Cellular mechanisms involved in the stenosis and obliteration of the cerebral aqueduct of hyh mutant mice developing congenital hydrocephalus

Two phases may be recognized in the development of congenital hydrocephalus in the hyh mutant mouse. During embryonic life the detachment of the ventral ependyma is followed by a moderate hydrocephalus. During the first postnatal week the cerebral aqueduct becomes obliterated and a severe hydrocephalus develops. The aim of the present investigation was to elucidate the cellular phenomena occurring at the site of aqueduct obliteration and the probable participation of the subcommissural organ in this process. Electron microscopy, immunocytochemistry, and lectin histochemistry were used to investigate the aqueduct of normal and hydrocephalic hyh mice from embryonic day 14 (E-14) to postnatal day 7 (PN-7). In the normal hyh mouse, the aqueduct is an irregularly shaped cavity with 3 distinct regions (rostral, middle, and caudal) lined by various types of ependyma. In the hydrocephalic mouse, these 3 regions behave differently; the rostral end becomes stenosed, the middle third dilates, and the caudal end obliterates. The findings indicate that the following sequence of events lead to hydrocephalus: 1) denudation of the ventral ependyma (embryonic life); 2) denudation of dorsal ependyma and failure of the subcommissural organ to form Reissner fiber (first postnatal week); 3) obliteration of distal end of aqueduct; and 4) severe hydrocephalus. No evidence was obtained that NCAM is involved in the detachment of ependymal cells. The process of ependymal denudation would involve alterations of the surface sialoglycoproteins of the ependymal cells and the interaction of the latter with macrophages.     

A programmed ependymal denudation precedes congenital hydrocephalus in the hyh mutant mouse.

Hydrocephalic hyh mice are born with moderate hydrocephalus and a normal cerebral aqueduct. At about the fifth postnatal day the aqueduct becomes obliterated and severe hydrocephalus develops. The aim of the present investigation was to investigate the mechanism of this hydrocephalus, probably starting during fetal life when the cerebral aqueduct is still patent. By use of immunocytochemistry and scanning electron microscopy, mutant (n = 54) and normal (n = 61) hyh mouse embryos were studied at various developmental stages to trace the earliest microscopic changes occurring in the brains of embryos becoming hydrocephalic. The primary defect begins at an early developmental stage (E-12) and involves cells lining the brain cavities, which detach following a well-defined temporo-spatial pattern. This ependymal denudation mostly involves the ependyma of the basal plate derivatives. There is a relationship between ependymal denudation and ependymal differentiation evaluated by the expression of vimentin and glial fibrillary acidic protein. The ependymal cells had a normal appearance before and after detachment, suggesting that their separation from the ventricular wall might be due to abnormalities in cell adhesion molecules. The process of detachment of the ventral ependyma, clearly visualized under scanning electron microscope, is almost completed before the onset of hydrocephalus. Furthermore, this ependymal denudation does not lead to aqueductal stenosis during prenatal life. Thus, the rather massive ependymal denudation appears to be the trigger of hydrocephalus in this mutant mouse, raising the question about the mechanism responsible for this hydrocephalus. It seems likely that an uncontrolled bulk flow of brain fluid through the extended areas devoid of ependyma may be responsible for the hydrocephalus developed by the hyh mutant embryos. The defect in these embryos also includes loss of the hindbrain floor plate and a delayed in the expression of Reissner fiber glycoproteins by the subcommissural organ.     

Patterned neuropathologic events occurring in hyh congenital hydrocephalic mutant mice

