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Isolation and culture of amoeboid microglial cells from the corpus callosum and cavum septum pellucidum in postnatal rats.

In phase contrast and scanning electron microscopy, diverse structural forms of cells tenaciously adherent to glass coverslips were observed in the culture of the corpus callosum and cavum septum pellucidum from postnatal rats. In day 1 culture, many of the cultured cells were round, with well spread peripheral cytoplasm which appeared homogeneous. Cell organelles aggregated mainly around the reniform or round nucleus. Some cells showed spinous projections. In day 3-5 culture, the cells became irregular, sending out long branching pseudopodial processes; often they displayed a vacuolated cytoplasm. The cultured cells were highly phagocytic, as shown by their uptake of colloidal carbon particles and latex beads, in light microscopy and scanning electron microscopy, respectively. Cytochemical studies have shown that the cells were peroxidase-negative but were strongly positive for non-specific esterase, similar to the amoeboid microglial cells in the postnatal corpus callosum. On the basis of their structural features, both in phase contrast and scanning electron microscopy, experimental as well as cytochemical properties, it is concluded that the cells in the present culture are in fact amoeboid microglial cells which are active macrophages in the developing corpus callosum.

Animals↗

[Characteristics of the neuronal composition of the central nervous system of nudibranch mollusks].

Studies have been made on the neuronal composition of the central nervous system of the Nudibranch molluscs from the White Sea -- Coryphella rufibranchialis, Dendronotus arborescens and Cadlina laevis -- at various stages of their postlarval development, Localization maps for the largest neurons were composed. For C. rufibranchialis and D. arborescens, about 65--78 neurons were mapped; some of the characteristics of these neurons were investigated, such as age changes in the body size, the shape, pigmentation and neurosecretory activity. Using impregnation of silver nitrate into slices, two types of branching of neuronal processes were revealed (nervous cells with "branching" and "bifurcational" axons).

Animals↗

Scanning electron microscopic study of lead effects on cerebral astrocytes in primary culture.

The effects of lead acetate on the cell surface of primary cultured astrocytes from dissociated neonatal rat cerebrums were studied using scanning electron microscopy. Astrocytes, grown for two weeks in a modified Eagle's minimal essential medium containing 20% calf serum, were exposed to lead (1, 10, 30, 50, or 100 micrograms Pb/ml) for four days. Astrocytes grown in lead-free medium showed membrane folds (ruffles) and microvilli on the surfaces of perikarya. Vesicular structures or blebs (gliosomes) were scattered on the surfaces of perikarya and branched cell processes. Cultures exposed to 1 microgram Pb/ml were indistinguishable from controls. After exposure to 10 and 30 micrograms Pb/ml, microvilli disappeared, membrane folds were less extensive, and the number of surface blebs increased. Astrocytes exposed to 50 micrograms Pb/ml showed no surface membrane folds or microvilli while the blebs increased sufficiently to cover the surface of the perikarya. Cells treated with the highest lead concentration (100 micrograms/ml) were shrunken and perikarya again were covered with blebs. Transmission electron microscopy of the cells exposed to 10-100 micrograms Pb/ml showed membrane bound vacuoles and areas of cytoplasmic clearing, some of which may correspond to the surface blebs seen by SEM. These studies suggest that the changes in the cell surface resulting from lead exposure represent a toxic effect of lead on primary cultured astrocytes.

Animals↗

Macrophages in the external muscle layers of mammalian intestines.

