Microscopic focus on ependymal cells of the spinal cord of the one-humped camel (Camelus dromedarius): Histological, immunohistochemical, and transmission microscopic study.

The current study was designed to give a complete microscopic description of the ependymal cells of the one-humped camel (Camelus dromedarius) using histological, immunohistochemical, and transmission microscopic descriptions of the ependymal cells of the fresh 35 spinal cord samples immediately after their slaughtering. In our findings, the central canal of the spinal cord was lined by multilayered stratified cuboidal or columnar ependymal cells. The ependymal cells had an irregular striated border at their free surface. The ependymal cells do not exhibit a basement membrane. The simple oval nucleus was occupied a large part of the cell with spherical mitochondria. The apical surface of the ependymal cells possesses long cilia; each cilium was bounded by an evagination of the luminal plasma membrane. Some ependymal cells had minute finger-like projections on their luminal plasma membrane. In the perinuclear zone of ependymal cells, many cristiform mitochondria, free ribosomes, and Golgi complexes usually occur. Vacuoles with homogenous and clear fluid were observed. The lateral surface of the adjacent ependymal cells exhibits several tight junctions represented by zonulae occludens and adherens. There were many desmosomes between the neighboring ependymal cells. A perinuclear whorl of filaments fills the lateral part of these ependymal cells. The ependymal cells revealed a clear immunohistochemical reaction with proliferating cell nuclear antigen and nestin stain. There were no obvious differences between the different segments of the spinal cord. Our data concluded that the ependymal cells display clear differences in anatomy as well as ultrastructure that may reflect their distinct functional activity.

Autoradiography and inmmunolabeling suggests that lizard blastema contains arginase-positive M2-like macrophages that may support tail regeneration.

BACKGROUND: The regenerating blastema of the tail in the lizard Podarcis muralis contains numerous macrophages among the prevalent mesenchymal cells. Some macrophages are phagocytic but others are devoid of phagosomes suggesting that they have other roles aside phagocytosis. METHODS: The presence of healing macrophages (M2-like) has been tested using autoradiographic, immunohistochemical and ultrastructural studies. RESULTS: Autoradiography shows an uptake of tritiated arginine in sparse cells of the blastema and in the regenerating epidermis. Bioinformatics analysis suggests that epitopes for arginase-1 and -2, recognized by the employed antibody, are present in lizards. Immunofluorescence shows sparse arginase immunopositive macrophages in the blastema and few macrophages also in the apical wound epidermis. The ultrastructural study shows that macrophages contain dense secretory granules, most likely inactive lysosomes, and small cytoplasmic pale vesicles. Some of the small vesicles are arginase-positive while immunolabeling is very diffuse in the macrophage cytoplasm. CONCLUSIONS: The presence of cells incorporating arginine and of arginase 1-positive cells suggests that M2-like macrophages are present among mesenchymal and epidermal cells of the regenerative tail blastema. M2-like macrophages may promote tail regeneration differently from the numerous pro-inflammatory macrophages previously detected in the scarring limb. The presence of M2-like macrophages in addition to hyaluronate, support the hypothesis that the regenerative blastema of the tail in lizards is an immuno-privileged organ where cell proliferation and growth occur without degenerating in a tumorigenic outgrowth.

Regional differences in the ependyma of the optic tectal ventricle of adult zebrafish with structures referring to brain hydrodynamics.

Classical electron microscopic morphological studies provide detailed ultrastructural information, which may lend insights into cellular functions. As a follow-up to our morphological investigation of the adult zebrafish (Danio rerio) optic tectum, in this study, we have analyzed the ependymal structures lining the surfaces of the tectal ventricle: the torus, tegmental surface of the valvula cerebelli and the periventricular gray zone of the optic tectal cortex. We used toluidine blue stained plastic (semithin) sections for light microscopy and scanning electron microscopy. Our morphological findings of gated entrances and/or egresses indicate that, at least in the adult zebrafish brain, there may be a bidirectional direct flow communication between the ventricular cerebrospinal fluid and the parenchymal interstitial fluid.

