Distribution of intraventricularly injected horseradish peroxidase in cerebrospinal fluid compartments of the rat spinal cord

Summary The circulation of the cerebrospinal fluid along the central canal and its access to the parenchyma of the spinal cord of the rat have been analyzed by injection of horseradish peroxidase (HRP) into the lateral ventricle. Peroxidase was found throughout the central canal 13 min after injection, suggesting a rapid circulation of cerebrospinal fluid along the central canal of the rat spinal cord. It was cleared from the central canal within 2 h, in contrast with the situation in the brain tissue, where it remained in the periventricular areas for 4 h. In the central canal, HRP bound to Reissner's fiber and the luminal surface of the ependymal cells; it penetrated through the intercellular space of the ependymal lining, reached the subependymal neuropil, the basement membrane of local capillaries, and appeared in the lumen of endothelial pinocytotic vesicles. Furthermore, it accumulated in the labyrinths of the basement membrane contacting the basolateral aspect of the ependymal cells. In ependymocytes, HRP was found in single pinocytotic vesicles. The blood vessels supplying the spinal cord were classified into two types. Type-A vessels penetrated the spinal cord laterally and dorsally and displayed the tracer along their external wall as far as the gray matter. Type-B vessels intruded into the spinal cord from the medial ventral sulcus and occupied the anterior commissure of the gray matter, approaching the central canal. They represented the only vessels marked by HRP along their course through the gray matter. HRP spread from the wall of type-B vessels, labeling the labyrinths, the intercellular space of the ependymal lining, and the lumen of the central canal. This suggests a communication between the central canal and the outer cerebrospinal fluid space, at the level of the medial ventral sulcus, via the intercellular spaces, the perivascular basement membrane and its labyrinthine extensions.     

Morphological changes in the central canal of the cat after kaolin injection into the cisterna magna

40 adult cats were made hydrocephalic by intracisternal injection of 200 mg Kaolin. 34 survived between 24 hours to 4 months. In 19 cases a ventriculostomy was carried out, whereby in 13 animals a contrast filling of the central canal occurred. The contrast medium injected into the ventricles entered the external CSF-space in the lumbo-sacral junction of the filum terminale. Light- and electron-microscopic studies showed adaptive structural changes of the central canal epithelium in the early stages. In later stages massive destructions of ependyma and spinal cord parenchyma were found.     

Reissner’s fibre supports the survival of chick cortical neurons in primary mixed cultures

Reissner's fibre, a thread-like structure present in the central canal of the spinal cord, is a product of the condensation of specific glycoproteins that are released by specialized ependymal cells into the cerebrospinal fluid. These secretory ependymocytes constitute the subcommissural organ, a circumventricular organ that lines the roof of the third ventricle of the brain. The subcommissural organ/Reissner's fibre complex is a permanent structure in the vertebrate central nervous system. The addition of bovine Reissner's fibre itself or of soluble material released by Reissner's fibre to primary mixed cultures of chick cerebral cortical cells markedly enhances neuronal survival. The responsive cells have been identified as neurons by labelling them with antibodies to neurofilament proteins. This neuronal survival effect is dose-dependent and does not require the presence of serum in the culture medium. Affinity-purified polyclonal antibodies raised against bovine Reissner's fibre partially block the effect of Reissner's fibre on neuronal survival. These results suggest that Reissner's fibre is involved in developmental processes of the central nervous system.     

Method of prolonged perfusion of the spinal cord central canal in the adult cat in vivo in neurophysiological studies

A method for in-vivo long-lasting perfusion of the central canal of the cat lumbosacral spinal cord with artificial cerebrospinal fluid is described. The method provides for a stable continuous flow of perfusion fluid for many hours. The perfusion adjustment does not entail any injuries to the inferior lumbar and sacral segments of the spinal cord or their roots.     

