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PubMed1982

Scanning electron microscopy of isolated dog renal tubules.

Isolation of canine renal tubular segments, cell clusters and individual cells permits direct observation by scanning electron microscopy (SEM) of the three dimensional cell shapes and the complex lateral and basal labyrinth structures. These functionally vital areas of the various tubular segments have not been previously observed in such detail. The tubular segments were freed of the obscuring basement membranes by prolonged tryptic digestion of tissue perfusion-fixed with glutaraldehyde, followed by attachment to gelatine coated coverslips and preparation for SEM. The tubular basal labyrinth is found to consist of a mosaic of basal processes arising from the lateral processes and the base of the cell body. The lateral processes and basal processes interdigitate with adjacent cells. Each tubular segment exhibits its own characteristic patterns of basal processes. The basal processes of the proximal convoluted tubule (PCT) are shaped like an inverted anvil, often branching. The basal processes of the proximal straight tubule (PST) and thin limbs are short microvilli. The distal straight tubule (DST) have fishtail-like basal processes. The distal convoluted tubule (DCT) show a swirling mixture of anvil shaped and microvillous basal processes. The collecting ducts exhibit short basal microvilli, plicae and villi lined basal infoldings. The looser structure of the basal labyrinth of cell clusters and isolated cells produced by this isolation procedure facilitated recognition of these complex structures.

Animals↗
PubMed1988

Identification of motoneurons and interneurons in the spinal network for escapes initiated by the mauthner cell in goldfish.

We used intracellular recording and staining techniques to study the spinal circuitry of the escape behavior (C-start) initiated by the Mauthner axon (M-axon) in goldfish. Simultaneous intracellular recordings from one or both M-axons and a spinal neuron, followed by HRP labeling of the spinal cell, show that each M-axon makes monosynaptic, chemical excitatory synapses onto 2 populations of ipsilateral spinal neurons. The first consists of the large primary motoneurons that, based on earlier work (Fetcho, 1986), innervate exclusively the faster, white muscle fiber types in the myomeres. The second group of cells is formed by previously undescribed descending interneurons with ipsilateral axonal branches that have contacts with primary and secondary motoneurons spread over 2 or more body segments. Indirect evidence suggests that these descending interneurons are excitatory, and they may explain the polysynaptic activation of motoneurons observed in earlier studies of the spinal circuitry (Diamond, 1971). Both classes of neurons excited by the ipsilateral M-axon are disynaptically inhibited by the contralateral one. The morphology and physiology indicate that this inhibition is mediated by interneurons that are electrotonically coupled to one M-axon and have processes that cross the cord to inhibit contralateral neurons in the region where these postsynaptic cells receive excitatory input from the other M-axon. We have identified interneurons with the physiological and morphological features of these predicted crossed inhibitory interneurons. These cells are electrotonically coupled to the ipsilateral M-axon and receive a chloride-dependent disynaptic inhibitory input from the contralateral M-axon. Their very simple somata give rise to a process that crosses the spinal cord between the 2 M-axons. Once on the opposite side of the cord, the crossing process sends myelinated branches that run rostrally and caudally, roughly parallel to the contralateral M-axon. Processes that arise from these longitudinal branches terminate in a striking association with collaterals of the M-axon; nearly every M-axon collateral along the longitudinal course of an interneuron is met by a branch or branches of the interneuron whose terminals are apposed to neurons postsynaptic to the collateral.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

The effect of federal health policy on occupational medicine.

