Transitory cystic cavities in the developing mammalian brain - normal or anomalous?

Transitory cavities associated with the ventricular system represent probably one of the most unique features in the developing mammalian brain. In rodents, the cavities exist transiently in the developing brain and do not appear to be associated with any pathological events. Among the various cavities, the pyramidal-shaped cavum septum pellucidum (CSP) located beneath the corpus callosum and between the lateral ventricles is most well defined. In addition to the CSP are the bilateral subependymal cysts that are consistently associated with the third and fourth ventricles as well as the aqueduct. The cavities/cysts contain a large number of amoeboid microglia expressing surface receptors and hydrolytic enzymes common to tissue macrophages. The significance of these cavities in the developing brain remains a conjecture. Firstly, the cavity walls are free of an apparent epithelial lining; hence, it is speculated that the cavities that appear to communicate with the widened neighboring interstitial tissue spaces may have resulted from physical traction due to the rapid growth of the perinatal brain. Secondly, the cavities contain prominent clusters of amoeboid microglia that may be involved in clearing the debris of degenerating axons and cells resulting from the early brain tissue remodeling. With the increase in brain tissue compactness following the beginning of myelination in the second postnatal week, all cavities are obliterated; concomitantly, the number of amoeboid microglia in them diminishes and all this might signal further maturation of the brain.

[Effect of single or combined application of UDP-glucose, GDNF and memantine on improvement of white matter injury in neonatal rats assessed with light and electron microscopy pathologically].

OBJECTIVE: To evaluate pathologically the effect of the single or combined application of UDP-glucose, GDNF and memantine on the improvement of white matter injury in neonatal rats with periventricular leukomalacia (PVL) under light and electron microscopy. METHODS: A five-day-old neonatal rat model for PVL was established by ligation of the lateral common carotid artery following 120-minute hypoxia. Rats were randomly divided into six groups (30 rats in each group): sham-operated, PVL, UDP-glucose (UDP-glucose 2000 mg/kg intraperitoneally after PVL), GDNF (GDNF 100 µg/kg intracerebrally after PVL), tmemantine (memantine 20 mg/kg intraperitoneally after PVL), and a combination administration of three drugs (UDP-glucose, GDNF and memantine). The rats were sacrificed 7 or 21 days after PVL for assessment of pathological changes in the white matter under both light and electron microscopy. The number and thickness of the myelin sheath in the white matter were measured under electron microscopy, and both of pathological grading and scoring were undertaken under light microscopy. RESULTS: There was rare and sparse myelinogenesis with a loose arrangement of nerve fibers in the white matter under electron microscopy in the PVL group at 7 and 21 days after PVL. The number and thickness of the myelin sheath in the PVL group were significantly less than in the sham-operated, UDP-glucose, GDNF, memantine and combination administration groups (P<0.01). The results of pathological grading of white matter under light microscopy showed that all rats in the PVL group manifested either mild injury (38%-50%) or severe injury (50%-62%) at 7 and 21 days after PVL. The majority of rats (50%-88%) in the four drug administration groups had normal white matter at 7 and 21 days after PVL. The pathological scores at 7 and 21 days after PVL in the PVL group were the highest, and they were significantly higher than in the other five groups (P<0.05). CONCLUSIONS: The single or combined application of UDP-glucose, GDNF and memantine may significantly improve pathological changes in the white matter of rats with PVL. The favorable effect is inferred to be closely correlated with the improvement of brain microenvironment and the enhancement of nerve regeneration promoted by the three drugs.

Molecular heterogeneity in the choroid plexus epithelium: the 22-member γ-protocadherin family is differentially expressed, apically localized, and implicated in CSF regulation.