The literature on macrophages in the muscularis externa of mouse, rat, guinea pig, cat, dog and human gut is reviewed. In smaller mammals macrophages are regularly situated in two locations: in the serosa and at the level of Auerbach's plexus between the longitudinal and circular muscle layers. In addition a few solitary cells are present at the level of the deep muscular plexus. At the level of Auerbach's plexus the macrophages occur as a constant and regularly distributed cell population with intimate associations between macrophages and interstitial cells of Cajal. Morphologically they differ from most resident macrophages in being irregular in shape with 4-6 primary cytoplasmic processes, which branch and give a stellate appearance. They have been demonstrated with endocytotic markers (trypan red, FITC-dextran, cholera toxin), immunocytochemically with macrophage antibodies (F4/80, M1/70) and antibodies against MHC class-II antigen, GABA and cGMP. In muscularis externa of the human gut a regularly distributed cell population of macrophages is not obvious. However, a phenotypically distinct subgroup is identified by light microscopy with the pan macrophage antibodies (EBM11, C3b1 and partly by p150.95), and shows MHC class-II antigen. By electron microscopy muscularis externa macrophages, in all species investigated, appear to be endocytically downregulated, and since they are lysozyme, prostaglandine H synthase (both constitutive and activated) and acid phosphatase negative, they appear to be inactivated cells. Both origin and function of these cells are unknown. They may be immuno-competent, participate in a neuroimmune axis, tissue growth and modulation or other regulations of specific cell functions.

Animals↗

Ultrastructural features in the structure of blood capillaries of the Haversian canal.

Electron microscopy was used to study osteon capillaries of the compact tissue of human growing long bones. The capillaries were found to be of the continuous type. On the lumenal surface of endothelial cells of the arterial capillaries there were a great number of micropinocytotic vesicles as well as a lot of branching cytoplasmic processes forming a characteristic spongioid layer. Vesicles and filopodia were considerably fewer on the basal surface of the cells. The endothelial cells of the venous capillaries had a small number of micropinocytotic vesicles and single cytoplasmic projections on their lumenal and basal surfaces. The capillary wall contained a great amount of pericytes. There were a lot of micropinocytotic vesicles in the cytoplasm.

Capillaries↗

[Cellular ultrastructure of various species of the genus Bifidobacterium].

Morphologic heterogeneity of cells from developing populations of bifidobacteria correlates with ultrastructure peculiarities. Active proliferating cells in exponential phase are characterized by formation of intracytoplasmatic membrane complex represented by lamellar, myelinoform, vesicular structures. Nucleoid is localized as the central polybranched or disperse osmophobic zone. Nucleoid distribution is determined by morphogenesis processes--exobudding, branching or multiseptation. Electronograms reveal multiple polyphosphate and polysaccharide inclusions. Ageing of bifidobacterial populations is accompanied with ultrastructural changes: cell wall hypersynthesis, reorganization and increased size of intracytoplasmatic membrane complex, altered morphology and compactness of nuclei, formation and dissimilation of inclusions.

Bifidobacterium↗

Intrahepatic branching patterns of portal vein. A study by corrosion cast.

Eighty-five corrosion casts of human portal veins were studied. The following observations were made: (1) In 88% of the cases, the portal vein divided into right and left branches, and in 12% of the cases, it divided into the left branch of the portal vein and anterior and posterior segmental n:veins; (2) Anterior and posterior segmental veins were branches of the right branch of the portal vein in 88% of the cases and arose directly from the portal trunk in 12% of the cases. In all cases, the anterior and posterior segmental veins in turn divided into superior and inferior area veins. (3) The left branch of the portal vein underwent a sharp kink leading into two parts: pars transversa nad pars umbilicalis. (4) The lateral superior area vein either sprang from the left side of the kink (69%), or from pars transversa near the kink (16%), or from the left side of pars umbilicalis near the kink (14%). The lateral inferior area vein arose from the left side of pars umbilicalis in all cases. (5) The medical segmental veins originated from the right side of pars umbilicalis. The number of branches varied from two to four. Further division of these into area branches were not fixed. (6) The right portion of the caudate lobe received portal blood either from the pars transversa (68%), or from the right branch of portal trunk (14%), or from the portal trunk (18%). The left portion of the caudate lobe received portal blood always from the pars transversa of the left trunk. (7) The branches to the caudate process arose from right branch of the portal vein in the majority of cases (72%). In the rest of the cases, these arose either from portal trunk (16% cases) or from pars transversa of the left trunk (12% cases).

Humans↗

Mechanisms and molecules controlling the development of retinal maps.