All three branches of the federal government affect occupational medicine. Notable examples include: 1) the Department of Transportation ruling (1988) requiring drug testing in diverse areas of the transportation industry (executive branch); 2) the Workplace Drug Act (1988) calling for organizations to have a policy towards drug and alcohol abuse (legislative branch); and 3) the Supreme Court ruling on the constitutionality of drug testing in the transportation industry (1989) and that infectious diseases are a handicap in accordance with the 1973 Federal Rehabilitation Act (1987). The executive branch plays a major role in occupational medicine primarily through the Occupational Safety and Health Administration (OSHA), which issues standards based on a rule making process; the executive branch can also affect occupational medicine indirectly, as evidenced by President Reagan's Executive Order 12291 calling for Office of Management and Budget oversight of regulatory initiatives. The legislative branch enacts laws, conducts hearings, and requests reports on the operations of federal agencies. The judicial branch addresses occupational health issues when people affected by an executive ruling want to challenge the ruling; or in the case of the Supreme Court, when deliberating an issue over which two circuit courts of appeal have come to divergent opinions. The Occupational Medicine profession can participate in the political process through awareness of proposed legislation and by responding accordingly with letters, resolutions, or testimony. Similar options exist within the executive branch by participating in the rule-making process. A representative of the Governmental Affairs Committee, through periodic visits with key Washington representatives, can keep members of the American College of Occupational Medicine informed about federal legislative and regulatory activities. In appropriate cases, the organization can then take a formal position on governmental activities that affect the speciality.

Health Policy↗
PubMed1981

Periarterial macrophage sheaths (ellipsoids) in cat spleen--an electron microscope study.

Periarterial macrophage sheaths (PAMS), a term we introduce to replace "ellipsoids," surround arterial capillaries in the red pulp of the spleen and are major sites for clearance of blood-borne particles. PAMS and their arterial capillaries in cat spleens in various states of congestion and contraction were studied by transmission electron microscopy. Thorotrast, a colloidal suspension of thorium dioxide, was injected to label macrophages. A PAMS consisted of a fine meshwork of reticular cells and reticular fibers which held macrophages and formed a cylindrical sheath around an arterial capillary lying in its longitudinal axis. Some PAMS were spongy due to loosening of cell associations by plasma infiltration; others were tightly compressed. Blood cells were both free in the interstices of the PAMS and attached to macrophages. Reticular cells formed a closely applied but incomplete layer adventitial to the arterial capillary and extended branches which contributed to the meshwork. Small villous processes on the major branches of reticular cells approached each other, sometimes forming intercellular junctions, and fit into complementary indentations in the surfaces of macrophages and endothelial cells. Thin filaments within reticular cells filled the villous processes and formed a border beneath the plasmalemma; intermediate filaments ran through the centers of the branches. Reticular fibers lay between reticular cells. Basement membrane fabricated of the same material as reticular fibers lay between the endothelium and reticular cells. Macrophages contained Thorotrast and abundant debris of phagocytized cells and were joined by extensive interdigitation of micropseudopodia. Endothelial cells were long rods which lay parallel and were joined along their bases by interdigitating lateral processes. Intercellular junctions were present at some points, but at others lateral processes were everted to form open interendothelial slits through which blood cells could pass. Endothelial cells possessed great numbers of randomly oriented intermediate filaments and small patches of thin filaments scattered along the basal plasmalemma and in lateral processes. Thin filaments may function to attach cells to one another and to the basement membrane and may assist in closing interendothelial slits. We believe that the endothelium responds to changes in arterial blood pressure and blood flow. It stretches to allow dilatation and recoils, probably due to the intermediate filaments, squeezing blood cells through interendothelial slits.

Animals↗
PubMed2000

Growth cones are not required for initial establishment of polarity or differential axon branch growth in cultured hippocampal neurons.

Hippocampal neurons developing in culture exhibit two types of differential, seemingly competitive, process outgrowth in the absence of external cues. During the initial acquisition of polarity, one of several equivalent undifferentiated minor neurites preferentially grows to become the axon. Once the axon has formed, it typically branches, and the branches grow differentially rather than concurrently. In axons with only two branches, growth alternates between branches. In both axon establishment and branch growth alternation, growth among sibling processes or branches must be differentially regulated. We found that elaborate and dynamic growth cones were associated with growth, whereas diminished growth cones were associated with nongrowing processes or branches. To test whether growth cones were necessary for differential growth, growth cone motility was eliminated by application of cytochalasin E. Although cytochalasin treatment before axon formation yielded longer processes overall, a similar percentage of both treated and untreated neurons had one process that grew more rapidly and became much longer than its sibling processes. Immunostaining to visualize dephospho-tau, an axonal marker, demonstrated that these single dominant processes were axons. Axons that formed in cytochalasin were thicker and showed more intense anti-tubulin staining than their sibling processes. Branched axons deprived of growth cones retained a pattern of differential growth and often included alternation. These results indicate that neither formation of a single axon nor differential growth of branches are dependent on growth cone motility and suggest that the neuron can regulate neurite elongation at sites other than at the growth cone.

