammalian retina late in development, have been postulated either to originatein situ from Müller cells or extra-retinally from the optic stalk epithelium, only subsequently invading the eye. The site of origin and the developmental characteristics of retinal astrocytes were examined in the mouse, a species not previously studied for this purpose. Sections of normal eyes and stalks at different ages were examined. Cells positive for glial fibrillary acidic protein (GFAP) were first observed at post-conceptional day 17 at the optic disc end of the stalk. From this site, the GFAP-positive cells migrated into and across the retina at a rate of 290 m per day, reaching its edge by post-conceptional day 28. While migrating across the retina, the astrocytes progressively increased in size and morphological complexity, observations confirmed by measurement of their fractal dimension. Over the same period, a wave of differentiation swept along the stalk in the cranial direction. Further evidence that retinal astrocytes are born outside the retina emerged when foetal hemiretinae with or without optic stalks were explanted to the chorioallantoic membrane of the chick. When examined one to twelve days later, no expiant cultured without the optic stalk contained GFAP-positive astrocytes, while expiants with the stalk left attached contained relatively normal numbers of astrocytes. We observed, using fluorescence confocal microscopy, that retinal astrocytes in the mouse as in the rat, associate predominantly with blood vessels, not axonal bundles. It was of interest to determine whether this class of glia is essential to the normal cytoarchitectural development of the neural retina. Morphological analysis of the expiants revealed no observable differences in cytoarchitecture or in the timing of developmental events between retinae maturing with or without astrocytes. It was therefore concluded that astrocytes may not be essential to the normal structural development of the murine retina.     

Is Thy-1 expressed only by ganglion cells and their axons in the retina and optic nerve?

Summary The distribution of Thy-1 in the retina and optic nerve has been examined immunohistochemically, and compared to that of the astrocytic marker glial fibrillary acidic protein. The axons and cell bodies of ganglion cells were found to be Thy-1 positive as were processes within the inner plexiform layer. Transection of the optic nerve in the neonatal rat results in the rapid degeneration of the ganglion cells but some Thy-1 staining remains in the inner plexiform layer. We have estimated using an immunoassay of normal and optic nerve transected retinae that about 70% of the Thy-1 in the retina is on ganglion cells and their axons and the remainder is on cells which contribute processes to the inner plexiform layer, presumably amacrine, bipolar or Müller cells.In the optic nerve the Thy-1 was found to be limited to the fascicles of optic nerve fibres and the intrafascicular spaces, containing astrocytes and their processes, were not stained. Axotomy of the adult nerve, which produced axonal degeneration and astrocytic proliferation, led to a loss of over 95% of the Thy-1 from the nerve. We found no evidence that the astrocytes of the retina or optic nerve were Thy-1 positive in normal animals or during degeneration.     

Effects of neonatal transection on glial cell development in the rat optic nerve: evidence that the oligodendrocyte-type 2 astrocyte cell lineage depends on axons for its survival

Summary We have previously provided evidence that the rat optic nerve contains three types of macroglial cells that develop as two distinct lineages: one lineage comprises type 1 astrocytes, which develop before birth, while the other comprises oligodendrocytes and type 2 astrocytes, which develop after birth from a common, bipotential glial progenitor cell. In the present study we have examined the influence of axons on the development of these two glial cell lineages by cutting the optic nerve at birth so that the retinal ganglion cell axons in the nerve degenerate. Using antibodies to distinguish the different types of glial cells in suspensions and semithin frozen sections of cut and uncut optic nerves, we show that neonatal transection results in a striking decrease in the total number of oligodendrocytes, type 2 astrocytes and their progenitor cells but has much less effect on the number of type 1 astrocytes. Since the [³H]thymidine labelling indices of oligodendrocytes and their progenitor cells were not significantly decreased in cut nerves, our results suggest that the progenitor cells and/or their progeny die in large numbers following neonatal nerve transection. We conclude that axons are required for the survival of cells of the oligodendrocyte-type 2 astrocyte lineage, at least during postnatal development.     

