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Journal of Comparative Physiology. A,... Jan 2017The facet lenses of the compound eyes of long-legged flies (Dolichopodidae) feature a striking, interlaced coloration pattern, existing of alternating rows of...
The facet lenses of the compound eyes of long-legged flies (Dolichopodidae) feature a striking, interlaced coloration pattern, existing of alternating rows of green-yellow and orange-red reflecting facets, due to dielectric multilayers located distally in the facet lenses (Bernard and Miller. Invest Ophthalmol 7:416-434 (1968). We investigated this phenomenon in the dolichopodid Dolichopus nitidus by applying microspectrophotometry, electron microscopy and optical modeling. The measured narrow-band reflectance spectra, peaking at ~540 and ~590 nm with bandwidth ~105 nm, are well explained by a refractive index oscillating sinusoidally in six periods around a mean value of about 1.44 with amplitude 0.6. The facet lens reflectance spectra are associated with a spectrally restricted, reduced transmittance, which causes modified spectral sensitivities of the underlying photoreceptors. Based on the modeling and electroretinography of the dolichopodid Condylostylus japonicus we conjecture that the green and orange facets narrow the spectral bandwidths of blue and green central photoreceptors, respectively, thus possibly improving color and/or polarization vision.
Topics: Animals; Compound Eye, Arthropod; Diptera; Electroretinography; Female; Insect Proteins; Iridescence; Male; Microscopy, Electron, Transmission; Microspectrophotometry; Models, Biological; Photoreceptor Cells, Invertebrate; Retinal Pigments
PubMed: 27873005
DOI: 10.1007/s00359-016-1131-y -
Acta Crystallographica. Section D,... Oct 2014X-ray-radiation-induced alterations to protein structures are still a severe problem in macromolecular crystallography. One way to avoid the influence of radiation...
X-ray-radiation-induced alterations to protein structures are still a severe problem in macromolecular crystallography. One way to avoid the influence of radiation damage is to reduce the X-ray dose absorbed by the crystal during data collection. However, here it is demonstrated using the example of the membrane protein bacteriorhodopsin (bR) that even a low dose of less than 0.06 MGy may induce structural alterations in proteins. This dose is about 500 times smaller than the experimental dose limit which should ideally not be exceeded per data set (i.e. 30 MGy) and 20 times smaller than previously detected specific radiation damage at the bR active site. To date, it is the lowest dose at which radiation modification of a protein structure has been described. Complementary use was made of high-resolution X-ray crystallography and online microspectrophotometry to quantitatively study low-dose X-ray-induced changes. It is shown that structural changes of the protein correlate with the spectroscopically observed formation of the so-called bR orange species. Evidence is provided for structural modifications taking place at the protein active site that should be taken into account in crystallographic studies which aim to elucidate the molecular mechanisms of bR function.
Topics: Bacteriorhodopsins; Catalytic Domain; Crystallography, X-Ray; Dose-Response Relationship, Radiation; Fourier Analysis; Models, Molecular; Protein Conformation; Proteins; X-Rays
PubMed: 25286851
DOI: 10.1107/S1399004714017295 -
Scientific Reports Mar 2016Monitoring the effect of the substrate on the local surface plasmon resonance (LSPR) of metallic nanoparticles is key for deepening our understanding of light-matter...
Monitoring the effect of the substrate on the local surface plasmon resonance (LSPR) of metallic nanoparticles is key for deepening our understanding of light-matter interactions at the nanoscale. This coupling gives rise to shifts of the LSPR as well as changes in the scattering pattern shape. The problem requires of high-throughput techniques that present both high spatial and spectral resolution. We present here a technique, referred to as Spatially Multiplexed Micro-Spectrophotometry (SMMS), able to perform polarization-resolved spectral and spatial analysis of the scattered light over large surface areas. The SMMS technique provides three orders of magnitude faster spectroscopic analysis than conventional dark-field microspectrophotometry, with the capability for mapping the spatial distribution of the scattered light intensity with lateral resolution of 40 nm over surface areas of 0.02 mm(2). We show polarization-resolved dark-field spectral analysis of hundreds of gold nanoparticles deposited on a silicon surface. The technique allows determining the effect of the substrate on the LSPR of single nanoparticles and dimers and their scattering patterns. This is applied for rapid discrimination and counting of monomers and dimers of nanoparticles. In addition, the diameter of individual nanoparticles can be rapidly assessed with 1 nm accuracy.
PubMed: 26953042
DOI: 10.1038/srep22836 -
Journal of Fish Biology Sep 2022The main goal of this study was to clarify whether the spectral properties of retinal photoreceptors reflect the features of behaviour of closely related fish species...
