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1.
Cell Tissue Res ; 395(1): 53-62, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37985496

RESUMO

Glomerular epithelial protein-1 (Glepp1), a R3 subtype family of receptor-type protein tyrosine phosphatases, plays important role in the activation of Src family kinases and regulates cellular processes such as cell proliferation, differentiation, and apoptosis. In this study, we firstly examined the functional evaluation of Glepp1 in tooth development and morphogenesis. The precise expression level and developmental function of Glepp1 were examined by RT-qPCR, in situ hybridization, and loss and gain of functional study using a range of in vitro organ cultivation methods. Expression of Glepp1 was detected in the developing tooth germs in cap and bell stage of tooth development. Knocking down Glepp1 at E13 for 2 days showed the altered expression levels of tooth development-related signaling molecules, including Bmps, Dspp, Fgf4, Lef1, and Shh. Moreover, transient knock down of Glepp1 revealed alterations in cellular physiology, examined by the localization patterns of Ki67 and E-cadherin. Similarly, knocking down of Glepp1 showed disrupted enamel rod and interrod formation in 3-week renal transplanted teeth. In addition, due to attrition of odontoblastic layers, the expression signals of Dspp and the localization of NESTIN were almost not detected after knock down of Glepp1; however, their expressions were increased after Glepp1 overexpression. Thus, our results suggested that Glepp1 plays modulating roles during odontogenesis by regulating the expression levels of signaling molecules and cellular events to achieve the proper structural formation of hard tissue matrices in mice molar development.


Assuntos
Proteínas Tirosina Fosfatases Classe 3 Semelhantes a Receptores , Dente , Animais , Camundongos , Regulação da Expressão Gênica no Desenvolvimento , Morfogênese , Odontogênese , Proteínas Tirosina Fosfatases/metabolismo , Proteínas Tirosina Fosfatases Classe 3 Semelhantes a Receptores/metabolismo , Transdução de Sinais , Dente/metabolismo
2.
Sensors (Basel) ; 24(6)2024 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-38544049

RESUMO

In this work, we propose our newly developed wafer-type plasma monitoring sensor based on a floating-type double probe method that can be useful for two-dimensional (2D) in situ plasma diagnosis within a semiconductor processing chamber. A key achievement of this work is the first realization of an ultra-thin plasma monitoring sensor with a system thickness of ~1.4 mm, which supports a fully automated robot arm transfer capability for in situ plasma diagnosis. To the best of our knowledge, it is the thinnest accomplishment among all wafer-type plasma monitoring sensors. Our proposed sensor is assembled with two Si wafers and SiO2-based probes; accordingly, it makes it possible to monitor the actual dynamics of processing plasmas under electrostatic chucking (ESC) conditions. Also, it allows for the prevention of chamber contamination issues after continuously exposing the radio frequency (RF) to various processing gases. Using a test-bed chamber, we successfully demonstrated the feasibility and system performance of the proposed sensor, including robot arm transfer capability, vacuum and thermal stress durability, and data integrity and reproducibility. Consequently, compared with the conventional plasma diagnostic tools, we expect that our proposed sensor will be highly beneficial for tool-to-tool matching (TTTM) and/or for studying various plasma-related items by more accurately providing the parameters of processing plasmas, further saving both time and manpower resources required for preventive maintenance (PM) routines as well.

3.
J Cell Physiol ; 238(7): 1520-1529, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37098720

RESUMO

To understand the mechanisms underlying tooth morphogenesis, we examined the developmental roles of important posttranslational modification, O-GlcNAcylation, which regulates protein stability and activity by the addition and removal of a single sugar (O-GlcNAc) to the serine or threonine residue of the intracellular proteins. Tissue and developmental stage-specific immunostaining results against O-GlcNAc and O-GlcNAc transferase (OGT) in developing tooth germs would suggest that O-GlcNAcylation is involved in tooth morphogenesis, particularly in the cap and secretory stage. To evaluate the developmental function of OGT-mediated O-GlcNAcylation, we employed an in vitro tooth germ culture method at E14.5, cap stage before secretory stage, for 1 and 2 days, with or without OSMI-1, a small molecule OGT inhibitor. To examine the mineralization levels and morphological changes, we performed renal capsule transplantation for one and three weeks after 2 days of in vitro culture at E14.5 with OSMI-1 treatment. After OGT inhibition, morphological and molecular alterations were examined using histology, immunohistochemistry, real-time quantitative polymerase chain reaction, in situ hybridization, scanning electron microscopy, and ground sectioning. Overall, inhibition of OGT resulted in altered cellular physiology, including proliferation, apoptosis, and epithelial rearrangements, with significant changes in the expression patterns of ß-catenin, fibroblast growth factor 4 (fgf4), and sonic hedgehog (Shh). Moreover, renal capsule transplantation and immunolocalizations of Amelogenin and Nestin results revealed that OGT-inhibited tooth germs at cap stage exhibited with structural changes in cuspal morphogenesis, amelogenesis, and dentinogenesis of the mineralized tooth. Overall, we suggest that OGT-mediated O-GlcNAcylation regulates cell signaling and physiology in primary enamel knot during tooth development, thus playing an important role in mouse molar morphogenesis.


