The collicular recess organ: Evidence for structural and secretory specialization of the ventricular lining in the collicular recess

Summary The collicular recess organ and adjacent portions of the collicular recess were studied by light microscopy, scanning electron microscopy and transmission electron microscopy. In the collicular recess, the ventricular wall contains folds and is well vascularized. The adluminal ependymal cells generally bear kinocilia and microvilli on their ventricular surface. Among the cilia, many secretory droplets, some axons, and few supraependymal cells are seen. Various stages of apocrine ependymosecretion are observed. In addition to tanycytes, coelocytes are found scattered throughout the ependymal lining of the collicular recess. Coelocytes, characterized by lumina containing cilia and a few microvilli, are accumulated in ependymal and hypependymal positions of the collicular recess organ at the roof of the collicular recess.Supported by PHS grants NS 09914 and T32 CA09156. We thank Dean Wyrick and John McNeill, Jr., for their technical assistanceNRSA Postdoctoral Trainee     

Ultrastructural analysis of the human cerebral ventricular system

Summary The ultrastructural organization of the human fetal choroid plexus was assessed with scanning electron microscopy. The membranous modifications of choroidal ependymal cells differ remarkably between 11 and 20 weeks of intrauterine development and suggest a variable functional capacity at different times of ontogenesis. Based upon existing data coupled with the ultra-architectural organization of cilia, clavate and linear microvilli are seen with scanning electron microscopy, a multiple functional role is hypothesized for choroidal ependymal cells.supported by USPH grant NS 08171.career development awardee K04 GM 70001     

Ependyma and ependymal protrusions of the lateral ventricles of the rabbit brain

Summary The ependymal lining of the lateral ventricles of the rabbit brain was studied by means of scanning (SEM) and transmission electron microscopy (TEM). There exist cells devoid of cilia in the anterior horn over the region of the caudate nucleus, in the inferior horn over the hippocampus and on the opposite side over cortical regions. On the surface of some of these ependymal cells, accumulations of cytoplasmic folds and globules can be found. They bulge at different height over the ependymal cells. Clots of these cell particles are tied off from the cell, coming to lie as globules either on or between the cilia of the ependyma. TEM reveals that these tissue protrusions are cell debris consisting of different sized vesicles, cell organelles, tubuli and filaments. They originate from the ependymal layer but may reach down to subependymal cells. Multivesicular protrusions into the ventricular lumen are also observed. Possible causes of these protrusions are discussed; they are likely to be related to the age of the animals.On the ependyma of the caudate nucleus cilia, microvilli, microblebs and supraependymal neuronal cell processes are distributed unevenly over the surface. Within regions where cilia predominate there are cells which are tightly covered with microvilli. A certain direction of the course of the supraependymal neuronal fibers could not be found.The author is pleased to acknowledge useful discussions with Prof. Dr. med. E. van der Zypen. This study was partly supported by the Stanley Thomas Johnson Foundation     

The ependymal surface of the fourth ventricle of the rat: a combined scanning and transmission electron microscopic study.

The morphological features of the ependymal surface and supraependymal elements of the fourth ventricle of the rat were examined by scanning electron microscopy (SEM) and by the transmission electron microscopy (TEM). The results confirm the following aspects: 1) The presence of supraependymal elements and microvilli in the ependymal territories, including the sites where the cilia completely cover the ependymal surface; 2) The existence of cilia with oval or spherical thickenings together with supraependymal bulbs similar in size to those of the larger ciliary swellings; 3) Identification of the long supraependymal fibres with intermittent fusiform dilations observed under the SEM with the nerve fibres seen under the TEM; 4) The existence of intraventricular axodendritic synapses.     

The ventricular surface of the area postrema and the adjacent area subpostrema in the rabbit brain]

The ependymal surface of the area postrema (rabitt) was examined by scanning and transmission electron microscopy. The flattened ependymal cells show few microvilli. Towards the central canal, the ependymal cells change gradually to a columnar shape; the number of microvilli increases concomitantly. The area postrema ependymal cell surface mostly bears a single cilia. In contrast, a region immediately adjacent to the area postrema, which has been named area subpostrema (Gwyn and Wolstencroft 1968), shows cilia arranged in bunches. These cilia are regularly covered with colloid -- like droplets. A period-acid-bisulfit-aldehydthionine method (Specht 1970) permits to identify these droplets with glyproteids.it has been suggested that the droplets might derive from the area subpostrema ependymal cells. Above the ependymal surface of the area postrema, a great number of fine unmyelinated neuronal processes and thicker processes are observed. Some of them show bulb-like endings. These terminals contain small vesicles, dense cored vesicles (400...800 A), and mitochondria which are mostly characterized by a single central prismatic tubule. The plasmalemma of some bulbs is in a synaptic contact with the apical plasmalemma of the ependyma, while other bulbs see to end freely in the ventricle. Some neuronal processes penetrate between ependymal cells of the area postrema into the ventricular lumen.     