Hyh mutant mice develop long-lasting hydrocephalus and represent a good model for investigating neuropathologic events associated with hydrocephalus. The study of their brains by use of lectin binding, bromodeoxyuridine labeling, immunochemistry, and scanning electron microscopy revealed that certain events related to hydrocephalus followed a well-defined pattern. A program of neuroepithelium/ependyma denudation was initiated at embryonic day 12 and terminated at the end of the second postnatal week. After the third postnatal week the denuded areas remained permanently devoid of ependyma. In contrast, a selective group of ependymal areas resisted denudation throughout the lifespan. Ependymal denudation triggered neighboring astrocytes to proliferate. These astrocytes expressed particular glial markers and formed a superficial cell layer replacing the lost ependyma. The loss of the neuroepithelium/ependyma layer at specific regions of the ventricular walls and at specific stages of brain development would explain the fact that only certain brain structures had abnormal development. Therefore, commissural axons forming the corpus callosum and the hippocampal commissure displayed abnormalities, whereas those forming the anterior and posterior commissures did not; and the brain cortex was not homogenously affected, with the cingular and frontal cortices being the most altered regions. All of these telencephalic alterations developed at stages when hydrocephalus was not yet patent at the lateral ventricles, indicating that abnormal neural development and hydrocephalus are linked at the etiologic level, rather than the former being a consequence of the latter. All evidence collected on hydrocephalic hyh mutant mice indicates that a primary alteration in the neuroepithelium/ependyma cell lineage triggers both hydrocephalus and abnormalities in telencephalic development.     

EXPERIMENTAL OBSTRUCTIVE HYDROCEPHALUS IN THE RAT: A SCANNING ELECTRON MICROSCOPIC STUDY

Hydrocephalus was induced in 12-day old rats by the cisternal infusion of a concentrated kaolin suspension. The animals were killed at day 20 and the ependymal lining of all the ventricles prepared for scanning electron microscopy. The dilation of the ventricles was moderate to gross in all cases. The ependyma of the lateral ventricles was similar in both control and experimental animals. Ependymal damage was present in six out of the twelve hydrocephalic rats. Two had fibres visible on the ependymal surface. Four had tears covered with small round cells, believed to be responsible for the repair of the ependyma. The third ventricle, cerebral aqueduct and fourth ventricle enlarged by incorporating folds of ependyma, present in control animals, into the ventricular walls. The circumventricular organs present in the third and fourth ventricles were not damaged by the dilation of the ventricles, even in severe hydrocephalus.     

INHERITED PRENATAL HYDROCEPHALUS IN THE H–Tx RAT: A MORPHOLOGICAL STUDY

The H-Tx rat has inherited hydrocephalus which is present at birth. In order to investigate the onset and early stages of hydrocephalus, the heads of fetuses from 16 to 21 days gestation and at 1 day after birth, were serially sectioned using conventional wax histology. Lateral and third ventricle volumes were measured with a graphics tablet and microcomputer. Hydrocephalus was first detected at 18-20 days gestation by enlarged lateral ventricles and it was sometimes accompanied by a large third ventricle. Most hydrocephalics had a non-patent cerebral aqueduct between the third ventricle and the posterior collicular recess and the remainder (about 25%) had an aqueduct which was patent but with a smaller lumen than in non-hydrocephalic littermates. Some fetuses prior to 18 days gestation with normal lateral ventricles also had non-patent aqueducts. Abnormal aqueducts were lined by ependymal cells which were ventrally displaced by thickening of the overlying midbrain; also the subcommissural organ was foreshortened. Infusion with fluorescent markers confirmed that the flow pathway through the aqueduct was obstructed in many hydrocephalic rats. It is concluded that the hydrocephalus may be the result of abnormal brain development in the midline region of the dorsal mesencephalon, leading to aqueduct closure.     

Immunohistochemical localization of superoxide dismutase in congenital hydrocephalic rat brain

The effects of active oxygen species in the development of congenital hydrocephalus have been investigated. Superoxide dismutase (SOD) is one of the scavengers of active oxygen species and there have been many recent reports on the relationship between neurological disorders by active oxygen species following reperfusion for ischemic brain and SOD. In this study, the localization of Cu-SOD and Zn-SOD in WIC-Hyd congenitally hydrocephalic rat brains was identified by the enzyme unlabeled antibody method. We examined the localization of SOD in the choroid plexus, hippocampus, and ependymal cells of the lateral ventricle and aqueduct of WIC-Hyd rats. SOD was hardly observed in the choroid plexus and faintly localized in the hippocampus and ependymal cells of the congenitally hydrocephalic brain, but was observed equally in the cytoplasm of the choroid plexus, hippocampus, and ependymal cells in control animals. In the hippocampus, less SOD was found in hydrocephalic rats than in controls. The SOD was slightly observed in the CA1 pyramidal cells in hydrocephalic rats. In the lateral ventricle and aqueductal ependyma, less SOD was found in hydrocephalic than in control rats. The amount of Cu, Zn-SOD in the congenitally hydrocephalic rat brain was less than in the control, especially in the choroid plexus. Therefore, we suspect that the production of SOD is congenitally reduced in the congenitally hydrocephalic rat brain, and this may promote the impairment of the function of choroid plexus and cilia due to increased active oxygen species. The reduction of SOD in the choroid plexus, hippocampus and ependymal cells of ventricles or aqueduct may promote the development of hydrocephalus in the congenitally hydrocephalic rat.     

