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PubMed1999

The oligosaccharidic component of the glycoconjugates in lichen planus, granuloma annulare, seborrheic keratosis and plamoplantar keratoderma: lectin histochemical study.

It is well known that cell surface glycoconjugates play an important role in cell proliferation, adhesion and differentiation. The aim of this investigation was to define the changes of the glycoconjugate saccharidic moieties in the epidermis and derma of patients affected by several skin pathologies such as seborrheic keratosis, lichen planus, granuloma annulare and palmoplantaris keratoderma. Bioptical specimens from skin lesions as well as from normal skin were fixed in Carnoy's fluid and routinely processed. The sections were treated with HRP-lectins (PNA, DBA, SBA, WGA, ConA, LTA and UEAI). Cytochemical controls were performed for specificity of lectin-sugar reaction. Some sections were pre-treated with neuraminidase prior to staining with HRP lectins. In comparison with normal human skin, epidermal lectin binding pattern in the considered diseases showed considerable qualitative and quantitative variations. In general, in all the considered pathologies, a lack and/or a decrease in lectin binding at the epidermal layers was observed; among the various diseases, differences in cellular localisation of the sugar residues were also noted. In such respect, an exception was represented by seborrheic keratosis, where the cells of the basal layer showed PNA reactivity, which was absent in the basal layer of the normal skin. Although seborrheic keratosis and lichen planus have been studied by others authors, our findings are not in total accordance concerning lectin binding; this is probably due to the different fixatives employed. Our findings seem to reveal significant changes in keratinocyte glycoconjugate oligosaccharides in the previously mentioned diseases, providing clues to their pathogenesis.

Concanavalin A↗
PubMed1997

Development of interglobular dentine in rat molars and its relation to maturation of enamel.

The development of interglobular dentine in the first upper and lower molars of Wistar rats aged 3, 7, 14, 21, 42 days was examined histochemically using a lectin, succinyl wheat germ agglutinin (sWGA), which is specific for N-acetyl-D-glucosamine. sWGA stained the interglobular dentine, predentin and Golgi area of odontoblasts. Interglobular dentine was not formed in the first molars of 3-day rats, but appeared in those of 7-day rats near the enamel-free area. In 14-day rats, interglobular dentine was present in most areas of the coronal dentine except the cervical area. At the interface between dentine and predentin, numerous sWGA-negative calcospherites were seen, suggesting that the interglobular dentine is formed actively there. In 21-day rats, the interlobular dentine was more numerous than in 14-day rats. Interglobular dentine was present in the cervical root dentine as well as in the coronal dentine, including the cervical area. The distribution of interglobular dentine in 42-day rats was similar to that in 21-day rats, but fluorescence of sWGA binding was less intense in the former. Because the development of interglobular dentine appeared to be time and position specific its relation to the stages of ameloblasts was analysed. Thin enamel matrix was formed at cusps in molars of 3-day rats and thickness of enamel matrix increased in 7-day rats. In these teeth, the ameloblasts were at the differentiating or secretory stage. The Golgi area and Tomes' processes of the secretory ameloblasts, the cells of intermediate layer and the enamel matrix were weakly positive with sWGA. The epithelial cells at the enamel-free area were also stained with sWGA. In 14-day rats, most of the ameloblasts in the first maxillary molars were at the maturative stage except in the cervical area, where the ameloblasts were at the transitional stage. sWGA stained the distal border and the Golgi area of the maturative ameloblasts as well as the cells of the papillary layer. The distal border of the maturative ameloblasts appeared either thick or thin, suggesting a ruffle-end and smooth-end of the cells. Ameloblasts were absent in the first molars of 21-day rats and the cervical part of the enamel was covered with the stratified epithelium like that of 42-day rats. The present study has demonstrated that interglobular dentine contains sWGA-binding glycoconjugates and the formation of the interglobular dentine is largely associated with the enamel maturation. These results suggest that matrix-to-cell interaction is important for the development of interglobular dentine.

