Transformation of cells of astrocyte lineage into macrophage-like cells in organotypic cultures of mouse spinal cord tissue

Phagocytic cells on the surface of the explants and their relationships to the surface were examined morphologically and immunocytochemically in organotypic cultures of mouse spinal cord tissue. Phagocytic cells were rounded, had smooth cytoplasmic surfaces and were occasionally closely apposed to underlying cells by junctional complexes. These cells contained dense bodies, vacuoles, smooth and coated vesicles, a few microtubules and bundles of intermediate filaments similar to astroglial filaments. The superficial layer of the explant which usually consisted of astroglial cell bodies and their processes, sometimes contained immature neuroepithelial cells with numerous free ribosomes, centrioles, Golgi apparatus, microtubules and infrequently, intermediate filaments. Overall, the cells resembled poorly differentiated astrocytes. Numerous dense bodies and coated vesicles were observed in some of these immature cells as well as in astrocytes in the surface layer of the explant. Cytoplasmic bridges between immature cells within the explant and phagocytic cells on the surface were observed. Immunocytochemistry revealed the presence of glial fibrillary acidic protein within these surface phagocytic cells. It thus appears that immature neuroepithelial cells of astrocytic lineage are capable of transforming into macrophage-like cells in organotypic culture.     

Effects of methylmercury on neuroepithelial germinal cells in the developing telencephalic vesicles of mice

Summary Methylmercury (MeHg) poisoning (20 mg/kg body weight) in embryonic mice resulted in significant reductions of mitotic indices in the neuroepithelial germinal cells of the telencephalon at the ventricular surface 4 to 12 h following intoxication. After 24 h, no significant difference in the mitotic indices was observed as compared to controls. However, after 48 h there was an increase in mitotic indices of MeHg group as compared to controls. Analysis of the mitotic figures revealed features suggestive of early-phase mitotic arrest in MeHg-exposed animals. Radioautographic studies suggest a disturbance in the interkinetic nuclear migration of proliferating ventricular cells in the MeHg group. Acute degenerative changes in scattered ventricular cells characterized by edema and spongy changes of cytoplasm associated with dissolution of ribosomes, clearing of the cytoplasmic matrix and loss of organelles including microtubules were observed by electron microscopy. Loss of microtubules was also evident within mitotic figures in MeHg-poisoned animals. It is suggested that reduction and arrest of mitotic activity and disturbances in the interkinetic nuclear migration of neuroepithelial germinal cells are related to cytotoxic effects of MeHg on ventricular cells, including effects on microtubules. These findings suggest in that MeHg severely affects proliferating neuroepithelial germinal cells during the acute phases of MeHg poisoning, and that these changes may eventually affect the architectonic makeup of the cortical plate as the brain matures.Supported by NIH grant no. ES 02928     

Bipotent cortical progenitor cells process conflicting cues for neurons and glia in a hierarchical manner.

Neurons and glia of the cerebral cortex are thought to arise from a common, multipotent progenitor cell that is instructed toward alternate fates by extracellular cues. How do these cells behave when confronted with conflicting cues? We show here that nestin-positive neuroepithelial (NE) cells from embryonic day 14 rat cortex coexpress surface receptor proteins for ciliary neurotrophic factor (CNTF) and platelet-derived growth factor (PDGF). Both sets of these receptor proteins are functional in NE cells, as shown by ligand-dependent activation of downstream signal-generating proteins. Transient (30') exposure to CNTF instructs NE cells toward an astrocyte fate. Brief exposure to PDGF initiates neuronal differentiation. However, when challenged with conflicting cues, PDGF is dominant to CNTF. Moreover, CNTF-treated NE cells can be "redirected" by a subsequent exposure to PDGF to form neurons instead of astrocytes, whereas the converse is not true. The asymmetric relationship between CNTF and PDGF indicates that these two growth factors act on a common progenitor cell that has, at a minimum, two fates available to it rather than separate populations of precommitted neuroblasts and astroblasts. This bipotent progenitor cell processes conflicting cues for neurons and glia in a hierarchical manner.     

Polarity and intracellular compartmentalization of Drosophila neurons

Background

The choice of a stem cell to divide symmetrically or asymmetrically has profound consequences for development and disease. Unregulated symmetric division promotes tumor formation, whereas inappropriate asymmetric division affects organ morphogenesis. Despite its importance, little is known about how spindle positioning is regulated. In some tissues cell fate appears to dictate the type of cell division, whereas in other tissues it is thought that stochastic variation in spindle position dictates subsequent sibling cell fate.

