Nagelhus E A, Veruki M L, Torp R, Haug F M, Laake J H, Nielsen S, Agre P, Ottersen O P
Department of Anatomy, Institute of Basic Medical Sciences, University of Oslo, N-0317 Oslo, Norway.
J Neurosci. 1998 Apr 1;18(7):2506-19. doi: 10.1523/JNEUROSCI.18-07-02506.1998.
The water permeability of cell membranes differs by orders of magnitude, and most of this variability reflects the differential expression of aquaporin water channels. We have recently found that the CNS contains a member of the aquaporin family, aquaporin-4 (AQP4). As a prerequisite for understanding the cellular handling of water during neuronal activity, we have investigated the cellular and subcellular expression of AQP4 in the retina and optic nerve where activity-dependent ion fluxes have been studied in detail. In situ hybridization with digoxigenin-labeled riboprobes and immunogold labeling by a sensitive postembedding procedure demonstrated that AQP4 and AQP4 mRNA were restricted to glial cells, including MHller cells in the retina and fibrous astrocytes in the optic nerve. A quantitative immunogold analysis of the MHller cells showed that these cells exhibited three distinct membrane compartments with regard to AQP4 expression. End feet membranes (facing the vitreous body or blood vessels) were 10-15 times more intensely labeled than non-end feet membranes, whereas microvilli were devoid of AQP4. These data suggest that MHller cells play a prominent role in the water handling in the retina and that they direct osmotically driven water flux to the vitreous body and vessels rather than to the subretinal space. Fibrous astrocytes in the optic nerve similarly displayed a differential compartmentation of AQP4. The highest expression of AQP4 occurred in end feet membranes, whereas the membrane domain facing the nodal axolemma was associated with a lower level of immunoreactivity than the rest of the membrane. This arrangement may allow transcellular water redistribution to occur without inducing inappropriate volume changes in the perinodal extracellular space.
细胞膜的水通透性在数量级上存在差异,这种差异大多反映了水通道蛋白水通道的差异表达。我们最近发现中枢神经系统含有水通道蛋白家族的一个成员——水通道蛋白4(AQP4)。作为理解神经元活动期间细胞对水的处理的前提,我们研究了AQP4在视网膜和视神经中的细胞及亚细胞表达,其中对活动依赖性离子通量已进行了详细研究。用地高辛标记的核糖探针进行原位杂交以及通过灵敏的包埋后程序进行免疫金标记表明,AQP4和AQP4 mRNA仅限于神经胶质细胞,包括视网膜中的米勒细胞和视神经中的纤维型星形胶质细胞。对米勒细胞的定量免疫金分析表明,就AQP4表达而言,这些细胞呈现出三个不同的膜区室。终足膜(面向玻璃体或血管)的标记强度比非终足膜高10 - 15倍,而微绒毛则没有AQP4。这些数据表明,米勒细胞在视网膜的水处理中起重要作用,并且它们将渗透驱动的水通量导向玻璃体和血管,而不是视网膜下间隙。视神经中的纤维型星形胶质细胞同样显示出AQP4的差异区室化。AQP4的最高表达出现在终足膜中,而面向节段性轴突膜的膜结构域的免疫反应性水平低于膜的其余部分。这种排列可能允许跨细胞水重新分布发生,而不会在节段周围细胞外空间引起不适当的体积变化。