Characteristic features of a nerve primo-vessel suspended in rabbit brain ventricle and central canal.

Bonghan theory was proposed by Bonghan Kim to illustrate the anatomy and physiology of the acupuncture meridian system. One of his astonishing claims was the physical presence of the nerve primo-vessel, which can be involved with a regenerating system of nerves. Our previous work has shown that there is a nerve primo-vessel in brain ventricles and the central canal of the spine of a rabbit. In this study, confocal laser scanning microscopy, transmission electron microscopy, and high voltage electron microscopy demonstrated that a nerve primo-vessel comprised DNA particles, other microparticles, and rod-shaped nuclei encircled by helix-shaped actins. The nerve primo-vessel had acridine orange-stained DNA particles that varied in size and were in parallel. These characteristics of the nerve primo-vessel are crucial for a comprehensive understanding of their function in the central nervous system, which may be associated with nerve regeneration.

Glutamate-immunoreactive neurons and their gonadotropin-releasing hormone-neuronal interactions in the monkey hypothalamus.

Glutamate (Glu) is the most prevalent excitatory neurotransmitter in the brain and has been implicated in the regulation of GnRH secretion in several mammalian species, including the monkey. To investigate the neuroanatomical basis for Glu-GnRH interactions, we performed an immunocytochemical study at both the light and electron microscopic levels on the brains of four female and five male macaques. Initially, we determined the location of Glu-immunoreactive (-ir) elements using a monoclonal antibody specific for glutaraldehyde-fixed Glu (Glu-2) and 3,3'-diaminobenzidine-4-HCl (DAB). Glu-ir was observed in the cytoplasm and to a variable degree in the nuclei of neurons in the diencephalon. Cytoplasmic staining was particularly intense in numerous neurons in the arcuate nucleus, supraoptic nucleus, and many paraventricular nucleus neurons. Short Glu-ir processes were evident in these and other hypothalamic regions and were extremely dense in the infundibular stalk and median eminence. Prior absorption of the Glu-2 antibody with a Glu-glutaraldehyde-BSA conjugate completely abolished all immunostaining in both neuronal nuclei and cytoplasm. Double label Glu-GnRH immunostaining for light microscopy was performed using Glu-2 and DAB without enhancement, and a polyclonal antibody (LR1 or LR2) with silver-enhanced DAB for Glu and GnRH, respectively. Glu-ir interactions with GnRH-ir cell bodies were not apparent, but a few Glu-ir axons seemed to contact GnRH-ir dendrites in the organum vasculosum of the lamina terminalis, medial septum, and arcuate nucleus regions. Reciprocal interactions occurred more frequently, however, in which GnRH-ir axons and dendritic fibers engaged Glu-ir cell bodies en passant, particularly toward the medial and posterior hypothalamus. For ultrastructural analyses, Glu-ir elements were stained with the Glu-2 antibody and 15 nm immunogold or DAB. Electron microscopy demonstrated that Glu-ir was associated with clear microvesicles within the neuronal cytoplasm. Glu-ir processes made classical asymmetrical synapses with one another and received asymmetrical synapses from unlabeled afferents. In sections double labeled for Glu with immunogold and for GnRH with DAB, axo-somatic interactions were not observed. However, axo-dendritic Glu-GnRH synapses were seen, which usually exhibited Glu-ir labeling of terminal vesicles and inconsistent postsynaptic densities, with GnRH-ir neurosecretory granules sometimes congregated in the apposing dendrite or spine. Surprisingly, reverse GnRH-Glu interactions were observed more frequently.(ABSTRACT TRUNCATED AT 400 WORDS)

Cerebral implants of histotypic hypothalamic cultures.

In the present report, the implantation of hypothalamic histotypic cultures within the cerebral hemisphere is described. The cultures were prepared from 10-day-old mice and rotary incubated during 6 or 10 days. When the cells reached their phenotypic characteristics at the end of the incubation period, the cultures were implanted in the posterior area of the lateral ventricle. 30 days postoperatively, the whole implanted area was removed and processed for light and electron microscopy observations. The implanted cultures completed maturation in the host tissue and no rejection signs were noticed; on the contrary, we were able to observe a favorable reaction from the host tissue, such as the formation of secondary blood vessels which penetrated the mass of the implanted culture. Well-developed magno- and parvocellular neurons were seen to contain neurosecretory vesicles in their terminals, and the neuroglial relationships established within these implants were homologable to those normally found.