Acetylglucosamine↗
PubMed2002

Conservation of Brachyury, Mef2, and Snail in the myogenic lineage of jellyfish: a connection to the mesoderm of bilateria.

One major difference between simple metazoans such as cnidarians and all the bilaterian animals is thought to involve the invention of mesoderm. The terms diploblasts and triploblasts are therefore, often used to group prebilaterian and bilaterian animals, respectively. However, jellyfish contain well developed striated and smooth muscle tissues that derive from the entocodon, a mesoderm-like tissue formed during medusa development. We investigated the hypothesis, that the entocodon could be homologous to the third germ layer of bilaterians by analyzing the structures and expression patterns of the homologues of Brachyury, Mef2, and Snail in the jellyfish Podocoryne carnea. These are regulatory genes from the T-box, MADS-box and zinc finger families known to play important roles in bilaterian mesoderm patterning and muscle differentiation. The sequence and expression data demonstrate that the genes are structurally and functionally conserved and even more similar to humans or other deuterostomes than to protostome model organisms such as Drosophila or Caenorhabditis elegans. Based on these data we conclude that the common ancestor of the cnidarians and bilaterians not only shared genes that play a role in regulating myogenesis but already used them to develop and differentiate muscle systems similar to those of triploblasts.

Animals↗
PubMedDate not supplied

The distribution and morphology of LGN K pathway axons within the layers and CO blobs of owl monkey V1.

The lateral geniculate nucleus (LGN) of primates contains three classes of relay cells, the magnocellular (M), parvocellular (P), and koniocellular (K) cells. At present, very little is known about either the structure or function of the K relay cells in New or Old World monkeys (simian primates). In monkeys, K cells are located between the main LGN layers and adjacent to the optic tract. For convenience, these intercalated cell layers are numbered K1-K4 starting closest to the optic tract with K1. The objective of this study was to examine the details of K axon morphology in the primary visual cortex (V1) of owl monkeys and to determine if different K layers give rise to distinct axon types. For this purpose, injections of WGA-HRP or PHA-L were made into specific K LGN layers and the distribution and morphology of the resulting labeled axons were analyzed. Injections of fluorescent tracers also were made within the superficial layers of V1 to further document connections via analysis of the patterns of retrogradely labeled cells in the LGN. Our main finding is that K axons in owl monkeys terminate as delicate focused arbors within single cytochrome oxidase (CO) blob columns in cortical layer III and within cortical layer I. Overall, the morphology of the K axons in these monkeys is quite similar to what we described previously for K geniculocortical axons in the distantly related bush baby (prosimian primate), suggesting that the basic features of this pathway are common to all primates. Our results also provide evidence that the axon arbors from different K layers are morphologically distinct; axons from LGN layer K1 project mainly to cortical layer I, while axons from LGN layer K3 chiefly terminate in cortical layer III. Taken together, these results imply that the basic features of axons within the K pathway are conserved across primates, and that the K axons from different K layers are likely to differ in function based upon their different morphologies.

Animals↗
PubMedDate not supplied

[T-Box genes and developmental decisions that cells make].

During gastrulation in vertebrate embryos, three definitive germ layers (ectoderm, mesoderm, and endoderm) are formed by organized and coordinated cell movements. In zebrafish, further subdivision of the mesoderm gives rise to the axial, adaxial and paraxial mesoderm. The axial mesoderm contributes to the prechordal plate and notochord whereas the adaxial and paraxial cells give rise to slow and fast muscles, respectively (Devoto et al., 1996; Blagden et al., 1997; Currie and Ingham, 1998). An inductive interaction in which the notochord plays an essential role will also provide an input in forming other specialized types of tissue contributing to the axial structures: the floor plate located dorsally to the notochord in the ventral spinal cord and the hypochord located ventrally of the notochord and deriving probably from the endoerm. It is known that despite the difference in developmental roles (Strähle et al., 1993; Krauss et al., 1993), the floor plate and hypochord co-express a number of common molecular markers (Jan et al., 1995; our unpublished results) that may illustrate a certain similarity of their origin. Their close proximity to the notochord determines specialized features of these structures that differ substantially from the rest of the neural tube and endoderm, correspondingly. Once formed under the influence of the notochordal signaling, the floor plate will acquire an ability, similar to the notochord, to express genes of the Hedgehog family and several other groups of genes and to induce specification of ventral cell types in the neural tube during later development (for review, see Korzh, 1998). The biology of the hypochord is much less understood. It seems that the hypochord develops slightly later than the floor plate. It may be required for proper positioning of the dorsal aorta as well as induction of some other endoderm derivatives.

