Hillenkamp Jost, Hussain Ali A, Jackson Timothy L, Constable Paul A, Cunningham Joanna R, Marshall John
Department of Ophthalmology, The Rayne Institute, St. Thomas' Hospital, London, UK.
Invest Ophthalmol Vis Sci. 2004 Nov;45(11):4099-105. doi: 10.1167/iovs.04-0624.
To assess the relative resistance presented individually by Bruch's membrane-choroid (BC) and the retinal pigment epithelium (RPE) to movement of taurine between the choroidal circulation and the outer retina. To quantify the effect of light-evoked changes in subretinal potassium concentration on the transepithelial transport of taurine across bovine RPE.
Transport studies were performed in Ussing chambers with intact and RPE-denuded specimens of BC. RPE viability was monitored by recording transepithelial potential (TEP) and transepithelial resistance (TER). Taurine transport with substrate concentrations in the micro- and millimolar range, reflecting physiological taurine concentrations in plasma, retina, and subretinal space was quantified by high-performance liquid chromatography (HPLC) and radiotracer techniques. Taurine transport was also assessed after apical potassium concentration was lowered from 6.0 to 2.2 mM to mimic the effects of light.
Transport of taurine across RPE-BC at a 10-mM substrate concentration increased from 32.92 before to 111.72 nanomoles/4 mm per hour after removal of the RPE. Similarly, at 50 microM taurine, transport rates increased from 0.158 to 0.439 nanomoles/4 mm per hour after removal of the RPE. At both high (10 mM) and low (50 microM) substrate concentrations, lowering of apical potassium was associated with decreased transport of taurine across the RPE. For taurine concentrations greater than 42 microM, the rate-limiting compartment for transport of taurine to the outer retina was the RPE monolayer. Similar rates were observed across each compartment for concentrations <42 microM.
The magnitude and directionality of taurine transport across the RPE is determined solely by the driving taurine concentration gradient and is modulated by subretinal levels of potassium. Such modulation may provide a mechanism for conserving retinal taurine. Processes that increase the resistance to diffusion across Bruch's membrane such as human ageing and increased thickening and deposition of debris associated with age-related macular degeneration (AMD) are likely to affect transport across the RPE, culminating in a secondary retinal taurine deficiency.
评估布鲁赫膜-脉络膜(BC)和视网膜色素上皮(RPE)分别对牛磺酸在脉络膜循环与视网膜外层之间移动所呈现的相对阻力。量化光诱发的视网膜下钾离子浓度变化对牛磺酸跨牛眼RPE上皮转运的影响。
在尤斯灌流小室中对完整的和去除RPE的BC标本进行转运研究。通过记录跨上皮电位(TEP)和跨上皮电阻(TER)来监测RPE的活力。采用高效液相色谱(HPLC)和放射性示踪技术对微摩尔和毫摩尔范围内的底物浓度(反映血浆、视网膜和视网膜下间隙中的生理性牛磺酸浓度)下的牛磺酸转运进行量化。在将顶端钾离子浓度从6.0 mM降至2.2 mM以模拟光照效应后,也对牛磺酸转运进行了评估。
在10 mM底物浓度下,去除RPE后,牛磺酸跨RPE-BC的转运从之前的32.92增加到111.72纳摩尔/4毫米·小时。同样,在50 μM牛磺酸时,去除RPE后转运速率从0.158增加到0.439纳摩尔/4毫米·小时。在高(10 mM)、低(50 μM)底物浓度下,顶端钾离子浓度降低均与牛磺酸跨RPE的转运减少有关。对于牛磺酸浓度大于42 μM时,牛磺酸向视网膜外层转运的限速区是RPE单层。对于浓度<42 μM时,各区间的转运速率相似。
牛磺酸跨RPE转运的大小和方向性仅由驱动性牛磺酸浓度梯度决定,并受视网膜下钾离子水平调节。这种调节可能为保存视网膜牛磺酸提供一种机制。诸如人类衰老以及与年龄相关性黄斑变性(AMD)相关的碎片增厚和沉积增加等增加跨布鲁赫膜扩散阻力的过程,可能会影响跨RPE的转运,最终导致继发性视网膜牛磺酸缺乏。