Antimyeloperoxidase-associated proliferative glomerulonephritis: an animal model.

To develop an animal model for antimyeloperoxidase (MPO)-associated necrotizing crescentic glomerulonephritis (NCGN), we immunized Brown Norway rats with MPO and localized a neutrophil lysosomal enzyme extract, primarily consisting of MPO and elastinolytic enzymes, plus H2O2, the substrate of MPO, to the glomerular basement membrane (GBM). Upon immunization rats developed antibodies and positive skin tests to MPO. After unilateral perfusion of the left kidney with the lysosomal enzyme extract and H2O2, MPO and immunoglobulin (Ig)G localized transiently along the GMB. At the time of maximal inflammation, at 4 and 10 d after perfusion, MPO, IgG, and C3 could not be detected anymore. MPO-immunized rats perfused with the lysosomal enzyme extract and H2O2, in contrast to control-immunized and/or control-perfused rats, developed a proliferative GN characterized by intra- and extracapillary cell proliferation, ruptured Bowman's capsule, periglomerular granulomatous inflammation, and formation of giant cells. Monocytes, polymorphonuclear leukocytes (PMN), and to a far lesser extent T cells were found in the glomeruli. Interstitial infiltrates consisted of monocytes, PMN, and T cells. Granulomatous vasculitis of small vessels was found at 10 d after perfusion. The proliferative NCGN in this rat model closely resembles human anti-MPO-associated pauci-immune NCGN, and enables the study of the pathophysiology of anti-MPO-associated NCGN.

Estrous cycle-dependent neural plasticity in the caudal brainstem in the female golden hamster: ultrastructural and immunocytochemical studies of axo-dendritic relationships and dynamic remodeling.

UNLABELLED: During the short four-day estrous cycle of the female hamster various behavioral (lordosis, vocalization and aggression) and autonomic adaptations occur. Presumably, these changes are under ovarian control. Recently, we described a distinct estrogen receptor-alpha immunoreactive (ER-alpha-IR) cell group, now called nucleus para-retroambiguus (NPRA), in the caudal ventrolateral medulla (Gerrits et al., 2008). Neurons of this group project to the ipsilateral intermediolateral cell column in the thoracic and upper lumbar cord. Clearly, the NPRA is part of an estrogen-sensitive neuronal network and the same applies to the region containing the commissural part of the solitary tract nucleus (NTScom) and the A2 group, here called NTScom/A2. Estrogen is known to modulate neuronal ultrastructure in various brain areas and spinal cord, but not in the caudal brainstem. Because we assumed that the NPRA plays a role in estrous cycle related adaptations, we hypothesized the occurrence of plasticity in this nucleus. In the present study we examined morphological changes of axo-dendritic relationships in NPRA and NTScom/A2 in estrous, diestrous and ovariectomized (OVX) hamsters, using immuno-electron microscopy and the 1D5 anti-ER-alpha antibody. Ultrastructural analysis revealed that the ratio "axon terminals surface/dendrite surface" was significantly increased in both the NPRA and NTScom/A2 during the estrous phase compared to the OVX and diestrous conditions. Remodeling of axon terminals due to axonal sprouting into large terminal fields filled up with pleomorphic vesicles resulted in contacts with more dendrites, and was the main cause for the "axonal terminal-dendritic-ratio" shift. IN CONCLUSION: Estrous cycle-induced axonal and dendritic plasticity is present in the NPRA, and in the NTScom/A2 group. Our findings support our hypothesis that estrogen-sensitive neuronal networks in the caudal brainstem display structural plasticity, probably to modulate steroid hormone dependent behaviors or autonomic adaptations.

Review of environmental enrichment for broiler chickens.

