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Adaptor protein complex 2mediated, clathrin-dependent endocytosis, and related gene activities, are a prominent feature during maturation stage amelogenesis

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单位: [1]Univ So Calif, Herman Ostrow Sch Dent, Ctr Craniofacial Mol Biol, Los Angeles, CA 90605 USA [2]Univ Leeds, Leeds Dent Inst, Dept Oral Biol, Leeds, W Yorkshire, England [3]Univ So Calif, Keck Sch Med, Dept Biochem & Mol Biol, Los Angeles, CA 90605 USA [4]Huazhong Univ Sci & Technol, Tongji Med Coll, Tongji Hosp, Ctr Stomatol, Wuhan 430074, Peoples R China [5]Univ Calif Los Angeles, Sch Dent, Los Angeles, CA 90024 USA [6]Univ Oslo, Fac Dent, Dept Biomat, Oslo, Norway [7]Univ So Calif, Sch Pharm, Dept Pharmacol & Pharmaceut Sci, Los Angeles, CA 90605 USA [8]McGill Univ, Dept Anat & Cell Biol, Facil Elect Microscopy Res, Montreal, PQ, Canada [9]McGill Univ, Fac Dent, Montreal, PQ, Canada
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关键词: AMELOGENESIS CLATHRIN EMDOGAIN ENAMEL ENDOSOMES ENDOCYTOSIS LYSOSOMES pH HOMEOSTASIS

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Molecular events defining enamel matrix removal during amelogenesis are poorly understood. Early reports have suggested that adaptor proteins (AP) participate in ameloblast-mediated endocytosis. Enamel formation involves the secretory and maturation stages, with an increase in resorptive function during the latter. Here, using real-time PCR, we show that the expression of clathrin and adaptor protein subunits are upregulated in maturation stage rodent enamel organ cells. AP complex 2 (AP-2) is the most upregulated of the four distinct adaptor protein complexes. Immunolocalization confirms the presence of AP-2 and clathrin in ameloblasts, with strongest reactivity at the apical pole. These data suggest that the resorptive functions of enamel cells involve AP-2 mediated, clathrin-dependent endocytosis, thus implying the likelihood of specific membrane-bound receptor(s) of enamel matrix protein debris. The mRNA expression of other endocytosis-related gene products is also upregulated during maturation including: lysosomal-associated membrane protein 1 (Lamp1); cluster of differentiation 63 and 68 (Cd63 and Cd68); ATPase, H+ transporting, lysosomal V0 subunit D2 (Atp6v0d2); ATPase, H+ transporting, lysosomal V1 subunit B2 (Atp6v1b2); chloride channel, voltage-sensitive 7 (Clcn7); and cathepsin K (Ctsk). Immunohistologic data confirms the expression of a number of these proteins in maturation stage ameloblasts. The enamel of Cd63-null mice was also examined. Despite increased mRNA and protein expression in the enamel organ during maturation, the enamel of Cd63-null mice appeared normal. This may suggest inherent functional redundancies between Cd63 and related gene products, such as Lamp1 and Cd68. Ameloblast-like LS8 cells treated with the enamel matrix protein complex Emdogain showed upregulation of AP-2 and clathrin subunits, further supporting the existence of a membrane-bound receptor-regulated pathway for the endocytosis of enamel matrix proteins. These data together define an endocytotic pathway likely used by ameloblasts to remove the enamel matrix during enamel maturation. (c) 2013 American Society for Bone and Mineral Research.

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出版当年[2012]版:
大类 | 1 区 医学
小类 | 2 区 内分泌学与代谢
最新[2025]版:
大类 | 1 区 医学
小类 | 1 区 内分泌学与代谢
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出版当年[2011]版:
Q1 ENDOCRINOLOGY & METABOLISM
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Q1 ENDOCRINOLOGY & METABOLISM

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第一作者单位: [1]Univ So Calif, Herman Ostrow Sch Dent, Ctr Craniofacial Mol Biol, Los Angeles, CA 90605 USA
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通讯机构: [1]Univ So Calif, Herman Ostrow Sch Dent, Ctr Craniofacial Mol Biol, Los Angeles, CA 90605 USA [*1]Univ So Calif, Herman Ostrow Sch Dent, Ctr Craniofacial Mol Biol, 2250 Alcazar St,CSA103, Los Angeles, CA 90605 USA
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