Center for Biochemistry, Medical Faculty, University of Cologne, Cologne, Germany.
Ophthalmic Res. 2010;43(2):108-12. doi: 10.1159/000247595. Epub 2009 Oct 15.
To determine the distribution of major basement membrane constituents, particularly nidogen 1 and 2, in young and aging mouse retinae.
The specificity of antibodies against basement membrane proteins was ascertained by immunoblotting with proteins extracted from mouse retinae. The same antibodies were used in indirect immunofluorescence microscopy to localize basement membrane proteins in paraffin sections of retinae from 1-, 12- and 18-month-old C57BL/6 mice.
At a young age, laminin, perlecan and collagen IV were most abundant in Bruch's membrane. Later, the proteins were clearly detected in capillary basement membranes and the inner limiting membrane. In both of these basement membranes, a massive increase in protein amount was seen upon aging, whereas in Bruch's membrane the staining intensity was less drastically changed. Both nidogen 1 and 2 were present in vascular basement membranes and Bruch's membrane throughout the age periods studied. In the inner limiting membrane, the nidogens were more strongly expressed at higher ages, with an earlier and more extensive deposition of nidogen 1.
All major basement membrane constituents are present in the mouse retina, but the onset of deposition differs among the different proteins and between the various retinal basement membranes. In general, basement membrane protein deposition increases with age.
确定主要基底膜成分(尤其是巢蛋白 1 和 2)在年轻和衰老小鼠视网膜中的分布。
通过用从小鼠视网膜中提取的蛋白质进行免疫印迹,确定针对基底膜蛋白的抗体的特异性。相同的抗体用于间接免疫荧光显微镜,以定位来自 1 个月、12 个月和 18 个月龄 C57BL/6 小鼠视网膜石蜡切片中的基底膜蛋白。
在年轻时,层粘连蛋白、perlecan 和胶原 IV 在布鲁赫膜中最为丰富。之后,这些蛋白质在毛细血管基底膜和内界膜中清晰检测到。在这两种基底膜中,随着年龄的增长,蛋白质的含量明显增加,而在布鲁赫膜中,染色强度的变化则不那么剧烈。巢蛋白 1 和 2 均存在于血管基底膜和整个研究期间的布鲁赫膜中。在内界膜中,随着年龄的增长,巢蛋白表达更强,巢蛋白 1 的沉积更早且更广泛。
所有主要的基底膜成分都存在于小鼠视网膜中,但不同蛋白质之间以及不同视网膜基底膜之间的沉积起始时间不同。一般来说,基底膜蛋白的沉积随年龄增长而增加。