Biomedical subjects

H Takahashi-Iwanaga

Publications and source records attributed to H Takahashi-Iwanaga.

At least 19 recordsLinked to original sources

PubMed2005

Calcium signaling in terminal Schwann cells associated with lanceolate sensory endings in rat vibrissae.

Transient elevations of the intracellular Ca2+ concentration ([Ca2+]i) in glia mediate various cell activities to regulate neuronal functions. The present study focuses on spatiotemporal dynamics of Ca2+ signaling in terminal Schwann cells, glial elements of the lanceolate sensory endings innervating the rat vibrissa. Arrays of lanceolate endings were enzymatically isolated from the vibrissal follicle, and subjected to [Ca2+]i image recording by time-lapse confocal microscopy. Each terminal Schwann cell displayed a round cell body, and extended long cytoplasmic stalks, which branched into two to five lamellae resting on different axon endings. Each axon ending, on the other hand, was covered on both flattened sides of the lancet by two Schwann lamellae of different cell origin. Thus the peripheral glia constituted an extensive network connecting the sensory endings. Image analyses of [Ca2+]i characterized the Schwann lamellae as functional compartments that can independently generate Ca2+ signals: these cell portions primarily responded to local mechanical stimuli with a [Ca2+]i spike, and individually initiated [Ca2+]i oscillations during bath application of the sensory modulator adenosine 5'-triphosphate (ATP). The stimulus-induced signals sometimes propagated along the glial network to activate neighboring lamellae after a delay of 2-4 sec. These findings suggest that the terminal Schwann cells contribute both to individual regulation and total coordination of the arrayed sensory endings.

Adenosine Triphosphate↗

PubMed2001

Early anchoring collagen fibers at the bone-tendon interface are conducted by woven bone formation: light microscope and scanning electron microscope observation using a canine model.

To clarify the early process of recovery at the bone-tendon interface, we used light microscopy and SEM to examine the process of anchoring of collagen fibers to bone in a canine model. At two weeks, tendon, scar tissue, woven bone and lamellar bone were present at the insertion site. SEM revealed anchoring of collagen fibril bundles of the scar to the woven bone. By 4 weeks, the number of anchoring fibers had increased and a parallel arrangement of fibers was observed. SEM demonstrated deep penetration of fibers into the woven bone layer. In addition, the fibers were observed to project into and intermingle with the scar tissue. By 6 weeks, the anchoring fibers had developed fully and were distributed densely over the interface. SEM also revealed that the collagen fibril bundles in the scar tissue had connected with the collagen fibrils of the woven bone by way of the anchoring bundles. The woven bone was identifiable throughout the early stages of recovery as the interface between soft tissue and hard tissue. Throughout all experimental periods, no staining was observed at the interface of the tendon and bone by Saffranin-O. The formation of woven bone was important during early recovery of the tendon-bone interface prior to the completion of fibrocartilage-mediated insertion.

Animals↗

PubMed2001

Scanning electron microscopic observation of Merkel cells in the lamprey epidermis.

The Merkel cell in the epidermis has generally been regarded as a mechanoreceptor which detects tissue deformations with its microvilli, and subsequently releases certain transmitters to nerve endings. In order to analyze the mechanism of mechanoreception, the fine structure of lamprey Merkel cells and their relationships with surrounding tissue were examined by scanning electron microscopy (SEM) after exposure of the cells by NaOH maceration, as well as by transmission electron microscopy (TEM) according to a conventional method. By SEM, lamprey Merkel cells revealed small round cell bodies bearing numerous microvilli on the upper and lower poles in accord with previous TEM reports. Combined SEM and TEM observations showed that the Merkel cell bodies were tightly held in corresponding concavities of other epidermal cells, with some desmosomes connecting the cells with each other. On the other hand, microvilli of the Merkel cells extended freely in intercellular spaces bound with complex microplicae of epidermal cells. The regional difference in mechanical anchorage of the Merkel cells probably leads to transient deflection and subsequent recovery of their microvilli during a given mechanical stimulation, suggesting rapidly adapting mechanoreception by the cells.

Animals↗

PubMed2001

Three-dimensional microanatomy of mechanoreceptors and their possible mechanism of sensory transduction.

