首页> 美国卫生研究院文献>The Journal of Neuroscience >Honeybee Blue- and Ultraviolet-Sensitive Opsins: Cloning Heterologous Expression in Drosophila and Physiological Characterization
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Honeybee Blue- and Ultraviolet-Sensitive Opsins: Cloning Heterologous Expression in Drosophila and Physiological Characterization

机译:蜜蜂对蓝光和紫外线敏感的视蛋白:克隆果蝇中的异源表达和生理学表征

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摘要

The honeybee (Apis mellifera) visual system contains three classes of retinal photoreceptor cells that are maximally sensitive to light at 440 nm (blue), 350 nm (ultraviolet), and 540 nm (green). We performed a PCR-based screen to identify the genes encoding the Apis blue- and ultraviolet (UV)-sensitive opsins. We obtained cDNAs that encode proteins having a high degree of sequence and structural similarity to other invertebrate and vertebrate visual pigments. The Apis blue opsin cDNA encodes a protein of 377 amino acids that is most closely related to other invertebrate visual pigments that are thought to be blue-sensitive. The UV opsin cDNA encodes a protein of 371 amino acids that is most closely related to the UV-sensitive Drosophila Rh3 and Rh4 opsins. To test whether these novel Apis opsin genes encode functional visual pigments and to determine their spectral properties, we expressed them in the R1–6 photoreceptor cells of blindninaE mutant Drosophila, which lack the major opsin of the fly compound eye. We found that the expression of either the Apis blue- or UV-sensitive opsin in transgenic flies rescued the visual defect of ninaEmutants, indicating that both genes encode functional visual pigments. Spectral sensitivity measurements of these flies demonstrated that the blue and UV visual pigments are maximally sensitive to light at 439 and 353 nm, respectively. These maxima are in excellent agreement with those determined previously by single-cell recordings fromApis photoreceptor cells and provide definitive evidence that the genes described here encode visual pigments having blue and UV sensitivity.
机译:蜜蜂(Apis mellifera)视觉系统包含三类视网膜感光细胞,它们对440 nm(蓝色),350 nm(紫外线)和540 nm(绿色)的光最大敏感。我们进行了基于PCR的筛选,以鉴定编码Apis蓝和紫外线(UV)敏感视蛋白的基因。我们获得了编码与其他无脊椎动物和脊椎动物视觉色素具有高度序列和结构相似性的蛋白质的cDNA。 Apis蓝色视蛋白cDNA编码一个377个氨基酸的蛋白质,该蛋白质与被认为对蓝光敏感的其他无脊椎动物可见色素最密切相关。紫外线视蛋白cDNA编码一个371个氨基酸的蛋白质,与紫外线敏感果蝇Rh3和Rh4视蛋白密切相关。为了测试这些新颖的Apis视蛋白基因是否编码功能性视觉色素并确定其光谱特性,我们将它们在缺乏果蝇复眼的主要视蛋白的blindinaE突变果蝇的R1-6感光细胞中表达了它们。我们发现,转基因果蝇中Apis蓝敏感蛋白或UV敏感视蛋白的表达挽救了ninaEmutants的视觉缺陷,表明这两个基因均编码功能性视觉色素。这些果蝇的光谱灵敏度测量结果表明,蓝色和UV视觉颜料分别对439和353 nm的光最大敏感。这些最大值与先前通过Apis感光细胞的单细胞记录确定的最大值非常吻合,并提供了确凿的证据表明此处描述的基因编码具有蓝色和紫外线敏感性的视觉色素。

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