Neuroblast proliferation on the surface of the adult rat striatal wall after focal ependymal loss by intracerebroventricular injection of neuraminidase

The subventricular zone of the striatal wall of adult rodents is an active neurogenic region for life. Cubic multiciliated ependyma separates the subventricular zone from the cerebrospinal fluid (CSF) and is involved in the control of adult neurogenesis. By injecting neuraminidase from Clostridium perfringens into the right lateral ventricle of the rat, we provoked a partial detachment of the ependyma in the striatal wall. The contralateral ventricle was never affected and was used as the experimental control. Neuraminidase caused widening of the intercellular spaces among some ependymal cells and their subsequent detachment and disintegration in the CSF. Partial ependymal denudation was followed by infiltration of the CSF with macrophages and neutrophils from the local choroid plexus, which ependymal cells never detached after neuraminidase administration. Inflammation extended toward the periventricular parenchyma. The ependymal cells that did not detach and remained in the ventricle wall never proliferated. The lost ependyma was never recovered, and ependymal cells never behaved as neural stem cells. Instead, a scar formed by overlapping astrocytic processes sealed those regions devoid of ependyma. Some ependymal cells at the border of the denudated areas lost contact with the ventricle and became located under the glial layer. Concomitantly with scar formation, some subependymal cells protruded toward the ventricle through the ependymal breaks, proliferated, and formed clusters of rounded ventricular cells that expressed the phenotype of neuroblasts. Ventricular clusters of neuroblasts remained in the ventricle up to 90 days after injection. In the subventricular zone, adult neurogenesis persisted.     

On the pathology of experimental hydrocephalus.

Acute hydrocephalus was produced in newborn rabbits by injection of kaolin into the cisterna magna. The light microscopic changes which occurred in the ependyma and periventricular brain tissue were studied. Some animals also received intraventricular injection of Evans blue albumin (EBA) at various times after the kaolin injection to study the permeability of the ependyma. There was a progressive dilatation of the lateral ventricles from the second day after the kaolin injection. Marked hydrocephalus was seen after 2 weeks. The white matter of the cerebral hemispheres showed increasing reduction in volume with the degree of hydrocephalus. Neither destruction of brain tissue nor macrophage response or inflammation were seen. The ependyma adapted remarkably well to the increased intraventricular pressure by extensive flattening and stretching. No convincing breaks or ruptures were seen. There was a patchy spongy zone beneath the ependyma, probably indicating oedema of the periventricular white matter due to transventricular absorption of the cerebrospinal fluid (CSF). Denudement of the ependymal lining is not necessary for the concept of transventricular flow of CSF. No difference was seen in the penetration of EBA into the periventricular tissue between hydrocephalic and control animals. The reduction of the cerebral mantle thickness was probably caused by simple pressure atrophy. This indicates that the morphological changes may to a large extent be reversible if the hydrocephalus is properly treated within reasonable time. The role of morphological changes in the pathophysiology of hydrocephalus if briefly commented upon in relation to certain aspects of human hydrocephalus.     