All mature vertebrates exhibit precise topographic mapping from the retina to the tectum, or its mammalian homologue, the superior colliculus (SC). In frogs and fish the development of this projection is precise from the outset; in avians retinal axon targeting is more diffuse but respects a coarse topographic matching; and in rodents early projections show no topographic specificity. Topography in avians and rodents emerges from a process of branch extension, arborization, and elimination of aberrant axonal projections. Despite these differences, the basic mechanisms controlling the development of this retinotopy are conserved. It has been hypothesized that molecules distributed in a position-dependent manner in the retina and the tectum or SC control the development of these maps. A number of candidate molecules have been identified on the basis of their distribution, or their ability to influence axonal growth in vitro. In addition, transcription factors and signaling molecules are expressed in a position-dependent manner and may regulate the expression of molecules involved in retinotopic map formation.

Animals↗

Use of transgenic mice to study autocrine-paracrine signaling in lung morphogenesis and differentiation.

Transgenic mice have been useful in demonstrating critical autocrine-paracrine interactions that are mediated by polypeptide hormones and associated receptors. These systems control interactions between the pulmonary mesenchyme and epithelium that have profound influences on lung morphogenesis, cell proliferation, and differentiation and include both inhibitory and stimulatory pathways. These pathways underlie the process of branching, sacculation, and alveolar development and may be served by several redundant polypeptides and receptors that work in concert to ensure normal pulmonary development. It is also likely that the same systems are involved in the proliferation and differentiation that accompany repair following injury. It is hoped that these transgenic models will provide insight into growth factors and receptors involved in cell proliferation, differentiation, and function during pulmonary organogenesis and also will serve to clarify the role of growth factors that may influence repair following lung injury, as seen in bronchopulmonary dysplasia and pulmonary fibrosis following severe lung injury in clinical settings.

Animals↗

Termination in the stellate ganglion of axons arising from the hilar vegetative plexus of the lung. Peripheral reflex arcs.

In cats the right lung was removed by thoracotomy. After 24-48 h the right stellate ganglion, the communicating branches between the right stellate ganglion and the right vagus, as well as the portion of both vagus nerves below and above the communicating branches, were processed for electron-microscopic investigation. Degeneration of synaptic axon terminals in the anteromedial part of the stellate ganglia, occurring after removal of the ipsilateral lung, shows that processes of ganglion cells located in the terminal ganglia of the bronchial tree establish synapses in the stellate ganglia. These results speak in favour of the existence of peripheral reflex arcs in the autonomic nervous system. For the time being, the data are not sufficient to define a new pathway related to the autonomic ganglia.

Animals↗

Malignant meningioma with a new intracytoplasmic inclusion body.

A 55-year-old man presented with a frontal mass lesion, which histologically consisted of pleomorphic tumor cells with necrotic foci and abundant mitotic figures, showing a meningothelial pattern focally. In addition, many eosinophilic round- or irregular-shaped bodies were seen not only in tumor cell cytoplasm but also in between the tumor cells. By electron microscopy, the tumor cells showed a minimal adherence to each other and the intercellular spaces were wide. Tumor cells had numerous branching cytoplasmic processes and some of them were united by desmosomal junctions. These features suggested that the tumor was a malignant variety of arachnoid trabecular cell meningioma. The inclusion bodies consisted of homogeneous fine granular material and were not surrounded by microvilli. This type of inclusion body has not been reported in meningiomas in the past. The significance and possible pathogenesis of the inclusions are discussed.

Cell Division↗

The ultrastructure of oligodendroglioma: personal experience and the review of the literature.

Ultrastructurally, oligodendrogliomas were tumors of rather monotonous appearance, with round and oval nuclei without indentations and cytoplasm of an electron-density lower than that of astrocytic tumors. Cisterns of endoplasmic reticulum and those of Golgi apparatus were occasionally distended. In addition to the usual subcellular organelles, the cytoplasm was rich in ribosomes, microtubules and glycogen granules. Some cellular processes contained branching cisterns, similar to those encountered in human neuroaxonal dystrophies. There are no discernible intermediate filaments. Blood vessels are apparently normal in oligodendrogliomas. Probably, the most characteristic features of oligodendrogliomas are concentric arrays of membranes (so called membrane laminations, whorls or scrolls). A fragment of the cytoplasm sequestrated within a particular whorl may contain mitochondria, lysosomes or abundant glycogen granules. The apparent continuity of membranes of whorls and the channels of endoplastic reticulum is rarely observed.