Labelling of individual ependymal areas in lateral ventricles of human brain: ependymal tables.

Different types of ependymal areas were studied and labelled in the human brain lateral ventricle. Periventricular structures were included in coining the names of the ependymal areas because they represent a basic and stable part of brain nerve structures suitable for the sake of clarity of localization of the ependyma. The labelling of individual ependymal areas was composed from letters: "Lv" (lateral ventricle); "E" (ependymal area) and letters for abbreviations of the closest periventricular structure, e.g. the septum pellucidum is "sp". The labelling for ependymal area over the septum pellucidum is thus "LvE-sp". The studied types of ependymal areas were arranged in so­called ependymal tables for cornu anterius, pars centralis, cornu inferius and cornu posterius of the human lateral ventricle. Labelling of individual ependymal areas allows for better localization and characterisation of these areas in future studies carried out by various methods (e.g. morphological, biological, molecular) and will prevent from using misnomers with different types of ependymal areas in norm as well as in pathology (Tab. 5, Fig. 6, Ref. 22). Text in PDF www.elis.sk.

On the origin of the term ependyma.

The term ependyma is considered as a translation of the expression integumentum ventriculorum cerebri or Überzug der Hirnhöhlen [the lining of the brain ventricles] in German used by the Wenzel brothers (1812). The first documented usage of this term is found in the work of the German anatomist Karl Ernst Bock from the year 1839, but nobody has ever claimed authorship of the word. Formulations such as "so-called" are used in connection with the term, avoiding any reference to a specific originator. The term first started being used in anatomical literature written in German. In its subsequent history, various interpretations of the meaning of the term have emerged, and certain attempts have also been made to change its formal aspect.

[STRUCTURAL ORGANIZATION OF THE PROCESSES OF EPENDYMO- CYTES LINING THE LATERAL VENTRICLES OF THE RAT BRAIN].

The aim of this study was to examine the structural organization of processes of ependymocytes lining the lateral ventricles of the rat brain using vimentin immunocytochemistry and confocal laser microscopy. The study was performed on adult male rats (n = 3). It was found that most typical ependymocytes had basal processes, while 1/3 of these cells had none. Some vimentin-immunopositive tanycyte-like cells with long processes appoaching blood vessels, were found inside the ependymal lining In some typical ependymocytes, cytroskeleton wa s formed by intermediate filaments of mixed type containing both vimentin and glial fibrillary acidic protein.

3D Modeling of the Lateral Ventricles and Histological Characterization of Periventricular Tissue in Humans and Mouse.

The ventricular system carries and circulates cerebral spinal fluid (CSF) and facilitates clearance of solutes and toxins from the brain. The functional units of the ventricles are ciliated epithelial cells termed ependymal cells, which line the ventricles and through ciliary action are capable of generating laminar flow of CSF at the ventricle surface. This monolayer of ependymal cells also provides barrier and filtration functions that promote exchange between brain interstitial fluids (ISF) and circulating CSF. Biochemical changes in the brain are thereby reflected in the composition of the CSF and destruction of the ependyma can disrupt the delicate balance of CSF and ISF exchange. In humans there is a strong correlation between lateral ventricle expansion and aging. Age-associated ventriculomegaly can occur even in the absence of dementia or obstruction of CSF flow. The exact cause and progression of ventriculomegaly is often unknown; however, enlarged ventricles can show regional and, often, extensive loss of ependymal cell coverage with ventricle surface astrogliosis and associated periventricular edema replacing the functional ependymal cell monolayer. Using MRI scans together with postmortem human brain tissue, we describe how to prepare, image and compile 3D renderings of lateral ventricle volumes, calculate lateral ventricle volumes, and characterize periventricular tissue through immunohistochemical analysis of en face lateral ventricle wall tissue preparations. Corresponding analyses of mouse brain tissue are also presented supporting the use of mouse models as a means to evaluate changes to the lateral ventricles and periventricular tissue found in human aging and disease. Together, these protocols allow investigations into the cause and effect of ventriculomegaly and highlight techniques to study ventricular system health and its important barrier and filtration functions within the brain.