Welfare problems are commonly found in both conventional and organic production of broiler chickens. In order to reduce the extent of welfare problems, it has been suggested to provide stimulating, enriched environments. The aim of the present paper is to provide a review of the effect on behavior and welfare of the different kinds of environmental enrichments in the production of broilers that have been described in the scientific literature. Environmental enrichment is defined as an improvement of the environment of captive animals, which increases the behavioral opportunities of the animal and leads to improvements of the biological function. This definition has been broadened to include practical and economic aspects, as any enrichment strategy that adversely affects the health of animals or that has too many economic or practical constraints will never be implemented on commercial farms and thus never benefit animals. Environmental enrichment for broilers often has the purpose of satisfying behavioral needs and/or stimulating the broilers to an increased level of activity, which among others will reduce the occurrence of leg problems. Potentially successful environmental enrichments for broiler chickens are elevated resting-places, panels, barriers, and bales of straw ("point-source enrichment"), as well as covered verandas and outdoor ranges ("complex enriched environments"). Many of the ideas for environmental enrichment for broilers need to be further developed and studied, preferably in commercial trials, with respect to the use, the effect on behavior and on other welfare aspects such as leg health, and the interaction with genotype, production system, stocking density, light, and flock size. In addition, information on the practical application and the economics of the production system is often lacking, although it is important for application in practice.

Novel Algorithms for Improved Sensitivity in Non-Invasive Prenatal Testing.

Non-invasive prenatal testing (NIPT) of cell-free DNA in maternal plasma, which is a mixture of maternal DNA and a low percentage of fetal DNA, can detect fetal aneuploidies using massively parallel sequencing. Because of the low percentage of fetal DNA, methods with high sensitivity and precision are required. However, sequencing variation lowers sensitivity and hampers detection of trisomy samples. Therefore, we have developed three algorithms to improve sensitivity and specificity: the chi-squared-based variation reduction (χ2VR), the regression-based Z-score (RBZ) and the Match QC score. The χ2VR reduces variability in sequence read counts per chromosome between samples, the RBZ allows for more precise trisomy prediction, and the Match QC score shows if the control group used is representative for a specific sample. We compared the performance of χ2VR to that of existing variation reduction algorithms (peak and GC correction) and that of RBZ to trisomy prediction algorithms (standard Z-score, normalized chromosome value and median-absolute-deviation-based Z-score). χ2VR and the RBZ both reduce variability more than existing methods, and thereby increase the sensitivity of the NIPT analysis. We found the optimal combination of algorithms was to use both GC correction and χ2VR for pre-processing and to use RBZ as the trisomy prediction method.

Ultrastructural evidence for direct excitatory retroambiguus projections to cutaneous trunci and abdominal external oblique muscle motoneurons in the cat.

The nucleus retroambiguus (NRA) is a group of neurons, located laterally in the caudal medulla oblongata. The NRA is thought to modulate abdominal pressure in the framework of respiration, vomiting, vocalization, probably parturition, and, in all likelihood mating behavior. The NRA exerts this control through its projections to motoneurons to the nucleus ambiguus in the lateral medulla (innervating pharynx, larynx), and spinal cord (innervating cutaneous trunci, intercostal, abdominal, pelvic floor, and lower limb muscles). The nature of these NRA-motoneuronal projections is unknown. In this study we have determined the ultrastructure of the NRA-motoneuronal projections, and especially those to the abdominal external oblique and cutaneous trunci muscles. In four cats 0.1% cholera toxin subunit b was injected in the external oblique and cutaneous trunci muscles to retrogradely label their motoneurons in the spinal cord. Wheat germ agglutinin-conjugated horseradish peroxidase was injected into the NRA to anterogradely label its contralaterally descending fibers to the motoneurons of both muscles. In order to prevent anterograde labeling of ipsilaterally descending systems not originating from the NRA, a hemisection was made at the level of C2 prior to the NRA injection. The ultrastructural results indicate that the majority (60-74%) of the anterogradely labeled NRA-terminals made monosynaptic contacts with retrogradely labeled dendrites of the external oblique and the cutaneous trunci muscle motoneurons. The majority (86-95%) of the NRA terminals made asymmetric synaptic contacts and 79-84% contained round vesicles. These results demonstrate the existence of direct, presumably excitatory, projections from NRA to external oblique and cutaneous trunci muscle motoneurons.