Brain Neoplasms↗

Astrocytes in adult rat brain express type 2 inositol 1,4,5-trisphosphate receptors.

Astrocytes respond to neuronal activity by propagating Ca(2+) waves elicited through the inositol 1,4,5-trisphosphate pathway. We have previously shown that wave propagation is supported by specialized Ca(2+) release sites, where a number of proteins, including inositol 1,4,5-trisphosphate receptors (IP(3)R), occur together in patches. The specific IP(3)R isoform expressed by astrocytes in situ in rat brain is unknown. In the present report, we use isoform-specific antibodies to localize immunohistochemically the IP(3)R subtype expressed in astrocytes in rat brain sections. Astrocytes were identified using antibodies against the astrocyte-specific markers, S-100 beta, or GFAP. Dual indirect immunohistochemistry showed that astrocytes in all regions of adult rat brain express only IP(3)R2. High-resolution analysis showed that hippocampal astrocytes are endowed with a highly branched network of processes that bear fine hair-like extensions containing punctate patches of IP(3)R2 staining in intimate contact with synapses. Such an organization is reminiscent of signaling microdomains found in cultured glial cells. Similarly, Bergmann glial cell processes in the cerebellum also contained fine hair-like processes containing IP(3)R2 staining. The IP(3)R2-containing fine terminal branches of astrocyte processes in both brain regions were found juxtaposed to presynaptic terminals containing synaptophysin as well as PSD 95-containing postsynaptic densities. Corpus callosum astrocytes had an elongated morphology with IP(3)R2 studded processes extending along fiber tracts. Our data suggest that PLC-mediated Ca(2+) signaling in astrocytes in rat brain occurs predominantly through IP(3)R2 ion channels. Furthermore, the anatomical arrangement of the terminal astrocytic branches containing IP(3)R2 ensheathing synapses is ideal for supporting glial monitoring of neuronal activity.

Amino Acid Sequence↗

Delivering the message: epimorphin and mammary epithelial morphogenesis.

The mammary gland consists of a highly branched tubular epithelium surrounded by a complex mesenchymal stroma. Epimorphin is an extracellular protein that is expressed by mammary mesenchymal cells that directs epithelial morphogenesis. Depending upon the context of presentation--polar versus apolar--epimorphin can selectively direct two key processes of tubulogenesis: branching morphogenesis (processes involved in tubule initiation and extension) and luminal morphogenesis (required for enlargement of tubule caliber). Here, we outline the fundamentals of mammary gland development and describe the function of epimorphin in these processes. We conclude with a review of recent studies that suggest similar morphogenic roles for epimorphin in other glandular organs.

Animals↗

cAMP promotes branching of laminin-induced neuronal processes.

Laminin is a potent stimulator of neurite outgrowth. We have examined the signal transduction events involved in the neuronal cell response to laminin. Cyclic nucleotides, calcium, and sodium-proton exchange do not appear to be required for the transduction of the laminin signal during neurite outgrowth. Direct measurement of cAMP and cGMP levels shows no changes in NG108-15 cells when cultured on laminin. Exogenous cAMP alone had no effect on either the rate of process formation or process length, but did alter the morphology of laminin-induced neurites. A four-fold increase in the number of branches per neurite and a two-to-three-fold increase in the number of neurites per cell were observed in both NG108-15 and PC12 cells cultured on laminin when either 8-BrcAMP or forskolin was added. The cAMP-induced branching was also observed when PC12 cells were cultured on a laminin-derived synthetic peptide (PA22-2), which contains the neurite-promoting amino acid sequence IKVAV. By immunofluorescence analysis with axonal or dendritic markers, the PC12 processes on laminin and PA22-2 were axonal, not dendritic, and the cAMP-induced morphological changes were due to axonal branching. These data demonstrate that changes in cAMP are not involved in laminin-mediated neurite outgrowth, but cAMP can modulate the effects of laminin.

8-Bromo Cyclic Adenosine Monophosphate↗

Separate branches of the uvr gene-dependent excision repair process in ultraviolet-irradiated Escherichia coli K-12 cells; their dependence upon growth medium and the polA, recA, recB, and exrA genes.