Ultrastructure and movement of the ependymal and tracheal cilia in congenitally hydrocephalic WIC-Hyd rats

The aim of the present investigation is to determine whether or not hydrocephalus occurring in hydrocephalic Wistar-Imamichi strain rats (WIC-Hyd) is caused by functional and structural disorders of ependymal cilia. Ultrastructures and movement of cilia in the ependyma of the lateral, III and IV ventricles and aqueduct of Sylvius and in the trachea walls of the animals were examined by means of scanning electron microscopy (SEM), transmission electron microscopy (TEM), and light microscopy using a phase-contrast microscope equipped with a high-speed video recording system. SEM revealed that a marked decrease in the length and number of cilia in the ependymal and tracheal walls occurred in affected male WIC-Hyd. This finding was noted even before the development of ventricular dilatation and was not related to the degree of ventricular enlargement after development of hydrocephalus. A moderate decrease in length and number of cilia was also seen among the normal ciliary tufts in affected female rats which developed a mild degree of hydrocephalus. TEM cilia findings included abnormal axonemal structures such as a lack of dynein arms and displacement of microtubules. The incidence of these ultrastructural abnormalities was found to be greater in affected male rats than in affected female rats. All cilia in affected male rats before and after development of hydrocephalus were immotile. A variety of movement disorders such as immobile, rotatory, and vibratory cilia were observed beside normally beating cilia (motile cilia) in affected female rats which never developed hydrocephalus as severe as that seen in affected male rats. These results seem to indicate that there is a correlation between cilia movement disorder and the degree of ultrastructural abnormalities. Consequently, hydrocephalus developing in affected male and female WIC-Hyd appears to be caused by a motility disorder of ependymal cilia which is part of the primary ciliary dyskinesia (PCD) affecting these animals. The present study appears to indicate that the movement of ependymal cilia may play a role in cerebrospinal fluid circulation, and that dysfunction of ependymal ciliary movement may contribute to development of hydrocephalus in WIC-Hyd rats.     

Overexpression of nestin and vimentin in the ependyma of spinal cords from hydrocephalic infants

The ependyma of the spinal central canal in cases of hydrocephalus shows abnormalities which vary with the aetiology of ventricular dilatation. To determine whether these ependymal changes are developmental or reactive in nature, immunohistochemical findings were compared between nine normal controls and 12 cases of hydrocephalus (three each of congenital aqueductal stenosis, Dandy‐Walker malformation, Chiari type II malformation, and post‐haemorrhagic hydrocephalus) using antisera to nestin, vimentin and glial fibrillary acidic protein. The main pathological findings were disruption of ependymal layer, apparent pseudostratification of ependyma, expansion, cleft or syrinx formation in relation to the central canal, and ependymal rosette formation. In normal developing fetal spinal cord, nestin and vimentin were expressed mainly in pseudostratified ependymal cells and radial fibres in the median septum. In cases with congenital hydrocephalus (congenital aqueductal stenosis, Dandy‐Walker malformation, and Chiari type II malformation), nestin was overexpressed in immature ependymal cells, and strong vimentin immunoreactivity was detected in the long tract of radial fibres in the median septum. Nestin and vimentin were also expressed in small cells and their fibres which covered areas denuded of ependymal cells in cases of Chiari type II malformation and post‐haemorrhagic hydrocephalus. Two conclusions are suggested by this report. First, the ependyma of the spinal central canal in congenital hydrocephalus shows a delay in maturation of radial glial cells into mature astrocytes and ependymal cells. Second, areas of ependymal denudation may be repaired by the immature glial cells derived from subependymal cells.     

THE PATHOLOGY OF EXPERIMENTAL OBSTRUCTIVE HYDROCEPHALUS

Acute hydrocephalus was produced in newborn rabbits by the injection of kaolin into the cisterna magna. The light microscopic changes which occurred in the ependyma and periventricular brain tissue were studied. Some animals also received intraventricular injection of Evans blue albumin (EBA) at various times after the kaolin injection to study the permeability of the ependymal lining. There was a progressive dilatation of the lateral ventricles from the second day after the kaolin injection. Marked hydrocephalus was seen after 2 weeks. The white matter of the cerebral hemispheres showed increasing reduction in volume with the degree of hydrocephalus. No destruction of brain tissue, macrophage response, or inflammation were seen. The ependyma adapted remarkably well to the increased intraventricular pressure by extensive flattening and stretching. Apart from occasional and inconstant ruptures at the sites of the ventricular coarctations, no convincing breaks or defects of the ependymal lining were seen. There was a patchy spongy zone beneath the ependyma, possibly indicating oedema of the periventricular white matter due to transventricular movement of cerebrospinal fluid (CSF). Denudation of the ependymal lining is not necessary for the concept of transventricular flow of CSF. No difference was seen in the penetration of EBA into the periventricular tissue between hydrocephalic and control animals. The reduction of the cerebral mantle thickness was probably caused by simple pressure atrophy. This indicates that the changes may to a large extent be reversible if the hydrocephalus is treated within reasonable time.     

