β1 integrin as the integrating component in cell-cell cooperation for maintenance of lens transparency.
β1 integrin as the integrating component in cell-cell cooperation for maintenance of lens transparency.
复制标题
β1 整合素作为细胞间合作的整合成分，用于维持晶状体透明度。
DOI：
10.1134/s1607672913060069
复制
发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Suchkov,SV
中科院分区：
文献类型：
--
作者：
Simirskii,VN;Duncan,MK;Paltsev,MA;Suchkov,SV
SIMIRSKII et al. of development and degree of lens opacity. Multiple inbreeding crosses and the breeding of mice with a well expressed cataract yielded a subline of mice with a pronounced cataract (Figs. 1a, 1b). The lenses with cataracts contained disorganized fibers and multiple cavities, unlike the normal lens, in which the fibers were tightly packed and formed concentric layers (Figs. 1c, 1d). The most characteristic feature was the lack of contact of the majority of the fibers with the capsule and the presence of numerous small cavities along the lateral surfaces of the fibers. The efficiency and specificity of the β1 integrin gene knockout in mice was confirmed by PCR analysis of genomic DNA isolated from the lens fibers. Basing on the location of loxP sites, the deletion of exon 3 in the β1 integrin gene could be expected. In addition, αАcrystallin gene promoter, which we have used to control the Cre recombinase expression, does not function in the epithelial cells of the lens and is only activated in differentiated lens fibers [6]. With this in mind, we expected that the β1 integrin gene knockout will be limited to the lens fibers. Indeed, PCR analysis of the genomic DNA isolated from the lens fibers of mice with the βflox/flox/Cre+ genotype confirmed the deletion of exon 3 in the β1 integrin gene. It should be noted that the size of the amplification products for the fibers of knockout and wild type mice differed by the value corresponding to the estimated size of the DNA region between the loxP sites (exon 3 with sur rounding intron regions)(Fig. 2). Corresponding analysis of DNA from the lens epithelium of the mice showed that exon 3 in the knockout mice was not deleted. β1 Integrin was immunochemically detected in the epithelium; in the fibers, it was localized only in the superficial layers in the equatorial region of the lens at all stages of embryonic development. In the deeper layers of the fibers, β1 integrin was not detected starting from 15.5–16.5 days post conception (dpc)(Fig. 3). This agrees with the start of differentiation of secondary fibers of the lens and subsequent activation of the Cre recombinase on 13.5 dpc [6, 8]. The delay in the pro tein loss was apparently due to the fact that β1 integrin synthesized in the epithelium is not degraded immedi ately and is stored for some time in the early fibers dif ferentiated from the epithelial cells. The preferential localization of β1 integrin in the epithelium (with its
