Trichromatic visual system in an insect and its sensitivity control by blue light

Summary The spectral sensitivity of the visual cells in the compound eye of the mothDeilephila elpenor was determined by electrophysiological mass recordings during exposure to monochromatic adapting light. Three types of receptors were identified. The receptors are maximally sensitive at about 350 nm (ultraviolet), 450 nm (violet), and 525 nm (green). The spectral sensitivity of the green receptors is identical to a nomogram for a rhodopsin with _max at 525 nm. The spectral sensitivity of the other two receptors rather well agrees with nomograms for corresponding rhodopsins. The recordings indicate that the green receptors occur in larger number than the other receptors. The ultra-violet and violet receptors probably occur in about equal number.The sensitivity after monochromatic adapting illumination varies with the wavelength of the adapting light, but is not proportional to the spectral sensitivity of the receptors. The sensitivity is proportional to the concentration of visual pigment at photoequilibrium. The equilibrium is determined by the absorbance coefficients of the visual pigment and its photoproduct at each wavelength. The concentration of the visual pigment, and thereby the sensitivity, is maximal at about 450 nm, and minimal at wavelengths exceeding about 570 nm.The light from a clear sky keeps the relative concentration of visual pigment in the green receptors, and the relative sensitivity, at about 0.62. The pigment concentration in the ultra-violet receptors is about 0.8 to 0.9, and that in the violet receptors probably about 0.6. At low ambient light intensities a chemical regeneration of the visual pigments may cause an increase in sensitivity. At higher intensities the concentrations of the visual pigments remain constant. Due to the constant pigment concentrations the input signals from the receptors to the central nervous system contain unequivocal information about variations in intensity and spectral distribution of the stimulating light.The work reported in this article was supported by the Swedish Medical Research Council (grant no B 73-04X-104-02B), by Karolinska Institutet, and by a grant (to G. Höglund) from Deutscher Akademischer Austauschdienst, and by the Deutsche Forschungsgemeinschaft, Schwerpunktsprogramm Rezeptorphysiologie HA 258-10, and SFB 114.     

The ultrastructure of the uterine epithelium of the pig during the estrous cycle and early pregnancy

Summary In the compound eye of the moth Antheraea polyphemus, three types of visual pigments were found in extracts from the retina and by microspectrophotometry in situ. The absorption maxima of the receptor pigment P and the metarhodopsin M are at (1) P 520–530 nm, M 480–490 nm; (2) P 460–480 nm, M 530–540 nm; (3) P 330–340 nm, M 460–470 nm. Their localization was investigated by electron microscopy on eyes illuminated with different monochromatic lights. Within the tiered rhabdom, constituted of the rhabdomeres of nine visual cells, the basal cell contains a blue-and the six medial cells have a greenabsorbing pigment. The two distal cells of most ommatidia also have the blue pigment; only in the dorsal region of the eye, these cells contain a UV-absorbing pigment, which constitutes a portion of only 5% of the visual pigment content within the entire retina. The functional significance of this distribution is discussed.     

Scanning electron microscopy and microspectrophotometry of the photoreceptors of ictalurid catfishes

The retinal photoreceptors from larval channel catfish (Ictalurus punctatus) were studied using single cell, in situ microspectrophotometry. Rods appear at 5 days after hatch; cones are present from day one. The rods contain a visual pigment which absorbs light maximally at 540 nm. The cones contain either a green sensitive visual pigment with peak absorbance at 535 nm or a red sensitive visual pigment with peak absorbance at 608 nm. All pigments are based on vitamin A₂. Visual pigment complement does not change with age, as photoreceptors from adultI. punctatus, I. catus andI. melas contain visual pigments virtually identical to those of the larvalI. punctatus. Regardless of age, no visual pigment with peak absorbance in the short wavelength region of the spectrum was ever observed. Scanning electron microscopy of adultI. punctatus retinas showed large rods with long, cylindrical outer segments and smaller cones with short, tapered outer segments. The myoids of both rods and cones are extensable. The rods, embedded in a granular tapetal material, comprise from 50 to 60% of the photoreceptors. Only single cones are present. The data are consistent with the idea that the ictalurid catfishes spend their entire lives in an environment deficient in blue light.     