Animals↗
PubMedDate not supplied

Localization of androgen and estrogen receptors in adult male mouse reproductive tract.

There is considerable variation, both within and between species, in reports of nuclear steroid receptor localizations in the male reproductive tract. In this study, androgen receptor (AR) and estrogen receptors ERalpha and ERbeta were visualized by immunohistochemistry in adult male mice reproductive tracts, including testes, efferent ductules; initial segment, caput, corpus, and cauda epididymides; and vas deferens. Antibody specificity was demonstrated by Western blot and antibody competition. In testis, AR was expressed in Leydig cells, Sertoli cells, and most peritubular cells, but not in germ cells; Sertoli cells showed more intense staining in stages VI-VII; ERalpha was present in Leydig and some peritubular cells; ERbeta was in Leydig, some peritubular, all Sertoli and germ cells except in spermatids and meiotic spermatocytes. In efferent ductules, AR was strongly expressed in ciliated and nonciliated epithelial cells and in stromal cells; ERalpha was strongly expressed in ciliated and nonciliated epithelial cells; stromal cells were negative; and ERbeta was strongly expressed in ciliated and nonciliated epithelial cells and also in stromal cells. In epididymis, AR was strongly expressed in all epithelial cells (not in intraepithelial lymphocytes); ERalpha was strongly expressed in apical, narrow, and some basal cells of the initial segment, and in caput, principal cells of the caput, clear cells of the distal caput through cauda; stromal cells were negative in the initial segment, but more stromal cells were stained from caput to cauda; ERbeta was strongly expressed in most of epithelial cells of the epididymis, but stromal cells were inconsistently stained. In vas deferens, AR was weakly expressed or absent in principal cells but moderately stained in basal cells, smooth muscle cells of stroma were stained intensely, ERalpha was absent in epithelial cells but present in a subepithelial smooth muscle layer, and ERbeta was strongly expressed in all epithelial cells and most stromal cells. This study demonstrates that the reproductive tracts of male mice differ considerably from those of rats in expression of ARs and ERs and that caution is needed when extrapolating nuclear steroid receptor data across mammalian species.

Animals↗
PubMed1993

Heterogenous distribution of glycoconjugates in the kidney of dogfish Scyliorhinus caniculus (L.) with reference to changes in the glycosylation pattern during ontogenetic development of the nephron.

Eight fluorochrome-coupled lectins with different sugar specificities were applied to cryosections of dogfish kidney. Despite profound differences in renal architecture between elasmobranch fish and other vertebrates, the sequence of nephron segments as revealed by the lectin-binding pattern was rather similar to that of tetrapodes. Wheat germ agglutinin (WGA) bound to cell membranes of epithelial cells of glomeruli, proximal and distal tubules, their basement membranes, the collecting tubule, and epithelial cells. Among other broadly binding lectins were Ricinus communis agglutinin I (RCA-I), soybean agglutinin (SBA), peanut agglutinin (PNA), Lycopersicon esculentum agglutinin (LEA), and Jacalin, all of which marked proximal as well as distal portions of the renal tubule. Dolichos biflorus agglutinin (DBA) did not react with any renal structure. Ulex europaeus agglutinin I (UEA-I), which indicates the presence of alpha-L-fucose, very strongly and specifically marked single epithelial cells of the early distal nephron, all epithelial cells of the late distal tubule, the beginning of the collecting tubule in the mesial tissue zone, and single cells in the end portion of the collecting tubule in the lateral bundles. Binding of UEA-I to receptors of distal nephron cells could be useful for the identification of these cells in functional studies employing teased tubule and/or isolated cell preparations. Binding of UEA-I to dogfish kidney structures resembles staining with UEA-I conjugates of late distal tubules and collecting tubules in the kidneys of frog and other, higher vertebrates. Epithelial cells of early developmental stages showed, very rarely, binding sites for most lectin-fluorochrome conjugates. A large number of lectin binding sites was observed in the extracellular matrix of fibroblast layers surrounding the early anlage and the S-shaped body. Lectin binding sites of the nephron epithelia appeared in a sequential manner in the next stages of development of the nephron. Ontogenetic and phylogenetic aspects of the merging region between nephron proper (late distal tubule) and collecting system (collecting tubule) are discussed.