Assuntos
N-Acetilglucosaminiltransferases , Dente , Animais , Camundongos , Apoptose/fisiologia , Proteínas Hedgehog/genética , Proteínas Hedgehog/metabolismo , N-Acetilglucosaminiltransferases/genética , N-Acetilglucosaminiltransferases/metabolismo , Processamento de Proteína Pós-Traducional , Dente/crescimento & desenvolvimento , Dente/metabolismo
4.
Histochem Cell Biol ; 159(6): 477-487, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36814002

RESUMO

Mechanically activated factors are important in organogenesis, especially in the formation of secretory organs, such as salivary glands. Piezo-type mechanosensitive ion channel component 1 (Piezo1), although previously studied as a physical modulator of the mechanotransduction, was firstly evaluated on its developmental function in this study. The detailed localization and expression pattern of Piezo1 during mouse submandibular gland (SMG) development were analyzed using immunohistochemistry and RT-qPCR, respectively. The specific expression pattern of Piezo1 was examined in acinar-forming epithelial cells at embryonic day 14 (E14) and E16, which are important developmental stages for acinar cell differentiation. To understand the precise function of Piezo1 in SMG development, siRNA against Piezo1 (siPiezo1) was employed as a loss-of-function approach, during in vitro organ cultivation of SMG at E14 for the designated period. Alterations in the histomorphology and expression patterns of related signaling molecules, including Bmp2, Fgf4, Fgf10, Gli1, Gli3, Ptch1, Shh, and Tgfß-3, were examined in acinar-forming cells after 1 and 2 days of cultivation. Particularly, altered localization patterns of differentiation-related signaling molecules including Aquaporin5, E-cadherin, Vimentin, and cytokeratins would suggest that Piezo1 modulates the early differentiation of acinar cells in SMGs by modulating the Shh signaling pathway.


Assuntos
Mecanotransdução Celular , Glândula Submandibular , Camundongos , Animais , Glândula Submandibular/metabolismo , Glândulas Salivares , Morfogênese/fisiologia , Diferenciação Celular , Canais Iônicos/metabolismo
5.
Sensors (Basel) ; 22(18)2022 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-36146384

RESUMO

With the development of maritime technology and equipment, most ships are equipped with an automatic identification system (AIS) to store navigation information. Over time, the size of the data increases, rendering its storage and processing difficult. Hence, it is necessary to transform the AIS data into trajectories, and then simplify the AIS trajectories to remove unnecessary information that is not related to route shape. Moreover, topographic information must be considered because otherwise, the simplified trajectory can intersect obstacles. In this study, we propose an AIS trajectory simplification algorithm considering topographic information. The proposed algorithm simplifies the trajectories without the intersection of the trajectory and obstacle using the improved Douglas-Peucker algorithm. Polygon map random (PMR) quadtree was used to consider topographic information on the coast, and the intersection between topographic information and simplified trajectories was efficiently computed using the PMR quadtree. To verify the effectiveness of the proposed algorithm, experiments were conducted on real-world trajectories in the Korean sea. The proposed algorithm yielded simplified trajectories with no intersections of the trajectory and obstacle. In addition, the computational efficiency of the proposed algorithm with the PMR quadtree was superior to that without the PMR quadtree.