[The ependymal lining of the infundibular recess in males and hermaphrodites in small ruminants in scanning electron microscopy]. / Ependýmová výstelka infundibulárneho recesu samcov a hermafrodita malých prezúvavcov v riadkovacom elektrónovom mikroskope.

A scanning electron microscope was used to study the surface of the ependymal epithelium of the lower part of the third cerebral ventricle of rams, he-goats, and in one goat with a developmental sexual disorder (testicular hermaphroditism). During mating, the apical membranes of the ependymal tanycytes of the rams and billy goats were found to be covered by numerous blebs (ependymosecretion). It was only in the billy goats in the mating season (not in the rams) that individual supraependymal cells with protrusions forming "ruffled membranes" were present on the surface of the infundibular recession. Although the finding on the sexual organs of the hermaphrodite was closer to the male, the presence of the large number of supraependymal cells was rather a female trait. As suggested by the results, it is also in the males of small ruminants that the activity of the sexual organs is associated with the structure and function of the ependymal epithelium in the infundibular recess.

Macrophages of hypothalamic third ventricle. I. Functional characterization of supraependymal cells in situ.

Cells on the ependymal surface of hypothalamic third ventricle of hamsters exhibit surface, ultrastructural, and phagocytic features of macrophages. The object of this investigation was to identify functions that a resident phagocytic system of the ventricles of the brain may have normally. We found that supraependymal macrophages in two- to three-week-old hamsters were associated with intact and degenerating intraventricular neuronal processes, ependymal cytoplasmic blebs, and degenerating intraventricular cells. These macrophages frequently contained cellular debris within phagosomal vacuoles. In the vicinity of neuronal ganglion on the floor of the third ventricle, there were numerous structures identified as growth cones, with organelles in varying stages of alteration and degeneration. Supraependymal macrophages apparently remove cellular debris resulting from ependymal metabolic processes and from cellular and neuritic degeneration. Such activity may be part of a normal process of remodeling, involving cell growth, renewal, or cell death within the third ventricle of developing animals.

Ultrastructural effects of anteroventral third ventricle lesions on supraoptic nuclei and neural lobes of rats.

Small lesions of the tissue surrounding the anterior ventral third ventricle (AV3V) cause adipsia, but there is no compensatory antidiuretic response. Therefore, the fine structure of the supraoptic nucleus and neural lobe, the major sites of synthesis and release of antidiuretic hormone (ADH), were compared in rats rendered adipsic by AV3V lesions 3 days earlier, rats deprived of water for 3 days and rats drinking normally. In sham-lesioned rats which were deprived of water, neuronal somas in the supraoptic nucleus show signs of stimulated secretory activity. However, the neuronal somas of supraoptic nuclei of rats which did not drink because they were made adipsic by AV3V lesions resemble those of normally hydrated controls. Neural lobes of water deprived animals contain a sharply reduced number of neurosecretory granulated vesicles and reduced apposition of glial processes with the perivascular connective tissue compared to those of normally hydrated rats. In contrast, neural lobes of rats with AV3V lesions contain large accumulations of densely packed neurosecretory vesicles, as well as abundant dense bodies and multilamellar bodies which may be evidence of increased crinophagy, and they have increased interposition of glial processes between axon endings and the perivascular connective tissue. In rats with AV3V lesions the severe dehydration due to adipsia was unable to stimulate release of ADH. The accumulation of neurosecretory vesicles in the neural lobe indicates that transport of ADH to the neural lobe was not impaired in this time period, but that exocytosis of ADH-containing neurosecretory vesicles in the neural lobe was blocked.

Histological study of the ependyma of the hypothalamic third ventricle in the water snake Natrix maura (L).