Animals↗
PubMed1998

The specificity of gap junctional channels in early fish embryos and its significance for pattern formation in development.

The data obtained support the hypothesis that cells of developing embryos use cell-to-cell communication via gap junctions (GJs) for the transmission of signals participating in the pattern formation of gene expression. It was revealed that a ring area of the early mesoderm gene activation in loach (Misgurnus fossilis) embryos, which lies in the proximity of the yolk cell (YC), is simultaneously a region of the increased dye transfer between the blastomeres and the YC for fluorescein (F) and Lucifer Yellow (LY). In search for the reason of such a heterogeneity of dye transfer, it was found that the permeability of GJs between the blastomeres increased from the early through mid-blastula stage. In parallel, the adhesiveness of the blastomeres also increased, that, apparently, directs the GJ uncoupling in the central region of the blastoderm basal layer. More important is that the changes in the permeability of the GJs between the YC and the blastomeres located in the presumptive mesoderm area were accompanied by changes in GJ selectivity. By comparing the kinetics of the transfer of the fluorescent substances of opposite charges (LY and F as anions and 4', 6-diamidino-2-phenylindole (DAPI) as a cation), we found that all GJs at the mid-blastula stage and GJs of the blastomeres located in the central area of the basal layer at the early gastrula stage were permeable for each dye, but the permeability for the cationic dye was slightly lower than for the anionic ones. At the early gastrula stage, GJs of a germ ring area (prospective mesoderm region) acquire much higher anionic selectivity; they become more permeable for LY and F but virtually impermeable for DAPI. These results suggest that embryonic cells can regulate the selectivity of GJ channels and control the transport of the secondary messengers based on their electrochemical properties.

Animals↗
PubMedDate not supplied

Glutamate containing neurons in the cat superior colliculus revealed by immunocytochemistry.

Glutamate is the probable neurotransmitter of both retinal and cortical afferents to the cat superior colliculus (SC). The present study shows that glutamate is also contained in many postsynaptic neurons in SC. The distribution, morphology, and ultrastructure of neurons in SC were examined using glutamate antibody immunocytochemistry. Labeled cells were widely distributed throughout, but a specific laminar pattern was evident. Relatively few cells were found in the zonal and upper superficial gray layers (SGL). A dense band of intensely labeled neurons was found within the deep superficial gray and upper optic layers. Many cells were also labeled in the deeper layers. Labeled cells had varied sizes and morphologies. Soma diameters ranged from 9-67 microns, with a mean of 22 microns. Cells with stellate, vertical fusiform, and multipolar morphologies were labeled. Cells in the deep subdivision all had morphologies and sizes typical of projection neurons. To determine if labeled cells in the dense band were also projection neurons, WGA-HRP was injected into the lateral posterior nucleus and these sections were double-labeled with the glutamate antibody. Over one-half of cells in the dense band that were labeled by HRP were also obviously labeled by antibody. At the electron-microscope level, both medium- and large-sized neurons were also labeled by glutamate antibodies. These cells had different but characteristic morphologies.

Animals↗
PubMed1983

Variation in corn (Zea mays L.) for fatty acid compositions of triglycerides and phospholipids.