6.
Int J Mol Sci ; 21(21)2020 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-33138041

RESUMO

FUSE binding protein 1 (Fubp1), a regulator of the c-Myc transcription factor and a DNA/RNA-binding protein, plays important roles in the regulation of gene transcription and cellular physiology. In this study, to reveal the precise developmental function of Fubp1, we examined the detailed expression pattern and developmental function of Fubp1 during tooth morphogenesis by RT-qPCR, in situ hybridization, and knock-down study using in vitro organ cultivation methods. In embryogenesis, Fubp1 is obviously expressed in the enamel organ and condensed mesenchyme, known to be important for proper tooth formation. Knocking down Fubp1 at E14 for two days, showed the altered expression patterns of tooth development related signalling molecules, including Bmps and Fgf4. In addition, transient knock-down of Fubp1 at E14 revealed changes in the localization patterns of c-Myc and cell proliferation in epithelium and mesenchyme, related with altered tooth morphogenesis. These results also showed the decreased amelogenin and dentin sialophosphoprotein expressions and disrupted enamel rod and interrod formation in one- and three-week renal transplanted teeth respectively. Thus, our results suggested that Fubp1 plays a modulating role during dentinogenesis and amelogenesis by regulating the expression pattern of signalling molecules to achieve the proper structural formation of hard tissue matrices and crown morphogenesis in mice molar development.


Assuntos
Proteínas de Ligação a DNA/metabolismo , Embrião de Mamíferos/citologia , Regulação da Expressão Gênica no Desenvolvimento , Morfogênese , Odontogênese , Proteínas de Ligação a RNA/metabolismo , Dente/embriologia , Animais , Proliferação de Células , Proteínas de Ligação a DNA/genética , Embrião de Mamíferos/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Proteínas de Ligação a RNA/genética , Transdução de Sinais , Dente/metabolismo
7.
J Cell Physiol ; 234(11): 20354-20365, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-30963569

RESUMO

To understand the role of endoplasmic reticulum (ER)-stress in mice molar development, we studied Tmbim6 that antagonizes the unfolded protein response, using Tmbim6 knockout (KO) mice and in vitro organ cultivation with knocking down using small interfering RNA. During molar development, Tmbim6 is expressed in developing tooth at E14-E16, postnatal0 (PN0), and PN6. Mineral content in Tmbim6 KO enamel was reduced while dentin was slightly increased revealing ultrastructural changes in pattern formation of both enamel and dentin. Moreover, odontoblast differentiation was altered with increased Dspp expression at PN0 followed by altered AMELX localizations at PN5. These results were confirmed by in vitro organ cultivation and showed altered Bmp signaling, proliferation, and actin rearrangement in the presumptive ameloblast and odontoblasts that followed the altered expression of differentiation and ER stress-related signaling molecules at E16.5. Overall, ER stress modulated by Tmbim6 would play important roles in patterned dental hard tissue formation in mice molar within a limited period of development.


Assuntos
Diferenciação Celular/genética , Estresse do Retículo Endoplasmático/genética , Proteínas de Membrana/genética , Dente Molar/metabolismo , Odontoblastos/metabolismo , Ameloblastos/metabolismo , Animais , Proteínas da Matriz Extracelular/metabolismo , Camundongos Knockout , Sialoglicoproteínas/genética , Transdução de Sinais/fisiologia
8.
J Clin Periodontol ; 46(1): 96-104, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30372547

RESUMO

OBJECTIVES: To evaluate the histologic and volumetric changes of gingival tissues following grafting with collagen-based matrices at labial aspect of teeth in canines. MATERIALS AND METHODS: Gingival augmentation was performed in the mandibular incisor area using two types of xenogeneic cross-linked collagen matrices (CCMs), bovine CCM for BCCM group and porcine CCM for PCCM group, whereas the contralateral sides remained untreated (B-control group and P-control group). Descriptive histology, histometric and volumetric analyses were performed after 12 weeks. For statistical comparison between each test group and respective control group, paired t test was used for histometric analysis, and repeated-measured analysis of variance was used for volumetric analysis (p < 0.05). RESULTS: An increased number of rete pegs and an enhanced formation of new blood vessels were observed at both grafted sites compared to the corresponding control sites. There was statistically significant gain of horizontal thickness only in BCCM group (1.36 ± 0.27 mm vs. 1.26 ± 0.34 mm; p < 0.05) compared to the B-control groups. CONCLUSION: BCCM was effective for gingival augmentation in terms of horizontal thickness at the labial aspect of teeth at 12 weeks post-surgery.