Animals↗
PubMed1979

An electron microscope study of synaptic contacts in the abdominal ganglion of Aplysia californica.

The fine structure of the abdominal ganglion of Aplysia californica has been studied in preparations fixed by immersion in aldehydes, either directly or after a survival of a few hours in artificial sea water. The central core of neuropil is surrounded by a rind of neuronal cell bodies floating in a subcapsular space containing a loose meshwork of neuronal and glial processes, separated by wide extracellular spaces. Large primary processes with deeply infolded membranes leave the neuronal perikarya and enter the neuropil where they branch into smaller processes containing either neurofilaments, neurotubules or both. Some have the appearance of initial segments. The neuropil is not a homogeneous structure. Rather, four types of zones can be distinguished: (1) zones of fibers of passage coursing together in the neuropil and making few synaptic contacts: (2) zones of neurosecretory fibers containing large granules and dense-core vesicles, again making few synaptic contacts: (3) zones with a great variety of synaptic contacts between medium size and small profiles; and (4) glomerular zones. The differentiated membranes of the synapses are characterized by a slight increase in density and by being regularly parallel to each other. Presynaptic densities are sometimes quite prominent but specialized dense cytoplasmic opacities have never been seen bordering the postsynaptic membranes, i.e., all synapses are of the symmetrical type. Interlemmal opacities vary considerably in density. In zone 3, the synaptic vesicles are of several sizes, are round, oval or flat, and are either clear or filled with different types of dense material. The population of vesicles within a single profile may consist either of a homogeneous group of similar vesicles or of various mixtures of two or three kinds of vesicles. In profiles with mixtures of clear and large dense-core vesicles, it is often only the clear vesicles which agglomerate towards the differentiated membranes. In such cases the large dense-core vesicles lie as a peripheral halo around the clear vesicles. Here, and especially in other large neuronal profiles not forming contact in the plane of section, they can be seen to associate specifically with mitochondria and glycogen. It is proposed that they do not contain neurotransmitters but are related to mitochondrial activities such as the storage of ATP or the movement of calcium ions. In profiles with mixtures of clear and small dense-core vesicles, both types of vesicles often touch the presynaptic membrane, suggesting the release of two transmitters or of a modulator or neurohormone with a transmitter, by a single terminal. Serial synapses are present in this zone. The glomerular zones contain small profiles forming many synaptic contacts, some of which are arranged in such a way as to suggest the existence of "reciprocal" serial synapses.

Animals↗
PubMed1991

Arborization of afferent fibers in ectostriatum centrale. Golgi study.

The study demonstrates one kind of the afferent fibers in ectostriatum in Golgi preparates. These fibers enter the ventro-medial border of the area. The primary section and further course and branches, as well as the terminal arborization of these fibers were observed. Also the maturation process of the branchings of the fibers was studied in the Golgi preparates after hatching. The fiber-networks found in one-day-old and 28-day-old chicken ectostriatum centrale were compared.

Afferent Pathways↗
PubMed1978

[Synaptoarchitectonics of the ganglia of the celiac plexus in white rats].

