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Biomedical subjects

F I Hárosi

Publications and source records attributed to F I Hárosi.

At least 19 recordsLinked to original sources

Visual pigments of African cichlid fishes: evidence for ultraviolet vision from microspectrophotometry and DNA sequences.

We have found evidence for ultraviolet visual capabilities in a Lake Malawi cichlid fish, Metriaclima zebra. Microspectrophotometry of single cones revealed a visual pigment with peak sensitivity at 368+/-4 nm. M. zebra also expresses a putative ultraviolet opsin gene whose sequence is closely related to the SWS-1 opsin for other fishes. Several other African cichlids have a functional copy of this UV gene in their genomic DNA, but do not appear to express this gene as adults. These results suggest that ultraviolet vision is important for some cichlid fishes. UV wavelengths should therefore be included in future studies of cichlid vision, behavior and color patterns.

Amino Acid Sequence↗

Double-cone internal reflection as a basis for polarization detection in fish.

Some species of fish are able to discriminate, in addition to intensity and wavelength (color), the direction of polarization of visible light. Optical experiments on axially oriented retinal cones from trout and sunfish with use of two types of polarization microscope indicate anisotropic light transmission through paired cones. The measured linear birefringence of paired cone ellipsoids is consistent with the presence of membranous partitions. It is proposed that the partition between the two members of a paired cone, which often appears extensive and flat, functions as a dielectric mirror and that polarization-dependent reflection and refraction at this partition constitutes the underlying mechanism in the transduction of polarization into intensity variation at the photoreceptor's outer segments. We support this hypothesis with linear birefringence and linear dichroism measurements, histological evidence, large-scale optical model measurements, and theoretical calculations based on Fresnel's formulas.

Animals↗

Sickling of anoxic red blood cells in fish.

The occurrence of the mutant hemoglobin Hb S in human red blood cells results in sickle cell anemia. This disease, including its genetic and molecular bases, has been extensively investigated and is well understood (1,2). The presence of deoxy-induced sickling of animal erythrocytes is largely unknown, however. We examined red blood cells (RBCs) from several fish species in vitro under aerated and anoxic conditions. Our polarized light microscopic techniques were aimed at establishing correlations between erythrocyte morphology, state of oxygenation, spectral absorbance, linear dichroism, and linear birefringence. We found no fish with intracellular HbO2 polymerization; but there were intraerythrocytic aggregations of deoxy Hb with a high degree of either molecular order or disorder. The ordered aggregates in the RBCs of Atlantic cod, haddock, and toadfish were remarkably similar in dichroic ratio magnitudes and birefrigence to those in human RBCs that contain HbS. Therefore, fish hemoglobins appear to be good models of sickling disorders and polymerization-related phenomena. The consequences of sickling on animal health and fish aquaculture remain to be studied.

Animals↗

Visual pigment types and quantum-catch ratios: implications from three marine teleosts.

Experimental data on photoreceptor cells and visual pigments are the basis for model calculations performed to assess photoreceptor quantum catches under disparate irradiance conditions. Three unrelated species of fish--black sea bass (Centropristis striata), sea raven (Hemitripterus americanus), and adult winter flounder (Pseudopleuronectes americanus)--are considered. In each case, receptor type quantum catches are compared for various water types and depths. By associating the habit and habitat of an organism with the physical properties of its photoreceptors, quantum-catch ratios are found as possible criteria in the selection of pigment peaks (lambda max). In addition to integrated ("total") quantum catches by the receptor types, rates of quantum catches are determined as a function of wavelength. The latter functions are replotted as pairwise difference spectra. These, in turn, are used to assess the ability of receptor types to participate in wavelength discrimination.

Animals↗

An analysis of two spectral properties of vertebrate visual pigments.

Spectral data are scrutinized on two properties: (1) the chromophore-induced wavelength shift, i.e. the variation in absorbance maximum (lambda max) upon exchanging the vitamin A1-based chromophore with one based on vitamin A2 in the same opsin; (2) the half-band width (HBW) variation as a function of the reciprocal of the peak absorbance (lambda max)-1. It is shown that in an extended spectral range that includes the UV and when data are plotted in wavenumbers, the chromophore-induced shifts can be approximated with a parabola, whose minimum occurs near 23,000 cm-1 (approximately 430 nm). Similarly, HBW variations can also be fitted with parabolas, however, these show maxima near 430 nm. The theoretical implications of the two phenomena concerning lambda max tuning in vertebrate visual pigments are discussed.

Amphibians↗

Spectral characteristics of visual pigments in rainbow trout (Oncorhynchus mykiss).