Experimental hydrocephalus

Summary SD-JCL rats were injected with 25 mg/kg of ethylnitrosourea on day 9 1/2 of gestation. Fetuses were excised near term and examined under dissecting microscope. 78% of viable fetuses were found to be hydrocephalic. On the histologic study of the serial sections of the brain, a stenosis of the Sylvian aqueduct without subependymal gliosis, edematous choroid plexus with engorged vessels, scanty connective tissue and swollen epithelium, thinning of the telencephalic wall with poorly developed granular layers and the neuroblasts remaining in the ependymal zone, were the characteristic findings in the hydrocephalic fetuses.The natural course of the treated young rats was studied. 75% of the newborn rats showed some clinical evidence of hydrocephalus. Severely affected rats became progressively lethargic and/or paralytic, and died usually before the end of 4th week. Protein content of the cerebrospinal fluid was significantly increased. Subdural, subarachnoid and intraventricular hemorrhages were observed and suspected to be the cause of death.On the histological examination of hydrocephalic youngs, flattening of the ependymal cells and engorgement of the cortical vessels were observed as early as in 2-week-old youngs. In more advanced stages, there was noted the thinning of the periventricular white matter, especially around the occipital horn. The thin pallium showed vacuolization of the perineural tissue, enlargement of the Virchow-Robin space and the status spongiosus-like degeneration. Various types of ependymal proliferation was seen.The present study represents the first postnatal observation of the transplacentally induced congenital hydrocephalus in the rat. The present technique proved to be a simple and reproducible model of producing a congenital hydrocephalus in the rat.Supported in part by a grant from the Ministry of Education (1970–1971).     

Overexpression of nestin and vimentin in ependymal cells in hydrocephalus

In order to elucidate the immunohistochemical features of hydrocephalic ependyma, immunohistochemical examination was undertaken in 11 normal, post-mortem brains (age range, 11 weeks’ postconception to 6 months after birth) and 12 hydrocephalic brains (three cases each of congenital aqueductal stenosis, Dandy-Walker malformation, Arnold-Chiari type II malformation and posthemorrhagic hydrocephalus) by using antisera to nestin, vimentin and glial fibrillary acidic protein (GFAP). In normal brains, nestin was predominantly expressed in neuroepithelial cells and radial glial fibers during the period of neuronal migration. Vimentin immunoreactivity was principally detected in immature ependymal cells and their basal fibers after the period of neuronal migration, then partly replaced by GFAP reactivity during late gestation. In hydrocephalus, the areas of ependymal disruption were covered with nestin- or vimentin-positive cells. Nestin and vimentin were also expressed in immature ependymal cells or their basal processes in anatomical regions such as the roof or floor plate of the fourth ventricle or the cerebral aqueduct, and the ventral part of the third ventricle. These results suggest that the overexpression of nestin and vimentin in hydrocephalus follows two patterns: a reactive pattern of proliferating immature glial cells associated with ependymal cell loss and an abnormal developmental pattern of immunopositivity associated with anatomical regions in the midline mesencephalon. Received: 27 November 1995 / Accepted: 29 December 1995     

Hydrocephalus following prenatal methylmercury poisoning

Summary Prenatal methylmercury poisoning of C57BL/6J mice was followed by the development of communicating hydrocephalus in 15% to 25% of surviving offspring. Although examination of the serially sectioned cerebral aqueduct in hydrocephalic animals revealed the presence of stenosis, complete occlusion of the lumen was not observed. The ependymal epithelium of the cerebral aqueduct was preserved, and there was no evidence of periaqueductal inflammation or reactive gliosis. Edema and vacuolar change were, however, observed subependymally. The cerebral white matter, which bore the brunt of the degenerative changes seen in hydrocephalic brains, showed edema, spongy degeneration, gross cystic change and loss of parenchyma. In addition, ependymal cells and choroid plexus epithelial cells in Hg-treated animals contained large amounts of mercury within their cytoplasm, and it is possible that this may have contributed to the development of hydrocephalus by causing disturbances of CSF homeostasis. We believe that the appearance of aqueductal stenosis in Hg-intoxicated animals represents the result rather than the cause of the hydrocephalus.Supported in part by USPHS grant no. ES 02928     