Noninvasive monitoring of pneumococcal meningitis and evaluation of treatment efficacy in an experimental mouse model

Noninvasive real-time in vivo bioluminescent imaging was used to assess the spread of Streptococcus pneumoniae throughout the spinal cord and brain during the acute stages of bacterial meningitis. A mouse model was established by lumbar (LP) or intracisternal (IC) injection of bioluminescent S. pneumoniae into the subarachnoid space. Bacteria replicated initially at the site of inoculation and spread progressively from the spinal cord to the brain or from the brain down to the cervical part of the spinal column and to the lower vertebral levels. After 24 hr, animals showed strong bioluminescent signals throughout the spinal canal, indicating acute meningitis of the intracranial and intraspinal meninges. A decline in bacterial cell viability, as judged by a reduction in the bioluminescent signal, was observed over time in animals treated with ceftriaxone, but not in untreated groups. Mice treated with the antibiotic survived infection, whereas all mice in untreated groups became moribund, first in the IC group then in the LP group. No untreated animal survived beyond 48 hr after induction of infection. Colony counts of infected cerebrospinal fluid (CSF) correlated positively with bioluminescent signals. This methodology is especially appealing because it allows detecting infected mice as early as 3 hr after inoculation, provide temporal, sequential, and spatial distribution of bacteria within the brain and spinal cord throughout the entire disease process and the rapid monitoring of treatment efficacy in a nondestructive manner. Moreover, it avoids the need to sacrifice the animals for CSF sampling and the potential manipulative damage that can occur with other conventional methods.     

Scrapie I. Transmission and Pathogenesis

Homogenized brain, extracts, and residues of brain from normal and scrapie sheep were inoculated into 116 sheep. Of 72 sheep inoculated with scrapie material 27 developed the disease, whereas four of 44 inoculated with ;normal' brain material showed symptoms similar to those of scrapie. The scrapie agent survived extraction for 18 hours with diethyl ether followed by water for 24 hours. Similarly the agent survived extraction in a Soxhlet apparatus with 95 per cent ethanol containing 2.0 per cent 4 M HC1 for 24 hours. There was no evidence of demyelination in sections of brain and spinal cord supporting the view that allergic encephalomyelitis is not responsible for the scrapie symptoms. Liver function, as measured by the bromsulfalein test, remained normal until just before death. These results preclude that liver dysfunction contributes to the disease but do not exclude the possibilities of other metabolic derangements.     

The spinal cord supports of vertebrae in the crown‐group salamanders (Caudata,Urodela)

The development of spinal cord supports (bony thickenings which extend into the vertebral canal of vertebrae) in primitive (Salamandrella keyserlingii) and derived (Lissotriton vulgaris) salamanders were described. The spinal cord supports develop as the protuberances of periostal bone of the neural arches in the anteroproximal part of the septal collagenous fibers which connect a transverse myoseptum with the notochord and spinal cord, in the septal bundle inside the vertebral canal. Spinal cord supports were also found in some teleostean (Salmo salar, Oncorhynchus mykiss) and dipnoan (Protopterus sp.) fishes. The absence of the spinal cord supports in vertebrates with cartilaginous vertebrae (lampreys, chondrichthyan, and chondrostean fishes) corresponds to the fact that the spinal cord supports are bone structures. The absence of the spinal cord supports in frogs correlates with the lack of the well developed septal bundles inside the vertebral canal. The spinal cord supports are, presumably, a synapomorphic character for salamanders which originated independently of those observed in teleostean and dipnoan fishes. J. Morphol. 2012. © 2012 Wiley Periodicals, Inc.     

Ornithodiplostomum ptychocheilus: migration to the brain of the fish intermediate host, Pimephales promelas.