The fine structure of sensory nerve endings and their topographical relationships with surrounding tissues were examined by a combination of scanning and transmission electron microscopy in order to analyze the mechanism of mechanoreception. Observations were reported on Ruffini endings in periodontal ligaments of rat incisors, and on longitudinal lanceolate endings surrounding rat sinus hairs. Both types of receptors exhibited the triplet structure known as the axon-Schwann cell complex; a flattened axon terminal was sandwiched between two Schwann cell lamellae. The two receptor types additionally revealed their specific modifications at each distal end, where fine tuft-like processes of Schwann cells projected into surrounding tissues with finger-like projections of an axon terminal attached to their bases. In the Ruffini endings of the periodontal ligament, the terminal tufts coiled about collagen bundles in favor of continuous transmission of tissue distortions to their accompanying axon fingers. In the lanceolate endings of sinus hair follicles, the Schwann cell tufts were suspended in an amorphous matrix with only their end feet anchored to rigid connective tissue elements. Terminal axon fingers associated with these Schwann cell processes were supposed to transiently deflect during acceleration and deceleration phases of a given hair movement because of inertia. The present study proposes the terminal tuft of Schwann cell processes and their accompanying axon fingers as a structural complex which potentially contributes to mechano-electric transduction.

Animals↗

PubMed2000

Three-dimensional microanatomy of longitudinal lanceolate endings in rat vibrissae.

The longitudinal lanceolate endings are ubiquitous sensory terminals in the sinus and nonsinus hairs of mammals that form a palisade around the hair follicle. To analyze how the nerve endings detect hair movements, the present study re-examined their fine structure and relationships with surrounding connective tissue in rat vibrissae by using a combination of three methods: immunohistochemistry for S-100 protein, scanning electron microscopy of NaOH-macerated specimens, and transmission electron microscopy of serial sections. Observations showed the lanceolate endings to be represented by triplet units with a flattened axon terminal flanked on each side by a Schwann cell lamella, as reported previously. Two distinct parts were discriminated in the lanceolate ending: a principal portion in which the axon terminal protruded numerous fine fingers from between the Schwann cell coverings, and an apical cone that enclosed a large axon finger in an attenuated Schwann sheath. Long foot processes of Schwann cells fanned out distally from each apical cone. The principal portions of the lanceolate endings were firmly linked to the surrounding connective tissue by the narrow edges equipped with axon fingers, suggesting their continuous deformation by sustained hair deflections. In contrast, the apical cones were freely suspended in an amorphous matrix with only the end feet of the Schwann cell projections attached to rigid tissue elements. This part of the ending was proposed as a possible transducer site to generate rapidly adapting receptor potentials, both retreating and overshooting during the acceleration and deceleration phases of a given vibrissal movement.

Animals↗

PubMed2000

Parasitic forms of a myxosporean in the kidney of the arctic lamprey, Lampetra japonica: an ultrastructural study.

We found frequent and unique parasitism by an unidentified myxosporean in the kidney of the arctic lamprey, Lampetra japonica, living in Japan. Trophozoites (pseudoplasmodia with or without sporoblasts) existed predominantly in the lumina of proximal urinary tubules, but were rarely found in any other regions of the kidney. Since no mature spores were produced in the trophozoites, exact identification of the species was impossible. Two parasitic forms were recognized in proximal urinary tubules: one adhering to the epithelial cells of renal tubules, and the other free-floating in the lumina of tubules. Ultrastructurally, the attaching trophozoites developed microvilli-like projections towards the apical surface of epithelial cells and vigorously interdigitated with microvilli of the brush border. In contrast, the whole surface of the floating trophozoite was smooth without any cell projections. The developed projections in the former type of trophozoite may contribute to their firm attachment to the epithelial cells and/or to absorption of nutrients via the epithelial cells. Against the myxosporean infection, the lamprey as the host exhibited a local immune reaction by disposition of numerous lymphocytes and macrophages into the epithelium of urinary tubules.

Animals↗

PubMed1999

Three-dimensional ultrastructure of synoviocytes in the horse joint as revealed by the scanning electron microscope.