The effects of hydrocephalus upon the developing brain. Histological and quantitative studies of the ependyma and subependyma in hydrocephalic rats

A suspension of kaolin was injected into the cisterna magna of 44 rats at 2 weeks of age. Animals killed at intervals from 5--19 weeks of age showed varying degrees of hydrocephalus. Light microscopy, scanning and transmission electron microscopy revealed stretching and flattening of the ependymal cells but no significant loss of cilia. Histological evidence of periventricular tissue damage in these chronically hydrocephalic animals was only present when the ventricular dilatation was extensive. A quantitative assessment was made of the ependymal and subependymal cell reactions around the lateral ventricles of the hydrocephalic animals. Although the ependymal cells were clearly stretched around the ventricles, there was no apparent proliferation of these cells. An increase in the total number of subependymal cells was observed in hydrocephalic animals when compared with a series of 39 aged-matched controls. The greatest proliferation was in the dorsal and lateral walls of the ventricles which were the regions most severely stretched by the ventricular dilatation. There is evidence that subependymal cells differentiate into astrocytes and microglia so that proliferation of these cells may be interpreted as a response to continuing and progressive brain damage in chronic hydrocephalus. Such progressive tissue damage could adversely affect the developing brain.     

The pathology of experimental obstructive hydrocephalus

Summary Obstructive hydrocephalus was produced in 10–14 day-old rabbits by injection of kaolin into the cisterna magna and the ependyma and subependymal tissue was studied by electron microscopy. Generally, the study confirmed recent light microscopic observations on similar models (Torvik et al., 1976). In contrast to most previous reports,it was found that the ependyma adapted remarkably well to ventricular dilatation. No true ependymal defects occurred even in extensive hydrocephalus except at the sites of the ventricular synechiae which sometimes ruptured. The specialized ependymal junctions remained intact but outside the junctions the intercellular clefts were widened, particularly along the lateral wall of the lateral ventricle. The density of the microvilli and cilia decreased, probably because of the increase in the surface area of the ependyma. Dense bundles of filaments developed in the ependymal cells of the hydrocephalic animals.The extracellular space of the subependymal white matter appeared increased but there was no evidence of destruction of fibres or cells. Thus, the reduction of the cerebral mantle thickness was probably mainly caused by pressure atrophy.     

Ultrastructural observations in experimental hydrocephalus in the rabbit

Changes in the ependyma and periventricular brain tissues of the lateral, 3rd and 4th ventricles and the cervical spinal canal were studied electron-microscopically in young rabbits on the 9th day after injecting kaolin into the cisterna magna. The ependyma of the lateral ventricle overlying the white matter was notably stretched causing increased egress of CSF and disorganisation of the normal architecture of the white matter and capillaries. The neurons and glial cells close to the white matter showed edematous changes. The changes in the ependymal lining and the underlying grey matter were less severe in the dorsal part of the 3rd and the 4th ventricle. The ventral part of the 3rd ventricle was the least affected. The height and the arrangement of the ependymal cells, the surrounding grey matter with narrow interstitial spaces and the absorbing tanycytes seemed to be factors which were responsible for the minimal changes in these regions. The changes appeared to be reversible if the CSF pressure was relieved at this stage. The spinal canal remained unaffected in the majority of our hydrocephalic animals, which could probably be attributed to the type of animal and the degree of hydrocephalus.     