Microvascular changes in aged rat forebrain. Effects of chronic nimodipine treatment.

In the present study the effects of long-term treatment with the 1,4-dihydropyridine calcium antagonist nimodipine on ultrastructural alterations of the microvascular morphology were examined in the frontoparietal cortex, entorhinal cortex and CA1 of the hippocampus in the aged rat. Qualitative observations of cerebral microvasculature of aged (30 months) Wistar rats revealed the presence of microvascular fibrosis, membranous inclusions within the basement membrane and basement membrane thickenings. In several cortical regions the percentage of aberrant microvessels was significantly reduced in the nimodipine-treated rats. The observed microvascular anomalies were classified into five distinct categories of which microvascular fibrosis type II, defined as collagen deposits up to 1 micron within the microvascular basement membrane, showed the strongest reduction in the nimodipine-treated cases. The decrement of the percentage of aberrant microvessels and the relative occurrence of several classes of microvascular deviations showed some variation in the various brain regions examined and was most pronounced in frontoparietal cortex layer III. These results may provide a morphological basis for the improved motor and cognitive performance in aged rats after long-term oral nimodipine administration.

Ultrastructural evidence for a paucity of projections from the lumbosacral cord to the pontine micturition center or M-region in the cat: a new concept for the organization of the micturition reflex with the periaqueductal gray as central relay.

Information concerning the rate of bladder filling is determined by receptors in the bladder wall and conveyed via afferent fibers in the pelvic nerve to sensory neurons in the lumbosacral cord. It was assumed that this information is relayed from the lumbosacral cord to a medial cell group in the dorsolateral pontine tegmentum, called the M-region, the pontine micturition center, or Barrington's nucleus. The M-region, in turn, projects via long descending pathways to the sacral parasympathetic motoneurons. In the present electron microscopic study, it was investigated in cats whether monosynaptic projections from lumbosacral neurons to the M-region indeed exist. Wheat-germ agglutinin-horseradish peroxidase injections were made into the lumbosacral cord. Many retrogradely labeled dendrites and somata were found in the M-region, but no labeled terminals were found on retrogradely labeled dendrites or somata. Only a small number of anterogradely labeled terminals, which were filled with mainly round vesicles, contacted unlabeled dendrites in the M-region. In contrast, many more anterogradely labeled terminals, which were filled with mainly round and, to a limited extent, dense core vesicles and with asymmetrical synapses, were found on dendrites in the lateral part of the periaqueductal gray (PAG). Previously (Blok and Holstege [1994] Neurosci. Lett. 166:93-96), it was demonstrated that the lateral part of the PAG contains neurons projecting to the M-region. A concept for the central organization of the micturition reflex is presented in which ascending projections from the lumbosacral cord convey information on bladder filling to the PAG.(ABSTRACT TRUNCATED AT 250 WORDS)

Ultrastructural evidence for direct projections from the pontine micturition center to glycine-immunoreactive neurons in the sacral dorsal gray commissure in the cat.