In 50 intact white rats at the age of 6, 15, 23 and 30 months synapsoarchitectonics of the celiac plexus nodes was studied by an electron microscopy method. Peculiarities in synapsoarchitectonics are stipulated by pericaryon processes in neurons, some of them have no contacts with the axonal terminals, while others have contacts with the axonal terminals. The former include small (about 0.5 mkm) drop-like and large (up to 1.5 mkm) polymorphous processes within the limits of perisomatic membrane, as well as processes penetrating the neuronal capsule. All of them contain, in different combinations, vesicles, ribosomas, fibrillae, and the largest processes--small cisterns of granular cytoplasmatic network and single mitochondria. The processes of the first group are considered as original stages for the development of the second group processes. The latter are represented by different in size (about 1.0--2.0 mkm) in form (digital, cone-, pin-, goblet-shaped, cylindrical, branching) and in content formations. There is, as a rule, one contact on the processes of an uncomplicated form, while on the branching processes there can be up to three and more contacting axonal terminals. Peculiar features in the composition of the processes taken as a whole (specific forms, absence of dendritic tubes, sometimes numerous contacts with axonal terminals in spite of small size) distinguish them from newly forming dendritic processes and these formations are considered as independent specialized receptor apparatus in the pericaryon of neurons of the celiac plexus.

Animals↗
PubMed2003

Dissociative increase of oligodendrocyte progenitor cells between young and aged rats after transient cerebral ischemia.

Oligodendrocyte progenitor cells (OPCs) mature to oligodendrocyte and remyelinate axons. OPCs express neuron-glial antigen 2 (NG2) chondroitin sulfate proteoglycan. We stained NG2 to investigate the effect of aging on the OPCs after cerebral ischemia. NG2 positive cells were examined at 1, 3, 7 days after 90 min of transient middle cerebral artery occlusion in young and aged brains. The number of NG2 positive cells increased in the ischemic penumbra at 3 and 7 days after reperfusion. At 7 day, the number of NG2 positive cells was significantly greater in the young than the aged brains, and the processes of NG2 positive cells enlarged and were more highly branched in the young than the aged brains. These results suggest that the young brain shows a higher potential of proliferation and process branching of OPCs than the aged brain.

Aging↗
PubMed2005

Side-branch growth in two-dimensional dendrites. II. Phase-field model.

The development of side-branching in solidifying dendrites in a regime of large values of the Peclet number is studied by means of a phase-field model. We have compared our numerical results with experiments of the preceding paper and we obtain good qualitative agreement. The growth rate of each side branch shows a power-law behavior from the early stages of its life. From their birth, branches which finally succeed in the competition process of side-branching development have a greater growth exponent than branches which are stopped. Coarsening of branches is entirely defined by their geometrical position relative to their dominant neighbors. The winner branches escape from the diffusive field of the main dendrite and become independent dendrites.

Journal Article↗
PubMed1998

Functional interactions between the holliday junction resolvase and the branch migration motor of Escherichia coli.

Homologous recombination generates genetic diversity and provides an important cellular pathway for the repair of double-stranded DNA breaks. Two key steps in this process are the branch migration of Holliday junctions followed by their resolution into mature recombination products. In E.coli, branch migration is catalysed by the RuvB protein, a hexameric DNA helicase that is loaded onto the junction by RuvA, whereas resolution is promoted by the RuvC endonuclease. Here we provide direct evidence for functional interactions between RuvB and RuvC that link these biochemically distinct processes. Using synthetic Holliday junctions, RuvB was found to stabilize the binding of RuvC to a junction and to stimulate its resolvase activity. Conversely, RuvC facilitated interactions between RuvB and the junction such that RuvBC complexes catalysed branch migration. The observed synergy between RuvB and RuvC provides new insight into the structure and function of a RuvABC complex that is capable of facilitating branch migration and resolution of Holliday junctions via a concerted enzymatic mechanism.

Bacterial Proteins↗
PubMed2000

Histologic and physiologic evaluation of skeletonized internal thoracic artery harvesting with an ultrasonic scalpel.