The synovial membrane displays a superficial cellular lining composed of two types of synoviocytes: "absorptive" macrophages (type A cells) and "secretory" fibroblast-like cells (type B cells). The types are intermingled and extend a variety of processes, rendering the cellular architecture of the synovial membrane difficult to visualize. Previous electron microscopic and histochemical studies failed to demonstrate the entire shape of synoviocytes, except our immunohistochemical study for protein gene product 9.5 in the horse joint. The present SEM study is the first to demonstrate the three-dimensional ultrastructure of synoviocytes as well as their distribution in the synovial membrane, using macerated samples from the horse carpal joints. The equine synovial membrane was largely covered by conspicuously developed synovial villi. Type A synoviocytes were closely similar to macrophages in regard to surface structure, and showed uneven distribution with the densest occurrence around the tips of the synovial villi. In the basal half of villi, type B synoviocytes, which were situated in close proximity to the synovial cavity, projected thick processes horizontally and intertwined to form a regular network of processes on the synovial surface. Those in the upper half of the villi were located in the abluminal layers and protruded an antenna-like process into the joint cavity with tips covered with long microvilli, in addition to forming the superficial plexus of processes. Type B cells were also provided with fine, membranous extensions that tended to cover the surface of synovial intima. The meshwork of horizontal processes, the antenna-like processes, and the membranous processes imply advantages in not only secretion but also sensation and regulation of the barrier function in the synovial membrane.

Animals↗

PubMed1999

Porosity of the epithelial basement membrane as an indicator of macrophage-enterocyte interaction in the intestinal mucosa.

The epithelial basement membrane of intestinal villi is perforated with numerous small pores, through which free cells in the lamina propria communicate with the enterocytes. This study was a comparative analysis of the pores in the basement membrane by SEM after removal of the gut epithelium with OsO4 maceration. The porosity as represented by the area fraction of the pores varied along the baso-apical axis of villi in patterns specific for each animal species examined: consistent scantiness along the entire length of villi in mice, acute elevation in the second and third distal one-sixths of villi in rats, and gradual augmentation toward the villus tips in guinea pigs. Size distribution analyses of the pores indicated their heterogeneous enlargement in the regions of elevated porosity. Concomitant observation of lamina propria macrophages by histochemical labelings and by conventional TEM showed that the cells specifically clustered beneath the hyperporous basement membrane, with their thick processes penetrating it. The spatially-regulated patterns of perforation of the epithelial basement membrane indicate phase-specific interventions of lamina propria macrophages in the maturation or aging of enterocytes, which steadily proliferate in crypts and exfoliate at the villus tips.

Animals↗

PubMed1998

Generation of intestinal T cells from progenitors residing in gut cryptopatches.

Cryptopatches (CPs) are part of the murine intestinal immune compartment. Cells isolated from CPs of the small intestine that were c-kit positive (c-kit+) but lineage markers negative (Lin-) gave rise to T cell receptor (TCR) alphabeta and TCR gammadelta intestinal intraepithelial T cells after in vivo transfer or tissue engraftment into severe combined immunodeficient mice. In contrast, cells from Peyer's patches and mesenteric lymph nodes, which belong in the same intestinal immune compartment but lack c-kit+Lin- cells, failed to do so. These findings and results of electron microscopic analysis provide evidence of a local intestinal T cell precursor that develops in the CPs.

Animals↗

PubMed1998

Three-dimensional microanatomy of perineuronal proteoglycan nets enveloping motor neurons in the rat spinal cord.

Spinal motor neurons possess reticular coats of extracellular matrix proteoglycans on their somata and proximal dendrites. In order to define the anatomical background of the network, spatial relationships of the perineuronal proteoglycans with synaptic boutons and astrocyte processes were analyzed in rat motor neurons by TEM after histochemical detection of the substances with cationic iron colloid, and by SEM after exposure of the cytoarchitecture with NaOH maceration. Narrow intercellular channels filled with proteoglycan were found to extend along the surface of the neurons to form a homogeneous network of a mesh size of about 1 microm. The system of perineuronal channels consisted of two parts: a primary intervaricose net which meandered among synaptic boutons on the surface of the motor neuron, and secondary subvaricose nets which irrigated interfaces between larger boutons and the neuron. No elements in the perineuronal cytoarchitecture coincided with the meshwork of proteoglycan, indicating the involvement of postsynaptic factors in the distribution of the substance. Thin astrocyte processes surrounding the neurons formed a distinct network with heterogeneous meshes corresponding to boutons of various sizes. The perineuronal glial nets extended their surface area in contact with the intervaricose nets of proteoglycan by complex cellular interdigitations. The subvaricose nets of proteoglycan compartmentalized multiple synapses on large boutons, suggesting an involvement in the division of the synapses during development.