Assuntos
Colágeno , Gengiva , Animais , Bovinos , Tecido Conjuntivo , Cães , Incisivo , Suínos
9.
Retina ; 39(1): 143-149, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29095356

RESUMO

PURPOSE: To investigate ocular perfusion pressure (OPP) in patients with central serous chorioretinopathy (CSC) and pigment epitheliopathy (PE). METHODS: In this retrospective study, we included patients with acute idiopathic unilateral CSC and age- and sex-matched normal controls. The patients with CSC were classified into two groups according to the PE presence on their fellow eyes. We compared OPP among CSC patients with or without PE and normal controls. RESULTS: Among a total of 47 patients with CSC, 21 were classified into the PE group and 26 into the non-PE group. The mean OPP (±SD) of CSC and fellow eyes in the PE group (50.02 ± 4.98 and 50.83 ± 4.12 mmHg, respectively) was greater than that in the non-PE group (43.69 ± 6.88 and 44.0 ± 6.57 mmHg, respectively) and normal controls (45.64 ± 8.73 mmHg) (all, P < 0.05). The mean macular choroidal thickness of CSC and fellow eyes in the PE group (344.4 ± 31.8 and 310.1 ± 37.0 µm, respectively) was thicker than that in the non-PE group (318.5 ± 43.8 and 282.8 ± 36.1 µm, respectively) (all, P < 0.05). Greater macular choroidal thickness and higher OPP were associated with PE (P = 0.002 and P = 0.003). CONCLUSION: Forty-five percent of the patients with acute unilateral CSC had PE in their fellow eyes. Increased OPP may influence the development of bilateral CSC characteristics.


Assuntos
Velocidade do Fluxo Sanguíneo/fisiologia , Pressão Sanguínea/fisiologia , Coriorretinopatia Serosa Central/fisiopatologia , Corioide/patologia , Pressão Intraocular/fisiologia , Epitélio Pigmentado da Retina/patologia , Vasos Retinianos/fisiopatologia , Adulto , Coriorretinopatia Serosa Central/diagnóstico , Corioide/irrigação sanguínea , Feminino , Angiofluoresceinografia , Seguimentos , Fundo de Olho , Humanos , Macula Lutea/patologia , Masculino , Fluxo Sanguíneo Regional/fisiologia , Vasos Retinianos/diagnóstico por imagem , Estudos Retrospectivos , Tomografia de Coerência Óptica , Acuidade Visual
10.
Oral Dis ; 25(4): 1158-1168, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30801855

RESUMO

OBJECTIVES: Ameloblastomas are the most common odontogenic epithelial tumors with high recurrence rate. The aim of this study was to identify apoptosis-related genes with recurrence of ameloblastomas and to evaluate its feasibility as a prognostic marker and as a target molecule preventing from recurrence. MATERIALS AND METHODS: Public microarray data were analyzed. To evaluate their expression in ameloblastoma patients, immunohistochemical staining was performed in 89 human ameloblastoma tissues. Quantitative PCR was performed by use of ameloblastoma cell line (AM-1). Fluorescence activated cell sorting analysis and western blotting were conducted following transfection with siRNA. Further, AM-1 cells were implanted in the renal subcapsular layer of immunodeficient mice. RESULTS: Microarray data analysis revealed that osteoprotegerin (OPG) and B-cell lymphoma 2 (Bcl-2) were the two most upregulated genes in ameloblastoma. Only Bcl-2 expression was significantly (p = 0.020) associated with recurrence in conservative treatment group (n = 17) among 89 patients. Silencing of Bcl-2 increased apoptosis in AM-1 cells in vitro and inhibited tumor nodule formation of AM-1 cells in vivo. CONCLUSION: These results suggest that Bcl-2 expression is a useful biomarker to predict recurrence of ameloblastomas, and as a therapeutic target molecule to prevent recurrence of ameloblastoma.


Assuntos
Ameloblastoma/patologia , Neoplasias Maxilomandibulares/patologia , Linfoma de Células B/genética , Osteoprotegerina/genética , Adulto , Ameloblastoma/genética , Animais , Apoptose , Feminino , Humanos , Neoplasias Maxilomandibulares/genética , Masculino , Camundongos , Pessoa de Meia-Idade , Recidiva Local de Neoplasia , Prognóstico , Proteínas Proto-Oncogênicas c-bcl-2/análise , Proteínas Proto-Oncogênicas c-bcl-2/sangue , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo
11.
Implant Dent ; 26(5): 664-673, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28719568

