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The Journal of Neuroscience, December 15, 1999, 19(24):10716-10726
Blue- and Green-Absorbing Visual Pigments of
Drosophila: Ectopic Expression and Physiological
Characterization of the R8 Photoreceptor Cell-Specific Rh5 and Rh6
Rhodopsins
Ernesto
Salcedo1,
Armin
Huber2,
Stefan
Henrich2,
Linda V.
Chadwell1,
Wen-Hai
Chou1,
Reinhard
Paulsen2, and
Steven G.
Britt1
1 Institute of Biotechnology and Department of
Molecular Medicine, University of Texas Health Science Center, San
Antonio, Texas 78245, and 2 University of Karlsruhe,
Institute of Zoology, Department of Cell and Neurobiology,
Kornblumenstrasse 13, D-76128 Karlsruhe, Germany
Color discrimination requires the input of different photoreceptor
cells that are sensitive to different wavelengths of light. The
Drosophila visual system contains multiple classes of
photoreceptor cells that differ in anatomical location, synaptic
connections, and spectral sensitivity. The Rh5 and Rh6 opsins are
expressed in nonoverlapping sets of R8 cells and are the only
Drosophila visual pigments that remain uncharacterized.
In this study, we ectopically expressed Rh5 and Rh6 in the major class
of photoreceptor cells (R1-R6) and show them to be biologically active
in their new environment. The expression of either Rh5 or Rh6 in
"blind" ninaE17 mutant flies, which lack
the gene encoding the visual pigment of the R1-R6 cells, fully rescues
the light response. Electrophysiological analysis showed that the
maximal spectral sensitivity of the R1-R6 cells is shifted to 437 or
508 nm when Rh5 or Rh6, respectively, is expressed in these cells.
These spectral sensitivities are in excellent agreement with
intracellular recordings of the R8p and R8y cells measured in
Calliphora and Musca. Spectrophotometric analyses of Rh5 and Rh6 in vivo by
microspectrophotometry, and of detergent-extracted pigments in
vitro, showed that Rh5 is reversibly photoconverted to a stable
metarhodopsin ( max = 494 nm), whereas Rh6 appears
to be photoconverted to a metarhodopsin (max = 468 nm) that is less thermally stable. Phylogenetically, Rh5 belongs to a
group of short-wavelength-absorbing invertebrate visual pigments, whereas Rh6 is related to a group of long-wavelength-absorbing pigments
and is the first member of this class to be functionally characterized.
Key words:
Drosophila melanogaster; fruit fly; rhodopsin; visual pigment; spectral tuning; green-absorbing rhodopsin; blue-absorbing rhodopsin; protein expression
Copyright © 1999 Society for Neuroscience 0270-6474/99/192410716-11$05.00/0
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