Histological changes in the midbrain around the aqueduct in congenital hydrocephalic rat LEW/Jms

Primary aqueductal stenosis is one of the main causes of congenital hydrocephalus in humans and experimental models. The congenitally hydrocephalic rat strain LEW/Jms is one such model. In this report, we describe further detailed histological features of periaqueductal structure, including the posterior commissure, subcommissural organ (SCO), and ependyma, and discuss the changes in these structures in relation to the cause of hydrocephalus. Coronal sections of the aqueduct in normal rats showed that the usual ependyma was absent in the center of the base facing the dorsal side, which was replaced by tall columnar cells. On the other hand, in hydrocephalic rats the ependyma encircled the aqueductal cavity. In midline sagittal sections, normal and hydrocephalic rats showed the SCO, although the SCO in hydrocephalic rats was shorter than in normal rats. There was also a marked difference between normal and hydrocephalic rats in the dorsoventral dimension of the rostral midbrain. In hydrocephalus, this dimension was large in comparison with normal rats. The superior collicular commissure located caudal to the posterior commissure ran along the ventral side of the midbrain in rats with hydrocephalus, and there was a cell-depleted area just dorsal to the superior collicular commissure. The same findings were observed from the 17th day of gestation until the postnatal period. Although the role of the SCO has been widely discussed from the viewpoint of secretory function, the present study indicated that this organ might be involved in the formation of the shape of the aqueduct.     

New ependymal cells are born postnatally in two discrete regions of the mouse brain and support ventricular enlargement in hydrocephalus

A heterogeneous population of ependymal cells lines the brain ventricles. The evidence about the origin and birth dates of these cell populations is scarce. Furthermore, the possibility that mature ependymal cells are born (ependymogenesis) or self-renewed (ependymal proliferation) postnatally is controversial. The present study was designed to investigate both phenomena in wild-type (wt) and hydrocephalic α-SNAP mutant (hyh) mice at different postnatal stages. In wt mice, proliferating cells in the ventricular zone (VZ) were only found in two distinct regions: the dorsal walls of the third ventricle and Sylvian aqueduct (SA). Most proliferating cells were monociliated and nestin+, likely corresponding to radial glial cells. Postnatal cumulative BrdU-labeling showed that most daughter cells remained in the VZ of both regions and they lost nestin-immunoreactivity. Furthermore, some labeled cells became multiciliated and GLUT-1+, indicating they were ependymal cells born postnatally. Postnatal pulse BrdU-labeling and Ki-67 immunostaining further demonstrated the presence of cycling multiciliated ependymal cells. In hydrocephalic mutants, the dorsal walls of the third ventricle and SA expanded enormously and showed neither ependymal disruption nor ventriculostomies. This phenomenon was sustained by an increased ependymogenesis. Consequently, in addition to the physical and geometrical mechanisms traditionally explaining ventricular enlargement in fetal-onset hydrocephalus, we propose that postnatal ependymogenesis could also play a role. Furthermore, as generation of new ependymal cells during postnatal stages was observed in distinct regions of the ventricular walls, such as the roof of the third ventricle, it may be a key mechanism involved in the development of human type 1 interhemispheric cysts.     

Regional differentiation of the human fetal ependyma: immunocytochemical markers.

The development of the ependyma from 6 weeks (wk) gestation to term was studied in 26 human fetuses and infants for immunocytochemical differentiation using antibodies against vimentin, several cytokeratins, glial fibrillary acidic protein (GFAP) and S-100 protein. Acridine orange-RNA fluorescence was uniform in all differentiated ependymal cells. Marked differences were demonstrated among various anticytokeratin antibodies. Vimentin was demonstrated in undifferentiated cells, particularly during mitosis, and persisted as the ependyma matured. It was strong in floor plate cells and processes forming the ventral median septum. Vimentin and cytokeratin CK-904 coexisted with other immunoreactive proteins but disappeared in a caudorostral gradient with maturation. At 8 wk gestation, GFAP was detected in roof plate cells and their processes forming the dorsal median septum. S-100 protein appeared as early as 6 wk and had a more restricted regional distribution than GFAP at all ages. It was strong in the basal plate ependyma of the spinal cord in young fetuses. The temporal and spatial distributions of the immunoreactive proteins studied correlate with evidence that fetal ependymal cells synthesize compounds that attract or repel axonal growth cones to prevent axons from entering the ventricles or deviating from programmed projection pathways. An additional role may be to induce the transformation of radial glial cells in the subventricular zone.