Guo Jing, Bervoets Theodore J M, Henriksen Kim, Everts Vincent, Bronckers Antonius L J J
Department Oral Cell Biology, Academic Center of Dentistry Amsterdam (ACTA), University of Amsterdam and VU-University of Amsterdam, MOVE Research Institute, Gustav Mahlerlaan 3004, 1081 LA, Amsterdam, The Netherlands.
School of Stomatology/Dental Clinic, Zhejiang Chinese Medical University, Mailbox 97, Binwen Road 548, Binjiang District, 310053, Hangzhou, China.
Cell Tissue Res. 2016 Feb;363(2):361-70. doi: 10.1007/s00441-015-2263-z. Epub 2015 Sep 8.
ClC-7, located in late endosomes and lysosomes, is critical for the function of osteoclasts. Secretion of Cl(-) by the ruffled border of osteoclasts enables H(+) secretion by v-H(+)-ATPases to dissolve bone mineral. Mice lacking ClC-7 show altered lysosomal function that leads to severe lysosomal storage. Maturation ameloblasts are epithelial cells with a ruffled border that secrete Cl(-) as well as endocytose and digest large quantities of enamel matrix proteins during formation of dental enamel. We tested the hypothesis that ClC-7 in maturation ameloblasts is required for intracellular digestion of matrix fragments to complete enamel mineralization. Craniofacial bones and developing teeth in Clcn7(-/-) mice were examined by micro-CT, immunohistochemistry, quantified histomorphometry and electron microscopy. Osteoclasts and ameloblasts in wild-type mice stained intensely with anti-ClC-7 antibody but not in Clcn7(-/-) mice. Craniofacial bones in Clcn7(-/-) mice were severely osteopetrotic and contained 1.4- to 1.6-fold more bone volume, which was less mineralized than the wild-type littermates. In Clcn7(-/-) mice maturation ameloblasts and osteoclasts highly expressed Ae2 as in wild-type mice. However, teeth failed to erupt, incisors were much shorter and roots were disfigured. Molars formed a normal dental crown. In compacted teeth, dentin was slightly less mineralized, enamel did not retain a matrix and mineralized fairly normal. We concluded that ClC-7 is essential for osteoclasts to resorb craniofacial bones to enable tooth eruption and root development. Disruption of Clcn7 reduces bone and dentin mineral density but does not affect enamel mineralization.
位于晚期内体和溶酶体中的氯离子通道蛋白7(ClC-7)对破骨细胞的功能至关重要。破骨细胞褶皱缘分泌氯离子可使液泡型质子ATP酶分泌质子,从而溶解骨矿物质。缺乏ClC-7的小鼠表现出溶酶体功能改变,导致严重的溶酶体贮积。成熟成釉细胞是具有褶皱缘的上皮细胞,在牙釉质形成过程中分泌氯离子,并内吞和消化大量釉质基质蛋白。我们验证了这样一个假说,即成熟成釉细胞中的ClC-7是细胞内消化基质片段以完成釉质矿化所必需的。通过显微CT、免疫组织化学、定量组织形态计量学和电子显微镜检查了Clcn7基因敲除小鼠的颅面骨和发育中的牙齿。野生型小鼠的破骨细胞和成釉细胞用抗ClC-7抗体染色强烈,而Clcn7基因敲除小鼠则不然。Clcn7基因敲除小鼠的颅面骨严重骨硬化,骨体积比野生型同窝小鼠多1.4至1.6倍,矿化程度低于野生型同窝小鼠。在Clcn7基因敲除小鼠中,成熟成釉细胞和破骨细胞与野生型小鼠一样高表达Ae2。然而,牙齿未能萌出,门齿短得多,牙根变形。磨牙形成了正常的牙冠。在致密的牙齿中,牙本质矿化程度略低,釉质没有保留基质,矿化基本正常。我们得出结论,ClC-7对于破骨细胞吸收颅面骨以实现牙齿萌出和牙根发育至关重要。Clcn7基因的破坏降低了骨和牙本质的矿物质密度,但不影响釉质矿化。