The serine racemase mRNA is expressed in both neurons and glial cells of the rat retina

D-Serine, an endogenous and obligatory coagonist for the glycine site of the N-methyl-D-aspartate receptor in mammals, is synthesized from L-serine by serine racemase. Serine racemase and D-serine have long been believed to occur predominantly in astrocytes, according to immunohistochemical studies. Recent studies have demonstrated, however, that both the mRNA and protein levels of serine racemase are considerably higher in neurons than in astrocytes in primary cultures of the rat brain and that the mRNA level of serine racemase predominates in neurons of the adult rat brain. Here we report the application of in situ hybridization based on tyramide signal amplification for the detection of serine racemase mRNA in sections of the adult rat retina and optic nerve head. The localization of serine racemase mRNA could be demonstrated in ganglion cells, amacrine cells, bipolar cells, horizontal cells, and Müller cells of the retina as well as in the astrocytes of the optic nerve head and the lamina cribrosa. This is the first study to demonstrate the exact localization of serine racemase mRNA at the cellular or tissue level in the retina and the optic nerve head. These results suggest that both the neuron- and glia-derived D-serine could modulate neurotransmission via the glycine site of the N-methyl-D-aspartate receptors in the retina.     

The rabbit retina: a suitable mammalian tissue for obtaining astroglia-free Müller cell cultures

Monolayer cultures were prepared from two distinct parts of early postnatal rabbit retinae. Cell suspensions obtained from the developing medullary ray (MR) region contained neurons, Müller (glial) cells, and astrocytes, cells obtained from the remainder (peripheral) part of the retina contained neurons and Müller cells, but no astrocytes. Müller cells lack glial fibrillary acidic protein (GFAP) immunolabeling in situ but some of them acquire faint GFAP labeling in both types of cultures. Strongly GFAP-labeled cells, most likely astrocytes, were seen in MR cultures only. We propose that the periphery of the rabbit retina is ideal for obtaining astroglia-free Müller cell cultures to study their functional properties in vitro.     

Acquisition of vimentin in astrocytes cultured from postnatal rat brain

Summary Vimentin and glial fibrillary acidic protein (GFAP) represent the principal constituents of intermediate filaments found in astrocytes. In contrast to vimentin—GFAP transition which occurs during glial developmentin situ, vimentin coexists with GFAP in cortical astrocytes allowed to differentiate in culture. To examine whether culture conditions or proliferative activity of the cells is responsible for the expression of vimentin, we generated cultures of GFAP-positive, vimentin-negative astrocytes isolated from 26-day postnatal rat brain cortices. Isolated astrocytes are characterized by a very thin rim of perinuclear cytoplasm and by numerous processes. Antiserum to GFAP labelled major processes and cell somata of some astrocytes, especially those with relatively short and large processes. Within 3 days in culture, all astrocytes accumulated GFAP in hypertrophic cell bodies and many began to express vimentin. Vimentin appeared primarily close to nuclei, and filaments of vimentin extended into proximal segments of the cell processes. In some astrocytes, however, vimentin was always absent. Combined double immunolabelling and histoautoradiography experiments demonstrated that the acquisition of vimentin was independent of the ability of astrocytes to incorporate tritiated thymidine. The results indicate that astrocytes isolated from 26-day postnatal rat brain are heterogeneous with respect to their ability to express vimentin and that vimentin synthesis is not correlated with the growth state of the cells as had been previously suspected.     

Generation of retinal cells in the wallaby, Setonix brachyurus (quokka)

We have examined the generation of retinal cells in the wallaby, Setonix brachyurus (quokka). Animals received a single injection of tritiated thymidine between postnatal days 1-85 and retinae were examined at postnatal day 100. Retinae were sectioned, processed for autoradiography and stained with Cresyl Violet. Ganglion cells were labelled by injection of horseradish peroxidase into the optic tracts and primary visual centres. Other cells were classified according to their morphology and location. Retinal cell generation takes place in two phases. During the first phase, which concludes by postnatal day 30, cells destined to lie in all three cellular layers of the retina are produced. In the second phase, which starts by postnatal day 50, cell generation is almost entirely restricted to the inner and outer nuclear layers. Cells produced in the first phase are orthotopic and displaced ganglion cells, displaced and orthotopic amacrine cells, horizontal cells and cones. Glia in the ganglion cell layer, orthotopic amacrine cells, bipolar and horizontal cells. Muller glia, and rods are generated in the second phase. Cells became heavily labelled with tritiated thymidine in the central retina before postnatal day 7, over the entire retina (panretinal) by postnatal day 7 and from postnatal day 18, only in the periphery. The second phase of cell generation is initiated at P50, in a region extending from the optic nerve head to mid-temporal retina. Subsequently, cells are generated in annuli, centred on mid-temporal retina, which are seen at progressively more peripheral locations. Therefore, cell addition to the inner and outer nuclear layers continues for longer in peripheral than in mid-temporal retina. We suggest that such later differential cell addition to the inner and outer nuclear layers contributes to an asymmetric increase in retinal area. This non-uniform growth presumably results in more expansion of the ganglion cell layer peripherally than in mid-temporal retina and may play a role in establishing density gradients of ganglion cells.     