The ependymal region of the adult human spinal cord differs from other species and shows ependymoma-like features.

Several laboratories have described the existence of undifferentiated precursor cells that may act like stem cells in the ependyma of the rodent spinal cord. However, there are reports showing that this region is occluded and disassembled in humans after the second decade of life, although this has been largely ignored or interpreted as a post-mortem artefact. To gain insight into the patency, actual structure, and molecular properties of the adult human spinal cord ependymal region, we followed three approaches: (i) with MRI, we estimated the central canal patency in 59 control subjects, 99 patients with traumatic spinal cord injury, and 26 patients with non-traumatic spinal cord injuries. We observed that the central canal is absent from the vast majority of individuals beyond the age of 18 years, gender-independently, throughout the entire length of the spinal cord, both in healthy controls and after injury; (ii) with histology and immunohistochemistry, we describe morphological properties of the non-lesioned ependymal region, which showed the presence of perivascular pseudorosettes, a common feature of ependymoma; and (iii) with laser capture microdissection, followed by TaqMan® low density arrays, we studied the gene expression profile of the ependymal region and found that it is mainly enriched in genes compatible with a low grade or quiescent ependymoma (53 genes); this region is enriched only in 14 genes related to neurogenic niches. In summary, we demonstrate here that the central canal is mainly absent in the adult human spinal cord and is replaced by a structure morphologically and molecularly different from that described for rodents and other primates. The presented data suggest that the ependymal region is more likely to be reminiscent of a low-grade ependymoma. Therefore, a direct translation to adult human patients of an eventual therapeutic potential of this region based on animal models should be approached with caution.

FLAIR images at 7 Tesla MRI highlight the ependyma and the outer layers of the cerebral cortex.

OBJECTIVES: Fluid-attenuated inversion recovery (FLAIR) imaging is an important clinical 'work horse' for brain MRI and has proven to facilitate imaging of both intracortical lesions as well as cortical layers at 7T MRI. A prominent observation on 7T FLAIR images is a hyperintense rim at the cortical surface and around the ventricles. We aimed to clarify the anatomical correlates and underlying contrast mechanisms of this hyperintense rim. MATERIALS AND METHODS: Two experiments with post-mortem human brain tissue were performed. FLAIR and T2-weighted images were obtained at typical in vivo (0.8mm isotropic) and high resolution (0.25mm isotropic). At one location the cortical surface was partly removed, and scanned again. Imaging was followed by histological and immunohistochemical analysis. Additionally, several simulations were performed to evaluate the potential contribution from an artifact due to water diffusion. RESULTS: The hyperintense rim corresponded to the outer - glia rich - layer of the cortex and disappeared upon removal of that layer. At the ventricles, the rim corresponded to the ependymal layer, and was not present at white matter/fluid borders at an artificial cut. The simulations supported the hypothesis that the hyperintense rim reflects the tissue properties in the outer cortical layers (or ependymal layer for the ventricles), and is not merely an artifact, although not all observations were explained by the simulated model of the contrast mechanism. CONCLUSIONS: 7T FLAIR seems to amplify the signal from layers I-III of the cortex and the ependyma around the ventricles. Although diffusion of water from layer I into CSF does contribute to this effect, a long T2 relaxation time constant in layer I, and probably also layer II-III, is most likely the major contributor, since the rim disappears upon removal of that layer. This knowledge can help the interpretation of imaging results in cortical development and in patients with cortical pathology.

Melanin-concentrating hormone regulates beat frequency of ependymal cilia and ventricular volume.

Ependymal cell cilia help move cerebrospinal fluid through the cerebral ventricles, but the regulation of their beat frequency remains unclear. Using in vitro, high-speed video microscopy and in vivo magnetic resonance imaging in mice, we found that the metabolic peptide melanin-concentrating hormone (MCH) positively controlled cilia beat frequency, specifically in the ventral third ventricle, whereas a lack of MCH receptor provoked a ventricular size increase.

The inferior medullary velum: anatomical study and neurosurgical relevance.