The structure of the hypothalamic inferior lobes of the blacktip reef shark: Scanning and transmission electron microscopic observations

The inferior lobes of the shark hypothalamus were examined with light, transmission and scanning electron microscopy. The cells bordering the floor of the lateral recess appear to be typical liquor-contacting neurons. With scanning electron microscopy (SEM) the apical ends of these cells are seen to bulge into the ventricular lumen. In contrast, the roof is lined by a more typical ependymal cell characterized by numerous cilia and microvilli. In addition, SEM reveals several kinds of supraependymal cells with processes that appear to penetrate the ventricular lining. A periventricular nucleus underlies the ependymal cells. Neurons of the periventricular nucleus contain numerous lipofuchsin granules. The rest of the inferior lobe consists of many neuronal fibers. The morphology of the hypothalamic inferior lobe is discussed in relation to its possible role in feeding and aggressive behavior in both elasmobranchs and teleosts.     

Dysfunctional cilia lead to altered ependyma and choroid plexus function, and result in the formation of hydrocephalus

Cilia are complex organelles involved in sensory perception and fluid or cell movement. They are constructed through a highly conserved process called intraflagellar transport (IFT). Mutations in IFT genes, such as Tg737, result in severe developmental defects and disease. In the case of the Tg737orpk mutants, these pathological alterations include cystic kidney disease, biliary and pancreatic duct abnormalities, skeletal patterning defects, and hydrocephalus. Here, we explore the connection between cilia dysfunction and the development of hydrocephalus by using the Tg737orpk mutants. Our analysis indicates that cilia on cells of the brain ventricles of Tg737orpk mutant mice are severely malformed. On the ependymal cells, these defects lead to disorganized beating and impaired cerebrospinal fluid (CSF) movement. However, the loss of the cilia beat and CSF flow is not the initiating factor, as the pathology is present prior to the development of motile cilia on these cells and CSF flow is not impaired at early stages of the disease. Rather, our results suggest that loss of cilia leads to altered function of the choroid plexus epithelium, as evidenced by elevated intracellular cAMP levels and increased chloride concentration in the CSF. These data suggest that cilia function is necessary for regulating ion transport and CSF production, as well as for CSF flow through the ventricles.     

Ultrastructural Study of the Filum Terminate and Caudal Ampulla of the Spinal Cord of Amphioxus (Branchiostoma lanceolatum Pallas)

The filum terminale and caudal ampulla of amphioxus were studied by electron microscopy. The filum terminale consists of ependymal cells whose cilia are directed caudally. Remarkably, nerve fibres course through the filum terminale and caudal ampulla and end on the basal lamina forming neuro-connective structures. Moreover, these nerve boutons are divisible into several classes according to their vesicle content. Boutons containing large dense-cored vesicles are very similar in appearance to the neurosecretory terminals found in the caudal spinal cord of some vertebrates. These observations on nerve fibres suggest that a primitive neurosecretory system similar to the fish urophysis is present in the amphioxus.     

A scanning electron microscopic survey of the brain ventricular system of the female armadillo

Summary The scanning electron microscope was used to survey the brain ventricular system of the female armadillo (Dasypus novemcinctus) with emphasis on the third ventricle. The walls of the lateral ventricles, aqueduct, and fourth ventricle are covered by long cilia. In the lateral ventricle, the cilia are arranged in groups; but in the aqueduct and fourth ventricle, they are evenly placed over the cellular surfaces. The ependymal cells of the third ventricle are densely ciliated except for the organum vasculosum and infundibular recess. The non-ciliated luminal surface of these areas has a pebblestone appearance punctuated by numerous microvilli and two types of supraependymal cells.Supported by Edward G. Schlieder Foundation GrantThe authors would like to thank Jacqueline Skaggs for her secretarial assistance and Garbis Kerimian for his photographic work     

Scanning electron microscopy of the wall of the third ventricle in the brain of Rana temporaria. Part IV.

