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衰老研究中的激光微束与光镊技术。

Laser microbeams and optical tweezers in ageing research.

作者信息

Grigaravicius Paulius, Greulich Karl Otto, Monajembashi Shamci

机构信息

Leibniz Institute of Age Research, Beutenbergstrasse 11, 07745 Jena, Germany.

出版信息

Chemphyschem. 2009 Jan 12;10(1):79-85. doi: 10.1002/cphc.200800725.

Abstract

We show how a technique developed within the framework of physics and physical chemistry-in a true interdisciplinary approach-can answer questions in life sciences that are not solvable by using other techniques. Herein, we focus on blood-pressure regulation and DNA repair in ageing studies. Laser microbeams and optical tweezers are now established tools in many fields of science, particularly in the life sciences. A short glimpse is given on the wide field of non-age-research applications in life sciences. Then, optical tweezers are used to show that exerting a vertical pressure on cells representing the inner lining of blood vessels results in bursts of NO liberation concomitant with large changes in cell morphology. Repeated treatment of such human umbilical vein endothelial cells (HUVEC) results in stiffening, a hallmark of manifest high blood pressure, a disease primarily of the elderly. As a second application in ageing research, a laser microbeam is used to induce, with high spatial and temporal resolution, DNA damages in the nuclei of U2OS human osteosarcoma cells. A pairwise study of the recruitment kinetics of different DNA repair proteins reveals that DNA repair starts with non-homologous end joining (NHEJ), a repair pathway, and may only after several minutes switch to the error-free homologous recombination repair (HRR) pathway. Since DNA damages-when incorrectly repaired-accumulate with time, laser microbeams are becoming well-used tools in ageing research.

摘要

我们展示了一种在物理学和物理化学框架内开发的技术——以真正的跨学科方法——如何能够回答生命科学中使用其他技术无法解决的问题。在此,我们聚焦于衰老研究中的血压调节和DNA修复。激光微束和光镊现在是许多科学领域,特别是生命科学领域中既定的工具。我们简要介绍了生命科学中广泛的非衰老研究应用领域。然后,用光镊表明,对代表血管内衬的细胞施加垂直压力会导致一氧化氮(NO)释放爆发,并伴随着细胞形态的巨大变化。对这种人脐静脉内皮细胞(HUVEC)进行反复处理会导致细胞硬化,这是明显高血压(一种主要发生在老年人中的疾病)的一个标志。作为衰老研究中的第二个应用,使用激光微束以高空间和时间分辨率在U2OS人骨肉瘤细胞核中诱导DNA损伤。对不同DNA修复蛋白募集动力学的成对研究表明,DNA修复始于非同源末端连接(NHEJ)(一种修复途径),并且可能仅在几分钟后才切换到无错误的同源重组修复(HRR)途径。由于DNA损伤(如果修复不正确)会随着时间积累,激光微束正成为衰老研究中常用的工具。

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