The choroid plexus (CP) epithelium develops from the ependyma that lines the ventricular system, and plays a critical role in the development and function of the brain. In addition to being the primary site of CSF production, the CP maintains the blood-CSF barrier via apical tight junctions between epithelial cells. Here we show that the 22-member γ-protocadherin (γ-Pcdh) family of cell adhesion molecules, which we have implicated previously in synaptogenesis and neuronal survival, is highly expressed by both CP epithelial and ependymal cells, in which γ-Pcdh protein localization is, surprisingly, tightly restricted to the apical membrane. Multi-label immunostaining demonstrates that γ-Pcdhs are excluded from tight junctions, basolateral adherens junctions, and apical cilia tufts. RT-PCR analysis indicates that, as a whole, the CP expresses most members of the Pcdh-γ gene family. Immunostaining using novel monoclonal antibodies specific for single γ-Pcdh proteins shows that individual epithelial cells differ in their apically localized γ-Pcdh repertoire. Restricted mutation of the Pcdh-γ locus in the choroid plexus and ependyma leads to significant reductions in ventricular volume, without obvious disruptions of epithelial apical-basal polarity. Together, these results suggest an unsuspected role for the γ-Pcdhs in CSF production and demonstrate a surprising molecular heterogeneity in the CP epithelium.

Sustained activation of mTOR pathway in embryonic neural stem cells leads to development of tuberous sclerosis complex-associated lesions.

Tuberous Sclerosis Complex (TSC) is a multisystem genetic disorder characterized by hamartomatous neurological lesions that exhibit abnormal cell proliferation and differentiation. Hyperactivation of mTOR pathway by mutations in either the Tsc1 or Tsc2 gene underlies TSC pathogenesis, but involvement of specific neural cell populations in the formation of TSC-associated neurological lesions remains unclear. We deleted Tsc1 in Emx1-expressing embryonic telencephalic neural stem cells (NSCs) and found that mutant mice faithfully recapitulated TSC neuropathological lesions, such as cortical lamination defects and subependymal nodules (SENs). These alterations were caused by enhanced generation of SVZ neural progeny, followed by their premature differentiation and impaired maturation during both embryonic and postnatal development. Notably, mTORC1-dependent Akt inhibition and STAT3 activation were involved in the reduced self-renewal and earlier neuronal and astroglial differentiation of mutant NSCs. Thus, finely tuned mTOR activation in embryonic NSCs may be critical to prevent development of TSC-associated brain lesions.

Proliferation in the human ipsilateral subventricular zone after ischemic stroke.

BACKGROUND: It is uncertain whether neurogenesis occurs in humans after stroke. We studied the morphologic changes that occurred in the subventricular zone (SVZ) in patients who died following an acute ischemic stroke. METHODS: We examined coronal brain slices from patients who died after a first-ever cerebral nonlacunar infarction in the middle cerebral artery territory. We evaluated the morphologic changes in the ipsilateral and contralateral SVZ by light and electron microscopy. Using immunochemistry with Ki-67 and PCNA, we detected cell proliferation. We used Tuj-1 for immature neurons and PSA-NCAM for migrating cells. RESULTS: The study included 7 patients with a mean age of 82 +/- 5 (mean +/- SD) years; 4 were men. They died a mean of 10 +/- 5 days after the ischemic stroke. Brain samples were obtained a mean of 4 +/- 2 hours after death. In comparison with the contralateral SVZ, the following changes were observed in the ipsilateral SVZ: an increase in the width of the gap and ribbon layers, as well as in the cell density of the ribbon layer, an enlargement of the cytoplasmic volume of astrocytes, and an increase of Ki-67-positive cells. In the ipsilateral SVZ, mitoses and cells that stained for either Tuj-1 or PSA-NCAM markers were observed more frequently than in the contralateral SVZ. CONCLUSION: We found unequivocal evidence of active cell proliferation in the ipsilateral subventricular zone following an acute ischemic stroke in our patients.

A comparison of epithelial and neural properties in progenitor cells derived from the adult human ciliary body and brain.