We investigated retina preparations of young rainbow trout (Oncorhynchus mykiss) with body wt 5-40 g. Rods, single and double cones were measured in side-on orientation by microspectrophotometry, identifying five spectrally distinct visual pigments (or photoreceptors containing mixtures of visual pigments). The mean wavelength of peak absorbance (lambda max) of the alpha-bands were 365 and 434 nm in single cones, 531 and 576 nm in double cones, and 521 nm in the rods. The half-band width (HBW) of the main absorption bands were broader than expected of retinal- (vitamin A1-) based visual pigments, and thus, they were indicative of a mixed chromophore pool derived from both the vitamin A1 and A2 forms. One consequence of the utilization of mixed chromophores is the broadening of the alpha-band absorption in each pigment type. And yet, we obtained exceptionally narrow HBW for the UV-type pigment, when compared with HBW values expected on the basis of the linear trend seen in visual pigments absorbing in the visible spectrum. We conclude that the UV pigment in rainbow trout has an unusually narrow HBW. Nevertheless, this species is not exceptional in this regard, for the UV-absorbing visual pigments in other vertebrate species also have narrow HBW.

Animals↗

Zebrafish ultraviolet visual pigment: absorption spectrum, sequence, and localization.

In many vertebrates, UV-sensitive photoreceptors have been identified by microspectrophotometry and UV-visual sensitivity has been identified by behavioral studies, but as yet no vertebrate UV-sensitive pigment gene has been isolated. We have sequenced a cDNA clone that hybridizes to short single cone cells in the zebrafish (Brachydanio rerio). These cells, which make up 25% of the cone population in zebrafish retinae, are UV-sensitive (lambda max approximately 360 nm). The visual pigment encoded by this gene is unusual in that its amino acid sequence is more homologous to the rod pigment rhodopsin (up to 89%) than to other cone pigments (35-83%). Like all other vertebrate visual pigments, it contains a lysine residue at position 296, the presumptive retinal binding site, and a glutamate residue at position 113. However, it is unique in possessing a lysine residue at position 126, which may account for the UV-sensitivity of the pigment.

Amino Acid Sequence↗

Visual pigments of the tree shrew (Tupaia belangeri) and greater galago (Galago crassicaudatus): a microspectrophotometric investigation.

Optical density, linear dichroism and bleaching difference spectra were measured in photoreceptors from the cone-dominated retina of the tree shrew (Tupaia belangeri) and from the rod-dominated retina of the greater galago (Galago crassicaudatus) using a single-beam, wavelength-scanning, dichroic microspectrophotometer. In Tupaia, we obtained spectral records from 272 cone receptors (from 10 eyes), of which 264 were long-wave sensitive (lambda max = 555 +/- 6 nm) and 8 were short-wave sensitive (lambda max = 428 +/- 15 nm). Also, one anatomically-recognizable rod receptor was encountered and showed a peak absorption at approx. 496 nm. No mid-wave sensitive cone pigment was found, as would be expected in deutan-type dichromats like the tree shrew. Pre-retinal absorption by the cornea and lens was maximal at 370 nm and negligible beyond 430 nm. In Galago, all outer segments measured were rod-like in appearance (lambda max near 501 nm). Measurements of pre-retinal absorption yielded a single-peaked function with a maximum at 363 nm.

Absorption↗

Cynomolgus and rhesus monkey visual pigments. Application of Fourier transform smoothing and statistical techniques to the determination of spectral parameters.

Microspectrophotometric measurements were performed on 217 photoreceptors from cynomolgus, Macaca fascicularis, and rhesus, M. mulatta, monkeys. The distributions of cell types, for rods and blue, green, and red cones were: 52, 12, 47, and 44, respectively, for the cynomolgus, and 22, 4, 13, and 13 for the rhesus. Visual cells were obtained fresh (unfixed), mounted in various media (some containing 11-cis-retinal), and then located visually under dim red (650 nm) illumination. Absolute absorbance (A), linear dichroism (LD), and bleaching difference (BD) absorbance spectra were recorded through the sides of outer segments. The spectra were subjected to rigorous selection criteria, followed by digital averaging and Fourier transform filtering. Statistical methods were also applied to the accepted samples in the estimation of population means and variances. The wavelength of mean peak absorbance (lambda max) and the standard error at 95% certainty of the rod and blue, green, and red cone pigments in cynomolgus were 499.7 +/- 2.5, 431.4 +/- 4.2, 533.9 +/- 2.4, and 565.9 +/- 2.8 nm, respectively. The rhesus pigments were statistically indistinguishable from the cynomolgus, having lambda max of approximately 500, 431, 534, and 566 nm. Statistical tests did not reveal the presence of a lambda max subpopulation (i.e., anomalous pigments). The bandwidth of each alpha-band was determined in two segments, giving rise to the longwave half-density (LWHDBW), shortwave half-density (SWHDBW), and total half-density (THDBW) bandwidths. The LWHDBW was found to have the smallest variance. Both the LWHDBW and the THDBW showed linear dependence on the peak wavenumber (lambda max)-1 for the four macaque pigments.

