Giebelhaus D H, Eib D W, Moon R T
Department of Pharmacology, School of Medicine, University of Washington, Seattle 98195.
Cell. 1988 May 20;53(4):601-15. doi: 10.1016/0092-8674(88)90576-4.
Plasmids expressing partial-length sense or antisense protein 4.1 RNA were microinjected into fertilized Xenopus eggs. Nuclease protection assays reveal that antisense protein 4.1 RNA lead to the specific loss of endogenous protein 4.1 transcripts after midblastula transition, with no effect on the levels of three unrelated transcripts. As a control, we show that this dramatic loss of endogenous protein 4.1 transcripts is blocked when fertilized eggs receive a second injection of plasmids that express partial-length sense protein 4.1 RNA. Immunocytochemistry of tadpole embryos with antibodies monospecific for protein 4.1 demonstrates that the antisense protein 4.1 RNA blocks the normal expression of protein 4.1 in embryos and interferes with the normal interdigitation of the photoreceptor outer segments with the pigment epithelium layer in the retina. These data suggest that reduced expression of a single membrane skeleton protein is sufficient to perturb normal cellular interactions of the retina.
将表达全长有义或反义蛋白4.1 RNA的质粒显微注射到非洲爪蟾的受精卵中。核酸酶保护分析显示，反义蛋白4.1 RNA导致囊胚中期过渡后内源性蛋白4.1转录本的特异性缺失，而对三种不相关转录本的水平没有影响。作为对照，我们发现当受精卵再次注射表达全长有义蛋白4.1 RNA的质粒时，内源性蛋白4.1转录本的这种显著缺失被阻断。用对蛋白4.1单特异性的抗体对蝌蚪胚胎进行免疫细胞化学分析表明，反义蛋白4.1 RNA阻断了胚胎中蛋白4.1的正常表达，并干扰了视网膜中光感受器外段与色素上皮层的正常交错。这些数据表明，单一膜骨架蛋白表达的降低足以扰乱视网膜的正常细胞相互作用。
