In Vivo Epithelial Wound Repair Requires Mobilization of Endogenous Intracellular and Extracellular Calcium
In Vivo Epithelial Wound Repair Requires Mobilization of Endogenous Intracellular and Extracellular Calcium
复制标题
DOI：
10.1074/jbc.m113.488098
复制
发表时间：
2013-11-22
影响因子：
4.8
通讯作者：
Montrose, Marshall H.
中科院分区：
文献类型：
--
作者：
Aihara, Eitaro;Hentz, Courtney L.;Montrose, Marshall H.
我们报道，在体内，细胞内和细胞外的局部钙离子动员发生在显微上皮损伤的部位，并且是介导组织修复所必需的。活体共聚焦/双光子显微镜连续成像手术暴露的麻醉小鼠的胃粘膜，而胃上皮表面细胞的光损伤造成了一个微观损伤，在15分钟内愈合。带有细胞内钙敏感蛋白(黄色骆驼3.0)的转基因小鼠报告说，在修复受损细胞附近的胃上皮细胞时，细胞内钙离子选择性地增加。U-73122、吲哚美辛、2-氨基乙氧基二苯基硼烷或异搏定可抑制损伤的修复，并抑制细胞内钙离子的增加。Fura-Red染料在腔内高融合液中的共聚焦成像显示，在细胞内钙动员后，损伤附近的胃表面有局限性的细胞外钙升高。吲哚美辛和维拉帕米也能抑制钙离子浓度的升高。细胞内钙离子螯合作用(1，2-二(邻氨基苯氧基)乙烷-N，N-四乙酸/乙酰氧甲酯，BAPTA/AM)可完全抑制细胞内和细胞内钙离子的增加，而细胞内钙离子的螯合作用(N-(2-羟乙基)-乙二胺-N，N，N-三乙酸三钠，HEDTA)可阻断细胞内钙离子的增加，并不均匀地抑制细胞内钙离子的晚期动员。这两种钙离子螯合模式都会减缓胃修复。在质膜Ca-ATPase 1(+/-)小鼠，而不是质膜Ca-ATPase 4(-/-)小鼠，存在缓慢的上皮修复和减少的胃表面钙增加。我们的结论是，内源性钙离子通过信号通路和跨膜钙离子转运而动员，导致上皮损伤部位的钙水平升高，这是在体胃上皮细胞重建所必需的。
We report that a localized intracellular and extracellular Ca2+ mobilization occurs at the site of microscopic epithelial damage in vivo and is required to mediate tissue repair. Intravital confocal/two-photon microscopy continuously imaged the surgically exposed stomach mucosa of anesthetized mice while photodamage of gastric epithelial surface cells created a microscopic lesion that healed within 15 min. Transgenic mice with an intracellular Ca2+-sensitive protein (yellow cameleon 3.0) report that intracellular Ca2+ selectively increases in restituting gastric epithelial cells adjacent to the damaged cells. Pretreatment with U-73122, indomethacin, 2-aminoethoxydiphenylborane, or verapamil inhibits repair of the damage and also inhibits the intracellular Ca2+ increase. Confocal imaging of Fura-Red dye in luminal superfusate shows a localized extracellular Ca2+ increase at the gastric surface adjacent to the damage that temporally follows intracellular Ca2+ mobilization. Indomethacin and verapamil also inhibit the luminal Ca2+ increase. Intracellular Ca2+ chelation (1,2-bis(o-aminophenoxy)ethane-N,N,N,N-tetraacetic acid/acetoxymethyl ester, BAPTA/AM) fully inhibits intracellular and luminal Ca2+ increases, whereas luminal calcium chelation (N-(2-hydroxyetheyl)-ethylendiamin-N,N,N-triacetic acid trisodium, HEDTA) blocks the increase of luminal Ca2+ and unevenly inhibits late-phase intracellular Ca2+ mobilization. Both modes of Ca2+ chelation slow gastric repair. In plasma membrane Ca-ATPase 1(+/-) mice, but not plasma membrane Ca-ATPase 4(-/-) mice, there is slowed epithelial repair and a diminished gastric surface Ca2+ increase. We conclude that endogenous Ca2+, mobilized by signaling pathways and transmembrane Ca2+ transport, causes increased Ca2+ levels at the epithelial damage site that are essential to gastric epithelial cell restitution in vivo.