During micturition, according to the concept of Blok, Holstege, and colleagues ([1997] Neurosci. Lett. 233:109-112), the pontine micturition center (PMC) elicits bladder contraction by way of direct excitation of the parasympathetic bladder motoneurons. At the same time, the PMC elicits relaxation of the external urethral sphincter (EUS) by excitation of gamma-aminobutyric acid (GABA)-ergic interneurons in the sacral dorsal gray commissure (DGC), which, in turn, inhibit EUS motoneurons. The question is whether the inhibitory neurotransmitter glycine is also involved in this system. The present study investigated, first, whether there are glycine immunoreactive interneurons in the sacral DGC and, second, whether they receive direct PMC afferents. Finally, it was determined whether glycine and GABA are colocalized in DGC interneurons. In two adult male cats, the PMC was identified by electrical stimulation. Subsequently, the identified region was injected with the anterograde tracer WGA-HRP. Sections of sacral cord segments were processed for light and electron microscopic detection of anterograde labeling, as well as for glycine and GABA, using postembedding immunogold labeling with antibodies. In total 128 labeled PMC terminals were found in the DGC, which contained many round vesicles and asymmetric synapses. About 31.3% (40 of 128) made contact with glycine-immunoreactive dendrites. Eleven of them were selected for serial sectioning, which showed that 54.6% (6 of 11) of the glycine-immunoreactive dendrites were also immunoreactive for GABA. The results demonstrate that the PMC projects directly to dendrites of interneurons in the sacral DGC, which are immunoreactive for both glycine and GABA. These interneurons are thought to inhibit the EUS motoneurons during micturition.

Thiamin, riboflavin, and vitamins B-6 and C: impact of combined restricted intake on functional performance in man.

A double-blind study of combined restriction of thiamin, riboflavin, and vitamins B-6 and C was carried out with 23 healthy males. During 8 wk of low vitamin intake, 12 deficient subjects consumed daily a diet of normal food products, providing maximally 32.5% of the Dutch Recommended Dietary Allowances (RDA) for thiamin, riboflavin, vitamins B-6 and C. Other vitamins were supplemented at twice the RDA. Eleven control subjects consumed the same diet but with a supplementation of twice the RDA of all vitamins. In deficient subjects blood vitamin levels, urinary vitamin excretion, and erythrocytic enzyme activities decreased; in vitro enzyme stimulation increased. Vitamin depletion had no ill effects on health, physical activity, and mental performance. A significant decrease was observed in aerobic power (VO2max) and onset of blood lactate accumulation (p less than 0.001) of 9.8 and 19.6%, respectively. A combined restricted intake of thiamin, riboflavin, and vitamins B-6 and C causes a decrease in physical performance within a few weeks.

A comparison of serum HIV-1 RNA levels as measured by two quantitative assays in zidovudine-treated, asymptomatic, HIV-infected individuals.

HIV-1 RNA levels as measured by two commercially available quantitative assays were compared before and during zidovudine treatment. HIV-1 RNA levels were measured in stored serum samples from 24 Dutch zidovudine-treated participants of a zidovudine efficacy study (European-Australian Collaborative Group Study 017) at weeks -3, 0, 4 and 8, using quantitative nucleic acid sequence-based amplification (NASBA; Organon Technika) and quantitative reverse transcriptase-polymerase chain reaction (Amplicor; Roche Molecular Systems). HIV-1 RNA copy numbers and changes from baseline as measured by each assay were compared. Individual responses to treatment were compared using definitions based on the within-subject variation of each assay. Before treatment, HIV-1 RNA levels as measured by NASBA were 0.49 logs higher than the levels measured by the Amplicor assay (95% confidence interval (CI) 0.32-0.66). During treatment, this difference decreased significantly to 0.27 logs (95% CI 0.01-0.53; difference 0.22 logs; 95% CI 0.05-0.37). The smaller difference between the results of the two assays during treatment was a consequence of a larger decline in RNA level as measured by NASBA compared with that measured by the Amplicor assay (mean change after 4 weeks 0.77 and 0.49 logs, respectively). At week 8, the mean HIV-1 RNA level was still significantly below baseline values as measured by NASBA, but not when measured by the Amplicor assay. Discrepancies in individual responses as measured by the two assays were also observed. In conclusion, marked differences exist between the NASBA and Amplicor quantitative assays, in both HIV-1 RNA copy numbers without treatment and changes in RNA level during treatment. These differences should be considered in interpreting analyses of clinical trials and relationships between HIV-1 RNA level and clinical outcome, as well as in the use of RNA level in the management of HIV-infected patients.