Summary The surface specializations of the wall of the third cerebral ventricle of Rana temporaria were investigated with the scanning electron microscope. These specializations can be divided into three types: cilia, large bulbous protrusions, and microvillus-like protrusions.Most parts of the ventricular surface are densely ciliated. In contrast, other regions are either scantily ciliated or devoid of cilia. Four areas of the ventricular surface are studded with numerous large bulbous protrusions. These large protrusions can be divided into two types: One type consists of intraventricular end bulbs of dendrites of secretory neurons. The other type is represented by large cytoplasmic extensions of ependymal cells.In the third ventricle of Rana, microvillus-like surface specializations of ependymal cells are ubiquitous structures. Generally, filiform protrusions of varying length are the predominant type. The microvillus-like specializations are transient structures, the number of which varies according to different physiological states of the ependymal cells.This investigation was supported by a grant from the Belgian Nationaal Fonds voor Geneeskundig Wetenschappelijk Onderzoek     

Fine structure of the ependymal cells in the area postrema of the domestic fowl

Summary The ultrastructure of the ependymal cells in the area postrema of the domestic fowl was studied by scanning and transmission electron microscopy. The ependymal surface of the area postrema is covered with many furrows and ridges. These ridges consist of ependymal cells aggregated in a fan-like shape. The ependymal cell lacks clustered cilia, microvilli are few, and a long basal process extends through the parenchymal layer of the area postrema. Within the cytoplasm as well as in the basal process, a spherical body with a diameter ranging from 1.5 to 2 gmm is occasionally observed.This work was supported by a Scientific Research Grant, No. 144017, from the Ministry of Education of Japan to Professor M. YasudaThe authors are grateful to Drs. T. Fujioka and T. Watanabe for their valuable advice     

Scanning electron microscopic analysis of the linings of the fourth ventricle in the domestic fowl

Summary Surface features of the ependymal linings of the fourth ventricle in the fowl were analyzed employing the scanning electron microscope (SEM). On the floor of the median sulcus, each ependymal cell has a solitary cilium, whereas on both sides of the sulcus, cilia are so densely distributed that the details of the underlying cell surface are usually obscured. On the roof of the fourth ventricle, except for the surface of the ciliated groove where numerous cilia are present, the ependymal cells are polygonal in shape, and the center of each cell possesses an aggregate of ten to twenty cilia. Cell surfaces of the choroid tela are entirely covered with delicate microvilli and possess clumped cilia. The ependymal cell surfaces of the area postrema are dome-like in shape. Each ependymal cell has a solitary cilium and shows a smooth surface free of microvilli.This work was supported by a Scientific Research Grant, No. 144017, from the Ministry of Education of Japan to Professor M. Yasuda     

Scanning Electron Microscope Observations on Some Life History Stages of Carchesium polypinum (Ciliata,Peritrichia)1

SYNOPSIS. Sessile zooids, and mobile telotrochs and microgamonts of Carchesium polypinum (Protozoa, Ciliata, Peritrichia), were examined by scanning electron microscopy. The results were compared to earlier light and electron microscope studies in order to investigate structural changes concerned with adaptation and differentiation. Telotrochs and microgamonts always had a contracted peristome and usually had a long phalange of cilia. Striae around the contracted buccal apparatus in all 3 stages were convoluted and often had thickened margins; those in telotrochs and microgamonts had oral-aboral ectoplasmic cross-connections. Nonbuccal striae of telotrochs and microgamonts varied in structure and height differences between epiplasmic peaks and alveoli surface membranes. The number of striae were constant in all 3 stages. Pellicular pore structure did not vary in any of the stages examined and resembled parasomal sacs located near buccal structures. Fully relaxed sessile zooids had ectoplasmic ridges coursing from polykinety kinetosomes and cilia to an area in front of the ciliated portion of the haplokinety; these ridges were interpreted to be the interkinetal fibers. Telotroch bands of sessile zooids consisted of 2 or 3 parallel ectoplasmic ridges which circled the aboral region and contained structures resembling pores. Telotroch bands in telotrochs and microgamonts had 2 enlarged, parallel ectoplasmic ridges circling the aboral region; telotroch band cilia were found between these ridges. In addition, a fold-like, ectoplasmic structure extended beyond the 2 ridges and was located between the telotroch band cilia and the aboral ridge. The epiplasmic shelf surrounding the stalk in sessile zooids was enlarged in telotrochs, and cilia were seen in the scopula depression. No scopula organelle was seen in any microgamont.     

Antennal Sensory Hairs in Talitrid Amphipods (Crustacea)

Sensory hairs from antennulae and antennae of the two species Orchestia platensis Krøyer and Talorchestia deshayesii Audouin were studied by means of light microscopy and scanning and transmission electron microscopy. The complicated cuticular structures of the hair wall are described and figured. The central hair cavity opens in a rather wide terminal pore. Dendritic cilia from sensory neurons with perikarya inside the antennae run the whole length of the hair and terminate in the pore opening. The distal parts of the cilia contain osmiophilic material. It is concluded on morphological grounds that the hairs are chemosensory organs.     

