Chen Chunhe, Jiang Yunhai, Koutalos Yiannis
Department of Physiology and Biophysics, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA.
Biophys J. 2002 Sep;83(3):1403-12. doi: 10.1016/S0006-3495(02)73911-8.
Eukaryotic cells use membrane organelles, like the endoplasmic reticulum or the Golgi, to carry out different functions. Vertebrate rod photoreceptors use hundreds of membrane sacs (the disks) for the detection of light. We have used fluorescent tracers and single cell imaging to study the properties of rod photoreceptor disks. Labeling of intact rod photoreceptors with membrane markers and polar tracers revealed communication between intradiskal and extracellular space. Internalized tracers moved along the length of the rod outer segment, indicating communication between the disks as well. This communication involved the exchange of both membrane and aqueous phase and had a time constant in the order of minutes. The communication pathway uses approximately 2% of the available membrane disk area and does not allow the passage of molecules larger than 10 kDa. It was possible to load the intradiskal space with fluorescent Ca(2+) and pH dyes, which reported an intradiskal Ca(2+) concentration in the order of 1 microM and an acidic pH 6.5, both of them significantly different than intracellular and extracellular Ca(2+) concentrations and pH. The results suggest that the rod photoreceptor disks are not discrete, passive sacs but rather comprise an active cellular organelle. The communication between disks may be important for membrane remodeling as well as for providing access to the intradiskal space of the whole outer segment.
真核细胞利用内质网或高尔基体等膜性细胞器来执行不同的功能。脊椎动物的视杆光感受器利用数百个膜囊(圆盘)来检测光线。我们使用荧光示踪剂和单细胞成像技术来研究视杆光感受器圆盘的特性。用膜标记物和极性示踪剂对完整的视杆光感受器进行标记,揭示了圆盘内空间与细胞外空间之间的通讯。内化的示踪剂沿着视杆外段的长度移动,这也表明圆盘之间存在通讯。这种通讯涉及膜相和水相的交换,时间常数约为几分钟。通讯途径占用了约2%的可用膜圆盘面积,不允许大于10 kDa的分子通过。可以用荧光钙和pH染料加载圆盘内空间,它们显示圆盘内钙浓度约为1微摩尔,pH为酸性6.5,这两者与细胞内和细胞外的钙浓度及pH均有显著差异。结果表明,视杆光感受器圆盘不是离散的、被动的囊泡,而是一种活跃的细胞器。圆盘之间的通讯对于膜重塑以及整个外段的圆盘内空间的物质进入可能很重要。