Cells isolated from the ciliary body (CB) of the adult human eye possess properties of retinal stem/progenitor cells and can be propagated as spheres in culture. As these cells are isolated from a non-neural epithelium which has neuroepithelial origin, they may have both epithelial and neural lineages. Since it is the properties of neural progenitor cells that are sought after in a future scenario of autotransplantation, we wanted to directly compare human CB spheres with neurospheres derived from the human subventricular zone (SVZ), which is the best characterized neural stem cell niche in the CNS of adults. The CB epithelium was dissected from donor eyes (n = 8). Biopsies from the ventricular wall were harvested during neurosurgery due to epilepsy (n = 7). CB and SVZ tissue were also isolated from Brown Norwegian rats. Dissociated single cells were cultivated in a sphere-promoting medium and passaged every 10-30 days. Fixed spheres were studied by immunohistochemistry, quantitative RT-PCR and scanning/transmission electron microscopy. We found that both CB and SVZ spheres contained a mixed population of cells embedded in extracellular matrix. CB spheres, in contrast to SVZ neurospheres, contained pigmented cells with epithelial morphology that stained for cytokeratins (3/12 + 19), were connected through desmosomes and tight-junctions and produced PEDF. Markers of neural progenitors (nestin, Sox-2, GFAP) were significantly lower expressed in human CB compared to SVZ spheres, and nestin positive cells in the CB spheres also contained pigment. There was higher expression of EGF and TGF-beta receptors in human CB spheres, and a comparative greater activation of the canonical Wnt pathway. These results indicate that adult human CB spheres contain progenitor cells with epithelial properties and limited expression of neural progenitor markers compared to CNS neurospheres. Further studies mapping the regulation between epithelial and neural properties in the adult human CB spheres are vital to fully utilize them as a clinical source of retinal progenitor cells in the future.

Myxopapillary ependymoma of the lateral ventricle with local recurrences: histopathological and ultrastructural analysis of a case.

An exceptional case of a recurrent intracranial ependymoma of myxopapillary type arising from the lateral ventricle is reported in a 37-year-old man. This distinctive morphological variant of ependymoma is virtually restricted to the region of cauda equina and filum terminale or occasionally to pre- or post-sacral soft tissue. The intracranial cases of myxopapillary ependymoma are extremely rare and are generally associated with the primary ependymal tumour at the typical lumbosacral site. This case of intraventricular myxopapillary ependymoma did not demonstrate any MRI evidence of a primary spinal cord tumour. Moreover, the initial diagnosis of this histologically benign tumour was followed by two tumour recurrences during the three-year follow-up period. To our knowledge, this is the third documented case of a primary intraventricular myxopapillary ependymoma and the first one of intracranial localisation associated with local recurrences.

Anatomy of the brain neurogenic zones revisited: fractones and the fibroblast/macrophage network.

Cytogenesis in adult peripheral organs, and in all organs during development, occurs nearby basal laminae (BL) overlying connective tissue. Paradoxically, cytogenesis in the adult brain occurs primarily in the subependymal layer (SEL), a zone where no particular organization of BL and connective tissue has been described. We have reinvestigated the anatomy of the area considered the most neurogenic in the adult brain, the SEL of the lateral ventricle, in zones adjacent to the caudate putamen, corpus callosum, and lateral septal nucleus. Here, we report structural (confocal microscopy using laminin as a marker) and ultrastructural evidence for highly organized extravascular BL, unique to the SEL. The extravascular BL, termed fractones because of their fractal organization, were regularly arranged along the SEL and consisted of stems terminating in bulbs immediately underneath the ependyma. Fractones contacted local blood vessels by means of their stems. An individual fractone engulfed in its folds numerous processes of astrocytes, ependymocytes, microglial cells, and precursor cell types. The attachment site (base) of stems to blood vessels was extensively folded, overlying large perivascular macrophages that belong to a fibroblast/macrophage network coursing in the perivascular layer and through the meninges. In addition, collagen-1, which is associated with BL and growth factors during developmental morphogenetic inductions, was immunodetected in the SEL and particularly regionalized within fractones. Because macrophages and fibroblasts produce cytokines and growth factors that may concentrate in and exert their effect from the BL, we suggest that the structure described is implicated in adult neurogenesis, gliogenesis, and angiogenesis.