Planar polarity of ependymal cilia

Ependymal cells, epithelial cells that line the cerebral ventricles of the adult brain in various animals, extend multiple motile cilia from their apical surface into the ventricles. These cilia move rapidly, beating in a direction determined by the ependymal planar cell polarity (PCP). Ciliary dysfunction interferes with cerebrospinal fluid circulation and alters neuronal migration. In this review, we summarize recent studies on the cellular and molecular mechanisms underlying two distinct types of ependymal PCP. Ciliary beating in the direction of fluid flow is established by a combination of hydrodynamic forces and intracellular planar polarity signaling. The ciliary basal bodies' anterior position on the apical surface of the cell is determined in the embryonic radial glial cells, inherited by ependymal cells, and established by non-muscle myosin II in early postnatal development.     

The ependyma of goat. II. Ependymal lining of the lateral ventricles, mesencephalic canal, and fourth ventricle: scanning electron microscopic study

The ependymal surface of the lateral cerebral ventricle, mesencephalic canal, and IVth ventricle of 13 male and female goats were studied by means of scanning electron microscopy. The lateral cerebral ventricle was covered mostly with thin cilia that might have club-like terminations on nucleus caudatus, the wide "naked" places with bleb-like protrusions occurred in the transition to cornu temporale during anestrous cycle. The ependymal surface of mesencephalic canal of female goats during anestrus was covered with abundant spherical protrusions. No other changes that could be related to the sexual differences were found.     

Ependymal cells along the lateral ventricle express functional P2X7 receptors

Ependymal cells line the cerebral ventricles and are located in an ideal position to detect central nervous system injury and inflammation. The signaling mechanisms of ependymal cells, however, are poorly understood. As extracellular adenosine 5′-triphosphate is elevated in the context of cellular damage, experiments were conducted to determine whether ependymal cells along the mouse subventricular zone (SVZ) express functional purinergic receptors. Using whole-cell patch clamp recording, widespread expression of P2X₇ receptors was detected on ependymal cells based on their antagonist sensitivity profile and absence of response in P2X₇ ^−/− mice. Immunocytochemistry confirmed the expression of P2X₇ receptors, and electron microscopy demonstrated that P2X₇ receptors are expressed on both cilia and microvilli. Ca²⁺ imaging showed that P2X₇ receptors expressed on cilia are indeed functional. As ependymal cells are believed to function as partner cells in the SVZ neurogenic niche, P2X₇ receptors may play a role in neural progenitor response to injury and inflammation.     

The surface fine structure of the bovine subcommissural organ

The bovine subcommissural organ was studied by using scanning electron microscopy. The most prominent findings was the existence of protruded and dilated endings of the ependymal cells. The majority of these cells were ciliated with two or more cilia; only a few unciliated cells were seen. Some pore-like structures were also seen on the surface. From the functional point of view, the most interesting finding was an amorphous heterogeneous material on the subcommissural ependyma. Especially in the caudal part of the organ this material accumulated in abundance. No real filamentous structures such as Reissner's fibre could be seen, however, it was assumed that the heterogeneous material corresponds to this formation. No supraependymal neurones were demonstrated.     

Spinous extensions on ciliary necklaces in ependymal cells

Summary In ependymal cells of the mouse the neck region of all cilia examined by means of transmission electron microscopy exhibited rows of electron-dense spines. These structures correspond to the ciliary necklace reported from freeze-etch studies, a structure presumed to serve as an energy-regulating system in motile cilia. Send offprint requests to: Institute of Neuropathology, Free University Berlin, Hindenburgdamm 30, 1 Berlin 45     

The PDZ Protein Na+/H+ Exchanger Regulatory Factor-1 (NHERF1) Regulates Planar Cell Polarity and Motile Cilia Organization

Directional flow of the cerebrospinal fluid requires coordinated movement of the motile cilia of the ependymal epithelium that lines the cerebral ventricles. Here we report that mice lacking the Na⁺/H⁺ Exchanger Regulatory Factor 1 (NHERF1/Slc9a3r1, also known as EBP50) develop profound communicating hydrocephalus associated with fewer and disorganized ependymal cilia. Knockdown of NHERF1/slc9a3r1 in zebrafish embryos also causes severe hydrocephalus of the hindbrain and impaired ciliogenesis in the otic vesicle. Ultrastructural analysis did not reveal defects in the shape or organization of individual cilia. Similar phenotypes have been described in animals with deficiencies in Wnt signaling and the Planar Cell Polarity (PCP) pathway. We show that NHERF1 binds the PCP core genes Frizzled (Fzd) and Vangl. We further show that NHERF1 assembles a ternary complex with Fzd4 and Vangl2 and promotes translocation of Vangl2 to the plasma membrane, in particular to the apical surface of ependymal cells. Taken together, these results strongly support an important role for NHERF1 in the regulation of PCP signaling and the development of functional motile cilia.