The extent of repair of single-strand breaks (incision breaks) induced in the deoxyribonucleic acid (DNA) of Escherichia coli K-12 cells by the uvr gene-dependent excision repair process after ultraviolet (UV) radiation was determined in the wild-type, polA1, recA56, recB21, and exrA strains. The wild-type strain repaired all incision breaks after incident doses of UV radiation (254 nm) of approximately 60 J m(-2) or less when incubated in growth medium, or approximately 15 J m(-2) or less when incubated in buffer. The polA1 strain repaired the incision breaks completely after incident doses of approximately 12 J m(-2) or less when incubated in growth medium, or after approximately 4 J m(-2) when incubated in buffer. The recA13, recB21, and exrA strains showed essentially complete repair after incident doses of 10 to 15 J m(-2) whether the cells were incubated in buffer or growth medium. These results suggest that the uvr gene-dependent excision repair process may be divided into two branches, one which is dependent on the presence of growth medium and also the rec(+)exr(+) genotype, and a second which can occur in buffer (growth medium-independent) and is largely dependent on DNA polymerase I. The presence of chloramphenicol in the growth medium resulted in an inhibition of the growth medium-dependent repair occurring in wild-type and polA1 cells and had little or no effect on the extent of repair observed in recA56, recB21, or exrA cells. The similarities between the growth medium-dependent and -independent branches of excision repair and two known processes for the repair of X-ray-induced single-strand breaks are discussed.

Buffers↗

Renal branching morphogenesis: concepts, questions, and recent advances.

The ureteric bud (UB) is an outgrowth of the Wolffian duct, which undergoes a complex process of growth, branching, and remodeling, to eventually give rise to the entire urinary collecting system during kidney development. Understanding the mechanisms that control this process is a fascinating problem in basic developmental biology, and also has considerable medical significance. Over the past decade, there has been significant progress in our understanding of renal branching morphogenesis and its regulation, and this review focuses on several areas in which there have been recent advances. The first section deals with the normal process of UB branching morphogenesis, and methods that have been developed to better observe and describe it. The next section discusses a number of experimental methodologies, both established and novel, that make kidney development in the mouse a powerful and attractive experimental system. The third section discusses some of the cellular processes that are likely to underlie UB branching morphogenesis, as well as recent data on cell lineages within the growing UB. The fourth section summarizes our understanding of the roles of two groups of growth factors that appear to be particularly important for the regulation of UB outgrowth and branching: GDNF and FGFs, which stimulate this process via tyrosine kinase receptors, and members of the TGFbeta family, including BMP4 and Activin A, which generally inhibit UB formation and branching.

Animals↗

Action potential reflection and failure at axon branch points cause stepwise changes in EPSPs in a neuron essential for learning.

In leech mechanosensory neurons, action potentials reverse direction, or reflect, at central branch points. This process enhances synaptic transmission from individual axon branches by rapidly activating synapses twice, thereby producing facilitation. At the same branch points action potentials may fail to propagate, which can reduce transmission. It is now shown that presynaptic action potential reflection and failure under physiological conditions influence transmission to the same postsynaptic neuron, the S cell. The S cell is an interneuron essential for a form of nonassociative learning, sensitization of the whole body shortening reflex. The P to S synapse has components that appear monosynaptic (termed "direct") and polysynaptic, both with glutamatergic pharmacology. Reflection at P cell branch points on average doubled transmission to the S cell, whereas action potential failure, or conduction block, at the same branch points decreased it by one-half. Each of two different branch points affected transmission, indicating that the P to S connection is spatially distributed around these branch points. This was confirmed by examining the locations of individual contacts made by the P cell with the S cell and its electrically coupled partner C cells. These results show that presynaptic neuronal morphology produces a range of transmission states at a set of synapses onto a neuron necessary for a form of learning. Reflection and conduction block are activity-dependent and are basic properties of action potential propagation that have been seen in other systems, including axons and dendrites in the mammalian brain. Individual branch points and the distribution of synapses around those branch points can substantially influence neuronal transmission and plasticity.

Action Potentials↗