Ultrastructural analysis of climbing fiber-Purkinje cell synaptogenesis in the rat cerebellum.

Previous reports have described the transient expression of the neuropeptides calcitonin gene-related peptide and neuropeptide Y in selected subsets of rat olivocerebellar compartments during embryonic and postnatal development. Using these neuropeptides as endogenous markers for olivocerebellar fibers, the aim of this electron microscopic analysis was to reveal the synaptogenetic processes occurring between climbing fibers and their target Purkinje cells, from embryonic day 19 to postnatal day 16, the period during which Purkinje cells undergo intense emission and retraction of dendrites, and climbing fibers translocate their synapses along Purkinje cell membrane surfaces. The present findings provide the first direct evidence that climbing fiber synaptogenesis starts on embryonic day 19 and that these first synapses mainly involve the Purkinje cell embryonic dendrite rather than the Purkinje cell soma. At the same age, the presence of unlabeled synapses resembling calcitonin gene-related peptide-labeled synapses in the Purkinje cell plate makes it possible to conclude that climbing fibers form a major synaptic investment on embryonic Purkinje cells, a finding that strongly supports the hypothesis of an early differentiating role of climbing fibers on cerebellar development. Furthermore, during the period of intense dendritic remodeling of Purkinje cells, 'myelin figures' were often detected in Purkinje cell dendrites suggesting that they may at least in part represent real ultrastructural markers of membrane turnover that identifies the sites where Purkinje cell dendritic rearrangement is taking place. Finally the finding that the climbing fiber terminals apposed to degenerating dendrites did not generally show signs of degeneration leads us to suggests that climbing fiber translocation from a perisomatic to a dendritic location may be driven by the Purkinje cell dendritic remodeling.

Dendro-dendritic connections between motoneurons in the rat spinal cord: an electron microscopic investigation.

The ultrastructural characteristics of identified dendrite bundles in the rat spinal cord were analyzed following retrograde tracing from the soleus muscles. Dendrite bundles are arranged in networks that are heavily interconnected by means of gap junctions. The bundles are formed by at least 10 crossing dendrites travelling in different focal planes. In between dendrites, elongated gap junctional complexes are frequently found. Dendrite lamellar bodies, recently described to occur in relation with gap junctions in the central nervous system were not observed in the present study.

Evidence for monosynaptic projections from the nucleus retroambiguus to hindlimb motoneurons in the cat.

The nucleus retroambiguus (NRA) is a group of premotor neurons at the transition between brainstem and spinal cord. It projects to certain motoneuronal cell groups, among which is a distinct set of motoneurons in the lumbar enlargement innervating muscles including iliopsoas, adductor longus, and hamstrings. To find out whether these NRA-motoneuronal projections are monosynaptic, injections of wheat germ-agglutinin horseradish peroxidase (WGA-HRP) into the NRA were combined with injections of cholera toxin subunit b (CTb) into the hamstring muscles. Electron microscopical examination revealed that the NRA terminal profiles make monosynaptic contacts with dendrites of motoneurons innervating these muscles. The NRA terminal profiles formed asymmetrical synapses, and contained spherical and a few dense core vesicles. These findings provide evidence of monosynaptic NRA-hindlimb motoneuronal projections which are likely to be excitatory.

The pontine micturition center projects to sacral cord GABA immunoreactive neurons in the cat.

Stimulation of the pontine micturition center (PMC) results in micturition, i.e. an immediate relaxation of the bladder sphincter and a contraction of the detrusor muscle of the bladder. Earlier studies have shown that the bladder contraction is brought about by a direct excitatory pathway from the PMC to the parasympathetic bladder motoneurons in the sacral cord. How the PMC produces the inhibition of the bladder sphincter is not known. The present study in two adult male cats demonstrates at the ultrastructural level a direct pathway from the PMC to the dorsal gray commissure of the sacral cord. More than half (55%) of these terminals made contact with gamma amino butyric acid (GABA) immunoreactive neurons or somata, the others with non-GABA immunoreactive profiles. The PMC terminals contained many round vesicles, some dense cored vesicles and exclusively asymmetric synaptic clefts, which correspond with an excitatory pathway. A concept is put forward in which this pathway produces the relaxation of the bladder sphincter during micturition.

