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The Journal of Neuroscience, April 1, 1998, 18(7):2412-2422

Honeybee Blue- and Ultraviolet-Sensitive Opsins: Cloning, Heterologous Expression in Drosophila, and Physiological Characterization

Steven M. Townson¹, Belinda S. W. Chang², Ernesto Salcedo¹, Linda V. Chadwell¹, Naomi E. Pierce², and Steven G. Britt¹

¹ Institute of Biotechnology and Department of Molecular Medicine, The University of Texas Health Science Center, San Antonio, Texas 78245, and ² Program In Neuroscience, Organismic and Evolutionary Biology Department, Harvard University, Museum of Comparative Zoology Labs, Cambridge, Massachusetts 02138

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 blind ninaE 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 ninaE mutants, 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 from Apis photoreceptor cells and provide definitive evidence that the genes described here encode visual pigments having blue and UV sensitivity.

Key words: Apis mellifera; honeybee; visual pigment; spectral tuning; ultraviolet-sensitive opsin; blue-sensitive opsin; electroretinogram; protein expression; Drosophila melanogaster

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Copyright © 1998 Society for Neuroscience  0270-6474/98/1872412-11$05.00/0

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