RESUMO

PURPOSE: This study evaluates the synergistic effect of serial application of bone morphogenetic protein 2 (BMP2) and fibroblast growth factor 2 (FGF2) on both new bone formation and periodontal tissue regeneration using 1-wall intrabony defect in mongrel dogs. MATERIALS AND METHODS: One-wall defects were created at the mesial aspect of the mandibular first molars of 6 male mongrel dogs. Each mandibular defect received 1 of the 2 experimental treatments randomly (BMP2 or BMP2 + FGF2), and it was allowed to heal for either 4 or 8 weeks postoperatively. Histologic and histomorphometric analyses were performed for the evaluation of the overall healing patterns of new bone formation and periodontal tissue regeneration. RESULTS: The results showed that after 8 weeks, serial application of BMP2 and FGF2 significantly improved the periodontal tissue regeneration, whereas application of BMP2 only showed greater new bone formation after 4 weeks. CONCLUSIONS: The serial application of BMP2 and FGF2 may have synergistic effects on periodontal tissue regeneration over time.


Assuntos
Proteína Morfogenética Óssea 2/uso terapêutico , Fator 2 de Crescimento de Fibroblastos/uso terapêutico , Regeneração Tecidual Guiada Periodontal/métodos , Animais , Proteína Morfogenética Óssea 2/administração & dosagem , Regeneração Óssea/efeitos dos fármacos , Cães , Sinergismo Farmacológico , Fator 2 de Crescimento de Fibroblastos/administração & dosagem , Masculino , Mandíbula/diagnóstico por imagem , Mandíbula/patologia , Periodonto/diagnóstico por imagem , Periodonto/patologia , Microtomografia por Raio-X
12.
Histochem Cell Biol ; 146(3): 289-300, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27166834

RESUMO

Intimate crosstalk occurs between the pulmonary epithelium and the vascular network during lung development. The transcription factor forkhead box f1 (Foxf1) is expressed in the lung mesenchyme and plays an indispensable role in pulmonary angiogenesis. Sonic hedgehog (Shh), a signalling molecule, is expressed in lung epithelium and is required to establish proper angiogenesis. It has been suggested that Foxf1, a downstream target of the Shh signalling pathway, mediates interaction between angiogenesis and the epithelium in lung. However, there has been no clear evidence showing the mechanism how Foxf1 is regulated by Shh signalling pathway during lung development. In this study, we investigated the lung-specific enhancers of Foxf1 and the Gli binding on the enhancers. At first, we found three evolutionarily conserved Foxf1 enhancers, two of which were long-range enhancers. Of the long-range enhancers, one demonstrated tissue-specific activity in the proximal and distal pulmonary blood vessels, while the other one demonstrated activity only in distal blood vessels. At analogous positions in human, these long-range enhancers were included in a regulatory region that was reportedly repeatedly deleted in alveolar capillary dysplasia with misalignment of pulmonary vein patients, which indicates the importance of these enhancers in pulmonary blood vessel formation. We also determined that Gli increased the activity of one of these long-range enhancers, which was specific to distal blood vessel, suggesting that Shh regulates Foxf1 transcription in pulmonary distal blood vessel formation.


Assuntos
Vasos Sanguíneos/metabolismo , Elementos Facilitadores Genéticos/genética , Fatores de Transcrição Forkhead/genética , Veias Pulmonares/metabolismo , Transcrição Gênica , Animais , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Análise de Sequência de DNA
13.
Arthroscopy ; 32(8): 1660-6, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27090722

RESUMO

PURPOSE: To validate whether the Bernard quadrant method, which was developed for application on simple lateral radiography, can be used with 3-dimensional computed tomography (3D CT) to localize the femoral insertion of the reconstructed anterior cruciate ligament (ACL). METHODS: We analyzed 32 knees with ACL tears that were reconstructed using a metal interference screw for fixation at the femoral tunnel between March 2012 and May 2013. Postoperative lateral radiographs and 3D CT images were obtained 7 days after the operation. By use of the Bernard quadrant method, the location of the femoral tunnel was measured by 2 orthopaedic surgeons by locating the position of the metal interference screw using 3D CT imaging and simple lateral knee radiography. The correlation between the femoral tunnels on the 2 radiographic images was compared using the MedCalc statistical analysis program. RESULTS: On the 3D CT image, the position of the femoral insertion of the ACL as measured by the position of the metal screw head was 36.3% ± 6.0% in the x-coordinate and 39.6% ± 9.1% in the y-coordinate compared with 37.6% ± 5.8% and 41.0% ± 11.6%, respectively, on the simple radiograph. The Pearson correlation coefficients between 3D CT and simple radiography were 0.840 for the x-coordinate and 0.858 for the y-coordinate. Intraobserver reliability and interobserver reliability for both coordinates were greater than 0.9 on 3D CT. CONCLUSIONS: Application of the Bernard quadrant method on 3D CT showed high correlation to the originally described method using lateral radiographs and can be used reliably for localizing the reconstructed ACL. LEVEL OF EVIDENCE: Level III, diagnostic study.