Direct projections from the nucleus retroambiguus to cricothyroid motoneurons in the cat.

Vocalization can be elicited by stimulation in the periaqueductal gray (PAG). Light-microscopical tracing and physiological studies have revealed that the PAG uses the nucleus retroambiguus (NRA) as a relay to excite the vocalization muscle motoneurons. Direct NRA projections have been demonstrated to pharyngeal and abdominal wall muscle motoneurons, but not to laryngeal motoneurons. In two cats 0.1% cholera toxin subunit b was injected in the cricothyroid muscle of the larynx to retrogradely label its motoneurons, and 2.5% wheat germ agglutinin-horseradish peroxidase was injected into the NRA to anterogradely label its fibers. The electronmicroscopical results indicate that the NRA fibers make monosynaptic contacts with cricothyroid motoneuronal dendrites. Almost all NRA terminal profiles had asymmetrical synapses and contained mostly round or pleiomorphic vesicles, which strongly suggests that the NRA-cricothyroid motoneuronal projection is an excitatory pathway.

Tract-tracing in the nervous system of vertebrates using horseradish peroxidase and its conjugates: tracers, chromogens and stabilization for light and electron microscopy.

Surprisingly the first generation of tract-tracing techniques based on intra-axonal transport, that is the methods utilizing the uptake and transport of horseradish peroxidase (HRP), still rank among the most widely used neuroanatomical tracing techniques. The success of these methods can be ascribed to several characteristics. They are fast and easy to implement. Complicated injection apparatus is unnecessary. The reaction products are visualized through simple histochemical reactions, and they are permanent or can be stabilized into permanency. Most usefully, the reaction products are visible in the light and electron microscopes. HRP (mol. wt. 44 kDa) is extracted from the roots of the horseradish plant (Cochlearia armoracia L.). Uptake of HRP into cells occurs via a passive process of endocytosis. Since lectins like wheat germ agglutinin (WGA) and bacterial toxin fragments (subunit B of cholera toxin (CTB)) strongly induce active, receptor-mediated uptake mechanisms, conjugates of these substances with HRP have been successfully applied in sensitive tract-tracing HRP and its conjugates are transported both in anterograde and retrograde direction. Retrograde transport occurs in small vesicles that are incorporated in lysosome-like vacuoles and in the Golgi apparatus. These vesicles differ in membrane properties from the anterogradely transported HRP vesicles. The retrogradely transported vesicles tend to fuse and thus accumulate HRP at high densities, facilitating the visibility of the final reaction product. The anterogradely transported HRP does not accumulate directly in lysosome-like bodies and is distributed diffusely and therefore often requires specific visualization methods. HRP and WGA-HRP may therefore be used in anterograde and retrograde transneuronal (multineuron) transport studies. Even in fixed material, labeling through diffusion of HRP can provide details on neural connections. Visualization of transported HRP is achieved by means of using the oxidative enzymatic activity of HRP to precipitate a chromogen according to the following reaction: [symbol: see text] The final reaction product may be soluble in buffer or ethanol and may require stabilization to prevent fading. In this protocol we discuss the widely used chromogens 3,3'-diaminobenzidine tetrahydrochloride (DAB), 3,3',5,5'-tetramethylbenzidine (TMB), benzidine dihydrochloride (BDHC) and p-phenylenediamine dihydrochloride with pyrocatechol (PPD-PC). Other possible chromogens, not discussed here, are 4-chloro-1-naphthol (4C1N), 3-amino-9-ethylcarbazole (AEC) and o-phenylenediamine (OPD). The visualization of the reaction product can be further improved by intensification with metal salts. At the light microscopic level (LM) this intensification enables color differentiation between distinct markers. In the present protocol we provide an up-to-date guideline for the application of HRP and its conjugates in tracing with special emphasis on electron microscopic (EM) visualization. Some modifications for stabilization and of metal intensification to enhance visibility are incorporated in conjunction with specific methods for multiple labeling in combined tract-tracing experiments.