OBJECTIVES: The safety and reliability of a method of skeletonized internal thoracic artery harvesting with an ultrasonic scalpel (Harmonic Scalpel; Ethicon Endo-Surgery, CVG, Cincinnati, Ohio) were evaluated. METHODS: The mural branches of the internal thoracic artery were cut by means of 3 methods, differentiated by distance from the site of application of the Harmonic Scalpel blade to the internal thoracic artery. A total of 15 branches were cut from the internal thoracic artery at (0 mm) the origin (group I) or at 1 mm (group II) or 2 mm (group III) distal to the origin. Tissue preparations were examined for successful vessel closure and severity of tissue damage. The length of stump (L) and the length of tissue damage from the stump (D) were determined by a computer image analysis system, and pressure testing was performed to evaluate the physical strength of vessel closure. RESULTS: In group I, 8 of the 15 branches exhibited discontinuity of the vascular wall structure, probably because of insufficient sealing of the divided section, and 12 of the 15 branches exhibited tissue denaturation on the internal thoracic artery wall adjacent to areas of origin, which was probably caused by the heat transferred from the branches during the process of coagulation. In groups II and III, continuity of wall structure of stumps suggestive of stable closure of branches was confirmed. The lengths of tissue damage from the stump (D) were 0.96, 0.58, and 0.63 mm in groups I, II, and III, respectively, and the lengths of intact area (L - D) in the corresponding groups were -0.78, 0.61, and 1.51 mm. The negative figure in group I indicates the presence of tissue damage in the internal thoracic artery itself. By contrast, in groups II and III the internal thoracic arteries were intact, with a safety margin of greater than 0.5 mm. On physiologic evaluation of vessel closure, 2 of the 24 (8.3%) branches burst under a pressure lower than 350 mm Hg because of insufficient vessel coagulation, but the remaining 22 branches (91.7%) remained intact under pressures up to 350 mm Hg. CONCLUSION: The internal thoracic artery skeletonization method with an ultrasonic scalpel (Harmonic Scalpel: output level 2) appears to be a safe and reliable method of skeletonized internal thoracic artery harvesting when branches are sectioned at least 1 mm distal to their origin at a sufficiently slow speed.

Animals↗
PubMed1982

The ultrastructure of sheath cells in developing rat vomeronasal nerve.

Maturation of the vomeronasal nerve was studied in fetal, newborn and 3 months old rats. Early in development, each nerve consists of large numbers of bare axons with clusters of polygonal sheath cells lying at the periphery. The latter insinuate themselves between the axons which they segregate into bundles. The sheath cells and their processes which first delineate axon bundles from one another form a network in the interstices of which lie the emergent axon bundles. Each sheath cell is not confined to the sleeve around a single bundle. Its perikaryon and processes commonly contribute to the septa between several adjacent bundles. Eventually, each bundle comes to be surrounded by its own proper sheath which consists of processes of more than one Schwann cell. These developmental trends, of a progressive increase in the number both of axons per bundle and of Schwann cells associated with each bundle, are the reverse of those found in the PNS generally, where bundle size decreases and axon size increases with maturation. As individual bundles separate from one another, interfascicular collagen appears between them and each comes to be surrounded by a basal lamina. Separation is rarely complete, however; even at the mature stage, processes are exchanged between adjacent sheaths at one or more points on their circumferences. Schwann cell processes surrounding individual bundles become increasingly complex with maturation. Where adjoining processes meet, they commonly branch profusely and interdigitate intricately, forming stacks of closely apposed layered processes. In other areas, the branches are covered by basal lamina and bound intricate labyrinths which commonly extend deeply into the bundle and contain collagen fibrils.

Animals↗
PubMed1999

On a branching model of division-within-division.

We consider a deterministic version of a stochastic model for division-within-division processes described by Kimmel (1997, In: Proceedings of the IMA Workshop 'Classical and Modern Branching Processes' (K. Arthreya and P. Jagers, eds.)???? :????). It is shown that the behaviour of the deterministic model can be analyzed by using an associated Markov chain, using the methods of Barbour et al.

Cell Division↗
PubMed1986

A possible differentiation of anterior pituitary cells in collagen gels into neurons.

