Hales K H, Meredith J E, Storti R V
Department of Biochemistry M/C536, University of Illinois College of Medicine, Chicago 60612.
Dev Biol. 1994 Oct;165(2):639-53. doi: 10.1006/dbio.1994.1282.
We have characterized maternal and zygotic cytoskeletal tropomyosin mRNA expression during Drosophila embryogenesis using in situ hybridization to endogenous cytoskeletal tropomyosin mRNA and a cytoskeletal tropomyosin promoter/beta-galactosidase-encoding fusion gene mRNA in transgenic flies. A 2.0-kb maternal cytoskeletal tropomyosin mRNA is synthesized in the nurse cells and transported into the oocyte during oogenesis. During early embryogenesis, this mRNA becomes localized to the pole cell region and then to the cortex during the cellular blastoderm stage. In later embryos it is localized to the ventral and cephalic furrows and extending germ band. The major zygotic mRNA is 2.4 kb and is first detected at gastrulation. In early embryos, this mRNA is expressed in the invaginating anterior and posterior midgut, the transverse furrows, and the amnioserosa, all regions of the embryo undergoing intense cellular movement, and in later embryos, predominantly in the gut, brain, and epidermis. A transgene construct containing 1.2 kb of 5' cytoskeletal tropomyosin promoter sequences driving expression of Escherichia coli beta-galactosidase and the cytoskeletal tropomyosin 3' untranslated region in transgenic flies has the same distribution of maternal and zygotic transcripts throughout oogenesis and embryonic development as the endogenous transcripts. A transgene containing the 3' untranslated region from either the hsp70 gene or the SV40 early genes, on the other hand, does not express maternal RNA. Furthermore, none of the transgenes expressed in the follicle cells, suggesting that expression in these cells is under different transcriptional control. Our results indicate that maternal and zygotic cytoskeletal tropomyosin mRNAs are localized to specific regions of the developing embryo, particularly in regions of the embryo undergoing cell movement and invagination. Furthermore, the synthesis, transport, and accumulation of this RNA is under transcriptional and post-transcriptional control.
我们利用原位杂交技术,针对果蝇胚胎发育过程中母体和合子细胞骨架原肌球蛋白mRNA的表达进行了特征分析。原位杂交的对象是转基因果蝇中内源性细胞骨架原肌球蛋白mRNA以及一个细胞骨架原肌球蛋白启动子/β-半乳糖苷酶编码融合基因mRNA。一个2.0 kb的母体细胞骨架原肌球蛋白mRNA在卵母细胞发生过程中于滋养细胞中合成,并在卵子发生期间转运至卵母细胞。在胚胎发育早期,这种mRNA定位于极细胞区域,然后在细胞胚盘阶段定位于皮层。在后期胚胎中,它定位于腹沟和头沟以及延伸的胚带。主要的合子mRNA为2.4 kb,在原肠胚形成时首次被检测到。在早期胚胎中,这种mRNA在正在内陷的前、中肠、横向沟以及浆膜中表达,这些都是胚胎中经历剧烈细胞运动的区域,而在后期胚胎中,主要在肠道、大脑和表皮中表达。一个转基因构建体包含1.2 kb的5'细胞骨架原肌球蛋白启动子序列,驱动大肠杆菌β-半乳糖苷酶的表达以及转基因果蝇中细胞骨架原肌球蛋白3'非翻译区的表达,其母体和合子转录本在整个卵子发生和胚胎发育过程中的分布与内源性转录本相同。另一方面,一个含有hsp70基因或SV40早期基因3'非翻译区的转基因不表达母体RNA。此外,没有一个转基因在卵泡细胞中表达,这表明这些细胞中的表达受到不同的转录控制。我们的结果表明,母体和合子细胞骨架原肌球蛋白mRNA定位于发育中胚胎的特定区域,特别是在胚胎中经历细胞运动和内陷的区域。此外,这种RNA的合成、转运和积累受到转录和转录后控制。