Yasutoku Kogaya

Immunohistochemical localisation of amelogenin-like proteins and type I collagen and histochemical demonstration of sulphated glycoconjugates in developing enameloid and enamel matrices of the larval urodele ( Triturus pyrrhogaster ) teeth

The presence of collagen in enameloid distinguishes it clearly from true enamel, but little is known about the phylogenetic relationship between these 2 tissues. It has previously been reported that amelogenins are the principal proteins of the enamel matrix, that type I collagen and chondroitin sulphates are the predominant matrices in dentine, and that amphibian and reptilian aprismatic enamels contain no sulphated glycoconjugates, although certain sulphated substances are secreted into mammalian prismatic enamel during matrix formation. The larval urodele (Triturus pyrrhogaster) teeth are known to be composed of enameloid, dentine, and enamel‐like tissue. To characterise the tooth matrices, the localisation of amelogenin‐like proteins, type I collagen, and sulphated glycoconjugates was investigated. Chondroitin sulphates and fine fibrils immunoreactive for type I collagen were elaborated as the enameloid matrix inside the dental basement membrane. After the matrix had been deposited in full thickness, coarse collagen fibrils also immunoreactive for type I collagen and chondroitin sulphates were deposited below as the first dentine matrix. Further, enamel‐like matrix with no collagen fibrils or sulphated glycoconjugates but strongly immunoreactive for amelogenins was deposited on the dentine. Although no immunolabelling for amelogenins was found over the enameloid matrix, at least at the formation stage, the zone of coarse collagen fibrils of dentine was partially immunoreactive as observed in mammalian mantle dentine. From the ontogeny and matrix constituents of larval urodele teeth, it is suggested that enameloid is originally a dentine‐like tissue.

Ultrastructural distribution of acidic glycosaminoglycans associated with matrix vesicle-mediated calcification in mouse progenitor predentine

SummaryThe ultrastructural localization of acidic glycosaminoglycans, presumed to be proteoglycans, was examined during initial matrix vesicle-mediated calcification in dentine, by using ruthernium red (RR) staining, high iron diamine thiocarbohydrazide silver proteinate (HID-TCH-SP) staining, and an enzymatic digestion method. Progenitor predentine 2–10 μm width of developing mouse molar tooth germs was used throughout the present study. The outer surface membrane of the intact matrix vesicles had a strong affinity for RR. The RR positive materials appeared beaded and extended perpendicularly from the vesicle membrane. They tended to disappear with the disruption of the vesicular membrane, which resulted from overextension due to needle-, crystallike structures. The HID-TCH-SP stain deposits, approximately 10 nm in diameter, were densely distributed around the intact matric vesicles, though few were found inside them. Some matrix vesicles that were presumably disrupted, however, contained smaller stain deposits. On the outer surface membrane of the disrupted vesicles, HID-TCH-SP stain deposits were fewer in number. The results obtained from enzymatic degradation studies showed that the anionic materials on the outer surface membrane of the matrix vesicles were represented by chondroitin-4-sulfate and/or chondroitin-6-sulfate. We suggest that chondroitin sulfates attached to the outer leaflet of the vesicular membrane play an important role during the incipient stage of the matrix vesicle-mediated calcification process.

True enamel matrix of the newt, Triturus pyrrhogaster, contains no sulfated glycoconjugates

SummaryThe ultrastructural distibution and histochemical properties of sulfated glycoconjugates were investigated in the developing enamel of the adult newt, Triturus pyrrhogaster, by use of the high-iron diamine thiocarbohydrazide silver proteinate (HID-TCH-SP) staining and enzymatic digestion methods. Development and ultrastructure of the enamel were also studied. After deposition of the mantle dentin matrix to a certain thickness, the first enamel matrix, globular in shape, appeared in juxtaposition to the dental basement membrane and tended to be intermixed with the previously deposited dentin matrix. Subsequently, enamel matrix was deposited outside (ameloblastic side) of the dental basal lamina and formed a true enamel layer. Thus, developing enamel of the newt consists of two layers: (1) an inner layer made up of a dentin-enamel mixed matrix and (2) an outer layer composed of only true enamel matrix. HID-TCH-SP precipitates resulting from the abovementioned studies were found in the mixed matrix and were identified as chondroitin sulfates; in contrast, the true enamel matrix contained no sulfated glycoconjugates.

