Lo W K, Shaw A P, Takemoto L J, Grossniklaus H E, Tigges M
Department of Anatomy, Morehouse School of Medicine, Atlanta, GA 30310, USA.
Exp Eye Res. 1996 Feb;62(2):171-80. doi: 10.1006/exer.1996.0021.
Gap junction structures and distribution patterns of immunoreactive connexin46 (Cx46) and connexin50 (Cx50) in normal lenses and lens regrowths of rhesus monkeys were studied using electron microscopy and immunofluorescence double-labeling. Lens regrowths were collected from aphakic eyes of young monkeys whose natural lenses had been surgically removed 11-34 months earlier to simulate monocular congenital cataract surgery in human infants. Approximately 90% of the lens regrowths examined was in the form of a doughnut-shaped Soemmerring's ring located behind the iris. The lens regrowth consisted of lens epithelium and lens fibers enclosed within hypertrophied capsular material. The superficial equatorial region usually contained nucleated young fibers of normal appearance. The other regions consisted of many swollen fibers. Gap junctions were readily observed between fiber cells of both normal and swollen configuration in the lens regrowth. In superficial fibers, gap junctions were not associated with cytoskeletal components. In the intermediate and the deeper cortical regions, actin filament bundles were found specifically associated with gap junctions along both of their cytoplasmic surfaces. An immunofluorescence double-labeling study showed that Cx46 and Cx50 were labeled in the same gap junctions in both superficial and deeper cortical fibers of the normal lens. In contrast, in the lens regrowth strong co-labeling of Cx46 and Cx50 was only observed in the superficial fibers. The labeling for Cx50 was very weak or absent in the deeper cortex, whereas the strong labeling for Cx46 persisted throughout the major portion of the deeper cortex. The labeling for Cx46 finally disappeared in the much deeper cortex. This study shows that (1) the same distribution pattern of actin bundle/gap junction association found in normal lenses is seen in the lens regrowth, and (2) the immunoreactive distribution of Cx46 and Cx50 differ in the lens regrowths as compared with those in the normal lenses of rhesus monkeys.
利用电子显微镜和免疫荧光双标记技术,研究了恒河猴正常晶状体和晶状体再生中缝隙连接结构以及免疫反应性连接蛋白46(Cx46)和连接蛋白50(Cx50)的分布模式。晶状体再生取自幼年猴子的无晶状体眼,这些猴子的天然晶状体在11 - 34个月前已通过手术摘除,以模拟人类婴儿的单眼先天性白内障手术。大约90%被检查的晶状体再生呈位于虹膜后方的甜甜圈状索默林环的形式。晶状体再生由晶状体上皮和包裹在肥大囊膜物质内的晶状体纤维组成。浅表赤道区域通常含有外观正常的有核年轻纤维。其他区域则由许多肿胀的纤维组成。在晶状体再生中,正常和肿胀形态的纤维细胞之间都很容易观察到缝隙连接。在浅表纤维中,缝隙连接与细胞骨架成分无关。在中间和更深的皮质区域,发现肌动蛋白丝束沿着其两个细胞质表面与缝隙连接特异性相关。免疫荧光双标记研究表明,在正常晶状体的浅表和更深皮质纤维中,Cx46和Cx50在相同的缝隙连接中被标记。相比之下,在晶状体再生中,仅在浅表纤维中观察到Cx46和Cx50的强共标记。在更深的皮质中,Cx50的标记非常弱或不存在,而Cx46的强标记在更深皮质的大部分区域持续存在。Cx46的标记最终在更深得多的皮质中消失。这项研究表明:(1)在晶状体再生中观察到与正常晶状体中相同的肌动蛋白束/缝隙连接关联分布模式;(2)与恒河猴正常晶状体相比,Cx46和Cx50的免疫反应性分布在晶状体再生中有所不同。