The percentage of linoleic acid in corn germ oil of three crosses, C103D x B73, C103D x B84, and T220 x H51, and their reciprocals was investigated. Corn germ oil from F2, F3, and backcrossed generations was also examined. More than one gene locus appeared to be involved in conditioning the linoleic acid content in these crosses. Strong maternal effects were exhibited in the F1's. Genotype also superimposed variations in fatty acid compositions within the characteristic lipid class patterns of the phospholipids, phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol. Fatty acid placements in triglycerides, digalactosyldiglycerides, and phospholipids of one inbred, H51, were determined by lipase and phospholipase hydrolysis. The overall pattern of placement showed that the fatty acids at the 1 position were predominantly saturated saturated and those at the 2 position were predominantly unsaturated, but the fatty acid distribution was different for each individual lipid class. The molecular species of the phosphatidylcholines and phosphatidylethanolamines were separated by silver nitrate thin-layer chromatography. The major differences in the molecular species were a higher level of the dienoic-dienoic species and a lower level of the monoenoic-monoenoic species in the phosphatidylethanolamines than in the phosphatidylcholines.

Crosses, Genetic↗
PubMed1996

Evidence for a thymus-dependent form of tolerance that is not based on elimination or anergy of reactive T cells.

The avian embryo has provided an appropriate model to study the ontogeny of the primary lymphoid organs, thymus and bursa of Fabricius. By using the quail-chick marker system the embryonic origin of the highly intricate cell components which form these organs could be traced back to the initial endodermal, mesodermal and ectodermal germ layers. The timing and dynamics of the incoming and outcoming flows of hemopoietic cells which characterize their lymphopoietic activity could be revealed in both quail and chick embryos. This knowledge served as a basis for an investigation on the role of the epithelial component of the thymus (derived from the pharyngeal endoderm) on tolerance to tissue graft and, by extension, tolerance to self. When this work was undertaken, the prevailing view was that exposure of the developing immune system to foreign antigens in the embryo allows them to be assimilated to self components in the mature animal. In fact, this was found to be true for allogeneic grafts between MHC-distinct chickens, of certain tissues, such as for instance wing tissues. However, in heterospecific transplantations, i.e. when a limb bud was grafted from quail to chick embryos, the chick host acutely rejected the foreign limb soon after birth. In contrast, grafts of the quail thymic epithelial (TE) rudiment resulted in the development of a chimeric thymus in which the foreign epithelial component was not only tolerated but able to induce full tolerance of the grafted wing from the same donor. By monitoring the amount of quail TE implanted we showed in addition that only part of the peripheral T-cell population had to differentiate in the context of the quail epithelial cells to induce tolerance to quail tissues. This pointed to the generation in the thymus of regulatory T cells, coexisting with specific anti-quail reactive T cells, but able to inhibit them from reacting against the quail wing antigenic determinants. A mammalian model was then devised to further study this mechanism of tolerance that we have qualified as "dominant" by opposition to the current model based on either clonal elimination or anergy which can be considered as recessive or passive. Nude mice of MHC type A were grafted with TE of E10 type B embryos. They became reconstituted for T-cell function but tolerant for B skin allografts. Spleen cells from such tolerant animals injected to naive A nude mice reconstituted T cell function in the recipient and transferred the tolerance to B skin grafts. Reducing the number of donor cells resulted in the segregation of the two phenomena. For low numbers the recipients were restored but not tolerant, thus showing the coexistence in the tolerant donor of anti-B reactive T cells together with regulatory cells able to abolish their reactivity against B determinants. Other experiments demonstrated that TE-induced tolerance does not rely on clonal deletion or anergy. This was shown on systems where elimination of cells directed toward superantigens was screened. It turned out that tolerance to skin grafts and superantigen T-cell deletion are unrelated phenomena. These observations strongly suggest that tolerance to self results at least in part from the interplay between cells potentially harmful for self component and others which exert a strong control on their reactivity. The latter cell type depends upon interactions of thymocytes with the endodermal component of the thymus.

