Dept of Oral Cell Biology ACTA, University of Amsterdam and VU University Amsterdam, Research Institute MOVE, Gustav Mahlerlaan 3004, Amsterdam, The Netherlands.
Bone. 2012 Apr;50(4):901-8. doi: 10.1016/j.bone.2011.12.019. Epub 2012 Jan 8.
Maturation stage ameloblasts of rodents express vacuolar type-H-ATPase in the ruffled border of their plasma membrane in contact with forming dental enamel, similar to osteoclasts that resorb bone. It has been proposed that in ameloblasts this v-H-ATPase acts as proton pump to acidify the enamel space, required to complete enamel mineralization. To examine whether this v-H-ATPase in mouse ameloblasts is a proton pump, we determined whether these cells express the lysosomal, T-cell, immune regulator 1 (Tcirg1, v-H-Atp6v(0)a(3)), which is an essential part of the plasma membrane proton pump that is present in osteoclasts. Mutation of this subunit in Tcirg1 null (or oc/oc) mice leads to severe osteopetrosis. No immunohistochemically detectable Tcirg1 was seen in mouse maturation stage ameloblasts. Strong positive staining in secretory and maturation stage ameloblasts however was found for another subunit of v-H-ATPase, subunit b, brain isoform (v-H-Atp6v(1)b(2)). Mouse osteoclasts and renal tubular epithelium stained strongly for both Tcirg1 and v-H-Atp6v(1)b(2). In Tcirg1 null mice osteoclasts and renal epithelium were negative for Tcirg1 but remained positive for v-H-Atp6v(1)b(2). The bone in these mutant mice was osteopetrotic, tooth eruption was inhibited or delayed, and teeth were often morphologically disfigured. However, enamel formation in these mutant mice was normal, ameloblasts structurally unaffected and the mineral content of enamel similar to that of wild type mice. We concluded that Tcirg1, which is essential for osteoclasts to pump protons into the bone, is not appreciably expressed in maturation stage mouse ameloblasts. Our data suggest that the reported v-H-ATPase in maturation stage ameloblasts is not the typical osteoclast-type plasma membrane associated proton pump which acidifies the extracellular space, but rather a v-H-ATPase potentially involved in intracellular acidification.
鼠类成釉细胞在形成牙釉质的过程中,其质膜的皱褶缘表达有空泡型 H+-ATP 酶(v-H-ATPase),类似于破骨细胞吸收骨组织。有人提出,在成釉细胞中,这种 v-H-ATP 酶作为质子泵,将牙釉质空间酸化,这是完成牙釉质矿化所必需的。为了研究鼠类成釉细胞中的这种 v-H-ATP 酶是否为质子泵,我们测定了这些细胞是否表达溶酶体、T 细胞免疫调节因子 1(Tcirg1,v-H-Atp6v(0)a(3)),这是破骨细胞中存在的质膜质子泵的重要组成部分。Tcirg1 缺失(或 oc/oc)小鼠中该亚基的突变导致严重的骨质增生。在鼠类成釉细胞的成熟阶段,免疫组织化学检测不到 Tcirg1。然而,在分泌期和成釉细胞期的成釉细胞中,v-H-ATP 酶的另一个亚基,即脑型亚基 b(v-H-Atp6v(1)b(2)),表现出强烈的阳性染色。鼠类破骨细胞和肾小管上皮细胞均强烈表达 Tcirg1 和 v-H-Atp6v(1)b(2)。在 Tcirg1 缺失的小鼠中,破骨细胞和肾小管上皮细胞对 Tcirg1 呈阴性反应,但对 v-H-Atp6v(1)b(2)仍呈阳性反应。这些突变小鼠的骨呈骨质增生,牙齿萌出受阻或延迟,牙齿常呈畸形。然而,这些突变小鼠的牙釉质形成正常,成釉细胞结构不受影响,牙釉质的矿化程度与野生型小鼠相似。我们得出结论,Tcirg1 是破骨细胞向骨内泵入质子所必需的,在鼠类成釉细胞的成熟阶段并不明显表达。我们的数据表明,在成熟阶段的成釉细胞中报道的 v-H-ATP 酶不是典型的破骨细胞型质膜相关质子泵,该质子泵酸化细胞外空间,而是一种可能参与细胞内酸化的 v-H-ATP 酶。