Santhiya S T, Abd-alla S M, Löster J, Graw J
Post-Graduate Institute of Basic Medical Sciences, University of Madras, Taramani, India.
Graefes Arch Clin Exp Ophthalmol. 1995 Dec;233(12):795-800. doi: 10.1007/BF00184093.
From previous experiments it is known that the murine dominant cataract mutants carrying the gene Cat2 have a decreased content of gamma-crystallin-specific transcripts in the juvenile lens, when the cataract is completely expressed. Moreover, the mutant locus has been mapped recently to chromosome 1, closely linked to the gamma E-crystallin gene (map distance 0.3 +/- 0.3 cM). In the present paper we describe the phenotypic changes and the gamma-crystallin expression in embryonic lenses of the Cat2nop mutants as an example for the Cat2 allelic series.
The technique of in situ hybridization was applied using a probe from the murine gamma D-crystallin gene, and, for control, from the murine alpha A-crystallin gene. Simultaneously, a series of lens sections was examined histologically.
The presence of gamma-crystallin mRNA was demonstrated from embryonic day 13.5 (E13.5) onward, but in the mutants to a lower extent than in the wild-type lenses. However, the first morphological abnormality in the mutant lenses was observed as swelling of lens fibers at day E15.5. Progressive degeneration of the lens core followed, leading to a cataracta immatura.
The reduced level of gamma-crystallin transcripts is the first alteration observable during the embryonic development of the Cat2 mutant lenses: it precedes the morphological changes. This result represents an additional line of argument that the gamma-crystallin genes may be the target of the mutation in the Cat2 mice.
从先前的实验可知,携带Cat2基因的小鼠显性白内障突变体在幼年晶状体中,当白内障完全表现时,γ-晶状体蛋白特异性转录本的含量会降低。此外,该突变位点最近已被定位到1号染色体,与γE-晶状体蛋白基因紧密连锁(图距0.3±0.3厘摩)。在本文中,我们描述了Cat2nop突变体胚胎晶状体中的表型变化和γ-晶状体蛋白表达,作为Cat2等位基因系列的一个例子。
使用来自小鼠γD-晶状体蛋白基因的探针进行原位杂交技术,并使用来自小鼠αA-晶状体蛋白基因的探针作为对照。同时,对一系列晶状体切片进行组织学检查。
从胚胎第13.5天(E13.5)起就证明了γ-晶状体蛋白mRNA的存在,但在突变体中的含量低于野生型晶状体。然而,在突变体晶状体中观察到的第一个形态异常是在E15.5天时晶状体纤维肿胀。随后晶状体核心逐渐退化,导致未成熟白内障。
γ-晶状体蛋白转录本水平降低是Cat2突变体晶状体胚胎发育过程中可观察到的第一个变化:它先于形态学变化。这一结果进一步证明γ-晶状体蛋白基因可能是Cat2小鼠突变的靶点。