Dispersed cells from anterior pituitary glands of male rats were cultured embedded in collagen gels or on a plastic surface. After 6 or more days in culture, cells in collagen produced more prolactin than those on plastic. The cultures in collagen had fewer fibroblasts than those on plastic. Many cells cultured in collagen developed processes, and a few cells in every culture had long extended processes that sometimes branched and resembled those of neurons. About 60% of the cells in culture including cells with well developed processes bound the monoclonal antibody A2B5, an antibody that binds to neuronal cells in culture. Fibroblasts did not have detectable binding of A2B5. Some cells with short processes reacted with antiserum to prolactin or ACTH, but the cells with neuronlike processes did not. Collagen gels may provide a matrix in which cells from the anterior pituitary gland can differentiate into neuronlike cells.

Animals↗
PubMed2001

Shape diversity among chick retina Müller cells and their postnatal differentiation.

It is currently believed that in each vertebrate species Müller cells in the central retina constitutes a fairly homogeneous population from the morphologic point of view and that particularly the chick Müller cell attains full shape differentiation at prenatal stages. However, in this study of the chick retina, from day 1 to day 55 of life, we show that there is a large variety of Müller cell shapes and that many of them complete shape differentiation postnatally. We used a cell dissociation method that preserves the whole shape of the Müller cells. Unstained living and unstained fixed cells were studied by phase-contrast microscopy, and fixed cells immunostained for intermediate filaments of the cytoskeleton were studied by fluorescence microscopy. Our results show that (1) Müller cell shapes vary in the origination of the hair of vitread processes, in the shape of the ventricular (outer or apical) process, in the presence or absence of an accessory process, as well as in the number and shape of processes leaving from the ventricular process at the level of the outer nuclear and outer plexiform layers (ONL/OPL); (2) during the first month of life, many Müller cells differentiate the portion of the ventricular process that traverses the ONL, most Müller cells differentiate the ONL/OPL processes, and all Müller cells differentiate the thin short lateral processes leaving from the vitread hair processes at the level of the inner plexiform layer (IPL). The number of cells differing in the shape of the ventricular process and that of cells with and without accessory process were estimated. The spatial relationship between the outer portion of the ventricular process of the Müller cell and the photoreceptor cells was also studied. Our results show that the branching of the ventricular process and the refinement of Müller cell shape is achieved without apparent participation of growth cones. We give a schematic view of how the branching of the ventricular process might take place and propose the size increase of photoreceptor soma as a factor responsible for this branching.

Aging↗
PubMed2001

The interplay of matrix metalloproteinases, morphogens and growth factors is necessary for branching of mammary epithelial cells.

The mammary gland develops its adult form by a process referred to as branching morphogenesis. Many factors have been reported to affect this process. We have used cultured primary mammary epithelial organoids and mammary epithelial cell lines in three-dimensional collagen gels to elucidate which growth factors, matrix metalloproteinases (MMPs) and mammary morphogens interact in branching morphogenesis. Branching stimulated by stromal fibroblasts, epidermal growth factor, fibroblast growth factor 7, fibroblast growth factor 2 and hepatocyte growth factor was strongly reduced by inhibitors of MMPs, indicating the requirement of MMPs for three-dimensional growth involved in morphogenesis. Recombinant stromelysin 1/MMP3 alone was sufficient to drive branching in the absence of growth factors in the organoids. Plasmin also stimulated branching; however, plasmin-dependent branching was abolished by both inhibitors of plasmin and MMPs, suggesting that plasmin activates MMPs. To differentiate between signals for proliferation and morphogenesis, we used a cloned mammary epithelial cell line that lacks epimorphin, an essential mammary morphogen. Both epimorphin and MMPs were required for morphogenesis, but neither was required for epithelial cell proliferation. These results provide direct evidence for a crucial role of MMPs in branching in mammary epithelium and suggest that, in addition to epimorphin, MMP activity is a minimum requirement for branching morphogenesis in the mammary gland.

Actins↗