Visual predation during springtime foraging of the North Atlantic right whale (Eubalaena glacialis)

To assess the role that vision plays in the ability of the North Atlantic right whale (Eubalaena glacialis) to detect its primary prey species, the calanoid copepod Calanus finmarchicus, we have compared the absorbance spectrum of the E. glacialis rod visual pigment, the transmittance spectra of C. finmarchicus carotenoid pigments, as well as the downwelling irradiance and horizontal radiance spectra collected during springtime at three locations in the western Gulf of Maine. The E. glacialis rod visual pigment absorbs light maximally at 493 nm, while microspectrophotometric measurements of the C. finmarchicus carotenoid pigments reveal transmission spectra with minima matching very well with the E. glacialis rod visual pigment absorbance spectra maximum. Springtime spectral downwelling irradiance and horizontal radiance values from the surface waters of Cape Cod Bay and at all depths in Great South Channel overlap the E. glacialis rod absorbance spectrum, allowing C. finmarchicus to appear as a high‐contrast dark silhouette against a bright background spacelight, thus facilitating visually guided contrast foraging. In contrast, spectral downwelling irradiance and horizontal radiance at depth in Cape Cod Bay, and all depths in Wilkinson Basin, do not overlap the E. glacialis rod absorbance spectrum, providing little if any useful light for contrast vision.     

Regulation of phototransduction in short-wavelength cone visual pigments via the retinylidene Schiff base counterion.

Short-wavelength visual pigments (SWS1) have lambda(max) values that range from the ultraviolet to the blue. Like all visual pigments, this class has an 11-cis-retinal chromophore attached through a Schiff base linkage to a lysine residue of opsin apoprotein. We have characterized a series of site-specific mutants at a conserved acidic residue in transmembrane helix 3 in the Xenopus short-wavelength sensitive cone opsin (VCOP, lambda(max) approximately 427 nm). We report the identification of D108 as the counterion to the protonated retinylidene Schiff base. This residue regulates the pK(a) of the Schiff base and, neutralizing this charge, converts the violet sensitive pigment into one that absorbs maximally in the ultraviolet region. Changes to this position cause the pigment to exhibit two chromophore absorbance bands, a major band with a lambda(max) of approximately 352-372 nm and a minor, broad shoulder centered around 480 nm. The behavior of these two absorbance bands suggests that these represent unprotonated and protonated Schiff base forms of the pigment. The D108A mutant does not activate bovine rod transducin in the dark but has a significantly prolonged lifetime of the active MetaII state. The data suggest that in short-wavelength sensitive cone visual pigments, the counterion is necessary for the characteristic rapid production and decay of the active MetaII state.     

The pigment system of the photoreceptor 7 yellow in the fly,a complex photoreceptor

Summary The 7y photoreceptor in the fly (Musca, Calliphora) retina harbours an unusually complex pigment system consisting of a bistable visual pigment (xanthopsin, X and metaxanthopsin, M), a blue-absorbing C₄₀-carotenoid (zeaxanthin and/or lutein) and a uv sensitizing pigment (3-OH retinol).The difference spectrum and photoequilibrium spectrum in single 7y rhabdomeres were determined microspectrophotometrically (Fig. 2).The extinction spectrum of the C₄₀-carotenoid has a pronounced vibrational structure, with peaks at 430, 450 and 480 nm (Fig. 3). The off-axis spectral sensitivity, determined electrophysiologically with 1 nm resolution shows no trace of this fine structure thus excluding the possibility that the C₄₀-carotenoid is a second sensitizing pigment (Fig. 4).The absorption spectra of X and M are derived by fitting nomogram spectra (based on fly R1–6 xanthopsin) to the difference spectrum. max for X is 425 nm, and for M 510 nm (Fig. 5). It is shown that the photoequilibrium spectrum and the difference spectrum can be used to derive the relative photosensitivity spectra of X and M using the analytical method developed by Stavenga (1975). The result (Fig. 6) shows a pronounced uv sensitivity for both, X and M, indicating that the uv sensitizing pigment transfers energy to both X and M. A value of 0.7 for, the relative efficiency of photoconversion for X and M, is obtained by fitting the analytically derived relative photosensitivity spectra to the absorption spectra at wavelengths beyond 420 nm.     