The differences in calcium distribution pattern between preodontoblasts and preameloblasts in developing rat molar tooth germs

SummaryUltrastructural localization of calcium in preodontoblasts and preameloblasts was investigated using the potassium pyroantimonate technique, and it was confirmed that there were clear differences in calcium distribution pattern between preodontoblasts and preameloblasts. In preodontoblasts, pyroantimonate reaction products were mainly observed in the Golgi region, lateral intercellular spaces, and secretory granules, especially in the distal portion of cell body; however, few were found in mitochondria and on the plasma membrane. In preameloblasts, on the other hand, the precipitates were located in mitochondria, nuclei, and on the inner face of the plasma membrane; however, few reaction products were observed in the intercellular spaces, lysosomelike granules, secretory granules, and stippled materials. Granular reaction product approximately 20–40nm in diameter adhered preferentially to the growing end of needlelike crystals in the initial enamel matrix.

Ultrastructural Localization of Calcium in Matrix Vesicles and Preodontoblasts of Developing Rat Molar Tooth Germs during Initial Dentinogenesis

We investigated the ultrastructural localization of calcium in progenitor predentine and preodontoblasts of developing rat molar tooth germs using the potassium pyroantimonate technique. At the precalcification stage, antimonate reaction product was sparsely, randomly distributed in the preodontoblasts and in the progenitor predentine but no significant reaction could be noticed associated with matrix vesicles. At the matrix vesicle calcification stage, large amounts of antimonate reaction product tended to be localized in the region adjacent to the distal, outer surface membrane of preodontoblasts in which moderate antimonate reaction activity could be observed in mitochondria. Strong antimonate reaction was detected preferentially on the outer surface membrane of some matrix vesicles at this stage. At the subsequent collagen calcification stage, definite antimonate reaction was no longer seen within mitochondria of the late preodontoblasts, instead precipitate was mainly distributed in Golgi area, secretory granules and lateral intercellular spaces. It is suggested that although matrix vesicles contain few calcium capable of reacting to antimonate immediately after their biogenesis, subsequently, large amounts of calcium are accumulated associated with the outer surface membrane of matrix vesicles in the extracellular matrix.

Partial albinism in the Japanese shrew mole, Urotrichus talpoides, from Aichi, Japan

Abstract We describe partial albinism (a white-belted coloration) in the Japanese shrew mole (Urotrichus talpoides) from Aichi Prefecture, Japan. This shrew-mole did not present symptoms of generalized types of vitiligo vulgaris. The white-belted coloration may have occurred because of a lack of melanocyte formation due to a deficit of or to abnormalities in neural crest cells during ontogeny.

The Ganoine Membrane of the Scales of Polypterus senegalus Includes a Labyrinth-Like Structure

The ganoine membrane is a basement membrane-like structure appearing between the maturation stage inner ganoine epithelium (IGE) and mineralized ganoine of polypterids and lepisosteids. In the present study, further ultrastructural investigation of the ganoine membrane of Polypterus senegalus was made and it was confirmed that it possessed a specialized labyrinth-like structure. The complete ganoine membrane was composed of six distinct zones: (1) a filamentous zone facing the distal plasmalemma of IGE, (2) lamella zone I, (3) vertical striation zone, (4) lamella zone II, (5) electron-dense zone, (6) electron-lucent zone directly facing the maturing ganoine surface. The ganoine membrane sometimes partially lacked some of these zones, but the electron-dense zone was usually present. Oblique or horizontal sections of the membrane showed that the vertical striation zone was made up of a typical labyrinth-like structure. Certain sulfated glycoconjugates were localized at the filamentous zone, especially adjacent to the lamella zone I and over the electron-dense zones.