OBJECT: Although it is often visualized surgically, details regarding the inferior medullary velum are lacking in the literature. The present study is intended to better elucidate this neuroanatomical structure using microsurgical and immunohistochemical analyses. METHODS: To study the inferior medullary velum, the authors performed microdissection in 15 adult cadavers. Following gross study, specimens were examined histologically. RESULTS: The inferior medullary velum extended from the flocculus to the middle cerebellar peduncle and stretched between the inferior cerebellar peduncle and the nodule and pyramid. The average thickness of the velum was found to be 0.5 mm (range 0.35-0.8 mm) and the average length was found to be 6 mm (range 5.5-7.2 mm). Arterial branches were identified in all specimens that arose from medullary branches of the posterior inferior cerebellar artery and supplied the inferior medullary velum. Histologically and from internal to external, a choroid plexus epithelium as a single cell layer was adjacent to a cuboidal layer of ependymal cells with no visible cilia. The next layer contained scattered glia in single cells or small clusters. The most external layer was composed of flat spindle cells resembling fibroblasts. No neurons of any type were identified. Only rare axons traversed the thin hypocellular zone that disappeared toward the midline. CONCLUSIONS: Based on this cadaveric study, the authors conclude that division of the inferior medullary velum should be relatively harmless as no neuronal cells were identified in this structure, which appears to be a vestigial bridge of tissue between the left and right sides of the cerebellum.

Negative BOLD-fMRI signals in large cerebral veins.

Reductions in blood oxygenation level dependent (BOLD)-functional magnetic resonance imaging (fMRI) signals below baseline levels have been observed under several conditions as negative activation in task-activation studies or anticorrelation in resting-state experiments. Converging evidence suggests that negative BOLD signals (NBSs) can generally be explained by local reductions in neural activity. Here, we report on NBSs that accompany hemodynamic changes in regions devoid of neural tissue. The NBSs were investigated with high-resolution studies of the visual cortex (VC) at 7 T. Task-activation studies were performed to localize a task-positive area in the VC. During rest, robust negative correlation with the task-positive region was observed in focal regions near the ventricles and dispersed throughout the VC. Both positive and NBSs were dependent on behavioral condition. Comparison with high-resolution structural images showed that negatively correlated regions overlapped with larger pial and ependymal veins near sulcal and ventricular cerebrospinal fluid (CSF). Results from multiecho fMRI showed that NBSs were consistent with increases in local blood volume. These findings confirm theoretical predictions that tie neural activity to blood volume increases, which tend to counteract positive fMRI signal changes associated with increased blood oxygenation. This effect may be more salient in high-resolution studies, in which positive and NBS may be more often spatially distinct.

Constitution of the ependyma in the chicken telencephalon.

The constitution of ependyma derived from the ventricular zone is different from that derived from other regions of the central nervous system. In the mammalian cerebrum, the ependyma is varied by the regions to cortex or basal ganglia (BG). In the avian telencephalon (Tc), previous studies about the constitution of the ependyma have not revealed clear findings. In the present study, we performed immunostaining of ependymal cells in the chicken Tc to confirm differences in the ependyma of various regions. As a result, 4 patterns of ependyma were defined in the outer side of the lateral ventricle. In the base of the lamina pallio-subpallialis (LPS), ependyma consisted of vimentin/glial fibrillary acidic protein (GFAP) double-positive cells, whereas in the base of the lamina frontalis superior, it consisted primarily of vimentin-positive cells and a small number of vimentin/GFAP double-positive cells. With the exception of the above, the pallial ependyma was a single layer containing vimentin single-positive cells. Lastly, the ependyma of the BG was rich in vimentin single-positive cells. The constitutional differences of the ependyma of the pallium and BG concerned differences in ependymal morphology and cell characteristics. These finding suggest that the bounder between pallium and BG is LPS at the point of ependyma.

Postnatal deletion of Numb/Numblike reveals repair and remodeling capacity in the subventricular neurogenic niche.