Electron microscopy of the early postnatal development of fibrous astrocytes

The early postnatal development of fibrous astrocytes in rat optic nerves, has been studied with the electron and light microscopes. At birth, immature astrocytes form approximately 85% of the total cell population, and they have many features in common with the fibrous astrocytes of adult optic nerve. They are stellate cells whose processes form both the glia limitans and the glial sheets which group the axons into fascicles. However, in respect to their cytoplasmic characteristics, immature and mature astrocytes differ considerably. Early postnatal astrocytes have a more electron-dense cytoplasmic matrix containing an extensive array of organelles. Many microtubules occur in the processes of immature astrocytes, but the characteristic filaments of adult fibrous astrocytes are very sparse. During development, the number of microtubules decreases while the filaments increase until all cytoplasmic areas not occupied by organelles are filled by filaments. This suggests that filaments may be derived from the breakdown of microtubules. This and other possible functions of microtubules in developing astrocytes are discussed.     

Anomalous axonal outgrowth at the retina caused by injury to the optic nerve or tectal ablation in adultXenopus

Summary The retina and optic nerve head have been examined by light and electron microscopy in adultXenopus laeuis after injury to optic nerve fibres. Intraorbital resection, transection or crush of the optic nerve all resulted in the appearance at the retina of a mass of actively growing axons which formed a ring around the intraretinal and adjacent choroidal portions of the optic nerve head. Formation of this heterotopic axon population was first noted at two weeks after nerve injury and fibres persisted for at least six months. The ectopic fibres were seperated from the optic nerve head by astrocytes within the retina or by blood vessels and fibroblasts of the leptomeninges at extraretinal locations. In general, the orientation of the ectopic fibres was perpendicular to the fibres of the optic nerve. Bundles of axons were found between the ring of ectopic fibres and the pigment epithelial layer of the retina or among the blood sinuses of the choroid. Similar ectopic fibres were seen following transection of the optic nerve at the chiasm and after tectal ablation although the onset of these changes was slower than that seen after nerve resection. It is concluded that damage to visual pathways in the frog induces dramatic morphological alterations in the optic nerve and retina far proximal to the site of injury in this regenerating system.     

Two classes of astrocytes in the adult human and pig retina in terms of their expression of high affinity NGF receptor (TrkA)

Astrocytes have been implicated in axon guidance and synaptic regeneration in the retina and these processes involve activation of the high affinity nerve growth factor receptor, known as the tyrosine kinase A (TrkA) receptor. The purpose of the present study was to characterize the expression of TrkA in astrocytes of the adult pig and human retina. To this end, sections of human and pig retinas were immunolabeled with a combination of antibodies to glial fibrillary acidic protein (GFAP) and TrkA. Our study revealed that most of the GFAP-positive cells express TrkA, whereas a rare, novel subpopulation of astrocytes was found to be devoid of TrkA. Our results support the idea that astrocytes play an important neurotrophic role in the retina.     

Glial cell differentiation in neuron-free and neuron-rich regions

Summary The development of the human fetal hippocampus and dentate gyros has been studied immunocytochemically. The first glial cells to appear are vimentin-positive radial glial cells. A gradual transition from vimentin to glial fibrillary acidic protein (GFAP) reactivity in the radial glial cells occurs at week 8. The GFAP-positive radial glial cells transform into astrocytes from week 14. A population of small S-100-positive somata which morphologically and spatially are distinct from GFAP-positive radial glial cells and their transformed progeny, are found as early as week 9.5 in the hippocampus during the period of peak neurogenesis. The well-defined immunoreactivity of the morphologically homogenous cell subpopulation for S-100 protein, which has been used as an astrocytic marker in the adult hippocampus, indicates that astrocytes may differentiate at very early gestational ages in human fetuses. The S-100-positive astrocytes are thought to be derived from ventricular zone cells, which at the time of their appearance do not express any of the applied astrocytic markers (S-100, GFAP, vimentin). It is suggested that the S-100-positive astrocytic cell population interacts with the first incoming projection fibers, so modulating the pattern of connectivity.