Liu C Y, Olsen B R, Kao W W
Department of Ophthalmology, University of Cincinnati, Ohio 45267-0527.
Dev Dyn. 1993 Oct;198(2):150-7. doi: 10.1002/aja.1001980208.
Northern blot hybridization, reverse-transcription polymerase chain reaction (RT-PCR), and RNase protection assays were used to examine the expression of two alpha 1(IX) collagen mRNA species (long and short form) in developing mouse tissues. Furthermore, in situ hybridization was used to identify cells expressing the Col9a1 gene during eye development. The results indicate that during embryonic development eye and heart preferentially express the short form; lung and cartilage express the long form; whereas liver expresses a very low level of long form alpha 1(IX) mRNA which can only be detected by RT-PCR. In situ hybridization demonstrated that at 10.5 day postcoitum (d.p.c.), the alpha 1(IX) collagen mRNAs were first expressed in optic cup (neural ectoderm) but not in lens vesicle (surface ectoderm). By 13.5 d.p.c., the cells that express the alpha 1(IX) mRNA progressively were concentrated toward the anterior part of the neural retina. By 16.5-18.5 d.p.c., the hybridization signals were found exclusively in the inner non-pigmented layer of the presumptive ciliary epithelium. As ciliary epithelial cells become well differentiated 3 weeks after birth, cells expressing the Col9a1 gene were limited to the junction between mature ciliary folds and the neural retina. No hybridization signal could be detected in ocular tissues of mouse older than 6 weeks. It is of interest to note that a hybridization signal was not detected in cornea at the various developmental stages examined, suggesting that mouse cornea does not significantly express alpha 1(IX) mRNA during embryonic development. This differs from that of chick cornea development. In summary, the expression of the Col9a1 gene shows a temporospatial pattern throughout mouse eye development.(ABSTRACT TRUNCATED AT 250 WORDS)
采用Northern印迹杂交、逆转录聚合酶链反应(RT-PCR)和核糖核酸酶保护分析来检测发育中小鼠组织中两种α1(IX)型胶原mRNA种类(长型和短型)的表达。此外,利用原位杂交来鉴定眼睛发育过程中表达Col9a1基因的细胞。结果表明,在胚胎发育过程中,眼睛和心脏优先表达短型;肺和软骨表达长型;而肝脏表达的长型α1(IX)mRNA水平极低,只能通过RT-PCR检测到。原位杂交显示,在胚胎10.5天(d.p.c.)时,α1(IX)型胶原mRNA首先在视杯(神经外胚层)中表达,而不在晶状体泡(表面外胚层)中表达。到13.5 d.p.c.时,表达α1(IX)mRNA的细胞逐渐向神经视网膜前部集中。到16.5 - 18.5 d.p.c.时,杂交信号仅在假定睫状体上皮的内非色素层中发现。随着出生后3周睫状体上皮细胞充分分化,表达Col9a1基因的细胞局限于成熟睫状襞与神经视网膜之间的连接处。在6周龄以上的小鼠眼组织中未检测到杂交信号。值得注意的是,在所检测的各个发育阶段的角膜中均未检测到杂交信号,这表明小鼠角膜在胚胎发育过程中不显著表达α1(IX)mRNA。这与鸡角膜发育情况不同。总之,Col9a1基因的表达在小鼠眼睛发育过程中呈现出时空模式。(摘要截短至250字)