Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, MA 02115, USA.
Proc Natl Acad Sci U S A. 2013 Feb 5;110(6):2163-8. doi: 10.1073/pnas.1206392110. Epub 2013 Jan 22.
Bone remodeling requires osteoclasts to generate and maintain an acidified resorption compartment between the apical membrane and the bone surface to solubilize hydroxyapatite crystals within the bone matrix. This acidification process requires (i) apical proton secretion by a vacuolar H(+)-ATPase, (ii) actin cytoskeleton reorganization into a podosome belt that forms a gasket to restrict lacunar acid leakage, and (iii) basolateral chloride uptake and bicarbonate extrusion by an anion exchanger to provide Cl(-) permissive for apical acid secretion while preventing cytoplasmic alkalinization. Here we show that osteoclast-targeted deletion in mice of solute carrier family 4 anion exchanger member 2 (Slc4a2) results in osteopetrosis. We further demonstrate a previously unrecognized consequence of SLC4A2 loss of function in the osteoclast: dysregulation of calpain-dependent podosome disassembly, leading to abnormal actin belt formation, cell spreading, and migration. Rescue of SLC4A2-deficient osteoclasts with functionally defined mutants of SLC4A2 indicates regulation of actin cytoskeletal reorganization by anion-exchange activity and intracellular pH, independent of SLC4A2's long N-terminal cytoplasmic domain. These data suggest that maintenance of intracellular pH in osteoclasts through anion exchange regulates the actin superstructures required for bone resorption.
骨重建需要破骨细胞在顶膜和骨表面之间产生并维持酸化的吸收隔室,以溶解骨基质内的羟基磷灰石晶体。这个酸化过程需要:(i)液泡 H(+)-ATP 酶在顶膜处的质子分泌;(ii)肌动蛋白细胞骨架重组为形成垫圈的足突带,以限制腔隙酸泄漏;(iii)阴离子交换器在基底外侧摄取氯离子和排出碳酸氢根离子,以提供允许顶膜酸分泌的 Cl(-),同时防止细胞质碱化。在这里,我们显示了在小鼠中针对溶质载体家族 4 阴离子交换体成员 2 (Slc4a2) 的破骨细胞靶向缺失导致了骨质硬化症。我们进一步证明了 SLC4A2 功能丧失在破骨细胞中的一个先前未被认识的后果:钙蛋白酶依赖性足突解体的失调,导致异常的肌动蛋白带形成、细胞铺展和迁移。用功能定义的 SLC4A2 突变体拯救 SLC4A2 缺陷型破骨细胞表明,阴离子交换活性和细胞内 pH 对肌动蛋白细胞骨架重组的调节,独立于 SLC4A2 的长细胞质 N 端结构域。这些数据表明,破骨细胞中通过阴离子交换维持细胞内 pH 调节了骨吸收所需的肌动蛋白超结构。