Animals↗

PubMed1997

Scanning and transmission electron microscopy of Ruffini endings in the periodontal ligament of rat incisors.

The Ruffini organ is an arborized axon ending categorized as a low-threshold stretch receptor. We have previously shown that the lingual periodontal ligament of rat incisors is densely innervated with Ruffini endings. In the present study, fine structures in the surface of the periodontal Ruffini endings and their topographical relationship with the surrounding collagen fibers were observed by a combination of scanning and transmission electron microscopy to analyze the mechanism of the stretch reception. The entire length of the branches of the Ruffini endings, excepting their terminal portions, corresponded well with those depicted by previous investigators in the following points: (1) their cylindrical appearance covered by Schwann cell processes; (2) the presence of numerous axon fingers protruding through gaps in the Schwann sheath and; (3) their isolation from collagen fibers by multilayered basal lamina. On the other hand, tips of the axon branches-together with their Schwann sheaths-became attenuated and projected into tight bundles of collagen, indicating their susceptibility to mechanical deformations of the surrounding tissue. Margins of the axon terminals were conspicuously ruffled with long tongue-like projections of Schwann cells. The Schwann cell tongues twined around collagen bundles in their distal portions, and associated closely with fine axon projections in their proximal portions, suggesting their involvement in the mechanical transmission of stimuli to axon terminals.

Animals↗

PubMed1997

Neonatal granulocytosis is a postpartum event which is seen in the liver as well as in the blood.

In a recent series of studies, we demonstrated that stress in humans and animals, with resultant sympathetic nerve strain, induces severe granulocytosis, because granulocytes carry adrenergic receptors on the surface. Because activated granulocytes produce free radicals and superoxides, they sometimes induce tissue damage if the stress is too strong or continuous. Human neonates are also known to show high levels of granulocytes in the peripheral blood. In this study, we investigated whether such neonatal granulocytosis are a stress-associated response at birth. Both human and mouse materials, before and after birth, were used. The number of leukocytes in the blood, as well as some other factors in the serum, were measured. Although levels of granulocytes were found to be low in fetal humans and mice, they increased sharply after birth. In parallel with this postpartal granulocytosis, transaminases in sera increased transiently. In reference to results of a transient elevation in the levels of catecholamines at birth in mice, all these phenomena resemble stress-associated responses. Indeed, fatty liver and hematopoietic destruction in the liver were also observed in mice and humans. At this time, the production of inducible nitric oxide synthase (iNOS) by granulocytes in the liver was evident. These results suggest that neonatal granulocytosis is a postpartum event which results from various stresses (e.g., oxygen stress) at birth. This event may be responsible for such well-known neonatal phenomena as the termination of fetal hematopoiesis in the liver and as neonatal jaundice.

Alanine Transaminase↗

PubMed1997

Hydra regeneration from recombined ectodermal and endodermal tissue. II. Differential stability in the ectodermal and endodermal epithelial organization.

Hydra tissue consists of the ectodermal and the endodermal layers. When the two layers were separated by procaine treatment and then recombined, the ectodermal epithelial cells spread as a single cell layer over the endoderm as in epiboly in vertebrate embryogenesis, and the resultant spherical structure subsequently regenerated into a complete hydra. In this study, light and electron microscopy were used to examine the structural changes which took place in the cells and tissue during this epibolic ectodermal spreading process. Within a few hours after tissue recombination, the endoderm underwent dramatic changes; it lost its epithelial sheet organization, and turned into a mass of irregularly shaped cells without the apical-basal cell polarity initially present. In contrast, the ectoderm maintained its basic epithelial sheet organization as it spread over the endoderm. Later, the endodermal epithelial cells reorganized themselves into a single-layered epithelial sheet underneath the spreading ectodermal layer. The resultant spherical structure consisted of a single layer of ectodermal epithelial cells outside, a single layer of endodermal epithelial cells inside, and an empty cavity in the center as in normal hydra tissue. This structure regenerated into hydra in the following days. These and other observations demonstrate that the two-layered epithelial sheet organization is highly dynamic, and that its stability is maintained by strong interactions between the two layers in normal hydra. It is suggested that this dynamic nature of the hydra tissue, particularly the high plasticity of the endodermal epithelial sheet organization, may be an important element for the high regenerative capacity of this organism.