The ependyma of the hypothalamic third ventricle of Natrix maura (L) has been investigated by light microscopy employing Nissl and Golgi stains. Electron microscopy was also used. Brains were fixed by intraventricular perfusion and embedded in paraffin, celloidin and Spurr resin. Light-microscopically the ependyma appears as a simple layer epithelium with round or enlarged nuclei. It sometimes shows basal processes extending into the subependymal tissue. These processes can be clearly seen in Richardson stained semithin sections. They often contact the subependymal capillaries. Electronmicroscopically, tanycytes contacting capillaries show numerous microvilli and few cilia as well as pinocytotic vesicles at their apical surfaces. There are zonulae adherentes and probably gap junctions as well as digitations and desmosomes. In the apical cytoplasm there are mainly mitochondria, endoplasmic reticulum and Golgi apparatus. Near the nuclei and specially in the basal process there are many microtubules. The tanycytes contact the capillaries by an end foot containing mitochondria and pinocytotic vesicles, which are also observed in the endothelian cytoplasm. These structures lead us to think, like several other authors, that the tanycytes of Natrix maura (L) may serve as a linkage between CSF and the capillaries and that they possess the morphological features which are necessary to effect intracellular transportation.

Age related changes in the endocrine hypothalamus: I. Tanycytes and the blood-brain-cerebrospinal fluid barrier.

The fine structural organization of the floor of the third cerebral ventricle (dorsum of the median eminence of the hypothalamus) of 2 normal adult mice Fisher 344 rats was compared and contrasted with that of 2 aged rats 30 months old. Closely juxtaposed tanycytes (specialized ependymal cells) of normal young adults in the lower walls and floor of the third ventricle. In contrast, tanycytes in aged rats demonstrated significant intracellular separations, with only fine cytoplasmic processes remaining to interlink them. The phenomenon of mechanical separation between tanycytes in aged animals is discussed with respect to a potential impairment in the integrity of the blood-brain-cerebrospinal fluid barrier.

The ventricular surface of the monkey mediobasal hypothalamus. A scanning electron microscopic study.

The ventricular lining of the infundibular recess of monkey brains of both sexes was investigated with SEM. Based on the distribution patterns of surface profiles regional differences in the tanycytic ependyma are described. Variations in the number of surface profiles were observed in females during the ovarian cycle. These variations may be an indication not only of absorptive and/or secretory activities, but may also be related to cell membrane redistribution and membrane turnover in dependence of endocrine factors. In the males such variations were not registered. An essental sexual dimorphism does not seem to exist in the tanycytic ependyma of the monkey, only the reaction pattern is different. Supraependymal macrophages (SEC) were more numerous in the ovulating females than in the males or menstruating females. In addition to these SEC some monkeys had a second SEC type which was arranged in sheets. These are interpreted to be glioses. A variable number of axons can also be found in the lower regions of the infundibular recess. Their morphology suggests the presence of several types of axons, but they do not appear to constitute an organized network. No significant sexual differences were registered in relation to the axons.

Fetal hypothalamic transplants in the third ventricle of the adult rat brain. Correlative scanning and transmission electron microscopy.

Blocks of anterior hypothalamus were transplanted from 19 day-old fetuses of Wistar/Lewis rats into the third ventricle of adult male Brattleboro rats. Physiological changes in graft recipients and in sham-operated animals were monitored daily. Twenty days after surgery, the graft recipients and sham-operated animals were killed and their brains examined by correlative scanning and transmission electron microscopy. Host animals that exhibited both decreased polydipsia and increased urine concentration were found to have viable grafts within the third ventricle. The observed physiological changes suggested that synthesis and release of vasopressin occurred in the transplanted neurons. Grafts were well vascularized by vessels arising from the host hypothalamus. Neurons, with perikarya ranging from 8 to 30 micron in diameter, glial cells, and neurites were located throughout the transplants. A neurohemal contact zone, similar to that normally seen in the median eminence, could not be demonstrated in the grafts. The absence of complete glial and ependymal barriers indicates a relatively close association between cells in the transplants and the cerebrospinal fluid. A large increase in supraependymal neurons and their processes, including an eruption of neurons through the floor of the third ventricle in one animal, was observed in graft recipients but not in sham-operated animals.

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

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.

[Nerve fiber interrelationships with the ependyma of the 3d ventricle in rats in the perinatal period]. / Vzaimootnosheniia nervnykh volokon s épendimoi III zheludochka u krys v perinatal'nom periode.