Migration of cercariae of the diplostomatid trematode, Ornithodiplostomum ptychocheilus, to the brain of the fathead minnow, Pimephales promelas, takes place via directed, nonrandom movement. Penetration of the fish epidermis is rapid and is essentially complete by 2 hr postinfection. Migration to the central nervous system occurs almost exclusively via the general body musculature and connective tissue, although a few cercariae gain direct access to the nervous system via the eyes. Cercariae enter either the neural canal and spinal cord, or the brain via the spinal or cranial nerves and their associated foramina, although cercariae appear to remain in (on) these peripheral nerves for only a short time. Cercariae associated with cranial nerves continue to the brain. Those becoming associated with spinal nerves travel up the neural canal and (or) spinal cord to the brain. Data suggest that most arrive at the brain via the neural canal and spinal cord. Within the brain, most developing metacercariae (neascus-type) occur in the optic lobes and cerebellum. Whether this is “selective localization” or merely the result of the larger space afforded by these brain regions could not be determined.     

Spinal cord central canal of the German shepherd dog: Morphological,histological, and ultrastructural considerations

This study deals with some macroscopical, microscopical, and ultrastructural aspects of the spinal cord central canal of the German shepherd dog. The caudal end of the spinal cord is constituted by the conus medullaris, which may extend to the first sacral vertebra, the terminal ventricle, and the filum terminale. The latter structure is considered as internum (second to third sacral vertebrae) or externum (fifth caudal vertebra), according to its relation to the dura mater. Occasionally, there is a second anchorage which is close to the level of the sixth caudal vertebra. The central canal is surrounded by a ciliated ependymal epithelium, which differs depending upon the levels. The most caudal part of the filum terminale bears a columnar ciliated ependymal epithelium surrounded by two layers of glia and pia mater, which separate the central canal from the subarachnoid space. Microfil injections show a communication between the cavity and the subarachnoid space, as the plastic is able to pass through the ependymal epithelium. At the level of the terminal ventricle there are real separations of the ependymal epithelium, which seem to connect the lumen of the spinal canal with the subarachnoid space. These structures probably constitute one of the drainage pathways of the cerebrospinal fluid. The diameter of the central canal is related to the age of the animal. However, even in very old animals the spinal cord central canal reaches the tip of the filum terminale and remains patent until death. At the ultrastructural level the ependymal cells present villi, located on cytoplasmic projections, cilia, dense mitochondria, and oval nuclei. © 1995 Wiley-Liss, Inc.     

Circulation of marker substances in the cerebrospinal fluid of an amphibian,Rana pipiens

Summary Solutions of fluorescein-labelled dextran or Evans blue-albumin were infused into the lateral cerebral ventricle of Rana pipiens. The subsequent distribution in the cerebrospinal fluid (CSF) was investigated between 2 and 24 h after infusion by freezing and examination of the cut blocks of the head and vertebral column of the stage of a freezing microtome. These marker substances move out of the ventricles into the subarachnoid space at the caudal end of the fourth ventricle and spread rapidly along the subarachnoid space of the spinal cord. The spreading of marker substances is slower into the brain subarachnoid space. When the marker is infused into the subarachnoid space of the forebrain, it becomes distributed throughout the subarachnoid space of the brain and spinal cord but not in the ventricles.Partial clearance of markers from the ventricles takes place within 5 h and total clearance within 8 h. Clearance from the brain and cord subarachnoid space is somewhat slower and can only be detected in experiments lasting 10 h or more. Absorption of the markers from the CSF occurs via the intervertebral foramina of the spinal cord. Fluorescence microscopy of sections of the cord show that the fluorescence leaves the subarachnoid space at the point where the spinal nerves traverse the arachnoid membrane.     

Immunohistochemical properties of the "floor plate glycogen body" of the human embryonic spinal cord and brain stem

Prominent glycogen accumulations have been found in the floor plate radial glial cells of the human spinal cord and brain stem during the 6th to 13th week of intrauterine life. These glycogen-rich cells are totally negative to indirect immunoperoxidase staining with an antibody to glial fibrillary acidic protein, a protein that is strongly expressed in the remaining radial glial cells that border the central canal of the spinal cord. The glycogen-rich floor plate radial glial cells are, on the other hand, heavily stained by a monoclonal antibody against a vimentin-related protein. The neighbouring lateral radial glial cells do not express this protein. These and other distinctive features of the floor plate radial glial cells indicate an organoid specialisation of the floor plate during limited periods of intrauterine life. The function(s) of this specialised tissue remains obscure, but it may be related to cortico-spinal fibres crossing the midline through the floor plate, or to the transport of substances in both directions between blood vessels and the central canal.     