Animals↗

PubMed1996

Macrophagic cells outgrowth from normal rat glomerular culture: possible metaplastic change from podocytes.

One representative of a number of severe lesions that occur outside the glomerular capillaries and involve podocytes is crescentic glomerulonephritis. The question of whether the crescent-forming cells are derived from glomerular epithelial cells or monocytes/macrophages is highly controversial and has not yet been clarified. To investigate the pathophysiology of podocytes in crescentic glomerulonephritis, we attempted to establish methods for culturing cells confirmed to be derived from podocytes, focusing particularly on the relationship between podocytes and macrophages. Nonadherent cells of unknown origin that grew from normal rat isolated glomerular cultures increased in number, reaching a total of 3.5 x 10(5)/ml on Day 11. They showed several characteristics of macrophages, the expression of specific antigens and enzymes, morphology, and production of H2O2. They expressed Fx1A but lacked the expressions of Thy1.1 or factor VIII. A morphologic kinetic study on Days 3 to 11 of culture showed that the cells with foot processes on the glomerular basement membrane changed into macrophagic cells (MC) and migrated from the glomeruli. Immunofluorescence double staining indicated that the cells that migrated from the glomerular surface on Day 8 were both anti-podocalyxin- and ED-1-positive. Furthermore, immunoelectron microscopy revealed that the ED-1-positive cells were located on the glomerular basement membrane. Pretreatment with anti-macrophages and -Thy1.1 antibodies, both with complement, did not reduce the number of MC, whereas pretreatment with puromycin aminonucleoside predominantly reduced the number of MC. A predominant decrease in the number of glomerular macrophages by gamma-irradiation did not result in a reduction of the number of MC. MC derived from glomerular cultures of bone marrow chimeric rats expressed the la antigen originated from recipient, which indicates that MC is not derived from bone marrow cells. Macrophage colony-stimulating factor accelerated the speed of the change into MC, and granulocyte-macrophage colony-stimulating factor dramatically enhanced its degree with increase of cell number on Day 8. We concluded that podocytes change into MC in normal rat glomerular culture and that the change is enhanced by colony-stimulating factors. The results provide a completely new insight into the origin of crescent-forming cells.

Animals↗

PubMed1995

Relationships between intermediate TCR cells and NK1.1+ T cells in various immune organs. NK1.1+ T cells are present within a population of intermediate TCR cells.

Experiments to date have revealed a population of T cells that carry intermediate (int) levels of TCR (or CD3) and express IL-2R beta-chain (IL-2R beta) in mouse liver. Such int TCR cells also reside in other immune organs, although in low numbers. On the other hand, NK1.1+ T cells with int TCR do reside in the thymus and other peripheral organs. To determine the relationship of two types of cells, we characterized int CD3 cells and NK1.1+ T cells throughout the organs in terms of the phenotype, V beta repertoire, and morphology. Although both IL-2R beta+ T cells and NK1.1+ T cells are classified as int CD3 cells, NK1.1+ T cells are present within int CD3 cells. The majority of int CD3 cells in the liver and thymus were NK1.1+, whereas the minority of such cells in the spleen, lymph nodes, and bone marrow were NK1.1+. Among int CD3 cells, double-negative (DN) CD4-8- cells and/or CD4+ were abundant in NK1.1+ subset, whereas CD8+ cells were generally abundant in NK1.1- subset. Self-reactive V beta+ clones estimated by the M1s system were distributed to both NK1.1+ and NK1.1- subsets. High CD3 cells in the thymus and other organs contained neither DN cells nor forbidden clones. Int CD3 cells had the morphology of granular or agranular lymphocytes carrying perforin. Among int CD3 cells, NK1.1+ subset had a higher level of perforin-positive cells than NK1.1- subset. These results clearly demonstrate the relationship between int TCR cells and NK1.1+ T cells in various organs.

