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NMR and Fluorescence Studies on Exocrine Membrane Transport

外分泌膜运输的核磁共振和荧光研究

基本信息

批准号：
03044153
负责人：
SEO Yoshiteru
金额：
$ 14.08万
依托单位：
Okazaki National Research Institutes National Institute for Physiological Sciences
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1993
项目状态：
已结题

项目摘要

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

利用核磁共振和荧光光谱研究了与外分泌腺分泌相关的上皮运输过程，这使我们能够在完整的灌注大鼠唾液腺中无创地研究许多细胞内参数。细胞内液中电解质和水的分子动力学研究取得了很大进展。核磁共振波谱（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)分离细胞内和细胞外信号的化学移位试剂，ii)检测分子相关时间(t)的多量子核磁共振，以及iii)检测分子扩散系数(D)的脉冲场梯度核磁共振。胞内阳离子（Na， K, Rb）的运动要慢得多。[) 0.2 cm<@D12@>D1/sec)比细胞外阳离子（tau） ([] y .apprxeq。[1]刘建平，刘建平。[) 2 cm<@D12@>D1/sec)。一小部分胞内Na<@D1+@>D1被认为与大分子(tau [] SY.apprxeq。[) 10<@D1-7@>D1 sec)，与“自由”Na<@D1+@>D1 (tau (] SY.apprxeq。[) 10<@D1-11@>D1 sec)胞内液。由于细胞内和细胞外的水在细胞膜上相对快速的交换，水的运动有点复杂。[) 3.10<@D1-3@>D1 cm/sec)。细胞内水的内在运动[D] . [j]。[) 0.2 cm<@D12@>D1/sec)估计比细胞外水慢十分之一。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*处测量。）另一方面，我们开发了一种荧光技术来测量灌注器官和分散细胞。建立了一套定量胞内Ca和ph的方案。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细胞所做的生化研究。少