Uusitalo Marita, Schlötzer-Schrehardt Ursula, Kivelä Tero
Department of Ophthalmology, Helsinki University Central Hospital, Helsinki, Finland.
Invest Ophthalmol Vis Sci. 2003 Mar;44(3):961-4. doi: 10.1167/iovs.02-0489.
To localize the cell adhesion-related HNK-1 carbohydrate epitope in the human retina at cellular and subcellular levels.
Retinas were obtained from seven normal human eyes at autopsy (age, 43-78 years). The specimens were embedded in medium-grade resin and studied by postembedding immunoelectron microscopy using the primary mouse mAb HNK-1 (Leu 7) to the HNK-1 epitope and secondary antibodies conjugated to 10-nm colloidal gold particles.
Prominent immunolabeling with mAb HNK-1 was observed on the outer surface of the entire plasma membrane of Müller radial glial cells, including their microvilli between the inner segments of rods and cones, on the plasma membranes of astrocytes in the ganglion cell layer, in bipolar cells in the inner nuclear layer, and in photoreceptor cells in the outer nuclear layer. Fewer gold particles were present on plasma membranes of other main types of retinal neurons, including ganglion cells. Only the outer segments of rods and cones and the endothelial cells of retinal capillaries were never labeled. In the ciliary epithelium, gold particles localized to the basement membrane of the nonpigmented and pigmented layers and to the cytoplasm of the pigmented epithelium.
Unlike in many other species, the HNK-1 epitope in the human retina is found on both glial and neuronal cells, including photoreceptors. This epitope potentially contributes to neuron-to-neuron and glia-to-neuron adhesion of human retinal cells.
在细胞和亚细胞水平上定位人视网膜中与细胞黏附相关的HNK-1碳水化合物表位。
从7例正常人类尸体眼部(年龄43 - 78岁)获取视网膜。将标本包埋于中等硬度树脂中,采用包埋后免疫电子显微镜技术进行研究,使用针对HNK-1表位的小鼠单克隆抗体HNK-1(Leu 7)作为一抗,与10纳米胶体金颗粒偶联的二抗。
在Müller放射状神经胶质细胞整个质膜的外表面观察到明显的HNK-1单克隆抗体免疫标记,包括其在视杆细胞和视锥细胞内节之间的微绒毛上;在神经节细胞层的星形胶质细胞质膜上、内核层的双极细胞中以及外核层的光感受器细胞中也有免疫标记。在包括神经节细胞在内的其他主要类型视网膜神经元的质膜上,金颗粒较少。仅视杆细胞和视锥细胞的外节以及视网膜毛细血管的内皮细胞从未被标记。在睫状体上皮中,金颗粒定位于非色素层和色素层的基底膜以及色素上皮的细胞质中。
与许多其他物种不同,人视网膜中的HNK-1表位在神经胶质细胞和神经元细胞(包括光感受器)上均有发现。该表位可能有助于人视网膜细胞间的神经元与神经元以及神经胶质细胞与神经元的黏附。