Results

Here we investigate the relationship between neural progenitor identity and spindle positioning in the Drosophila optic lobe. We use molecular markers and live imaging to show that there are two populations of progenitors in the optic lobe: symmetrically dividing neuroepithelial cells and asymmetrically dividing neuroblasts. We use genetically marked single cell clones to show that neuroepithelial cells give rise to neuroblasts. To determine if a change in spindle orientation can trigger a neuroepithelial to neuroblast transition, we force neuroepithelial cells to divide along their apical/basal axis by misexpressing Inscuteable. We find that this does not induce neuroblasts, nor does it promote premature neuronal differentiation.

Conclusion

We show that symmetrically dividing neuroepithelial cells give rise to asymmetrically dividing neuroblasts in the optic lobe, and that regulation of spindle orientation and division symmetry is a consequence of cell type specification, rather than a mechanism for generating cell type diversity.     

Quantitative analyses of neuroepithelial cell shapes during bending of the mouse neural plate

Despite a wealth of information about cell behaviors contributing to neurulation in chick embryos, similar behaviors in mouse embryos have yet to be well characterized. This study examines cell behaviors occurring during bending of the mouse neural plate, in particular, qualitative and quantitative changes in neuroepithelial cell shape. Our current results demonstrate that in mouse embryos (1) the median hinge point (MHP), a localized region of neural plate that becomes anchored to the underlying prechordal plate mesoderm or notochord/ notochordal plate and forms a midline longitudinal furrow around which folding of the remaining neural plate (i.e., the part of the neural plate not involved in MHP formation) occurs, develops during stages of neural fold elevation; (2) the MHP is enriched with wedge-shaped neuroepithelial cells but has significantly fewer spindle-shaped, inverted wedge-shaped, and globular neuroepithelial cells than do the adjacent paired lateral areas of the neuroepithelium (L); and (3) each L is enriched with spindle-shaped, inverted wedge-shaped, and globular neuroepithelial cells but has significantly fewer wedge-shaped neuroepithelial cells than does the MHP. Thus wedging of neuroepithelial cells occurs during bending of the mouse neural plate and is localized to the MHP during neural fold elevation. Similarly, previous studies in the chick have shown that neuroepithelial cells become wedge shaped during bending of the neural plate and that such cell wedging is localized to the MHP during neural fold elevation. Such studies also have shed light on the roles of MHP formation and localized wedging of neuroepithelial cells within the MHP in the chick; however, such roles have yet to be elucidated in the mouse. It is probable that the MHP in mouse embryos, like that in chick embryos, provides a locus for bending of the neural plate and that wedging of neuroepithelial cells within the MHP provides the force necessary to generate the longitudinal midline furrow around which subsequent folding of the neural plate (i.e., neural fold elevation) occurs. Further studies are necessary to define these roles more precisely. © 1994 Wiley-Liss, Inc.     

Angiocentric neuroepithelial tumor mimicking Ammon's horn sclerosis--case report

CASE REPORT: We report on a 46-year-old male patient with pharmacoresistant temporal lobe epilepsy (TLE). Based on ictal EEG patterns and MRI scans, Ammon's horn sclerosis (AHS) or an epilepsy-associated tumor was included in the differential diagnosis. RESULTS: Histopathological examination of the surgical specimen revealed the unusual finding of a monomorphous angiocentric neuroepithelial tumor composed of small round cells and bipolar processes with perivascular aggregation. Immunohistochemistry detected perivascular-oriented expression of GFAP and cytoplasmic immunoreactivity of EMA and vimentin. Mitotic or other signs of proliferative activity were lacking. During a 2-year follow-up, the patient was seizure-free. CONCLUSIONS: Albeit AHS is the most frequent finding in TLE specimens, uncommon neuroepithelial tumors with hippocampal growth pattern have to be considered in the differential diagnosis of mesial TLE. The present case meets the criteria of an angiocentric neuroepithelial tumor recently proposed as a new clinicopathological entity. These tumors may be compatible with a maldevelopmental origin during early brain development.     

Neural-tube-derived neuroepithelial stem cells: a new transplant resource for Parkinson's disease

To assess the feasibility of using neuroepithelial stem cells as a transplant source for Parkinson's disease, neuroepithelial cells were harvested from the neural tube, cultured and stereotactically transplanted into the striatum of a rat model of Parkinson's disease. In culture, neuroepithelial cells generated abundant neurospheres and differentiated into both neurons and glia. After transplantation, tyrosine-hydroxylase-positive cells were detected in the graft. Furthermore, an apomorphine-induced rotation test showed that the implanted cells successfully promoted functional recovery in animals that underwent this transplantation procedure. Our results demonstrate that neuroepithelial cells may be a new source of donor material for Parkinson's disease.     