Microspectrophotometric characterization and localization of three visual pigments in the compound eye ofNotonecta glauca L. (Heteroptera)

Summary The absorption maxima ( _max) of the visual pigments in the ommatidia ofNotonecta glauca were found by measuring the difference spectra of single rhabdomeres after alternating illumination with two different adaptation wavelengths. All the peripheral rhabdomeres contain a pigment with an extinction maximum at 560 nm. This pigment is sensitive to red light up to wavelengths > 700 nm. In a given ommatidium in the dorsal region of the eye, the two central rhabdomeres both contain one of two pigments, either a pigment with an absorption maximum in the UV, at 345 nm, or — in neighboring rhabdoms — a pigment with an absorption maximum at 445 nm. In the ventral part of the eye only the pigment absorbing maximally in the UV was found in the central rhabdomeres. The spectral absorption properties of various types of screening-pigment granules were measured.     

Visual pigment spectra of the comma butterfly,<Emphasis Type="Italic"> Polygonia c-album</Emphasis>, derived from in vivo epi-illumination microspectrophotometry

The visual pigments in the compound eye of the comma butterfly, Polygonia c-album, were investigated in a specially designed epi-illumination microspectrophotometer. Absorption changes due to photochemical conversions of the visual pigments, or due to light-independent visual pigment decay and regeneration, were studied by measuring the eye shine, i.e., the light reflected from the tapetum located in each ommatidium proximal to the visual pigment-bearing rhabdom. The obtained absorbance difference spectra demonstrated the dominant presence of a green visual pigment. The rhodopsin and its metarhodopsin have absorption peak wavelengths at 532 nm and 492 nm, respectively. The metarhodopsin is removed from the rhabdom with a time constant of 15 min and the rhodopsin is regenerated with a time constant of 59 min (room temperature). A UV rhodopsin with metarhodopsin absorbing maximally at 467 nm was revealed, and evidence for a blue rhodopsin was obtained indirectly.     

Fast Electrical Potential from a Long-Lived,Long-Wavelength Photoproduct of Fly Visual Pigment

A rapid electrical potential, which we have named the M-potential, can be obtained from the Drosophila eye using a high energy flash stimulus. The potential can be elicited from the normal fly, but it is especially prominent in the mutant norp A^P12 (a phototransduction mutant), particularly if the eye color pigments are genetically removed from the eye. Several lines of evidence suggest that the M-potential arises from photoexcitation of long-lived metarhodopsin. Photoexcitation of rhodopsin does not produce a comparable potential. The spectral sensitivity of the M-potential peaks at about 575 nm. The M-potential pigment (metarhodopsin) can be shown to photoconvert back and forth with a "silent pigment(s)" absorbing maximally at about 485 nm. The silent pigment presumably is rhodopsin. These results support the recent spectrophotometric findings that dipteran metarhodopsin absorbs at much longer wavelengths than rhodopsin. The M-potential probably is related to the photoproduct component of the early receptor potential (ERP). Two major differences between the M-potential and the classical ERP are: (a) Drosophila rhodopsin does not produce a rapid photoresponse, and (b) an anesthetized or freshly sacrificed animal does not yield the M-potential. As in the case of the ERP, the M-potential appears to be a response associated with a particular state of the fly visual pigment. Therefore, it should be useful in in vivo investigations of the fly visual pigment, about which little is known.     