The main goal of this study was to clarify whether the spectral properties of retinal photoreceptors reflect the features of behaviour of closely related fish species cohabiting shallow marine and fresh waters. The spectral sensitivity of photoreceptors was compared between two smelt species, Hypomesus japonicus and Japanese smelt Hypomesus nipponensis. The spectral absorption of the visual pigments was measured using microspectrophotometry. In H. japonicus, a mostly marine species, all photoreceptors contained visual pigments based on retinal and were distributed differently in specific retinal areas. The absorbance maxima (λ ) of rods and long-wave-sensitive members of double cones throughout the retina amounted to 507 and 573 nm, respectively, but the λ value of the short-wave-sensitive members of double cones and single cones in the temporal hemiretina showed a significant blue shift compared to the nasal hemiretina: 485 vs. 516 nm and 375 vs. 412 nm, respectively, thus enhancing the short-wave sensitivity of the temporal hemiretina. In H. nipponensis, an euryhaline species, the estimated λ value of both rods and cones significantly varied between the groups caught in different localities (sea, river or estuary) because of the presence of rhodopsin/porphyropsin mixtures. The long-wavelength shift in rod and cone photoreceptors was observed because of changes in the chromophore complement in closely related but ecologically different species dwelling in freshened bodies of water. Considering the data available in the literature, several putative common opsin genes have been suggested for species under study.
Topics: Animals; Opsins; Osmeriformes; Photoreceptor Cells, Vertebrate; Retinal Cone Photoreceptor Cells; Rhodopsin
PubMed: 35655413
DOI: 10.1111/jfb.15128 -
Journal of the Royal Society, Interface Oct 2015Vision is the primary sensory modality of birds, and its importance is evident in the sophistication of their visual systems. Coloured oil droplets in the cone...
Vision is the primary sensory modality of birds, and its importance is evident in the sophistication of their visual systems. Coloured oil droplets in the cone photoreceptors represent an adaptation in the avian retina, acting as long-pass colour filters. However, we currently lack understanding of how the optical properties and morphology of component structures (e.g. oil droplet, mitochondrial ellipsoid and outer segment) of the cone photoreceptor influence the transmission of light into the outer segment and the ultimate effect they have on receptor sensitivity. In this study, we use data from microspectrophotometry, digital holographic microscopy and electron microscopy to inform electromagnetic models of avian cone photoreceptors to quantitatively investigate the integrated optical function of the cell. We find that pigmented oil droplets primarily function as spectral filters, not light collection devices, although the mitochondrial ellipsoid improves optical coupling between the inner segment and oil droplet. In contrast, unpigmented droplets found in violet-sensitive cones double sensitivity at its peak relative to other cone types. Oil droplets and ellipsoids both narrow the angular sensitivity of single cone photoreceptors, but not as strongly as those in human cones.
Topics: Animals; Chickens; Color; Electromagnetic Radiation; Humans; Light; Microscopy, Electron, Scanning; Microspectrophotometry; Models, Biological; Models, Statistical; Oils; Optics and Photonics; Refractometry; Retina; Retinal Cone Photoreceptor Cells; Vision, Ocular
PubMed: 26423439
DOI: 10.1098/rsif.2015.0591 -
Molecular Biology and Evolution Apr 2024Visual systems adapt to different light environments through several avenues including optical changes to the eye and neurological changes in how light signals are...
Visual systems adapt to different light environments through several avenues including optical changes to the eye and neurological changes in how light signals are processed and interpreted. Spectral sensitivity can evolve via changes to visual pigments housed in the retinal photoreceptors through gene duplication and loss, differential and coexpression, and sequence evolution. Frogs provide an excellent, yet understudied, system for visual evolution research due to their diversity of ecologies (including biphasic aquatic-terrestrial life cycles) that we hypothesize imposed different selective pressures leading to adaptive evolution of the visual system, notably the opsins that encode the protein component of the visual pigments responsible for the first step in visual perception. Here, we analyze the diversity and evolution of visual opsin genes from 93 new eye transcriptomes plus published data for a combined dataset spanning 122 frog species and 34 families. We find that most species express the four visual opsins previously identified in frogs but show evidence for gene loss in two lineages. Further, we present evidence of positive selection in three opsins and shifts in selective pressures associated with differences in habitat and life history, but not activity pattern. We identify substantial novel variation in the visual opsins and, using microspectrophotometry, find highly variable spectral sensitivities, expanding known ranges for all frog visual pigments. Mutations at spectral-tuning sites only partially account for this variation, suggesting that frogs have used tuning pathways that are unique among vertebrates. These results support the hypothesis of adaptive evolution in photoreceptor physiology across the frog tree of life in response to varying environmental and ecological factors and further our growing understanding of vertebrate visual evolution.
