Guevorkian Karine, Valles James M
Department of Physics, Brown University, Providence, RI 02912, USA.
Proc Natl Acad Sci U S A. 2006 Aug 29;103(35):13051-6. doi: 10.1073/pnas.0601839103. Epub 2006 Aug 17.
Earth's gravity exerts relatively weak forces in the range of 10-100 pN directly on cells in biological systems. Nevertheless, it biases the orientation of swimming unicellular organisms, alters bone cell differentiation, and modifies gene expression in renal cells. A number of methods of simulating different strength gravity environments, such as centrifugation, have been applied for researching the underlying mechanisms. Here, we demonstrate a magnetic force-based technique that is unique in its capability to enhance, reduce, and even invert the effective buoyancy of cells and thus simulate hypergravity, hypogravity, and inverted gravity environments. We apply it to Paramecium caudatum, a single-cell protozoan that varies its swimming propulsion depending on its orientation with respect to gravity, g. In these simulated gravities, denoted by f(gm), Paramecium exhibits a linear response up to f(gm) = 5 g, modifying its swimming as it would in the hypergravity of a centrifuge. Moreover, experiments from f(gm) = 0 to -5 g show that the response is symmetric, implying that the regulation of the swimming speed is primarily related to the buoyancy of the cell. The response becomes nonlinear for f(gm) >5 g. At f(gm) = 10 g, many paramecia "stall" (i.e., swim in place against the force), exerting a maximum propulsion force estimated to be 0.7 nN. These findings establish a general technique for applying continuously variable forces to cells or cell populations suitable for exploring their force transduction mechanisms.
地球引力在生物系统中直接作用于细胞时,所施加的力相对较弱,范围在10 - 100皮牛之间。然而,它会使游动的单细胞生物的取向产生偏差,改变骨细胞分化,并影响肾细胞中的基因表达。为了研究其潜在机制,人们已经应用了多种模拟不同强度重力环境的方法,比如离心法。在此,我们展示了一种基于磁力的技术,该技术的独特之处在于能够增强、减弱甚至反转细胞的有效浮力,从而模拟超重、失重和反重力环境。我们将其应用于尾草履虫,这是一种单细胞原生动物,其游动推进方式会根据自身相对于重力g的取向而变化。在这些以f(gm)表示的模拟重力环境中,尾草履虫在f(gm) = 5g之前呈现线性响应,其游动方式的改变就如同在离心机超重环境中的变化一样。此外,从f(gm) = 0到 - 5g的实验表明,这种响应是对称的,这意味着游动速度的调节主要与细胞的浮力有关。当f(gm) > 5g时,响应变为非线性。在f(gm) = 10g时,许多草履虫“停滞”(即逆着力原地游动),施加的最大推进力估计为0.7纳牛。这些发现建立了一种通用技术,可对细胞或细胞群体施加连续可变的力,适用于探索它们的力转导机制。