Neural stem cells are retained in the postnatal subventricular zone (SVZ), a specialized neurogenic niche with unique cytoarchitecture and cell-cell contacts. Although the SVZ stem cells continuously regenerate, how they and the niche respond to local changes is unclear. Here we generated nestin-creER(tm) transgenic mice with inducible Cre recombinase in the SVZ and removed Numb/Numblike, key regulators of embryonic neurogenesis from postnatal SVZ progenitors and ependymal cells. This resulted in severe damage to brain lateral ventricle integrity and identified roles for Numb/Numblike in regulating ependymal wall integrity and SVZ neuroblast survival. Surprisingly, the ventricular damage was eventually repaired: SVZ reconstitution and ventricular wall remodeling were mediated by progenitors that escaped Numb deletion. Our results show a self-repair mechanism in the mammalian brain and may have implications for both niche plasticity in other areas of stem cell biology and the therapeutic use of neural stem cells in neurodegenerative diseases.

Role of androgens and glucocorticoids in the regulation of diazepam-binding inhibitor mRNA levels in male mouse hypothalamus.

In peripheral organs, gonadal and adrenal steroids regulate diazepam-binding inhibitor (DBI) mRNA expression. In order to further investigate the involvement of peripheral steroid hormones in the modulation of brain DBI mRNA expression, we studied by semiquantitative in situ hybridization the effect of adrenalectomy (ADX) and castration (CX) and short-term replacement therapy on DBI mRNA levels in the male mouse hypothalamus. Cells expressing DBI mRNA were mostly observed in the arcuate nucleus, the median eminence and the ependyma bordering the third ventricle. In the median eminence and the ependyma bordering the third ventricule, the DBI gene expression was decreased in ADX rats and a single injection of corticosterone to ADX rats induced a significant increase in DBI gene expression at 3 and 12 h time intervals without completely restoring the basal DBI mRNA expression observed in intact mice. In the arcuate nucleus, ADX and corticosterone administration did not modify DBI mRNA expression. CX down-regulated DBI gene expression in the ependyma bordering the third ventricle. The administration of dihydrotestosterone (3-24 h) completely reversed the inhibitory effect of CX. In the median eminence and arcuate nucleus, neither CX or dihydrotestosterone administration modified DBI mRNA levels. These results suggest that the effects of glucocorticoids on the hypothalamo-pituitary-adrenocortical axis and androgens on the hypothalamo-pituitary-gonadal axis are mediated by DBI.

Microscopic stucture of the lamina terminalis: implications for microsurgical third ventriculostomy.

OBJECTIVE: The neurosurgical approach through the lamina terminalis (LT) is a commonly used technique for management of the third ventricle region pathology. Furthermore, LT fenestration is a recommended procedure during surgery of ruptured intracranial aneurysms. Though the LT is a rudimentary structure in adult human brain, its neurosurgical significance is eliciting increasing interest. The aim of the presented study is to characterize the LT histologically, with special attention to the previously recommended area of LT fenestration and to the localization and structure of the organum vasculosum lamina terminalis (OVLT). METHODS: The study was performed on tissue sampled from eight formalin-fixed brains. Paraffin sections taken from various levels of the LT were routinely stained with hematoxylin and eosin (H&E) and by immunohistochemical methods. RESULTS: The LT in the inferior part bordering the optic recess and immediately above the optic chiasm exhibited paucicellular, mainly fibrillar, glial tissue with scanty neural elements and small vessels. At about halfway along the length of the LT an area of loose structure, with an increased number of glial cells, small neurons and thin-walled vessels corresponding to the OVLT was observed. In the majority of examined cases the OVLT was poorly developed and was therefore sometimes overlooked. The superior segment of the LT near the anterior commissure disclosed again paucicellular and slightly loosened fine fibrillar tissue. CONCLUSIONS: The results of the present microscopic study confirm the opinion that the inferior segment of the LT is the most convenient place for safe incision. Its thinnest middle part immediately above the optic recess is composed mainly of gliotic tissue. Above, prominent loosened tissue and the rather rudimental structure of the OVLT seem to be additional favorable factors for a safe fenestration of the LT.

Anatomy and physiology of the leptomeninges and CSF space.