The localization and ultrastructure of nerve fibers in the wall and lumen of ventral region of III ventricle was studied from the 16th day of prenatal period till the 9th day of postnatal period in the Wistar rats. The nerve fibers were first found in the subependyme zone; between the ependyme cells and in the lumen of ventricle on the 18th day of development. The nerve fibers occur constantly in the lumen of ventricle beginning from the 3rd day of postnatal period. The dilatations of nerve fibers contain granules of 50 to 90 nm in diameter with the electron-dense center (carriers of monoamines and/or releasing hormonnes) granules with moderately osmiophilic contents and light vesicles of the same size. It is suggested that these structures reflect different stages of the release of neurohormones in the cerebrospinal fluid. The synapse-like contacts of nerve fibers with the ependyme cells are observed beginning from the 3rd day of postnatal period. Their role in the regulation of absorption of substances by the ependyme cells functioning is discussed.

Changes in activity of the organon vasculosum laminae terminalis in the annual cycle in Rana temporaria.

In 70 sexually mature male and femal Rana temporaria frogs captured in natural habitat, mean nuclear volumes for the cells of the pars ependymalis and pars parenchymalis of the organon vasculosum laminae terminalis (OVLT) were determined in seven characteristic stages in life. The mean nuclear volume for the cells of the pars ependymalis and pars parenchymalis of the OVLT showed distinct annual fluctuation. Maximum nuclear volume of the cells in both investigated parts of the OVLT were observed during the breeding period (Ist decade of April), and minimum volume of the nuclei of the pars ependymalis at the beginning of hibernation (IIIrd decade of October), and in the pars parenchymalis near the end of active life (Ist decade of September).

Ultrastructure of the organum vasculosum of the lamina terminalis of the Japanese quail, Coturnix coturnix japonica.

The structure of the organum vasculosum of the lamina terminalis (OVLT) of the Japanese quail has been studied by light and electron microscopy. The OVLT has a palisade arrangement. It forms a part of the terminal plate extending from the optic chiasma to the anterior commissure and is characterized by a special vascular arrangement. The organ consists of ependymal, internal and external zones. The ventricular surface of the organ is covered by non-ciliated ependymal cells characterized by the presence of raspberry-like ventricular protrusions. The ependymal zone is composed of two types of ependymal cells, one being a large, cuboidal cell with columinous cytoplasmic protrusions suggesting secretion into ventricle, and the other a slender tanycyte with long balar plexus of the external surface. The internal zone contains two types of secretory neurons, parvocellular neurons containing a few dense-cored granules 1,000 A in diameter, and mediocellular neurons containing in their perikarya many larger granules 1,300-1,500 A in diameter and well developed granular endoplasmic reticulum and Golgi apparatus. The granular formations are usually observed in the Golgi area of both types of cells, thus indicating of their secretory activities. There are three types of nerve terminals in the neuropil: (1) nerve endings containing spherical, dense-cored granules 800 A in diameter, which display axosomatic synapses with perikarya of the neurons, (2) nerve endings containing dense-cored granules 1,000 A in diameter and clear vesicles 400 A in diameter, and (3) nerve endings containing dense granules 1,300 A in diameter and clear vesicles. Types 2 and 3 do not display axo-xomatic synapses but often show axo-dendritic synaptic contacts with dendrites in the neuropil; thus they are probably axons originating from the parvocellular and mediocellular neurons of this organ. The functional significance of the neurons and axons of the OVLT is discussed and it is suggested that two kinds of neurohormones may be secreted from the OVLT of the Japanese quail.

Morphology and distribution of supraependymal cells in the third ventricle of the albino rat.

The supraependymal cells (SEC) are a normal component of the wall of the cerebral ventricles. In the hypothalamic area of the third ventricle they are restricted, in healthy animals, to the ependymal projection of the hypophyseotropic area. Here the SEC show great polymorphism. In addition to bipolar, multipolar and stellate or spider-like cells, transitional forms between these types can be seen. Their perikarya and processes can either remain at some distance from the ependyma or be in close contact with it. The processes may protrude between the ependymal cells or show surface differentiations that resemble the thin cytoplasmic folds of the mesenchymal wandering cells. Considering this and the variations in the number of cells, for example during the ovarian cycle, the SEC can be interpreted as mesenchymal cells, probably related to microglial cells of the subependymal layer. It is suggested that the SEC have a phagocytotic function and may be involved in the normal renewal of the ependyma. A definitive explanation for the restriction of the SEC to the hypophyseotropic area as well as the elucidation of their function remain to be found.