The photoreceptors and visual pigments of two species of Acipenseriformes, the shovelnose sturgeon (Scaphirhynchus platorynchus ) and the paddlefish (Polyodon spathula )

Scanning electron microscopy, microspectrophotometry, and spectrophotometry of digitonin extracts were employed to characterize the photoreceptors and visual pigments of two freshwater Acipenseriformes. The retinas of the shovelnose sturgeon, Scaphirhynchus platorynchus (Acipenseridae), and the paddlefish, Polyodon spathula (Polyodontidae) are dominated by large rods with long, broad outer segments. A second rod, rare and much narrower than the dominant rod, is present in Scaphirhynchus but not seen in Polyodon. The absorbance maximum of the visual pigment in the rods of Polyodon is near 540 nm; that of Scaphirhynchus near 534 nm. The retinas of both species contain substantial numbers of large, single cones, about 33% of the photoreceptors in Scaphirhynchus; 37% in Polyodon. Scaphirhynchus cone pigments have absorbance maxima near 610 nm, 521 nm and 470 nm, respectively. Polyodon cone pigments absorb maximally near 607 nm and 535 nm, respectively. All visual pigments are based on vitamin A₂. The data are compared to those from other Acipenseriformes and are discussed in terms of lifestyle and behavior. Accepted: 7 October 1998     

Changes in the spectral absorption of cone visual pigments during the settlement of the goatfish Upeneus tragula: the loss of red sensitivity as a benthic existence begins

The goatfish Upeneus tragula undergoes an abrupt metamorphosis at settlement when the pelagic larvae begin a reef-associated benthic mode of life. A microspectrophotometric investigation of the retinal visual pigments was carried out on fish prior to, during, and following settlement. It was found that the visual pigment in the long wavelength-absorbing member of the double cones in the dorsal retina changed rapidly from a rhodopsin with a wavelength of maximum absorption (max) of 580 nm to that of 530 nm. The second member of the double cones always had a rhodopsin with the max absorbing at shorter wavelengths. Prior to settlement the average for this class of cones was 487 nm whereas during and immediately following the settlement period the max recorded from individual outer segments was found to vary between 480 nm and 520 nm, with two possible classes of cone absorbance emerging within this range. These two classes of absorbance had average max values of 487 and 515 nm. The average max of the paired cone classes in one larger wild-settled fish were found to be at 506 nm and 530 nm. No change was detected in the max of the single cones or the rods which were always found to have a max of about 400 nm and 498 nm respectively. The loss of the redabsorbing pigment occurred over the same time scale as the metamorphosis of morphological features associated with the settlement process. It is thought that the loss of this visual pigment is associated with the change in light environment of the fishes as they leave the surface waters to begin a benthic mode of life in deeper water.Abbreviations AIMS Australian Institute of Marine Science - ANOVA Analysis of variance - IR infra-red - max wavelength of maximum absorption - MSP microspectrophotometer - NA numerical aperture - SL standard length     

The visual pigments of four deep-sea crustacean species

Summary The visual pigments of four mesopelagic crustacean species were studied at sea by means of microspectrophotometry. The absorbance maxima obtained for the visual pigments and their metarhodopsins, respectively, were: 493 nm and 481 nm (Systellaspis debilis), 485 nm and 480 nm (Acanthephyra curtirostris), 491 nm and 482 nm (A. smithi), and 495 nm and 487 nm (Sergestes tenuiremis). The spectral characteristics of the rhodopsins and metarhodopsins permit high photosensitivity and facilitate photoregeneration in a nearly monochromatic environment. Photic regeneration of rhodopsins from the deep-sea environment was demonstrated, and data were obtained which are consistent with the occurrence of dark regeneration. Specific optical density of the observed visual pigments was calculated for two species.     

