Josephsen Kaj, Praetorius Jeppe, Frische Sebastian, Gawenis Lara R, Kwon Tae-Hwan, Agre Peter, Nielsen Søren, Fejerskov Ole
The Water and Salt Research Center, Faculty of Health Sciences, Department of Dental Pathology, Operative Dentistry and Endodontics, School of Dentistry, and Institute of Anatomy, University of Aarhus, DK-8000 Aarhus C, Denmark.
Proc Natl Acad Sci U S A. 2009 Feb 3;106(5):1638-41. doi: 10.1073/pnas.0811682106. Epub 2009 Jan 21.
Osteoclasts are multinucleated bone-resorbing cells responsible for constant remodeling of bone tissue and for maintaining calcium homeostasis. The osteoclast creates an enclosed space, a lacuna, between their ruffled border membrane and the mineralized bone. They extrude H(+) and Cl(-) into these lacunae by the combined action of vesicular H(+)-ATPases and ClC-7 exchangers to dissolve the hydroxyapatite of bone matrix. Along with intracellular production of H(+) and HCO(3)(-) by carbonic anhydrase II, the H(+)-ATPases and ClC-7 exchangers seems prerequisite for bone resorption, because genetic disruption of either of these proteins leads to osteopetrosis. We aimed to complete the molecular model for lacunar acidification, hypothesizing that a HCO(3)(-) extruding and Cl(-) loading anion exchange protein (Ae) would be necessary to sustain bone resorption. The Ae proteins can provide both intracellular pH neutrality and serve as cellular entry mechanism for Cl(-) during bone resorption. Immunohistochemistry revealed that Ae2 is exclusively expressed at the contra-lacunar plasma membrane domain of mouse osteoclast. Severe osteopetrosis was encountered in Ae2 knockout (Ae2-/-) mice where the skeletal development was impaired with a higher diffuse radio-density on x-ray examination and the bone marrow cavity was occupied by irregular bone speculae. Furthermore, osteoclasts in Ae2-/- mice were dramatically enlarged and fail to form the normal ruffled border facing the lacunae. Thus, Ae2 is likely to be an essential component of the bone resorption mechanism in osteoclasts.
破骨细胞是负责骨组织持续重塑和维持钙稳态的多核骨吸收细胞。破骨细胞在其褶皱边缘膜与矿化骨之间形成一个封闭空间,即腔隙。它们通过囊泡型H(+)-ATP酶和ClC-7交换体的共同作用将H(+)和Cl(-) 分泌到这些腔隙中,以溶解骨基质中的羟基磷灰石。碳酸酐酶II在细胞内产生H(+)和HCO(3)(-),H(+)-ATP酶和ClC-7交换体似乎是骨吸收的先决条件,因为这两种蛋白质中任何一种的基因破坏都会导致骨硬化。我们旨在完善腔隙酸化的分子模型,假设一种能分泌HCO(3)(-)并装载Cl(-)的阴离子交换蛋白(Ae)对于维持骨吸收是必要的。Ae蛋白既能维持细胞内pH值中性,又能在骨吸收过程中作为Cl(-)的细胞进入机制。免疫组织化学显示,Ae2仅在小鼠破骨细胞的腔隙对侧质膜结构域表达。在Ae2基因敲除(Ae2-/-)小鼠中出现了严重的骨硬化,其骨骼发育受损,X线检查显示弥漫性放射密度较高,骨髓腔被不规则的骨小梁占据。此外,Ae2-/-小鼠中的破骨细胞显著增大且无法形成面向腔隙的正常褶皱边缘。因此,Ae2可能是破骨细胞骨吸收机制的一个重要组成部分。