Assuntos
Lesões do Ligamento Cruzado Anterior/diagnóstico por imagem , Lesões do Ligamento Cruzado Anterior/cirurgia , Reconstrução do Ligamento Cruzado Anterior/métodos , Fêmur/diagnóstico por imagem , Adolescente , Adulto , Parafusos Ósseos , Feminino , Fêmur/cirurgia , Humanos , Imageamento Tridimensional/métodos , Articulação do Joelho/diagnóstico por imagem , Articulação do Joelho/cirurgia , Masculino , Pessoa de Meia-Idade , Variações Dependentes do Observador , Período Pós-Operatório , Radiografia , Reprodutibilidade dos Testes , Tomografia Computadorizada por Raios X/métodos , Adulto Jovem
14.
Histochem Cell Biol ; 144(4): 377-87, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26170146

RESUMO

Adenomatosis polyposis coli downregulated 1 (APCDD1), a negative regulator of Wnt signaling, was examined to understand detailed mechanisms underlying Wnt signaling tooth development. In situ hybridization showed that Apcdd1 was expressed in the condensed mesenchyme at the bud stage, and in the inner enamel epithelium (IEE), including enamel knot (EK) at the cap stage. In vitro organ cultivation by using Apcdd1 antisense oligodeoxynucleotides was performed at E13.5 for 2 days to define the developmental functions of APCDD1 during tooth development. Analysis of histogenesis and cellular events such as cell adhesion, proliferation, apoptosis and epithelial rearrangement after Apcdd1 knockdown showed altered morphogenesis of the tooth germ with decreased cell proliferation and altered localization of cell adhesion molecules. Actin filament staining and 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) labeling of IEE cells showed that Apcdd1 knockdown enhanced epithelial rearrangement in the IEE and EK. To understand the precise signaling regulations of Apcdd1, we evaluated the altered expression patterns of signaling molecules, related with Wnt and enamel knot signalings using RT-qPCR. Tooth germs at cap stage were transplanted into the kidney capsules and were allowed to develop into calcified teeth for 3 weeks. Apcdd1 knockdown increased the number of ectopic cusps on the mesial side of the tooth. Our results suggested that APCDD1 modulates the gene expression of Wnt- and EK-related signaling molecules at the cap stage of tooth development, and is involved in tooth cusp patterning by modulating the epithelial rearrangement in the IEE.


Assuntos
Células Epiteliais/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas de Membrana/metabolismo , Dente Molar/metabolismo , Odontogênese , Animais , Moléculas de Adesão Celular/metabolismo , Proliferação de Células , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Silenciamento de Genes , Idade Gestacional , Peptídeos e Proteínas de Sinalização Intracelular/genética , Proteínas de Membrana/genética , Camundongos , Dente Molar/embriologia , Morfogênese , Oligonucleotídeos Antissenso/genética , Oligonucleotídeos Antissenso/metabolismo , Técnicas de Cultura de Tecidos , Via de Sinalização Wnt
15.
Development ; 138(9): 1807-16, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21447550

RESUMO

Each vertebrate species displays specific tooth patterns in each quadrant of the jaw: the mouse has one incisor and three molars, which develop at precise locations and at different times. The reason why multiple teeth form in the jaw of vertebrates and the way in which they develop separately from each other have been extensively studied, but the genetic mechanism governing the spatial patterning of teeth still remains to be elucidated. Sonic hedgehog (Shh) is one of the key signaling molecules involved in the spatial patterning of teeth and other ectodermal organs such as hair, vibrissae and feathers. Sostdc1, a secreted inhibitor of the Wnt and Bmp pathways, also regulates the spatial patterning of teeth and hair. Here, by utilizing maternal transfer of 5E1 (an anti-Shh antibody) to mouse embryos through the placenta, we show that Sostdc1 is downstream of Shh signaling and suggest a Wnt-Shh-Sostdc1 negative feedback loop as a pivotal mechanism controlling the spatial patterning of teeth. Furthermore, we propose a new reaction-diffusion model in which Wnt, Shh and Sostdc1 act as the activator, mediator and inhibitor, respectively, and confirm that such interactions can generate the tooth pattern of a wild-type mouse and can explain the various tooth patterns produced experimentally.


