Conditional disruption of ubiquitous calpains in the mouse
Conditional disruption of ubiquitous calpains in the mouse
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DOI：
10.1002/dvg.20216
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发表时间：
2006-06-01
期刊：
影响因子：
1.5
通讯作者：
Greer, Peter A.
中科院分区：
文献类型：
--
作者：
Tan, Yinfei;Dourdin, Nathalie;Greer, Peter A.
广泛存在的u-和m-calain蛋白与发育和细胞凋亡密切相关。它们是由分别由capn1和capn2编码的80 kDa催化亚基和由capn4编码的共同的28 kDa调节亚基组成的杂二聚体。调节亚基是维持u-和m-calain的稳定性和活性所必需的；因此，可以预测，capn4的基因破坏会消除这两种calain的活性。Capn4的胚系破坏导致胚胎死亡，阻碍了使用这些小鼠模型来探索生理钙蛋白功能。在这里，我们描述了一种以loxP/cre条件性capn4为靶标的小鼠模型，该模型能够实现组织特异性和暂时删除calain活性。利用普遍存在的巨细胞病毒启动子驱动的Cre重组酶转基因来破坏花朵状的capn4基因会导致中期胚胎死亡。这些胚胎的成纤维细胞缺乏可检测到的调节亚单位的表达，降低了u-和m-calain催化亚基的水平，并且没有检测到的u-和m-calain活性。这些缺陷被编码了Capn4的慢病毒纠正。
Ubiquitous mu- and m-calpain proteases are implicated in development and apoptosis. They are heterodimers consisting of 80-kDa catalytic subunits encoded by capn1 and capn2, respectively, and a common 28-kDa regulatory subunit encoded by capn4. The regulatory subunit is required to maintain stability and activity of mu- and m-calpains; thus, genetic disruption of capn4 was predicted to eliminate both calpain activities. Germline disruption of capn4 caused embryonic lethality, hampering the use of those mouse models to explore physiological calpain functions. Here we describe a loxP/cre conditional capn4 targeted mouse model that enables tissue-specific and temporal deletion of calpain activity. Disruption of the floxed capn4 gene using a ubiquitous cytomegalovirus promoter driven Cre recombinase transgene led to midgestation embryonic lethality. Fibroblasts from these embryos lacked detectable regulatory subunit expression, had reduced levels of the mu- and m-calpain catalytic subunits, and had no detectable mu- and m-calpain activities. These defects were corrected with a capn4-encoding lentivirus.