Animals↗

First step in vision: proton transfer or isomerization?

Recent views on the photochemistry of (vertebrate) vision are examined. Problems related to the structure of the proton bridge at the Schiff base chromophore are considered, in particular the possible shapes of the section of the potential surface governing the motion of the proton in the bridge. This leads to the question as to whether proton translocation occurs in the initial step of visual transduction and if it precedes or follows cis-trans isomerisation. The related controversy could be solved through the assumption of the presence of water molecules that stabilize the ions in the proton bridge. The causes of the instability of bathorhodopsin are discussed and the importance of additional perturbations by polar groups is stressed.

Animals↗

Ellipsosomes: organelles containing a cytochrome-like pigment in the retinal cones of certain fishes.

Ellipsosomes are dense spherical bodies containing a very large concentration of a heme pigment spectroscopically resembling pure cytochrome c. They are located at the outer ends of the inner segments of the cones of certain fishes. Although, superficially, they resemble the similarly located oil droplets in the cones of birds and reptiles, their ultrastructure and staining properties resemble those of the neighboring mitochondria. However, like the oil droplets, they may serve as intracellular color filters.

Animals↗

Photoreceptors and visual pigments in a cichlid fish, Nannacara anomala.

Correlation of visual pigment content and photoreceptor cell structure is studied in the cichlid fish Nannacara anomala the retina of which is characterized by simple organization of the outer plexiform layer. The visual cell types consist of long and bulky rods and only two types of cones: equal double cones and short single cones. Both types of cones differ form other known teleost cones in that their outer segments are very slender yet twice as long as the inner segments. The double cones form a square mosaic with four doubles along the sides an a single cone in the center. Four rods usually surround the single cone and a few others lie in the corners of the square. The population densities of photoreceptor cells in the central part of the retina is 45,600 cones and 52,600 rods per mm2. The cone to rod ratio is 1:1.15; the double cone to single cone ratio is 4.07:1. The outer segments of photoreceptors contain four spectroscopic types of pigment: one type in rods with lambda max = 498 +/- 5 nm, one type in single cones with lambda max = 460 +/- 5nm, and one in each member of the double cones with lambda max of 555 +/- 5 and 600 +/- 10 nm. Single cones are found to contain only the blue-absorbing pigment. The double cones, whenever both members are successfully recorded from, appear to use two spectrally different pigments. Based on spectral bandwidth determination, the four pigments are probably of the prophyropsin class, although some admixture (10--20%) of rhodopsin-class pigments is also indicated, especially in double cones.

Animals↗

Photochemistry of visual pigments: an interpretation of spectral changes in terms of molecular associations and isomerization.

A unified view of the photochemical part of the visual process is presented. It is proposed that both conformational changes and changes in intermolecular interactions in the sequence that leads from rhodopsin through batho-lumi- and meta-I to meta-II- rhodopsin have to be considered in order to elucidate the mechanism of the visual process. The main intermolocular associations are assumed to be the hydrogen bond involving the nitrogen atom of the Schiff base and the interaction between a negative group and the beta-ionone ring. The two together can be used to explain the absorption wavelength of rhodopsin without actual protonation. The main line of thought is as follows: when light is absorbed the basicity of the Schiff base increases significantly. This triggers proton transfer in the H-bond. At the same time cis-trans isomerization begins but it only reaches the coplanar all-trans stage at metarhodopsin-II. Lumi-, meta-I and meta-II are way stations in the stepwise isomerization whereby the energy of the photon is used together with thermal energy. Batho- is probably still close to 11-cis which then becomes successively strained 13-cis and 15-cis. In vertebrate rhodopsins at the meta-II stage both the H-bond and the beta-ionone interaction are severed and meta-II becomes exposed to attack by water molecules. The importance of syn-anti isomerization on the C=N bond is emphasized. The irreversibility necessary for the production of a signal requires that the proton does not return to its original donor. The possible identity of the donor is discussed: it might be an amino acid or the polar part of a lipid. Relevant observations made on bacteriorhodopsin, squid rhodopsin and chicken iodopsin are discussed.

Amino Acids↗