Assuntos
Padronização Corporal/genética , Proteínas Morfogenéticas Ósseas/fisiologia , Retroalimentação Fisiológica/fisiologia , Proteínas Hedgehog/fisiologia , Dente/embriologia , Proteínas Wnt/fisiologia , Proteínas Adaptadoras de Transdução de Sinal , Animais , Padronização Corporal/fisiologia , Proteínas Morfogenéticas Ósseas/genética , Proteínas Morfogenéticas Ósseas/metabolismo , Células Cultivadas , Simulação por Computador , Embrião de Mamíferos , Epistasia Genética/fisiologia , Proteínas Hedgehog/genética , Proteínas Hedgehog/metabolismo , Camundongos , Camundongos Knockout , Camundongos Nus , Modelos Teóricos , Odontogênese/genética , Odontogênese/fisiologia , Transdução de Sinais/genética , Transdução de Sinais/fisiologia , Dente/metabolismo , Proteínas Wnt/genética , Proteínas Wnt/metabolismo
16.
Cell Tissue Res ; 358(1): 109-21, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24865245

RESUMO

Teraspanin transmembrane protein, Perp (P53 apoptosis effector related to PMP22), which is found in the plasma membrane as a component of the desmosome, is reported to be involved in the morphogenesis of the epithelium and the enamel formation of the incisor. However, its expression pattern and signaling regulation during molar development have not been elucidated in detail. We have examined the precise expression patterns of Perp in developing lower molars and employed the knock-down of Perp by antisense oligodeoxynucleotide treatment during in vitro organ cultivation at embryonic day 13 to define the precise developmental function of Perp. Perp was expressed mainly in the dental lamina and stellate reticulum regions at the bud and cap stages. After Perp knock-down, the tooth germ showed disruption of the dental lamina and stellate reticulum with altered apoptosis and proliferation. The changed expression levels of related signaling molecules from the enamel knot and desmosome were evaluated by real-time quantitative polymerase chain reaction. A renal capsule transplantation method was employed to examine the effects of Perp knock-down on molar crown development. Ultrastructural observations revealed that enamel was deposited more densely in an irregular pattern in the cusp region, and that dentin was hypo-mineralized after Perp knock-down at the cap stage. Thus, Perp might play important roles in the formation and integration of stellate reticulum, dental lamina structure and enamel formation through signaling interactions with the enamel knot and desmosome-related signaling molecules at the cap stage of lower molar development.


Assuntos
Apoptose/fisiologia , Esmalte Dentário/embriologia , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Proteínas de Membrana/biossíntese , Dente Molar/embriologia , Morfogênese/fisiologia , Animais , Técnicas de Silenciamento de Genes , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos ICR
17.
Sci Rep ; 14(1): 13633, 2024 06 13.
Artigo em Inglês | MEDLINE | ID: mdl-38871845

RESUMO

Notum is a direct target of Wnt/ß-catenin signaling and plays a crucial role as a Wnt inhibitor within a negative feedback loop. In the tooth, Notum is known to be expressed in odontoblasts, and severe dentin defects and irregular tooth roots have been reported in Notum-deficient mice. However, the precise expression pattern of Notum in early tooth development, and the role of Notum in crown and root patterns remain elusive. In the present study, we identified a novel Notum expression in primary enamel knot (EK), secondary EKs, and dental papilla during tooth development. Notum-deficient mice exhibited enlarged secondary EKs, resulting in broader cusp tips, altered cusp patterns, and reduced concavity in crown outline. These alterations in crown outline led to a reduction in cervical tongue length, thereby inducing root fusion in Notum-deficient mice. Overall, these results suggest that the secondary EK size, regulated by the Wnt/Notum negative feedback loop, has a significant impact on the patterns of crown and root during tooth morphogenesis.