Immunohistochemical evidence for cholecystokinin-like peptides in neuronal cell bodies of the rat spinal cord

Summary Cholecystokinin-like immunoreactivity has been demonstrated by radioimmunoassay and immunocytochemistry in the spinal cord of various mammals, in particular in nerve fibers of the superficial layers of the posterior column, but had not been detected in neuronal cell bodies. We report immunohistochemical evidence for the presence of a group of cholecystokinincontaining neuronal cell bodies in the lumbar spinal cord of the rat. This group of cells is only visualized after direct injection of colchicine into the spinal cord and is located near the central canal in the intermedio-medial nucleus of area X of Rexed.     

Immunohistochemical investigation of neuropeptides in the central nervous system of the amphioxus,Branchiostoma belcheri

The immunohistochemical localization of nine different neuropeptides was studied in the central nervous system of the amphioxus, Branchiostoma belcheri. In the brain, perikarya immunoreactive for urotensin I and FMRFamide were localized in the vicinity of the central canal. One of the processes of each of these perikarya was found to cross the dorso ventral slit-like lumen of the central canal. Oxytocin-immunoreactive short fibers, but not perikarya, were detected in the ventral part of the brain. Perikarya immunoreactive for arginine vasopressin/vasotocin, oxytocin and FMRFamide were widely distributed in the spinal cord. Arginine vasopressin/vasotocin-immunoreactive fibers often made contacts with Rohde cell axons. Angiotensin II-immunoreactive perikarya were observed in the posterior half of the spinal cord, and urotensin I-immunoreactive perikarya were found in the caudal region of the spinal cord. Cholecystokinin/gastrin-immunoreactive fibers, but not perikarya, were detected in the spinal cord; some extended as far as the ependymal layer of the cerebral ventricle. No colocalization of the peptides examined was observed. No immunoreactivity for atrial and brain natriuretic peptides nor for urotensin II was detected. The present study indicates that there are at least six separate neuronal systems that contain different peptides, respectively, in the central nervous system of the amphioxus. Their functions remain to be determined.Part of this investigation has previously been presented in abstract form (Uemura et al. 1989)     

The filum terminale of the frog spinal cord

Summary The filum terminale, or terminal portion of the spinal cord, was studied in normal adult frogs (Rana pipiens) by means of light and electron microscopy. Astroglial cells are the predominant elements in this region. The rostral portion of the filum terminale consists mainly of (1) a peripheral dense ring of myelinated and some unmyelinated nerve fibers, and processes of astrocytes terminating at the subpial space; (2) an intermediate zone, in which astrocytes are the main cellular elements in addition to a few degenerated neurons; and (3) a central region where the central canal is lined by dark and light ependymal cells. In the caudal portion of the filum terminale, the amount of neuropil is greatly reduced. This region is formed mainly by astrocytic glial cells and very few neuronal elements. The central canal in the caudal portion is located ventrally and contains a lining consisting almost exclusively of dark ependymal cells.     

Catecholaminergic and serotoninergic brain structures in European green frogs: an autoradiographical study

Aminergic brain structures have been investigated by means of light microscopical autoradiography after injection of the tritiated catecholamines noradrenaline and dopamine and the indoleamine (or tryptamine) serotonin into the brain cavity of frogs of the Rana esculenta complex. These amines are fairly specifically taken up by catecholaminergic and serotoninergic neurons, respectively, which are located in structures like the catecholaminergic preoptic recess organ; the mixed catecholaminergic-serotoninergic paraventricular organ/nucleus infundibularis-complex and nucleus reticularis mesencephali; the telencephalic septal and striatal areas and the tectum opticum, which contain many catecholaminergic axon terminals; the habenular area, which contains serotoninergic axon terminals. The autoradiographical data on the location and the nature of these aminergic brain structures agree well with the mainly fluorescence microscopical and immunocytochemical data from the literature. The autoradiographical detection method can be combined at the light and the electron microscopical level with other histological, histochemical, or immunohistochemical techniques in one and the same preparation, and the results of the different treatments may eventually be made visible simultaneously.     

