The α isoform of protein kinase C is involved in signaling the response of desmosomes to wounding in cultured epithelial cells
The α isoform of protein kinase C is involved in signaling the response of desmosomes to wounding in cultured epithelial cells
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DOI：
10.1091/mbc.11.3.1077
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发表时间：
2000-03-01
影响因子：
3.3
通讯作者：
Garrod, D
中科院分区：
文献类型：
--
作者：
Wallis, S;Lloyd, S;Garrod, D
创伤后上皮再生的启动机制仍知之甚少。为了加强这种理解，我们在这里集中在培养的Madin-Darby犬肾细胞的桥粒的粘附状态的变化，以响应伤口的汇合细胞片。先前的研究结果表明，Madin-Darby犬肾细胞桥粒粘附从钙依赖性状态变化到融合细胞片中的钙独立性。我们表明，这种变化，这需要文化融合发展，是迅速逆转后伤口融合细胞片。此外，伤口边缘细胞中钙依赖性的变化传播到距离伤口边缘数百微米的细胞。当用激活PKC的佛波醇酯处理细胞时，也会发生从钙非依赖性到钙依赖性的快速转变。PKC抑制剂，包括传统的亚型抑制剂Go 6976，导致从钙依赖性到钙非依赖性的快速转变，即使在亚融合细胞中。PKC α亚型的细胞定位与桥粒的钙依赖性相关。在单层创伤后，PKC α迅速易位到细胞外周，变成Triton X-100不溶性，并且也变成浓缩在片状伪足中。损伤后PKC α a易位先于膜组分中PKC活性的增加和桥粒对钙依赖性的逆转。用反义寡核苷酸特异性消耗PKC α增加了具有钙非依赖性桥粒的细胞数量。这些结果表明，PKC α参与了一种新的信号通路，调节桥粒粘附在创伤。
Initiation of reepithelialization upon wounding is still poorly understood. To enhance this understanding, we focus here on changes in the adhesive state of desmosomes of cultured Madin-Darby canine kidney cells in response to wounding of confluent cell sheets. Previous results show that desmosomal adhesion in Madin-Darby canine kidney cells changes from a calcium-dependent state to calcium independence in confluent cell sheets. We show that this change, which requires culture confluence to develop, is rapidly reversed upon wounding of confluent cell sheets. Moreover, the change to calcium dependence in wound edge cells is propagated to cells hundreds of micrometers away from the wound edge. Rapid transition from calcium independence to calcium dependence also occurs when cells are treated with phorbol esters that activate PKC. PKC inhibitors, including the conventional isoform inhibitor Go6976, cause rapid transition from calcium dependence to calcium independence, even in subconfluent cells. The cellular location of the a isoform of PKC correlates with the calcium dependence of desmosomes. Upon monolayer wounding, PKC alpha translocates rapidly to the cell periphery, becomes Triton X-100 insoluble, and also becomes concentrated in lamellipodia. The PKC alpha a translocation upon wounding precedes both the increase in PKC activity in the membrane fraction and the reversion of desmosomes to calcium dependence. Specific depletion of PKC alpha with an antisense oligonucleotide increases the number of cells with calcium-independent desmosomes. These results show that PKC alpha participates in a novel signaling pathway that modulates desmosomal adhesion in response to wounding.