Objective: The aim of this immunocytochemical study was to characterize the expression and distribution of the progesterone receptor (PR) and estrogen receptor (ER) in gingival fibroblasts using culture cells derived from people at various ages. Background: The reaction of female hormones is tissue or cell specific, and receptor availability in the cell is one of the major causes for the different reactions. Gingiva is a target tissue for female hormones; however, the characteristics of PR and ER in both the fibroblasts and the other component cells remain largely unknown. Materials: Gingival tissue was obtained from six people at various ages and culture fibroblasts were established. At least three passages of each cell line were strained for PR and ER with monoclonal antibodies (Clone 1A6, Clone 1D5, respectively). Results: PR positive cells were detected in all six cell lines through early passages to late ones, but ER were only observed in two of six samples with faint reactions. The staining intensity for PR was greater than for ER, but less than that shown in the MCF‐7 breast cancer cells, positive control. In every positive control test, ER reactivity was equal to or higher than that of PR. During the interphase, significantly fewer positive fibroblasts occurred compared with negative fibroblasts, and positive nuclei were even fewer. Meanwhile, most of the mitotic cells were PR positive, showing intense localization around chromosomes and on microtubules. These findings suggest that gingival fibroblasts are fundamentally capable of expressing PR and transmit the signal to target genes. Conclusions: The present study may conclude that in either gender or at any age, gingival fibroblasts express PR rather low in level and do not necessarily localize PR in a nuclear dominant fashion, which is an essential feature for reproductive organ cells. The poor ER reactivity shown in the gingival fibroblasts was discussed in view of the receptor subtype.
Hypomyelinating leukodystrophy (HLD) is genetic demyelinating or dysmyelinating disease and is associated with at least 13 responsible genes. The mutations seem likely cause the functional deficiency of their gene products. HLD4- and HLD5-associated HSPD1 and FAM126A mutations affect biochemical properties of the gene products (Miyamoto et al. (2015,2014) [[1], [2]]). Herein we provide the data regarding the effects of HLD6-associated tubulin beta 4A (TUBB4A) mutations on the properties.
A monoclonal mouse IgG1 antibody was produced against the aminopropetide of dermatosparactic sheep procollagen type I by using the hybridoma technique. Radioimmunoassays demonstrated an apparent affinity constant of 10 8 1 · mol −1 . The antibody reacted with a 19‐amino‐acid‐long sequence spanning the procollagen N‐proteinase cleavage site with stronger binding to structures contributed by the aminopropeptide. The antibody showed strong cross‐reactions with similar antigens of bovine, human or chick origin but failed to react with the aminopropetide of procollagen type III. Incubation of chick or sheep procollagen type I with stoichiometric amounts of antibody blocked the release from procollagen molecules of the aminopropeptide by procollagen N‐proteinase. Thus, this antibody seems useful for studying various biological problems encountered in the conversion of procollagen.
Mature Schwann cells, the peripheral nervous system (PNS) glial cells, have two major roles for neuronal axons (Bunge, 1993) [1]. For large diameter axons, Schwann cells form myelin sheaths with multiple layers. For small diameter axons, they form Remak bundle composed only of single layer of the Schwann cell plasma membrane. In the PNS, ErbB3 forms a dimer with ErbB2 on the Schwann cell plasma membrane. ErbB3 plays a key role in myelination by myelinating Schwann cells, that is to say, its role in myelin thickness. Herein we provide the data regarding the effect of in vivo knockdown of ErbB3 on the thickness between an axon and a neighboring axon in Remak bundle, which is formed by non-myelinating Schwann cells. Since ErbB3 knockout mice are embryonically lethal, Schwann cell lineage-specific transgenic mice transcribing ErbB3 shRNA with an artificial miRNA backbone were generated and used in these experiments (Torii et al., 2014) [2].
To produce advanced dental technologists, researchers and educators in dental technology field, School of Oral Health Sciences in Hiroshima University teaches not only conventional manual dental technologies but also leading-edge ones including computer-aided fabrication technologies. First the students leam the theory of CAI)/CAM, rapid prototyping, and measurement of 3d-shape. Next they practice : 1) the fabrication of dental prostheses by using dental CAD/CAM systems ; 2) the fabrication of models for maxillofacial surgery by using 3d-software and rapid prototyping; and 3) measuring and 3d-modeling of human bodies.
During embryonic development of the peripheral nervous system (PNS), Schwann cell precursors migrate along neuronal axons to their final destinations, where they will myelinate the axons after birth. While the intercellular signals controlling Schwann cell precursor migration are well studied, the intracellular signals controlling Schwann cell precursor migration remain elusive. Here, using a rat primary cell culture system, we show that Dock8, an atypical Dock180-related guanine-nucleotide exchange factor (GEF) for small GTPases of the Rho family, specifically interacts with Nck1, an adaptor protein composed only of Src homology (SH) domains, to promote Schwann cell precursor migration induced by platelet-derived growth factor (PDGF). Knockdown of Dock8 or Nck1 with its respective siRNA markedly decreases PDGF-induced cell migration, as well as Rho GTPase activation, in precursors. Dock8, through its unique N-terminal proline-rich motif, interacts with the SH3 domain of Nck1, but not with other adaptor proteins composed only of SH domains, e.g. Grb2 and CrkII, and not with the adaptor protein Elmo1. Reintroduction of the proline-rich motif mutant of Dock8 in Dock8 siRNA-transfected Schwann cell precursors fails to restore their migratory abilities, whereas that of wild-type Dock8 does restore these abilities. These results suggest that Nck1 interaction with Dock8 mediates PDGF-induced Schwann cell precursor migration, demonstrating not only that Nck1 and Dock8 are previously unanticipated intracellular signaling molecules involved in the regulation of Schwann cell precursor migration but also that Dock8 is among the genetically-conservative common interaction subset of Dock family proteins consisting only of SH domain adaptor proteins.
The functions of Na(+) /H(+) exchangers (NHEs) during osteoclastic differentiation were investigated using the NHE inhibitor amiloride and a monoclonal antibody (MAb). Compared with sRANKL-stimulated control cells, amiloride decreased the number of large TRAP-positive osteoclast cells (OCs) with ≥10 nuclei and increased the number of small TRAP-positive OCs with ≤10 nuclei during sRANKL-dependent osteoclastic differentiation of RAW264.7 cells. NHE10 mRNA expression and OC differentiation markers were increased by sRANKL stimulation in dose- and time-dependent manners. NHEs 1-9 mRNA expression was not increased by sRANKL stimulation. Similar to amiloride, a rat anti-mouse NHE10 MAb (clone 6B11) decreased the number of large TRAP-positive OCs, but increased the number of small TRAP-positive OCs. These findings suggested that inhibition of NHEs by amiloride or an anti-NHE10 MAb prevented sRANKL-promoted cellular fusion. The anti-NHE10 MAb has the potential for use as an effective inhibitor of bone resorption for targeted bone disease therapy.
'%3e%3cpath fill='%23012169' d='M0 0v30h60V0z'/%3e%3cpath stroke='%23fff' stroke-width='6' d='m0 0 60 30m0-30L0 30'/%3e%3cpath stroke='%23C8102E' stroke-width='4' d='m0 0 60 30m0-30L0 30' clip-path='url(%23b)'/%3e%3cpath stroke='%23fff' stroke-width='10' d='M30 0v30M0 15h60'/%3e%3cpath stroke='%23C8102E' stroke-width='6' d='M30 0v30M0 15h60'/%3e%3c/g%3e%3c/svg%3e)