Early fate bias in neuroepithelial progenitors of hippocampal neurogenesis

Hippocampal adult neural stem cells emerge from progeny of the neuroepithelial lineage during murine brain development. Hippocampus development is increasingly well understood. However, the clonal relationships between early neuroepithelial stem cells and postnatal neurogenic cells remain unclear, especially at the single-cell level. Here we report fate bias and gene expression programs in thousands of clonally related cells in the juvenile hippocampus based on single-cell RNA-seq of barcoded clones. We find evidence for early fate restriction of neuroepithelial stem cells to either neurogenic progenitor cells of the dentate gyrus region or oligodendrogenic, non-neurogenic fate supplying cells for other hippocampal regions including gray matter areas and the Cornu ammonis region 1/3. Our study provides new insights into the phenomenon of early fate restriction guiding the development of postnatal hippocampal neurogenesis.     

The environment of axonal migration in the developing chick retina: a scanning electron microscopic (SEM) study.

We have made a SEM study of the basal intercellular spaces of the retina in chick embryos of different developmental stages. Since this is the environment where optic axons grow, the structural characteristics of this region might play some role in the orientation of axonal migration towards the choroid fissure. The basal region of undifferentiated retinas is formed by the vitreal expansions of neuroepithelial cells. In pre-axonal stages, the intercellular spaces between these expansions do not show any preferential orientation towards the fissure. The growth cones of ganglion cell axons appear in an apicobasal direction and turn towards the fissure immediately beneath the vitreal surface. Fasciculation is an early event during development and, in the more advanced stages, the vitreal expansions from retinal cells are placed in rows following the same orientation as the axon bundles. These observations are discussed in relationship to current hypotheses on axonal migration and orientation.     

Early development of the optic nerve in the turtle Mauremys leprosa

We show the distribution of the neural and non-neural elements in the early development of the optic nerve in the freshwater turtle, Mauremys leprosa, using light and electron microscopy. The first optic axons invaded the ventral periphery of the optic stalk in close relationship to the radial neuroepithelial processes. Growth cones were thus exclusively located in the ventral margin. As development progressed, growth cones were present in ventral and dorsal regions, including the dorsal periphery, where they intermingled with mature axons. However, growth cones predominated in the ventral part and axonal profiles dorsally, reflecting a dorsal to ventral gradient of maturation. The size and morphology of growth cones depended on the developmental stage and the region of the optic nerve. At early stages, most growth cones were of irregular shape, showing abundant lamellipodia. At the following stages, they tended to be larger and more complex in the ventral third than in intermediate and dorsal portions, suggesting a differential behavior of the growth cones along the ventro-dorsal axis. The arrival of optic axons at the optic stalk involved the progressive transformation of neuroepithelial cells into glial cells. Simultaneously with the fiber invasion, an important number of cells died by apoptosis in the dorsal wall of the optic nerve. These findings are discussed in relation to the results described in the developing optic nerve of other vertebrates.     

Neurochemical studies in a mouse teratoma with neuroepithelial differentiation. Presence of cyclic AMP, serotonin and enzymes of the serotonergic, adrenergic and cholinergic systems.

A transplantable mouse testicular teratoma (OTT 6050) which displays a spectrum of neuroepithelial differentiation was evaluated biochemically for concentrations of cyclic AMP (cAMP), serotonin (5-HT), and enzymes involved in the metabolism of the biogenic amines and acetylcholine. These values were compared between teratomas with neuroepithelial differentiation as the major or minor component and brains of neonatal and adult mice of related strains. cAMP, 5-HT, tryptophan hydroxylase (TPH), aromatic amino acid decarboxylase (AADC) and monoamine oxidase (MAO) were present. In addition, enzymes of the adrenergic system, i.e. tyrosine hydroxylase (TH) and dopamine-beta-hydroxylase (DBH), and of the cholinergic system, i.e. choline acetyltransferase and acetylcholinesterase, were studied. Biochemical differences in tumor groups probably reflected variations in the proportion of neuroepithelial components: trends suggested an increase of cAMP and an increased activity of TPH, AADC, TH and DBH in tumors with increased proportions of neuroepithelial cells. These findings indicate that the neuroepithelial component of the mouse teratoma may serve as a model for the study of neuronal differentiation in primitive neuroepithelial neoplasms.     