Reissner's fiber and the wall of the central canal in the lumbo-sacral region of the bovine spinal cord

Summary Reissner's fiber (RF) of the subcommissural organ (SCO), the central canal and its bordering structures, and the filum terminale were investigated in the bovine spinal cord by use of transmission electron microscopy, histochemical methods and light-microscopic immunocytochemistry. The primary antisera were raised against the bovine RF, or the SCO proper. Comparative immunocytochemical studies were also performed on the lumbo-sacral region of the rat, rabbit, dog and pig.At all levels of the bovine spinal cord, RF was strongly immunoreactive with both antisera. From cervical to upper sacral levels of the bovine spinal cord there was an increasing number of ependymal cells immunostainable with both antisera. The free surface of the central canal was covered by a layer of immunoreactive material. At sacral levels small subependymal immunoreactive cells were observed. From all these structures sharing the same immunoreactivity, only RF was stained by the paraldehyde-fuchsin and periodicacid-Schiff methods.At the ultrastructural level, ependymal cells with numerous protrusions extending into the central canal were seen in the lower lumbar segments, whereas cells displaying signs of secretory activity were principally found in the ependyma of the upper sacral levels. A few cerebrospinal fluid-contacting neurons were observed at all levels of the spinal cord; they were immunostained with an anti-tubulin serum.The lumbo-sacral segments of the dog, rat and rabbit, either fixed by vascular perfusion or in the same manner as the bovine material, did not show any immunoreactive structure other than RF.The possibilities that the immunoreactive ependymal cells might play a secretory or an absorptive role, or be the result of post-mortem events, are discussed.Supported by Grant I/38259 from the Stiftung Volkswagenwerk, Federal Republic of Germany, and Grant RS-82-18 from the Dirección de Investigaciones, Universidad Austral de ChileThe authors wish to thank Dr. Enrique Romeny from the Valdivia abattoir for kindly providing the bovine spinal cords     

Reissner's fiber in the central canal of primate spinal cord

The Reissner's fibre (RF) in the canalis centralis of the spinal cord was studied with the aid of transmission and scanning electron microscope in Tupaia, Cebus, Sarmiri, Macaca, Alouatta, Ateles, Callithrix and Callicebus. The RF of the primates is structurally not different from the corresponding structure of other chordates. The diameter of the RF is slightly increasing in the canalis direction. The increase of the diameter varies between 2 and 5 micron. It is assumed that the increase is brought about by the inclusion of cell debris into the fibre. Material which definitely does not belong to the RF is found in the cerebrospinal fluid (CSF) of the canalis centralis and occasionally it adheres to the surface of the fibre. This material appears either as amorphous flocculent substances, the granulation of which differs in both density and size of the granula or as vesicles. If the material makes contact with the RF, a membrane-like structure may appear which is a condensation restricted to the area of the contact between material of different viscosity and granulation. The origin of this variable material is still not clarified. According to our observations, it is probably sequestrated material from ependymal cells. It is not a secretion as sometimes assumed since the intracellular processes of the synthesis of the secretion have not been observed.     