The arachnoid membrane and pia mater are the two membranous layers that comprise the leptomeninges. Cerebrospinal fluid is made within the ventricular system by cells of the choroid plexus and ependyma. This chapter describes in detail the normal anatomic structure and physiologic interactions of the cerebrospinal fluid and leptomeningeal space that are critical to our understanding and treatment of leptomeningeal metastases.

Synthesis of transthyretin by the ependymal cells of the subcommissural organ.

Transthyretin (TTR) is a protein involved in the transport of thyroid hormones in blood and cerebrospinal fluid (CSF). The only known source of brain-produced TTR is the choroid plexus. In the present investigation, we have identified the subcommissural organ (SCO) as a new source of brain TTR. The SCO is an ependymal gland that secretes glycoproteins into the CSF, where they aggregate to form Reissner's fibre (RF). Evidence exists that the SCO also secretes proteins that remain soluble in the CSF. To investigate the CSF-soluble compounds secreted by the SCO further, antibodies were raised against polypeptides partially purified from fetal bovine CSF. One of these antibodies (against a 14-kDa compound) reacted with secretory granules in cells of fetal and adult bovine SCO, organ-cultured bovine SCO and the choroid plexus of several mammalian species but not with RF. Western blot analyses with this antibody revealed two polypeptides of 14 kDa and 40 kDa in the bovine SCO, in the conditioned medium of SCO explants, and in fetal and adult bovine CSF. Since the monomeric and tetrameric forms of TTR migrate as bands of 14 kDa and 40 kDa by SDS-polyacrylamide gel electrophoresis, a commercial preparation of human TTR was run, with both bands being reactive with this antibody. Bovine SCO was also shown to synthesise mRNA encoding TTR under in vivo and in vitro conditions. We conclude that the SCO synthesises TTR and secretes it into the CSF. Colocalisation studies demonstrated that the SCO possessed two populations of secretory cells, one secreting both RF glycoproteins and TTR and the other secreting only the former. TTR was also detected in the SCO of bovine embryos suggesting that this ependymal gland is an important source of TTR during brain development.

The sensory circumventricular organs of the mammalian brain.

The brain's three sensory circumventricular organs, the subfornical organ, organum vasculosum of the lamina terminalis and the area postrema lack a blood brain barrier and are the only regions in the brain in which neurons are exposed to the chemical environment of the systemic circulation. Therefore they are ideally placed to monitor the changes in osmotic, ionic and hormonal composition of the blood. This book describes their. General structure and relationship to the cerebral ventricles Regional subdivisions Vasculature and barrier properties Neurons, glia and ependymal cells Receptors, neurotransmitters, neuropeptides and enzymes Neuroanatomical connections Functions.

The distribution and morphological characteristics of serotonergic cells in the brain of monotremes.

The distribution and cellular morphology of serotonergic neurons in the brain of two species of monotremes are described. Three clusters of serotonergic neurons were found: a hypothalamic cluster, a cluster in the rostral brainstem and a cluster in the caudal brainstem. Those in the hypothalamus consisted of two groups, the periventricular hypothalamic organ and the infundibular recess, that were intimately associated with the ependymal wall of the third ventricle. Within the rostral brainstem cluster, three distinct divisions were found: the dorsal raphe nucleus (with four subdivisions), the median raphe nucleus and the cells of the supralemniscal region. The dorsal raphe was within and adjacent to the periaqueductal gray matter, the median raphe was associated with the midline ventral to the dorsal raphe, and the cells of the supralemniscal region were in the tegmentum lateral to the median raphe and ventral to the dorsal raphe. The caudal cluster consisted of three divisions: the raphe obscurus nucleus, the raphe pallidus nucleus and the raphe magnus nucleus. The raphe obscurus nucleus was associated with the dorsal midline at the caudal-most part of the medulla oblongata. The raphe pallidus nucleus was found at the ventral midline of the medulla around the inferior olive. Raphe magnus was associated with the midline of the medulla and was found rostral to both the raphe obscurus and raphe pallidus. The results of our study are compared in an evolutionary context with those reported for other mammals and reptiles.