Assuntos
Dente Molar , Coroa do Dente , Raiz Dentária , Animais , Camundongos , Regulação da Expressão Gênica no Desenvolvimento , Camundongos Knockout , Dente Molar/metabolismo , Dente Molar/crescimento & desenvolvimento , Odontogênese , Receptores Acoplados a Proteínas G , Coroa do Dente/crescimento & desenvolvimento , Coroa do Dente/metabolismo , Raiz Dentária/crescimento & desenvolvimento , Raiz Dentária/metabolismo , Via de Sinalização Wnt
18.
Dev Biol ; 365(1): 259-66, 2012 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-22387209

RESUMO

Retinoic acid receptors (RARs), which are involved in retinoic acid signal transduction, are essential for maintaining the differentiated state of epithelial tissues. Mammary glands are skin appendages whose development is initiated through continuous cell-cell interactions between the ectoderm and the adjacent mesenchyme. Considerable progress has been made in elucidating the molecular basis of these interactions in mammary gland formation in mouse embryos, including the network of initiating signals comprising Fgfs, Wnts and Bmps involved in gland positioning and the transcription factors, Tbx3 and Lef1, essential for mammary gland development. Here, we provide evidence that retinoic acid signaling may also be involved in mammary gland development. We documented the expression of gene-encoding enzymes that produce retinoic acid (Raldh2) and enzymes that degrade it (Cyp26a1, Cyp26b1). We also analyzed the expression of RAR-ß, a direct transcriptional target of retinoic acid signaling. Raldh2 and RAR-ß were expressed in E10-E10.5 mouse embryos in somites adjacent to the flank region where mammary buds 2, 3 and 4 develop. These expression patterns overlapped with that of Fgf10, which is known to be required for mammary gland formation. RAR-ß was also expressed in the mammary mesenchyme in E12 mouse embryos; RAR-ß protein was expressed in the mammary epithelium and developing fat pad. Retinoic acid levels in organ cultures of E10.5 mouse embryo flanks were manipulated by adding either retinoic acid or citral, a retinoic acid synthesis inhibitor. Reduced retinoic acid synthesis altered the expression of genes involved in retinoic acid homeostasis and also demonstrated that retinoic acid signaling is required for Tbx3 expression, whereas high levels of retinoic acid signaling inhibited Bmp4 expression and repressed Wnt signaling. The results of the experiments using RNAi against Tbx3 and Wnt10b suggested feedback interactions that regulate retinoic acid homeostasis in mammary gland-forming regions. We produced a molecular model for mammary gland initiation that incorporated retinoic acid signaling.


Assuntos
Glândulas Mamárias Animais/embriologia , Transdução de Sinais , Tretinoína/fisiologia , Animais , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Glândulas Mamárias Animais/fisiologia , Mesoderma/embriologia , Mesoderma/fisiologia , Camundongos , Modelos Moleculares , Interferência de RNA , Receptores do Ácido Retinoico/fisiologia , Transdução de Sinais/genética , Proteínas com Domínio T/fisiologia , Proteínas Wnt/fisiologia
19.
ACS Mater Au ; 3(1): 37-42, 2023 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-36647455

RESUMO

We report a series of adamantane-functionalized azobenzenes that store photon and thermal energy via reversible photoisomerization in the solid state for molecular solar thermal (MOST) energy storage. The adamantane unit serves as a 3D molecular separator that enables the spatial separation of azobenzene groups and results in their facile switching even in the crystalline phase. Upon isomerization, the phase transition from crystalline to amorphous solid occurs and contributes to additional energy storage. The exclusively solid-state MOST compounds with solid-solid phase transition overcome a major challenge of solid-liquid phase transition materials that require encapsulation for practical applications.

20.
J Bone Metab ; 30(1): 93-101, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36950845

RESUMO

BACKGROUND: The skull is a complex structure formed by the craniofacial bones' elaborate organization. The growth pattern in each craniofacial bone of the postnatal skull has been presented in wild-type mice. However, the skull's growth pattern, determined by the craniofacial bones' coordinated growth, is unfamiliar. This study aimed to examine the overall morphological change in the mid-sagittal plane of the postnatal mice's skulls and interaction between the craniofacial bones. METHODS: Geometric morphometric principal component analysis was performed in the mid-sagittal plane of 31 wild-type mice's skulls from postnatal days 28 to 98. The relationship between the cranial base and cranial vault was investigated by comparing skulls with early fusion and non-fusion of intersphenoid synchondrosis (ISS). RESULTS: The cranial vault flattening and sphenoid bone length increased with age. The cranial vault curvature and sphenoid base length showed a positive correlation that was confirmed by comparing the skulls with early fusion and non-fusion of ISS. The sphenoid bone length and cranial vault angle significantly decreased in the skulls with early fusion of ISS compared to non-fusion skulls. CONCLUSIONS: It is suggested that the cranial vault flattening is sphenoid bone length-induced but cranial vault length-independent during postnatal mice skull development.

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