Topics: Humans; Animals; Opsins; Retinal Pigments; Anura; Gene Duplication; Microspectrophotometry
PubMed: 38573520
DOI: 10.1093/molbev/msae049 -
The Journal of Physical Chemistry. B Dec 2018The fluorescence of Green Fluorescent Protein (wtGFP) and variants has been exploited in distinct applications in cellular and analytical biology. GFPs emission depends...
The fluorescence of Green Fluorescent Protein (wtGFP) and variants has been exploited in distinct applications in cellular and analytical biology. GFPs emission depends on the population of the protonated (A-state) and deprotonated (B-state) forms of the chromophore. Whereas wtGFP is pH-independent, mutants in which Ser65 is replaced by either threonine or alanine (as in GFPmut2) are pH-dependent, with a p K around 6. Given the wtGFP pH-independence, only the structure of the protonated form was determined. The deprotonated form was deduced on the basis of the crystal structure of the Ser65Thr mutant at basic pH, assuming that it corresponds to the conformation populated in solution. Here, we present an investigation where structures of the protonated and deprotonated forms of GFPmut2 were determined from crystals grown in either MPD at pH 6 or PEG at pH 8.5, and moved to either higher or lower pH. Both crystal forms of GFPmut2 were titrated monitoring the process via polarized absorption microspectrophotometry in order to precisely correlate the protonation process with the structures. We found that (i) in solution, chromophore titration is not thermodynamically coupled with any residue and Glu222 is always protonated independent of the protonation state of the chromophore; (ii) the lack of coupling is reflected in the structural behavior of the chromophore and Glu222 environments, with only the former showing variations with pH; (iii) titrations of low-pH and high-pH grown crystals exhibit a Hill coefficient of about 0.75, indicating an anticooperative behavior not observed in solution; (iv) structures where pH was changed in the crystal point to Glu222 as the ionizable group responsible for the outset of the anticooperative behavior; and (v) in GFPmut2 the canonical GFP proton wire involving the chromophore is not interrupted at the level of Ser205 and Glu222 at basic pH as in the Ser65Thr mutant. This allows proposing the structure of the deprotonated state of GFPmut2 as an alternative model for the analogous state of wtGFP.
Topics: Animals; Crystallography, X-Ray; Escherichia coli; Green Fluorescent Proteins; Hydrogen Bonding; Hydrogen-Ion Concentration; Hydrozoa; Imidazoles; Microspectrophotometry; Mutation; Protein Binding; Protons
PubMed: 30179482
DOI: 10.1021/acs.jpcb.8b07260 -
Radiolar Eyes of Serpulid Worms (Annelida, Serpulidae): Structures, Function, and Phototransduction.The Biological Bulletin Aug 2017Fan worms, represented by sabellid and serpulid polychaetes, have an astonishing array of unusual eyes and photoreceptors located on their eponymous feeding appendages....
Fan worms, represented by sabellid and serpulid polychaetes, have an astonishing array of unusual eyes and photoreceptors located on their eponymous feeding appendages. Here we organize the previous descriptions of these eyes in serpulids and report new anatomical, molecular, and physiological data regarding their structure, function, and evolution and the likely identity of their phototransduction machinery. We report that, as in sabellids, serpulids display a broad diversity of radiolar eye arrangements and ocellar structures. Furthermore, the visual pigment expressed in the eyes of Spirobranchus corniculatus, a species of the charismatic Christmas tree worms, absorbs light maximally at 464 nm in wavelength. This visual pigment closely matches the spectrum of downwelling irradiance in shallow coral reef habitats and lends support to the hypothesis that these radiolar photoreceptors function as a silhouette-detecting "burglar alarm" that triggers a rapid withdrawal response when the worm is threatened by potential predators. Finally, we report on the transcriptomic sequencing results for the radiolar eyes of S. corniculatus, which express invertebrate c-type opsins in their ciliary radiolar photoreceptors, closely related to the opsin found in the radiolar eyes of the sabellid Acromegalomma interruptum. We explore the potential for a shared evolutionary lineage between the radiolar photoreceptors of serpulids and sabellids and consider these unique innovations in the broader context of metazoan eye evolution.
Topics: Animals; Annelida; Light; Light Signal Transduction; Opsins; Photoreceptor Cells, Invertebrate; Transcriptome
PubMed: 29182501
DOI: 10.1086/694735 -
PloS One 2014The mesopelagic zone is a visual scene continuum in which organisms have developed various strategies to optimize photon capture. Here, we used light microscopy,...
