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The human blue opsin promoter directs transgene expression in short-wave cones and bipolar cells in the mouse retina.

机译:人蓝视蛋白启动子指导小鼠视网膜中短波视锥细胞和双极细胞中的转基因表达。

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摘要

Transgenic mouse lines were generated using either 3.8 or 1.1 kb of 5' upstream flanking sequence from the human blue opsin gene fused to the lacZ or human growth hormone reporter gene. Mice were analyzed for appropriate cell-specific and developmental expression patterns. In 13 independently derived lines of animals, transgene expression was limited to photoreceptor and inner nuclear layer cells. Photoreceptors were identified as cone cells based on morphological criteria and colocalization of transgene expression with the cone-associated marker, peanut agglutinin lectin. More specifically, transgene-positive photoreceptors were identified as short-wave cone cells (S-cones) by using the short-wave color opsin-specific antibody, OS-2. Reporter-gene-positive cells of the inner nuclear layer were identified as bipolar cells based on morphological criteria. Transgenes and the endogenous mouse short-wave opsin gene were transcriptionally coactivated at embryonic day 13. These results show that 3.8 or 1.1 kb of human blue opsin upstream flanking sequences are capable of directing expression in short-wave cone cells in a spatially and temporally appropriate fashion and that the human blue opsin gene is the homologue of the short-wave-sensitive pigment, S-opsin, in the short-wave cones of the mouse retina. Expression in the bipolar cells may reflect regulatory mechanisms that are common to these cells and to the cone photoreceptors.
机译:转基因小鼠品系是使用3.8或1.1 kb的5'上游侧翼序列从融合到lacZ或人生长激素报告基因的人蓝视蛋白基因中产生的。分析小鼠的适当的细胞特异性和发育表达模式。在13个独立衍生的动物品系中,转基因表达仅限于感光细胞和内核层细胞。根据形态学标准和转基因表达与视锥细胞相关标记花生凝集素凝集素的共定位,将感光细胞鉴定为视锥细胞。更具体地说,通过使用短波彩色视蛋白特异性抗体OS-2,将转基因阳性感光体鉴定为短波视锥细胞(S-锥体)。内核层的报告基因阳性细胞根据形态学标准被鉴定为双极细胞。转基因和内源性小鼠短波视蛋白基因在胚胎第13天被转录共激活。这些结果表明,3.8或1.1 kb的人蓝视蛋白上游侧翼序列能够指导短波视锥细胞在空间和时间上适当的表达以及人类蓝色视蛋白基因是小鼠视网膜短波视锥细胞中短波敏感色素S-视蛋白的同源物。双极细胞中的表达可能反映了这些细胞和视锥细胞感光细胞共有的调节机制。

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