Experimental schizencephaly induced by Kilham strain of mumps virus: pathogenesis of cleft formation

The pathogenesis of cleft formation in schizencephaly was analyzed by examining the brain lesions produced by the infection of the Kilham strain of mumps virus during the period of neuronal migration in hamsters. Mumps virus antigen was detected in the neuroepithelial cells within the ventricular zone, the choroid plexus in the lateral ventricles, and vimentin-immunoreactive radial glial fibers. The main pathological findings were cerebral hemorrhage, neuronal necrosis, microsulci on the cerebral cortex and cleft formation through the entire thickness of the cerebral mantle. The clefts seen in these experiments were lined by embryonal elements such as neuroepithelial cells and germinal cells. Based on these results and the original definition by Yakovlev and Wadsworth, the following two conclusions were suggested. First, the mumps virus localized to the neuroepithelial cells within the ventricular zone and the radial glial fibers may induce a destructive process and subsequent anomalous neuronal migration, resulting in cleft formation. Second, the formation of the ventricular cleft extending to the pial surface, which should be complete before the cortical infolding appears, is necessary in order to produce the characteristic cleft in schizencephaly which is associated with the pial-ependymal seam.     

Structural abnormalities develop in the brain after ablation of the gene encoding nonmuscle myosin II-B heavy chain

Ablation of nonmuscle myosin heavy chain II-B (NMHC-B) in mice results in severe hydrocephalus with enlargement of the lateral and third ventricles. All B(-)/B(-) mice died either during embryonic development or on the day of birth (PO). Neurons cultured from superior cervical ganglia of B(-)/B(-) mice between embryonic day (E) 18 and P0 showed decreased rates of neurite outgrowth, and their growth cones had a distinctive narrow morphology compared with those from normal mice. Serial sections of E12.5, E13.5, and E15 mouse brains identified developmental defects in the ventricular neuroepithelium. On E12.5, disruption of the coherent ventricular surface and disordered cell migration of neuroepithelial and differentiated cells were seen at various points in the ventricular walls. These abnormalities resulted in the formation of rosettes in various regions of the brain and spinal cord. On E13.5 and E15, disruption of the ventricular surface and aberrant protrusions of neural cells into the ventricles became more prominent. By E18.5 and P0, the defects in cells lining the ventricular wall resulted in an obstructive hydrocephalus due to stenosis or occlusion of the third ventricle and cerebral aqueduct. These defects may be caused by abnormalities in the cell adhesive properties of neuroepithelial cells and suggest that NMHC-B is essential for both early and late developmental processes in the mammalian brain.     

Immunohistochemical recognition of human neuroepithelial tumors by anti-Leu 7 (HNK-1) monoclonal antibody

Summary The immunoreactivity of the anti-Leu 7 (HNK-1) monoclonal antibody, a marker for natural killer cells, was evaluated with the peroxidase-antiperoxidase (PAP) method on sections of human paraffin-embedded tissues from 135 tumors of the central nervous system and five esthesioneuroblastomas. As shown independently by others, the antibody was found to react with most types of neoplastic neuroepithelial cells. Our findings indicate that the reaction is most often localized on the cytoplasmic membranes. The immunoreactive cell membranes were generally those of well-differentiated tumor cells and of neoplastic cells found in tumors that usually were not embryonal in nature. Parallel immunostaining either of the same or of successive sections with an anti-glial fibrillary acidic protein serum was of considerable assistance in discriminating between different immunoreactive cells, e.g., between astrocytes and cells presumed to be oligodendrocytes.Despite its cross-recognition of cells of various histogenesis, the anti-Leu 7 monoclonal antibody can, in well-defined circumstances, elucidate specific differential diagnostic problems involving neurogenic neoplasms that cannot be resolved with routine staining techniques.Supported by a grant from the Fondation Suisse de Bourses de Médecine et Biologie (EP) and by research grant CA 31271 from the National Cancer Institute, US Department of Health and Human Services (LJR)     

Spinal cord-notochord relationship in normal human embryos and in a human embryo with double spinal cord

Spinal cord-notochord relationship was analyzed histologically and immunohistochemically in normal human conceptuses between the 4–8 developmental weeks and in an 8-week embryo with double spinal the neural tube along the cranio-caudal body axis was paralleled by the differentiation of the median hindge point cells at the ventral midline of the tube and by its temporary close association with the notochord. During the 5th–8th developmental weeks, the neuroepithelium differentiating into three distinct layers was accompanied by a solid, ventromedially positioned notochord. In the abnormal 8-week embryo, the additional spinal cord was located ventrolaterally from the vertebral column. Both spinal cords appeared bilaterally asymmetric, with their floor and roof plates irregularly formed. An abnormally enhanced pattern of neuroepithelial differentiation characterized their dorsal parts. Furthermore, additional spinal nerves and ganglia and an abnormal bony structure were associated with the spinal cord positioned outside the vertebral column. The underlying vertebral bodies were misshaped and contained scattered supernumerary groups of notochord cells. Our investigation underlines the importance of the notochord-neural tube relationship in the morphogenesis of the spinal cord. We suggest that the double spinal cord was induced by the split notochord.     

