Sehy Jonathan V, Banks Alison A, Ackerman Joseph J H, Neil Jeffrey J
Program in Molecular Cell Biology, School of Medicine, Washington University, St. Louis, MO 63130, USA.
Biophys J. 2002 Nov;83(5):2856-63. doi: 10.1016/S0006-3495(02)75294-6.
The exchange of water across biological membranes is of fundamental significance to both animal and plant physiology. Diffusional membrane permeability (P(d)) for the Xenopus oocyte, an important model system for water channel investigation, is typically calculated from intracellular water pre-exchange lifetime, cell volume, and cell surface area. There is debate, however, whether intracellular water motion affects water lifetime, and thereby P(d). Mathematical modeling of water transport is problematic because the intracellular water diffusion rate constant (D) for cells is usually unknown. The measured permeability may be referred to as the apparent diffusional permeability, P, to acknowledge this potential error. Herein, we show that magnetic resonance (MR) spectroscopy can be used to measure oocyte water exchange with greater temporal resolution and higher signal-to-noise ratio than other methods. MR imaging can be used to assess both oocyte geometry and intracellular water diffusion for the same single cells. MR imaging is used to confirm the dependence of intracellular water lifetime on intracellular diffusion. A model is presented to relate intracellular lifetime to true membrane diffusional permeability. True water diffusional permeability (2.7 +/- 0.4 microm/s) is shown to be 39 +/- 6% greater than apparent diffusional permeability for 8 oocytes. This discrepancy increases with cell size and permeability (such as after water channel expression) and decreases with increasing intracellular water D.
水在生物膜间的交换对动植物生理学都具有根本意义。非洲爪蟾卵母细胞是水通道研究的重要模型系统,其扩散膜通透性(P(d))通常根据细胞内水预交换寿命、细胞体积和细胞表面积来计算。然而,关于细胞内水的运动是否会影响水的寿命,进而影响P(d),仍存在争议。水运输的数学建模存在问题,因为细胞的细胞内水扩散速率常数(D)通常是未知的。为了承认这种潜在误差,所测得的通透性可称为表观扩散通透性P。在此,我们表明,与其他方法相比,磁共振(MR)波谱可用于以更高的时间分辨率和更高的信噪比测量卵母细胞的水交换。MR成像可用于评估同一单个细胞的卵母细胞几何形状和细胞内水扩散。MR成像用于确认细胞内水寿命对细胞内扩散的依赖性。提出了一个模型,将细胞内寿命与真实的膜扩散通透性联系起来。结果表明,8个卵母细胞的真实水扩散通透性(2.7±0.4微米/秒)比表观扩散通透性大39±6%。这种差异随着细胞大小和通透性(如在水通道表达后)的增加而增加,随着细胞内水D的增加而减小。