Lentiviral gene delivery to CNS by spinal intrathecal administration to neonatal mice

BACKGROUND: Direct injection of lentivectors into the central nervous system (CNS) mostly results in localized parenchymal transgene expression. Intrathecal gene delivery into the spinal canal may produce a wider dissemination of the transgene and allow diffusion of secreted transgenic proteins throughout the cerebrospinal fluid (CSF). Herein, we analyze the distribution and expression of LacZ and SEAP transgenes following the intrathecal delivery of lentivectors into the spinal canal. METHODS: Four weeks after intrathecal injection into the spinal canal of newborn mice, the expression of the LacZ gene was assessed by histochemical staining and by in situ polymer chain reaction (PCR). Following the spinal infusion of a lentivector carrying the SEAP gene, levels of enzymatically active SEAP were measured in the CSF, blood serum, and in brain extracts. RESULTS: Intrathecal spinal canal delivery of lentivectors to newborn mice resulted in patchy, widely scattered areas of beta-gal expression mostly in the meninges. The transduction of the meningeal cells was confirmed by in situ PCR. Following the spinal infusion of a lentivector carrying the SEAP gene, sustained presence of the reporter protein was detected in the CSF, as well as in blood serum, and brain extracts. CONCLUSIONS: These findings indicate that intrathecal injections of lentivectors can provide significant levels of transgene expression in the meninges. Unlike intracerebral injections of lentivectors, intrathecal gene delivery through the spinal canal appears to produce a wider diffusion of the transgene. This approach is less invasive and may be useful to address those neurological diseases that benefit from the ectopic expression of soluble factors impermeable to the blood-brain barrier.     

The origins of syringomyelia: numerical models of fluid/structure interactions in the spinal cord

A two-dimensional axi-symmetric numerical model is constructed of the spinal cord, consisting of elastic cord tissue surrounded by aqueous cerebrospinal fluid, in turn surrounded by elastic dura. The geometric and elastic parameters are simplified but of realistic order, compared with existing measurements. A distal reflecting site models scar tissue formed by earlier trauma to the cord, which is commonly associated with syrinx formation. Transients equivalent to both arterial pulsation and percussive coughing are used to excite wave propagation. Propagation is investigated in this model and one with a central canal down the middle of the cord tissue, and in further idealized versions of it, including a model with no cord, one with a rigid cord, one with a rigid dura, and a double-length untapered variant of the rigid-dura model. Analytical predictions for axial and radial wave-speeds in these different situations are compared with, and used to explain, the numerical outcomes. We find that the anatomic circumstances of the spinal cerebrospinal fluid cavity probably do not allow for significant wave steepening phenomena. The results indicate that wave propagation in the real cord is set by the elastic properties of both the cord tissue and the confining dura mater, fat, and bone. The central canal does not influence the wave propagation significantly.     

Akhirin regulates the proliferation and differentiation of neural stem cells in intact and injured mouse spinal cord

Although the central nervous system is considered a comparatively static tissue with limited cell turnover, cells with stem cell properties have been isolated from most neural tissues. The spinal cord ependymal cells show neural stem cell potential in vitro and in vivo in injured spinal cord. However, very little is known regarding the ependymal niche in the mouse spinal cord. We previously reported that a secreted factor, chick Akhirin, is expressed in the ciliary marginal zone of the eye, where it works as a heterophilic cell‐adhesion molecule. Here, we describe a new crucial function for mouse Akhirin (M‐AKH) in regulating the proliferation and differentiation of progenitors in the mouse spinal cord. During embryonic spinal cord development, M‐AKH is transiently expressed in the central canal ependymal cells, which possess latent neural stem cell properties. Targeted inactivation of the AKH gene in mice causes a reduction in the size of the spinal cord and decreases BrdU incorporation in the spinal cord. Remarkably, the expression patterns of ependymal niche molecules in AKH knockout (AKH?/?) mice are different from those of AKH+/+, both in vitro and in vivo. Furthermore, we provide evidence that AKH expression in the central canal is rapidly upregulated in the injured spinal cord. Taken together, these results indicate that M‐AKH plays a crucial role in mouse spinal cord formation by regulating the ependymal niche in the central canal. © 2014 Wiley Periodicals, Inc. Develop Neurobiol 75: 494–504, 2015