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首页> 外文期刊>The Journal of investigative dermatology. >A transgenic mouse for imaging caspase-dependent apoptosis within the skin.
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A transgenic mouse for imaging caspase-dependent apoptosis within the skin.

机译：用于在皮肤内成像caspase依赖性细胞凋亡的转基因小鼠。

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Apoptosis is an essential process for the maintenance of normal physiology. The ability to noninvasively image apoptosis in living animals would provide unique insights into its role in normal and disease processes. Herein, a recombinant reporter consisting of beta-galactosidase gene flanked by two estrogen receptor regulatory domains and intervening Asp-Glu-Val-Glu sequences was constructed to serve as a tool for in vivo assessment of apoptotic activity. The results demonstrate that when expressed in its intact form, the hybrid reporter had undetectable beta-galactosidase activity. Caspase 3 activation in response to an apoptotic stimulus resulted in cleavage of the reporter, and thereby reconstitution of beta-galactosidase activity. Enzymatic activation of the reporter during an apoptotic event enabled noninvasive measurement of beta-galactosidase activity in living cells, which correlated with traditional measures of apoptosis in a dose- and time-dependent manner. Using a near-infrared fluorescent substrate of beta-galactosidase (9H-{1,3-dichloro-9,9-dimethylacridin-2-one-7-yl} beta-D-galactopyranoside), noninvasive in vivo imaging of apoptosis was achieved in a xenograft tumor model in response to proapoptotic therapy. Finally, a transgenic mouse model was developed expressing the ER-LACZ-ER reporter within the skin. This reporter and transgenic mouse could serve as a unique tool for the study of apoptosis in living cells and animals, especially in the context of skin biology.
机译：细胞凋亡是维持正常生理的重要过程。对活体动物凋亡进行无创成像的能力将为其在正常和疾病过程中的作用提供独特的见解。在本文中，构建了由两个侧翼的雌激素受体调节域和中间的Asp-Glu-Val-Glu序列两侧的β-半乳糖苷酶基因组成的重组报告基因，以用作体内评估细胞凋亡活性的工具。结果表明，当以完整形式表达时，杂交报告基因具有不可检测的β-半乳糖苷酶活性。响应凋亡刺激的胱天蛋白酶3激活导致报道基因的裂解，从而重建了β-半乳糖苷酶的活性。凋亡过程中报道分子的酶促活化使得能够对活细胞中的β-半乳糖苷酶活性进行非侵入性测量，这与剂量和时间依赖性方式与传统的细胞凋亡测量方法相关。使用β-半乳糖苷酶（9H- {1,3-二氯-9,9-二甲基ac啶-2-酮-7-基}β-D-吡喃半乳糖苷）的近红外荧光底物，实现了细胞凋亡的无创体内成像响应促凋亡疗法在异种移植肿瘤模型中的表达。最后，开发了在皮肤内表达ER-LACZ-ER报道基因的转基因小鼠模型。该报道基因和转基因小鼠可以作为研究活细胞和动物凋亡的独特工具，尤其是在皮肤生物学的情况下。

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