Three visual pigments inDeilephila elpenor (Lepidoptera,Sphingidae)

Summary Selective irradiation of digitonin extracts (pH 6.1) from dark-adapted eyes of the Sphingid mothDeihphila elpenor revealed three visual pigments with _max at about 345, 440 and 520 nm. Prom the reaction of the chromophoric group with hydroxylamine it is concluded that these pigments are based on retinaldehyde, and therefore can be classed with rhodopsins. At –15 °C, each of the rhodopsins can completely (P 520) or partially (P 440 and P 345) be converted by light to acid metarhodopsin absorbing maximally at 480 ± 10 nm, which decay at room temperature to retinaldehyde and protein moiety. Spectrophotometric measurements on isolated retinae (pH 7.4) indicate that the metarhodopsins, in contrast to the findings on extracts, are thermostable in the photoreceptor membrane at room temperature. The results are compared with electrophysiological and microspectrophotometrical data. The different properties of the metarhodopsins in detergent solution and in the isolated retina are discussed.This work was supported by grants of the Deutsche Forschungsgemeinschaft to Prof. Dr. K. Hamdori (Rezeptorphysiologie).We would like to thank Miss Gabriele Ziem for excellent technical assistance.     

A visual pigment from chicken that resembles rhodopsin: amino acid sequence, gene structure, and functional expression.

The amino acid sequence of a rhodopsin-like visual pigment from chickens has been determined by isolating and sequencing its gene. The predicted sequence is between 70% and 80% identical to bovine, human, and chicken rhodopsins and between 40% and 50% identical to human blue, green, and red cone pigments, the chicken red cone pigment, and cavefish long-wave cone pigments. The encoded pigment, produced by transfection of cDNA into cultured cells, absorbs maximally at 495 nm as determined from photobleaching difference spectra and reacts at 20 degrees C with 50 mM hydroxylamine with a half-time of 16 min. These properties, together with a high pI predicted from the amino acid sequence, suggest that this cloned gene encodes the chicken green pigment previously identified by biochemical and spectroscopic studies. This sequence defines a new branch of the visual pigment gene family.     

β-arrestin functionally regulates the non-bleaching pigment parapinopsin in lamprey pineal

The light response of vertebrate visual cells is achieved by light-sensing proteins such as opsin-based pigments as well as signal transduction proteins, including visual arrestin. Previous studies have indicated that the pineal pigment parapinopsin has evolutionally and physiologically important characteristics. Parapinopsin is phylogenetically related to vertebrate visual pigments. However, unlike the photoproduct of the visual pigment rhodopsin, which is unstable, dissociating from its chromophore and bleaching, the parapinopsin photoproduct is stable and does not release its chromophore. Here, we investigated arrestin, which regulates parapinopsin signaling, in the lamprey pineal organ, where parapinopsin and rhodopsin are localized to distinct photoreceptor cells. We found that beta-arrestin, which binds to stimulated G protein-coupled receptors (GPCRs) other than opsin-based pigments, was localized to parapinopsin-containing cells. This result stands in contrast to the localization of visual arrestin in rhodopsin-containing cells. Beta-arrestin bound to cultured cell membranes containing parapinopsin light-dependently and translocated to the outer segments of pineal parapinopsin-containing cells, suggesting that beta-arrestin binds to parapinopsin to arrest parapinopsin signaling. Interestingly, beta-arrestin colocalized with parapinopsin in the granules of the parapinopsin-expressing cell bodies under light illumination. Because beta-arrestin, which is a mediator of clathrin-mediated GPCR internalization, also served as a mediator of parapinopsin internalization in cultured cells, these results suggest that the granules were generated light-dependently by beta-arrestin-mediated internalization of parapinopsins from the outer segments. Therefore, our findings imply that beta-arrestin-mediated internalization is responsible for eliminating the stable photoproduct and restoring cell conditions to the original dark state. Taken together with a previous finding that the bleaching pigment evolved from a non-bleaching pigment, vertebrate visual arrestin may have evolved from a "beta-like" arrestin by losing its clathrin-binding domain and its function as an internalization mediator. Such changes would have followed the evolution of vertebrate visual pigments, which generate unstable photoproducts that independently decay by chromophore dissociation.     