Animals↗
PubMed1992

Anticonvulsant role of nigrotectal projection in the maximal electroshock model of epilepsy--II. Pathways from substantia nigra pars lateralis and adjacent peripeduncular area to the dorsal midbrain.

Lesion evidence suggests that the superior colliculus is essential for mediating the anticonvulsant properties of nigral suppression in the electroshock model of epilepsy. However, our companion paper [Redgrave et al. (1991) Neuroscience 46, 379-390] established that the region of dorsal midbrain where bicuculline was most effective in suppressing tonic hindlimb extension did not correspond well with the known distribution of nigrotectal terminals. The purpose of the present anatomical study was, therefore, to investigate in more detail ventral midbrain connections to the dorsal midbrain anticonvulsant zone in rat. Small injections (10-20 nl) of a 1% solution of wheatgerm agglutinin conjugated with horseradish peroxidase were made specifically into the region of dorsal midbrain where bicuculline was maximally effective. Numerous retrogradely labelled cells were found in substantia nigra pars lateralis and adjacent peripeduncular area but not in substantia nigra pars reticulata. Retrogradely labelled cells were also located in ventral zona incerta. When wheatgerm agglutinin-horseradish peroxidase injections were made into lateral substantia nigra, a region of anterogradely transported reaction product characteristic of nerve terminals was observed in the caudolateral deep layers and underlying reticular tissue; this area corresponded well to the dorsal midbrain anticonvulsant zone. These data suggest that, in the electroshock model of epilepsy, direct connections between substantia nigra pars lateralis and adjacent peripeduncular area and the dorsal midbrain anticonvulsant zone could be critical for mediating the anticonvulsant properties previously attributed to substantia nigra pars reticulata. During the course of this study, anterograde projections from substantia nigra pars lateralis and adjacent peripeduncular area to both superficial and intermediate layers of the ipsilateral superior colliculus were noted. Additional experiments using retrograde transport of the fluorescent tracer Fast Blue confirmed these projections.

Amidines↗
PubMed1997

Spatially distinct domains of cell behavior in the zebrafish organizer region.

To determine the sequence of cell behaviors that is involved in the morphogenesis of the zebrafish organizer region, we have examined the dorsal marginal zone of vitally stained zebrafish embryos using time-lapse confocal microscopy. During the late-blastula stage, the zebrafish dorsal marginal zone segregates into several cellular domains, including a group of noninvoluting, highly endocytic marginal (NEM) cells. The NEM cell cluster, which lies in a superficial location of the dorsal marginal zone, is composed of both enveloping layer cells and one or two layers of underlying deep cells. The longitudinal position of this cellular domain accurately predicts the site of embryonic shield formation and occupies a homologous location to the organizer epithelium in Xenopus laevis. At the onset of gastrulation, deep cells underneath the superficial NEM cell domain undergo involution to form the nascent hypoblast of the embryonic shield. Deep cells within the NEM cell cluster, however, do not involute during early shield formation, but instead move in front of the blastoderm margin to form a loose mass of cells called forerunner cells. Forerunner cells coalesce into a wedge-shaped mass during late gastrulation and eventually become overlapped by the converging lateral lips of the germ ring. During early zebrafish tail elongation, most forerunner cells are incorporated into the epithelial lining of Kupffer's vesicle, a transient teleostean organ rudiment long thought to be an evolutionary vestige of the neurenteric canal. Owing to the location of NEM cells at the dorsal margin of blastula-stage embryos, as well as their early segregation from other deep cells, we hypothesized that NEM cells are specified by an early-acting dorsalizing signal. To test this possibility, we briefly treated early-blastula stage embryos with LiCl, an agent known to produce hyperdorsalized zebrafish embryos with varying degrees of expanded organizer tissue. In Li(+)-treated embryos, NEM cells appear either within expanded spatial domains or in ectopic locations, primarily within the marginal zone of the blastoderm. These results suggest that NEM cells represent a specific cell type that is specified by an early dorsal patterning pathway.

