NMR and Fluorescence Studies on Exocrine Membrane Transport

利用核磁共振和荧光光谱技术研究了与分泌有关的外分泌腺上皮转运过程，使我们能够无创地研究完整灌流的大鼠唾液腺细胞内的许多参数，细胞内液电解质和水的分子动力学研究取得了很大进展。核磁共振谱（NMR）可以直接观察分子动力学。在这几十年中已经完成了三项技术和理论创新：i）化学位移试剂以分离细胞内和细胞外信号，ii）多量子NMR以检测分子相关时间（t），以及iii）脉冲场梯度NMR以检测分子扩散系数（D）。细胞内阳离子（Na、K、Rb）的运动要慢得多（D（）SY）。[）0.2 cm<@D12@>D1/sec）比细胞外阳离子（τ（）SY. [）10<@D1-12@>D1秒，D（] SY.apprxeq. [）2 cm<@D12@>D1/sec）。一小部分内部控制 关于我们 细胞Na<@D1+@>D1预期与大分子（tau（）SY）结合。[）10<@D1-7@>D1秒），并与“游离”Na<@D1+@>D1（τ（）SY. [）10<@D1-11@>D1秒）细胞内液。水的运动由于细胞内和细胞外水穿过细胞膜的相对快速的交换而有点复杂（Pd（OH）SY.apprxeq. [）3.10<@D1-3@>D1 cm/sec）。细胞内水的内在运动（D（）SY.apprxeq. [）0.2 cm<@D12@>D1/sec）估计比胞外水慢十分之一。因此，我们应该提醒电解质和水的微观环境与细胞外环境是完全不同的。（<@D1*@> D1所有值均在24* 下测量。）另一方面，我们开发了一种荧光技术来测量灌注的器官和分散的细胞。现在，我们可以比较两种实验条件下的结果，也可以很好地连接使用分离的细胞进行的生化研究。少

The epithelial transport processes associated with secretion in exocrine glands was studied by using NMR and fluorescence spectroscopies, which allow us to study many of the intracellular parameters noninvasively in the intact, perfused rat salivary gland.Investigations to the molecular dynamics of the electrolytes and water in the intracellular fluid got a big progress. Nuclear Magnetic Resonance Spectroscopy (NMR) can observe molecular dynamics directly. Three technical and theoretical innovations have been done in these decades : i)chemical shift reagent to separate the intra- and extracellular signal, ii)multi-quantum NMR to detect molecular correlation time (t), and iii)pulsed fielded gradient NMR to detect molecular diffusion coefficient (D). Motion of the intracellular cations (Na, K, Rb) is much slower (D (] SY.apprxeq.[) 0.2 cm<@D12@>D1/sec) than the extracellular cations (tau (] SY.apprxeq.[) 10<@D1-12@>D1 sec, D (] SY.apprxeq.[) 2 cm<@D12@>D1/sec). A small fraction of intrac … More ellular Na<@D1+@>D1 expected to be bound the macro-molecules (tau (] SY.apprxeq.[) 10<@D1-7@>D1 sec), and exchange rapidly with "free" Na<@D1+@>D1 (tau (] SY.apprxeq.[) 10<@D1-11@>D1 sec) intracellular fluid. Motion of water is a bit complicated by relatively rapid exchange of intra- and extracellular water across the cell membrane (Pd (] SY.apprxeq.[) 3.10<@D1-3@>D1 cm/sec). The intrinsic motion of the intracellular water (D (] SY.apprxeq.[) 0.2 cm<@D12@>D1/sec) is estimated one tenth slower than that of the extracellular water. Therefore, we should remind the microscopic environment of electrolytes and water is complete different from the extracellular milieu.(<@D1*@>D1All values are measured at 24*.)On the other hand, we had developed a fluorescence technique to measure the perfused organ and also the dispersed cells. A protocol is established to quantify the intracellular Ca and pH. Now, we can compare the results from the both experimental conditions, and also can a good connection between the biochemical studies have done using the isolated cells. Less