Nesting material as environmental enrichment has no adverse effects on behavior and physiology of laboratory mice.

Environmental enrichment may improve the quality of life of captive animals by altering the environment of animals so that they are able to perform more of the behavior that is within the range of the animal's species-specific repertoire. When enrichment is introduced into an animal's environment, it is important to evaluate the effect of the enrichment program and to assess whether the animal continues to use the enrichment in the long-term. Groups of mice were housed under either standard or enriched conditions for several weeks. Nesting material which was highly preferred in previous studies was used as enrichment. During the period of differential housing several behavioral parameters (behavioral tests and handling) and physiological parameters (urine and plasma corticosterone, food and water intake, body and adrenal weight) were monitored to determine the impact of environmental enrichment. Observations were made to determine whether or not the mice continued to use the enrichment. The results indicated that throughout the study all mice used the nesting material to build nests and that mice from enriched conditions weighed more than mice housed under standard conditions, although the latter consumed more food. No major differences for behavioral and physiological parameters were found between the groups of mice housed under different conditions. Therefore it is not likely that supply of nesting material will jeopardize the outcome of experiments.

Serotonergic intraventricular axons in the habenular region. Phagocytosis after induced degeneration.

Both intracerebroventricular injection of 5,7-dihydroxytryptamine and electrolytical midbrain-raphe lesions in rats induce degeneration of supraependymal axons (SEAs) normally occurring in large numbers upon the ependyma of the medial habenular nucleus and habenular commissure. It is concluded that the intraventricular axon plexus in the epithalamic region is comprised of serotonergic (5-HT) fibers originating in the dorsal and/or median raphe nuclei. Besides the elimination of SEAs, conspicious features were a marked reduction in the number of cilia, degenerative signs in the habenular ependyma, and the emergence of large numbers of supraependymal macrophages, being most probably involved in phagocytosis of the axonal debris. It is suggested that the nucleus habenulae medialis is influenced serotonergically by the midbrain raphe via (1) a direct projection upon its neurons and (2) an indirect projection by way of the intraventricular axon plexus. The origin of intraventricular macrophages is discussed in relation to recent data in the literature.

Supraependymal cell clusters in the rat hypothalamus.

The fine structure of supraependymal cell clusters of the median eminence was studied with TEM. The cluster cells were identified on the basis of ultrastructure and histochemical determination of glial fibrillar acidic protein (GFA). The phagocytic properties were also studied by means of intraventricular injections of HRP. Neurons, neuroglia cells and degenerating ependyma- and glial cells were found. The extrusion of degenerating infundibular elements into the ventricle is a constant phenomenon but its precise localization and intensity are variable. The close proximity of the clusters to capillary loops is stressed. Because of the broken ependyma at the neck of the cluster, the permeability of the infundibular lining for HRP is increased. Clusters may be seen as sites lacking a brain--CSF barrier.

Tanycytes in the medial habenular nucleus of the rat.

Tanycytes with foot processes contacting capillary basal membranes were identified in the rat medial habenular nucleus. They constitute a relatively small but constant population of cells among the conventional ependymal cells. In contrast to tanycytes in most circumventricular organs, habenular tanycytes possess cilia. Superimposed upon the cells are nerve fibers belonging to the serotonergic supraependymal axon plexus. Their ultrastructure differs in many respects from that of hypothalamic tanycytes.