Visual pigments in the individual rods of deep-sea fishes

Summary The visual pigments in the rods of 15 species of deep-sea fish were examined by microspectrophotometry. In 13 species a single visual pigment was found. The max of these pigments, which ranged from 475 nm to 488 nm, suggest they give the fish maximum sensitivity to the ambient light in the deep, blue ocean waters where they live. In two species two visual pigments were found in separate rods.Bathylagus bericoides had rhodopsins of max 466 nm and 500 nm andMalacocephalus laevis had two rhodopsins of max 478 nm and 485 nm. It is noted that the species with two visual pigments tend to be dark in colour and live in deeper, darker, water.     

Visual and lenticular pigments in the eyes of demersal deep-sea fishes

We report on the lens pigmentation and visual pigments of 52 species of demersal deep-sea fishes caught at depths ranging from 480 m to 4110 m in the Porcupine Seabight and Goban Spur area of the North-eastern Atlantic. Only one species, caught between 480 and 840 m, had a lens with large amounts of pigment, consistent with the hypothesis that heavily pigmented lenses in deep-sea fish serve to enhance the contrast of bioluminescent signals by removing much of the background radiance, which is only visible to fish living shallower than 1000 m. Low concentrations of lens pigmentation were also observed in a further two species (Rouleina attrita and Micromesisteus poutassou). The retinae of all species except five, contained only a single visual pigment, as determined by microspectrophotometry of individual rods, and/or spectrophotometry of retinal wholemounts and retinal extracts. Those fishes caught between 500 m and 1100 m had wavelengths of peak sensitivity (_max) ranging from 476 nm to 494 nm, while most fish living below 1100 m tended to be more conservative with (_max) values ranging from 475 nm to 485 nm. The only exceptions to this were three deep-living species caught between 1600 m and 2000 m whose retinae contain abnormally short-wave sensitive visual pigments (Cataetyx laticeps — _max 468 nm; Alepocephalus bairdii — _max 467 nm; Narcetes stomias _max 472 nm), suggesting adaptation for the detection of short-wave bioluminescence.     

Interspecific variation in the visual pigments of deep-sea fishes

Summary Visual pigments in the rods of 38 species of deep-sea fish were examined by microspectrophotometry. 33 species were found to have a single rhodopsin with a wavelength of maximum absorbance ( _max) in the range 470–495 nm. Such visual pigments have absorbance maxima close to the wavelengths of maximum spectral transmission of oceanic water. 5 species, however, did not conform to this pattern and visual pigments were found with _max values ranging from 451 nm to 539 nm. In 4 of these species two visual pigments were found located in two types of rod. Some 2-pigment species which have unusual red sensitivity, also have red-emitting photophores. These species have both rhodopsin and porphyropsin pigments in their retinae, which was confirmed by HPLC, and the two pigments are apparently located in separate rods in the same retinal area. In deep-sea fishes the occurrence of unusual visual pigments seems to be correlated with aspects of the species' depth ranges. In addition to ecological influences we present evidence, in the form of _max spectral clustering, that indicates the degree of molecular constraint imposed on the evolution of visual pigments in the deep-sea.     

Phototaxis in monochromatic light and microspectrophotometry of the cerebral eye of the rotifer Brachionus calyciflorus

The observed wavelength-dependent variations in the phototaxis of the rotifer Brachionus calyciflorus inform us only partially about the spectral characteristics of the sensory pigment of the eye, since these variations are also linked to the absorption spectrum of the accessory pigment(s).

The absence of phototaxis between 420 nm and 500 nm is due to the lack of sensitivity of the sensory pigments at these wavelengths

The absence of response between 650 nm and 700 nm is due to a drop in the absorbance of the accessory pigments, which consequently no longer play a screening role at these wavelengths

The existence of oriented responses between 350 nm and 420 nm and between 500 nm and 650 nm, is due to the joint intervention of the two types of pigments at these wavelengths