[Nucleolar apparatus of neuroepithelial cells and organization of the ventricular zone in human neocortical anlage]. / Iadryshkovyi apparat neiroépitelial'nykh kletok i organizatsiia ventrikuliarnoi zony v neokortikal'noi zakladke cheloveka.

Using silver nitrate impregnation nucleolar apparatus was studied in cells of ventricular zone of human neocortex in embryos at 6-13 wks of development. The number of nucleoli in cells grew gradually in the direction to ventricular surface in all cases studied. These data indicate characteristic localization of cell cycle phases in the neuroepithelium and correspond to the results of experiments performed in animals using DNA-labelled precursors. Specific cell population with scanty large nucleoli was found in inner portion of ventricular zone in 6-8 wks old embryos where mitotic figures are usually localized. Some hypotheses on these cells' origin and role are discussed.

Ultrastructural changes of the choroid plexus of the lateral ventricles of rats, rattus norvegicus, submitted to experimental chronic alcoholism, followed by alcohol withdrawal

Cuarenta ratas machos adultas se dividieron en dos grupos de 20 animales cada uno (grupos control y experimentales). El grupo experimental recibió alcohol de caña de azúcar comercial, marca "51 Pirassununga", diluida a 30º g/L como dieta líquida por un período de 240 días y agua potable por los siguientes 60 días. Los animales se sacrificaron después de 60, 120, 180, 240 y 300 días de tratamiento. Se observaron varios cambios morfológicos en el grupo experimental de animales de hasta 240 días de tratamiento, encontrándose dilatación de las cisternas del retículo endoplasmático rugoso, espacios intercelulares aumentados, espacios entre las interdigitaciones basolaterales de las células, inflamación del tejido conjuntivo y presencia de una gran cantidad de células pinocíticas, de vacuolas digestivas y de vesículas con contenido electrolúcido a lo largo del citoplasma. Después de 300 días de tratamiento, el grupo experimental de animales presentó una regresión importante de los cambios descritos. No se observó cambios en el grupo de animales control

Cellular composition and three-dimensional organization of the subventricular germinal zone in the adult mammalian brain.

The adult mammalian subventricular zone (SVZ) contains stem cells that give rise to neurons and glia. In vivo, SVZ progeny migrate 3-8 mm to the olfactory bulb, where they form neurons. We show here that the SVZ of the lateral wall of the lateral ventricles in adult mice is composed of neuroblasts, glial cells, and a novel putative precursor cell. The topographical organization of these cells suggests how neurogenesis and migration are integrated in this region. Type A cells had the ultrastructure of migrating neuronal precursors. These cells were arranged as chains parallel to the walls of the ventricle and were polysialylated neural adhesion cell molecule- (PSA-NCAM), TuJ1- (beta-tubulin), and nestin-positive but GFAP- and vimentin-negative. Chains of Type A cells were ensheathed by two ultrastructurally distinct astrocytes (Type B1 and B2) that were GFAP-, vimentin-, and nestin-positive but PSA-NCAM- and TuJ1-negative. Type A and B2 (but not B1) cells incorporated [3H]thymidine. The most actively dividing cell in the SVZ corresponded to Type C cells, which had immature ultrastructural characteristics and were nestin-positive but negative to the other markers. Type C cells formed focal clusters closely associated with chains of Type A cells. Whereas Type C cells were present throughout the SVZ, they were not found in the rostral migratory stream that links the SVZ with the olfactory bulb. These results suggest that chains of migrating neuroblasts in the SVZ may be derived from Type C cells. Our results provide a topographical model for the adult SVZ and should serve as a basis for the in vivo identification of stem cells in the adult mammalian brain.

Ultrastructural characteristics of central neurocytoma in cell culture.