Animals↗
PubMed1996

Occurrence of poly(alpha2,8-deaminoneuraminic acid) in mammalian tissues: widespread and developmentally regulated but highly selective expression on glycoproteins.

In tissues of higher organisms homopolymers of alpha2,8-linked N-acetylneuraminic acid can be found as a posttranslational modification on selected proteins. We report here the discovery of homopolymers of alpha2,8-linked deaminoneuraminic acid [poly(alpha2,8-KDN)] in various tissues derived from all three germ layers in vertebrates including mammals. The monoclonal antibody kdn8kdn in conjunction with a bacterial KDNase permitted the detection of poly(alpha2,8-KDN) by immunohistochemistry and immunoblotting. Further evidence for the existence of poly(alpha2,8-KDN) was obtained by gas/liquid chromatography. The poly(alpha2,8-KDN) glycan was detectable in all tissues studied with the exception of mucus-producing cells present in various organs, the extracellular matrix, and basement membranes. However, in certain organs such as muscle, kidney, lung, and brain its expression was developmentally regulated. Despite its widespread tissue distribution, the poly(alpha2,8-KDN) glycan was detected on a single 150-kDa glycoprotein except for a single >350-kDa glycoprotein in kidney, which makes it most distinctive among polysialic acids. The ubiquitous yet selective expression may be indicative of a general function of the poly(alpha2,8-KDN)-bearing glycoproteins.

Animals↗
PubMed1987

Specific binding sites for transferrin on ameloblasts of the enamel maturation zone in the rat incisor.

During enamel maturation in rodents, an iron-containing pigment is deposited into the surface layer of the enamel. Maturation zone ameloblasts presumably are responsible for this deposition. The presence of large amounts of ferritin in the cytoplasm of these cells suggests that they receive iron, presumably from circulating transferrin. An in vivo radioautographic binding assay using iodinated transferrin was used to determine if indeed maturation ameloblasts possess transferrin receptors at their cell surfaces. Experimental rats received systemic injections of labeled transferrin while control rats received injections of labeled transferrin plus a large excess of unlabeled transferrin in order to compete with the labeled transferrin for available specific receptors. Light microscope radioautography showed that ruffle-ended ameloblasts (RAs) of the enamel maturation zone had a high density of specific receptors for transferrin relative to smooth-ended ameloblasts (SAs). Electron microscopy and energy-dispersive X-ray spectroscopy confirmed the presence of ferritin and iron, respectively, within these cells. It is postulated that the iron responsible for enamel pigmentation is transported by transferrin to maturation ameloblasts and is bound to specific transferrin receptors found mostly on RAs and that the modulation of these cells into SAs results in a loss of most of these receptors.

Ameloblasts↗
PubMed1993

Calretinin immunoreactivity in the monkey hippocampal formation--II. Intrinsic GABAergic and hypothalamic non-GABAergic systems: an experimental tracing and co-existence study.

Our light and electron microscopic studies (Seress L., Nitsch R. and Leranth C. (1993) Neuroscience 55, 775-796.) indicated that in the hippocampus of the African Green monkey, calretinin is exclusively present in non-pyramidal cells. Calretinin-positive axons formed a prominent band at the border of the dentate molecular and granule cell layers and in the pyramidal layer of CA2, and established asymmetric synapses with different postsynaptic targets. The goal of this study is to determine the cells of origin of this presumably extrinsic innervation, and subsequently, the characterization of their neurochemical features. We were able to demonstrate that calretinin-immunoreactive axon terminals in the inner molecular layer of the dentate gyrus and in the pyramidal layer of CA2 disappear 10 days after fimbria-fornix transection. Retrograde tracing revealed their cells of origin to be in the supramammillary nucleus. Co-localization studies employing the cryostat consecutive, semithin section technique provided evidence that these large projecting neurons contained both calretinin and substance-P but lack GABA as an inhibitory transmitter. In contrast, co-localization studies revealed that almost all of the intrinsic calretinin-positive neurons in different areas of the primate hippocampus contained GAD or GABA. These results suggest that there are two separate calretinin-containing systems in the primate hippocampus, i.e. an intrinsic inhibitory and an extrinsic excitatory one, the latter deriving from the supramammillary nucleus of the hypothalamus.