The mesopelagic zone is a visual scene continuum in which organisms have developed various strategies to optimize photon capture. Here, we used light microscopy, stereology-assisted retinal topographic mapping, spectrophotometry and microspectrophotometry to investigate the visual ecology of deep-sea bioluminescent sharks [four etmopterid species (Etmopterus lucifer, E. splendidus, E. spinax and Trigonognathus kabeyai) and one dalatiid species (Squaliolus aliae)]. We highlighted a novel structure, a translucent area present in the upper eye orbit of Etmopteridae, which might be part of a reference system for counterillumination adjustment or acts as a spectral filter for camouflage breaking, as well as several ocular specialisations such as aphakic gaps and semicircular tapeta previously unknown in elasmobranchs. All species showed pure rod hexagonal mosaics with a high topographic diversity. Retinal specialisations, formed by shallow cell density gradients, may aid in prey detection and reflect lifestyle differences; pelagic species display areae centrales while benthopelagic and benthic species display wide and narrow horizontal streaks, respectively. One species (E. lucifer) displays two areae within its horizontal streak that likely allows detection of conspecifics' elongated bioluminescent flank markings. Ganglion cell topography reveals less variation with all species showing a temporal area for acute frontal binocular vision. This area is dorsally extended in T. kabeyai, allowing this species to adjust the strike of its peculiar jaws in the ventro-frontal visual field. Etmopterus lucifer showed an additional nasal area matching a high rod density area. Peak spectral sensitivities of the rod visual pigments (λmax) fall within the range 484-491 nm, allowing these sharks to detect a high proportion of photons present in their habitat. Comparisons with previously published data reveal ocular differences between bioluminescent and non-bioluminescent deep-sea sharks. In particular, bioluminescent sharks possess higher rod densities, which might provide them with improved temporal resolution particularly useful for bioluminescent communication during social interactions.
Topics: Animals; Luminescence; Pigmentation; Retinal Rod Photoreceptor Cells; Sharks
PubMed: 25099504
DOI: 10.1371/journal.pone.0104213 -
The Journal of Adhesive Dentistry 2016To assess the light irradiance (LI) delivered by two light-curing units and to measure the degree of conversion (DC) of three composite cements and one flowable...
PURPOSE
To assess the light irradiance (LI) delivered by two light-curing units and to measure the degree of conversion (DC) of three composite cements and one flowable composite when cured through zirconia or ceramic-veneered zirconia plates with different thicknesses.
MATERIALS AND METHODS
Three dual-curing composite cements (Clearfil Esthetic Cement, Panavia F2.0, G-CEM LinkAce) and one light-curing flowable composite (G-aenial Universal Flo) were investigated. Nine different kinds of zirconia plates were prepared from three zirconia grades (YSZ: Aadva and KATANA; Ce-TZP/Al2O3: NANOZR) in three different thicknesses (0.5- and 1.5-mm-thick zirconia, and 0.5-mm-thick zirconia veneered with a 1.0-mm-thick veneering ceramic). Portions of the mixed composite cements and the flowable composite were placed on a light spectrometer to measure LI while being light cured through the zirconia plates for 40 s using two light-curing units (n = 5). After light curing, micro-Raman spectra of the composite films were acquired to determine DC at 5 and 10 min, 1 and 24 h, and at 1 week.
RESULTS
The zirconia grade and the thickness of the zirconia/veneered zirconia plates significantly decreased LI. Increased LI did not increase DC. Only the Ce-TZP/Al2O3 (NANOZR) zirconia was too opaque to allow sufficient light transmission and resulted in significantly lower DC.
CONCLUSION
Although zirconia-based restorations attenuate the LI of light-curing units, the composite cements and the flowable composite could be light cured through the YSZ zirconia. LI is too low through Ce-TZP/Al2O3 zirconia, necessitating the use of self-/dual-curing composite cements.
Topics: Aluminum Oxide; Ceramics; Cerium; Composite Resins; Curing Lights, Dental; Dental Materials; Dental Veneers; Humans; Light; Light-Curing of Dental Adhesives; Materials Testing; Methacrylates; Microspectrophotometry; Polyethylene Glycols; Polymerization; Polymethacrylic Acids; Polyurethanes; Radiation Dosage; Resin Cements; Scattering, Radiation; Spectrum Analysis, Raman; Surface Properties; Yttrium; Zirconium
PubMed: 27022645
DOI: 10.3290/j.jad.a35842