The study was performed to determine the ultrastructural characteristics of central neurocytoma and its features in primary cell culture. Fresh tissue from three tumors was mechanically and enzymatically dissociated into individual cells, which were cultured onto poly-L-lysine precoated Aclar coverslips in the media. The tumor cells attached to the surface of the coverslips within 12 to 24 h and delicate cytoplasmic processes developed within 2 to 3 days. Electron microscopic examination of the cultured tumor cells and the tumor tissue supported neuronal origin. Combined tissue culture and electron microscopic study provides a rapid, reliable, and reproducible means for the diagnosis of central neurocytoma.

Histological and ultrastructural analysis of six colloid cysts in children.

Although colloid cysts of the third ventricle are unusual in children, we have recently encountered six examples. Histologically they were lined by cuboidal, pseudostratified or columnar ciliated and mucous-secreting epithelial cells. Two cases showed small microcysts within the fibrovascular stroma surrounding the main cyst. The outermost layer consisted of a glial-ependymal envelope, in keeping with the postulated supraventricular origin of colloid cysts. Scanning electron microscopy showed 10-40% ciliated cells, and no ballooning of non-ciliated cells. Aspiration of cyst contents was performed in three patients, two of whom subsequently required surgical resection 4 months and 8 years after drainage, respectively. In adults colloid cysts may be asymptomatic, whereas in children they have not been documented as incidental findings at autopsy. Two of our six cases died, both before a diagnosis was established. A colloid cyst of the third ventricle must be included in the evaluation of acute neurological deterioration in children, in whom they are more frequently lethal.

Central neurocytoma. A clinicopathological, immunohistochemical and ultrastructural study of 7 cases.

Characterised by distinctive clinicopathological features, the central neurocytoma (CN) is an uncommon and possibly under-recognised primary cerebral neuronal neoplasm. We present clinical and pathological details of seven patients with CN. Histological examination revealed a greater diversity of morphological appearances than is typically described in CN. No anaplastic features were identified. Cellular areas resembling both oligodendroglioma and ependymoma were present in all cases, but each tumour also contained stroma rich areas with hyalinised or aneurysmal vessels. Synaptophysin was expressed by all tumours and probably represents the immunohistochemical marker of choice for identifying CN. Distinguishing ultrastructural features included rounded cell bodies separated by numerous cell processes containing microtubules, pleomorphic neurosecretory granules and occasional synapses. Ki-67 immunostaining revealed a low cell proliferation index in each case. The distinction of CN from other pathological mimics can be reliably made using this multiparametric approach to diagnosis. The generally benign behaviour of CN is confirmed, though there was one patient death in the follow-up period of 10-122 months. Aggressive behaviour in this case was not associated with anaplastic histological features.

Enterogenous cyst in the fourth ventricle--case report.

A 53-year-old male presented with a rare enterogenous cyst in the fourth ventricle associated with repeated ventriculoperitoneal shunt malfunction. Surgical excision of the cyst resolved the shunt problems. Electron microscopy findings of the surgical sample were microvilli covered with electron-dense coating materials, basement membrane, and several intercellular junctional devices, suggesting the cyst was derived from the endodermal structure. The viscous and gelatinous contents of the cyst might be responsible for the shunt malfunction.

Adhesion and fusion of ependyma in rat brain.

Changes of the ependyma of the rat brain from 2 days after birth to 1 year were studied. In neonatal rats, the ependyma of the lateral ventricle extended medially to cover the hippocampus. The ependyma above the hippocampus disappeared with age in two ways: (1) a zipper-like withdrawal of the medial portion of the ventricle towards the lateral direction, and (2) adhesion and fusion of the ependyma in the lateral region. The zipper-like closure and fusion, which resulted in disappearance of the ependyma, took place in almost all rats within 2 or 3 weeks after birth. Initially, ependymal cells retained their cell polarity with regular organization at adhesion sites as a two-cell-layer seam. Subsequently, their polarity became disorganized in the fusion areas with convergence of the two-cell-layer seam into a one-cell-layer seam, followed by disruption. The ciliary bundle of individual ependymal cells became randomly oriented, sometimes in two or more directions. At the sites where fusion had occurred, fragmentary ependymal seams remained detectable (more than 50%) among the neuropil even in adults. These ectopic seams often contained cystic ependymal cells. In the third ventricle, adhesion was observed but fusion was not. The results indicate that adhesion and fusion of the ependyma occur in select areas during brain development, during which ependymal cells lose their cell polarity.