Animals↗

PubMed1995

Supportive cellular elements for hepatic T cell differentiation: T cells expressing intermediate levels of the T cell receptor are cytotoxic against syngeneic hepatoma, and are lost after hepatocyte damage.

Extrathymic T cells exist in the liver and are often seen in close contact with Kupffer cells in the hepatic sinusoids. Since selective depletion of Kupffer cells has become possible by using liposome-encapsulated clodronate, it was investigated whether elimination of Kupffer cells influences the level of extrathymic T cells in the liver. Extrathymic T cells were identified as interleukin-2 receptor beta-chain (IL-2R beta) intermediate TCR (TCRint) cells by two-color staining for CD3 or T cell receptor (TCR) and IL-2R beta. The elimination of Kupffer cells did not significantly affect levels of TCRint cells up to 7 days after treatment. We then examined monocyte colony stimulating factor (M-CSF)-deficient op/op mice (low levels of Kupffer cells). Extrathymic T cells both in the liver and spleen of these mice were detected at a level comparable to that of control mice. Since extrathymic T cells in the liver are sometimes located in the parenchymal space, the relationship between extrathymic T cells and hepatocytes was then examined. Electron microscopy revealed that some hepatic T cells adhered directly to hepatocytes. When hepatocytes were damaged by a single injection of CCl4, hepatocyte death and subsequent hepatic fibrosis were induced. Beginning 3 days after injection, CD3int cells, but not other type of cells, decreased prominently. Purified CD3int cells, as well as whole lymphocytes in the liver, were cytotoxic against syngeneic hepatoma. In parallel with the above-mentioned hepatic damage, the cytotoxic activity of lymphocytes against such targets was impaired in the liver. These results suggest that extra-thymic generation of TCRint cells and their acquisition of cytotoxic function are relatively independent of Kupffer cells, but are dependent on hepatocytes.

Animals↗

PubMed1995

Ultrastructural and time-lapse observations of intraepithelial lymphocytes in the small intestine of the guinea pig: their possible role in the removal of effete enterocytes.

In previous ultrastructural studies we have shown that at the tip of intestinal villi in guinea pigs, effete enterocytes are separated into two portions: a thin apical cytoplasm to be exfoliated into the lumen and a major basal portion to be ingested by lamina propria macrophages. During this process, intraepithelially disposed, large granular lymphocytes interdigitate with enterocytes in a complex manner. In the present study, the relation between the enterocytes and the lymphocytes in the villous epithelium of the guinea pig small intestine is described by use of transmission and scanning electron microscopy in an attempt to visualize the roles and activities of the lymphocytes more clearly. The lymphocytes project numerous pointed processes into effete enterocytes, even piercing them. Enterocytes are deeply indented or perforated, probably as a result of the encroaching lymphocyte processes. Some enterocytes are separated into apical and basal portions by numerous large excavations in the cytoplasm. These findings indicate that repeated perforating penetration of the lymphocytes induces cell cleavage. Supporting this supposition, our microcinematographic observations demonstrate the alternate protrusion and withdrawal of processes of lymphocytes. The processes advance with a pointed end, and subsequently, retract with a rounded end in a cycle of 8-18 seconds.

Animals↗

PubMed1994

Morphological analysis of multinucleated giant cells occurred in experimental autoimmune myocarditis.

Our previous study reported the rich existence of multinucleated giant cells in an autoimmune myocarditis experimentally induced in rats. The present study investigated the histochemical and ultrastructural characteristics of these giant cells. Histochemistry for an acid phosphatase clearly demonstrated multinucleated giant cells dispersed at the inflammatory foci. Ultrastructurally, the giant cells were shown to be single cells, but not clustered cells. Their ultrastructural characteristics were very similar to the basic features of macrophages, except that the giant cells were poor in lysosomes and phagosomes. It was noticeable that some macrophages possessed three or more nuclei, displaying an intermediate form between mononuclear macrophages and multinucleated giant cells. These findings suggest that the giant cell in the experimental autoimmune myocarditis is a single multinucleated cell, and possibly derived from macrophages by cell-to-cell fusion.

Acid Phosphatase↗