Animals↗
PubMed1995

Teeth and tooth nerves.

(1) Although our knowledge on teeth and tooth nerves has increased substantially during the past 25 years, several important issues remain to be fully elucidated. As a result of the work now going on at many laboratories over the world, we can expect exciting new findings and major break-throughs in these and other areas in a near future. (2) Dentin-like and enamel-like hard tissues evolved as components of the exoskeletal bony armor of early vertebrates, 500 million years ago, long before the first appearance of teeth. It is possible that teeth developed from tubercles (odontodes) in the bony armor. The presence of a canal system in the bony plates, of tubular dentin, of external pores in the enamel layer and of a link to the lateral line system promoted hypotheses that the bony plates and tooth precursors may have had a sensory function. The evolution of an efficient brain, of a head with paired sense organs and of toothed jaws concurred with a shift from a sessile filter-feeding life to active prey hunting. (3) The wide spectrum of feeding behaviors exhibited by modern vertebrates is reflected by a variety of dentition types. While the teeth are continuously renewed in toothed non-mammalian vertebrates, tooth turnover is highly restricted in mammals. As a rule, one set of primary teeth is replaced by one set of permanent teeth. Since teeth are richly innervated, the turnover necessitates a local neural plasticity. Another factor calling for a local plasticity is the relatively frequent occurrence of age-related and pathological dental changes. (4) Tooth development is initiated through interactions between the oral epithelium and underlying neural crest-derived mesenchymal cells. The interactions are mediated by cell surface molecules, extracellular matrix molecules and soluble molecules. The possibility that the initiating events might involve a neural component has been much discussed. With respect to mammals, the experimental evidence available does not support this hypothesis. In the teleost Tilapia mariae, on the other hand, tooth germ formation is interrupted, and tooth turnover ceases after local denervation. (5) Prospective dental nerves enter the jaws well before onset of tooth development. When a dental lamina has formed, a plexus of nerve branches is seen in the subepithelial mesenchyme. Shortly thereafter, specific branches to individual tooth primordia can be distinguished. In bud stage tooth germs, axon terminals surround the condensed mesenchyme and in cap stage primordia axons grow into the dental follicle.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed2004

The influence of different preservation and sterilisation steps on the histological properties of amnion allografts--light and scanning electron microscopic studies.

Despite thorough donor screening and preparation under aseptic conditions, conventional methods of preservation do not exclude the probability of a contamination with pathogenic germs. The purpose of this study was to investigate the changes of histological parameters of amnion transplants (ATs) through different methods of sterilisation and preservation. Therefore 10 different procedures for sterilisation and preservation of ATs were described. Specimens of each group were studied using different histological procedures such as light microscopy and scanning electron microscopy. General staining (Haematoxylin-eosin stain, periodic-acid-Schiff, Domack) and immunohistochemical methods have been applied in order to gain additional information concerning the structure of the amniotic epithelium and the basement membrane but also the distribution of collagens and intermediate filaments. Furthermore, the measurement of the ATs thickness was included in order to study the influence of the manufacturing procedures to this property. As a result we found that the histological appearance of the ATs is closely related to the applied sterilisation and preservation procedures. Although the basement membrane remained intact, especially the amniotic epithelium was partially destroyed by irradiation sterilisation. Further, the dissolution of the connective tissue layers into single fibre bundles was clearly visible. Procedures with and without peracetic acid sterilisation (PAA) preserved the tissue structure. Our results showed a significant variation in the tissue's thickness after different preservation procedures. Air- and freeze-dried ATs were found to be the thinnest tissues varying from 20 to 30 microm, the thickest ATs preserved in glycerol varied from 45 to 50 microm. Because ATs showed a preserved tissue structure after PAA sterilisation it can be recommended as an alternative for methods previously described in literature. Depending on the specific use of the AT one may choose from thinner or thicker allografts.

Amnion↗