The area postrema and vomiting.

The area postrema (AP) has been implicated as a chemoreceptor trigger zone for vomiting (emesis) for over 40 years. The AP is located on the dorsal surface of the medulla oblongata at the caudal end of the fourth ventricle. It is one of the so-called circumventricular organs that serve as an interface between the brain parenchyma and the cerebrospinal fluid (CSF)-containing ventricles. The AP lacks a specific blood-brain diffusion barrier to large polar molecules (i.e., a "blood-brain barrier") and is thus anatomically positioned to detect emetic toxins in the blood as well as in the CSF. The AP along with the nucleus of the solitary tract (NTS) and the dorsal motor nucleus of the vagus makes up the so-called dorsal vagal complex, which is the major termination site of vagal afferent nerve fibers. Lesions of the AP prevent vomiting in response to most, but not all, emetic drugs. However, the AP is not essential for vomiting induced by motion or by activation of vagal nerve afferents. The role of the AP in radiation-induced vomiting remains controversial. Electrophysiological studies have reported that neurons in the AP increase their firing in response to emetic drugs. Similarly, studies using the 2-deoxyglucose uptake and c-fos expression techniques have shown that the AP is excited by systemic administration of emetic drugs. Activation of the AP probably leads to nausea and vomiting through its projection to the neighboring NTS. The NTS may serve as the beginning of a final common pathway by which different emetic inputs trigger vomiting.

Electron-microscopic investigations of vasoactive intestinal peptide (VIP)-like immunoreactive terminal formations in the lateral septum of the pigeon.

Vasoactive intestinal peptide (VIP)-like immunoreactive terminal fields were examined in the lateral septum of the pigeon by means of immunocytochemistry. According to light-microscopic observations, these projections originated from VIP-like immunoreactive cerebrospinal fluid (CSF)-contacting neurons, which are located in the ependymal layer of the lateral septum and form a part of the lateral septal organ. The processes of these cells gave rise to dense terminal-like structures in the lateral septum. Pre-embedding immuno-electron microscopy revealed that VIP-like immunoreactive axon terminals had synaptoid contacts with perikarya of small VIP-immunonegative neurons of the lateral septum, which were characterized by an invaginated nucleus, numerous mitochondria, a well-developed Golgi apparatus, endoplasmic reticulum and a small number of dense-core vesicles (about 100 nm in diameter). VIP-like immunoreactive axons were also seen in contact with immunonegative dendrites in the lateral septum. In both axosomatic and axodendritic connections, VIP-like immunoreactive presynaptic terminals contained large dense-core vesicles, clusters of small vesicles and mitochondria. These findings suggest that VIP-immunoreactive neurons of the lateral septal organ project to small, presumably peptidergic nerve cells of the lateral septum and that the VIP-like neuropeptide serves as a neuromodulator (-transmitter) in this area.

Repercussion of craniopharyngioma on the morphology of the rostral wall of the III ventricle. A combined TEM and SEM study.

Using a combined scanning and electron microscope technique, the repercussion on the rostral wall of the III ventricle in a patient affected with a retrochiasmatic craniopharyngioma with growth in the ventricular direction were analysed. The apical surface of the ependymocytes was found to be free of cilia, although there were numerous microvilli. The most striking finding appreciated with the scanning electron microscope was the presence of bulbous protrusions towards the lumen of the ventricle; these were formed of cells with shapes, sizes and surface characteristics different from the rest of the ependyma, which appeared flattened. The ultrastructural study revealed the presence of large numbers of filaments and junction complexes both in the ependymal and subependymal cells. Additionally, the protruded zones corresponded to areas showing different degrees of cellular disorganization.