Ultrastructural analysis of chromosome translocation-induced neural tube defects

Neural tube closure defects occurred in 33% of the embryos obtained from matings of male mice heterozygous for a reciprocal chromosome translocation (T(2;4)1Sn) with normal female CFLP mice. Light and electron microscopic observations of neuroepithelium and mesenchyme in affected embryos indicated two distinct types of anomalies occurred. The first consisted of neuroepithelial hypertrophy and neural tube closure defects. These defects most frequently affected the midbrain and hindbrain, but occasional defects of the lumbosacral neural tube were also observed. Unlike the highly organized, pseudostratified neuroepithelium in control embryos, neuroepithelial cells became stratified and formed cell islands with secondary lumina within the wall of the neural tube. The second condition was associated with a reduction in neuroepithelial thickness, considerable neuroepithelial and neural crest cell death, basal lamina alterations and premature invasion of the neuroepithelium by subjacent endothelial cells. In both cases, the cephalic mesenchyme cells, rather than their normal stellate appearance, were markedly elongated in shape and reduced in area. The number of cell-cell contacts between mesenchymal cells was also reduced significantly. These results are discussed in light of recent theories regarding the role of mesenchyme and extracellular matrix in neurulation.     

Cellular and subcellular distribution of HNK-1 immunoreactivity in the neural tube of Xenopus

The HNK-1 antigen, a carbohydrate moiety bound to many cell adhesion and recognition molecules, is implicated in cell-cell and cell-substrate interactions during neural development. HNK-1 immunoreactivity (HNK1-IR) appears on neurons of the Xenopus neural tube very early in their development (Nordlander, Devel. Brain Res., 50:147-153, 1989). The distribution and onset of expression of the HNK-1 epitope on and within individual neurons is examined in this study. HNK-1 labels developing neurons and their processes, and focal areas of other structures which are directly contacted by neurons, such as neuroepithelial cell surfaces, basal lamina, and culture surfaces. HNK1-IR first appears in the Golgi apparatus and subsequently on the cell surface and in streams of punctate material directed toward the site of axon initiation and into the developing axon and its growth cone. The entire neuron is coated with a thin (20-30 nm) surface layer of HNK1-IR. In addition, the surface is dotted with small (100-250 nm) boluses of HNK1-IR material. Such boluses also occur within cytoplasmic vesicles, and extracellularly on basal lamina and culture substrata in proximity to neurons or their processes. The subcellar distribution of HNK1-IR in this tissue is compatible with a role for the HNK-1 epitope in axonal outgrowth and guidance. © 1993 Wiley-Liss, Inc.     

Cell surface changes accompanying the neural differentiation of an embryonal carcinoma cell line

The murine embryonal carcinoma cell line P19S18O1A1 develops into neuronlike cells after treatment with retinoic acid (Edwards and McBurney, 1983). We have analyzed the expression of cell surface carbohydrate antigens and intracellular cytoskeletal antigens in differentiating O1A1 cells in order to identify the cell types present in the cultures and to characterize the differentiation process. Undifferentiated O1A1 cells express the SSEA-1 antigen, GD3 ganglioside, and the D1.1 ganglioside antigen, carbohydrate markers that are found on early embryonic cells and neuroepithelial germinal cells in vivo. The cells also bind tetanus toxin, cholera toxin, and monoclonal antibody A2B5, probes that bind to gangliosides found on the surfaces of neurons and immature astrocytes in vivo and in vitro. They contain vimentin-type intermediate filament antigens but have no detectable neurofilament or glial filament protein antigens. After aggregation of the cells in medium containing retinoic acid followed by growth in a serum-free chemically defined medium, over 80% of the cells differentiate into neurons as determined by immunofluorescent labeling with antibodies against neurofilament protein antigens. The differentiated cells no longer express either the embryonic or neuroepithelial carbohydrate antigens, but they continue to express the cell surface markers characteristic of neurons. These changes in the expression of cell surface antigens are accompanied by changes in ganglioside metabolism, including a shift towards the synthesis of more complex gangliosides. Thus, the retinoic acid-induced changes in O1A1 cells in vitro resemble the in vivo development of neurons. This establishes the O1A1 cell line as a relevant model system for studies of the molecular basis of neuronal differentiation and development.