Victor Benno Meyer-Rochow

Selective photoreceptor damage in four species of insects induced by experimental exposures to UV-irradiation

Damage to photoreceptive cells of insect compound eyes exposed to abnormally high doses of UV-radiation of 350nm peak wavelength manifests itself in at least two different ways. In the butterflies Papilio xuthus and Pieris napi from Japan and northern Finland, respectively, only the cell bodies of retinula cells 1 and 2, (identified as short wavelength receptors), but not their corresponding rhabdomeres, exhibit damage with apoptotic features. In the eye of UV-irradiated adult crickets, however, cell bodies and cytoplasm remain normal, while the rhabdomeres of cells 7 and 8 exhibit signs of severe membrane disruptions. No signs of damage whatsoever occurred in the eyes of northern Finnish bumblebees exposed to UV. It is suggested that metabolic shortfalls in the UV-sensitive cells of the butterfly eyes result in cellular shut-down, but that in the cricket receptors UV-induced changes of the membrane lipids dominate, leading to membrane instability without concomittant cell death. The strong resistance of the bumblebee eye to UV-induced damage requires further investigation, but since preconditioning to light can reduce photic damage in the rat eye, the 24h daylight experienced by northern Finnish bumblebees during the summer season could be involved.

Fatty acid composition and ultrastructure of photoreceptive membranes in the crayfish Procambarus clarkii under conditions of thermal and photic stress

Abstract The ultrastructural state of the crayfish visual membrane is correlated with its fatty acid composition during times of photic and thermal stress and the period over which the dynamic events occur is investigated. Crayfish kept at 4 °C under constant darkness contain in their rhabdoms significantly increased amounts of unsaturated fatty acids such as 16:1, 18:1, 20:5, and 22:6 compared with individuals kept at 25 °C. The ratio of unsaturated/saturated fatty acids (UFA/SFA-ratio) amounts to 2.17 in the cold-water- and 1.46 in the warm water-acclimated animals. The visual membranes of crayfish suddenly transferred from 4 °C to 25 °C exhibited ultrastructural modifications such as membrane collapse and disappearance of microvillar dense␣core-filaments most clearly 3 h post-transfer. Parallel to the structural changes a significant increase in fatty acid 18:0 was observed, while the amounts of 16:1 and 20:1 decreased. When 4 °C, dark-adapted crayfish were exposed to light alone and not a temperature increase, only fatty acid 22:6 showed a significant reduction to 10% of its pre-experimental level within 2 h of exposure. Thereafter, it slowly increased again. In cold water-acclimated crayfish that had been exposed to light of 5000 lx for 3␣weeks no significant change of the UFA/SFA ratio was observed, although fatty acid species 18:0, 20:4, and 20:5 had increased at the expense of fatty acids 14:0, 16:0, 16:1, 18:1, 20:1, and 22:6. The total amount of fatty acids, however, had become significantly smaller (from 0.058 ng g−1 body weight in the dark-adapted to 0.048 ng g−1 in the light-adapted crayfish). Morphologically the rhabdom volume had decreased by approx. 20%, but ultrastructurally rhabdom microvilli remained almost unchanged. The amount of peroxidized lipids in the retina following irradiation with bright white light in the cold-adapted crayfish fell during the first 2 h of exposure from 0.4 nmol g−1 to 0.32 nmol g−1, but after 12 h of exposure had reached a level of 0.48 nmol g−1. Greatest structural abnormalities to the visual membranes occurred when dark-adapted, cold-acclimated crayfish were suddenly subjected to bright light and an increase in water temperature. Under such conditions the microvillar arrangement was disrupted and membrane collapse and disappearance of core-filaments were apparent. Our results provide evidence that the fatty acid composition of the membranes determines to a considerable extent the structural integrity of the photoreceptor, but that it is too simplistic a model to think that peroxidation of membrane lipids alone is responsible for the disintegration of the photoreceptive membranes in the crayfish eye following exposure to bright light.

Daily variation of melatonin content in the optic lobes of the crab Neohelice granulata

Melatonin is a biogenic amine, known from almost all phyla of living organisms. In vertebrates melatonin is produced rhythmically in the pinealocytes of the pineal gland, relaying information of the environmental light/dark cycle to the organism. With regard to crustaceans only a handful of studies exist that has attempted to identify the presence and possible daily variation of this substance. We set out to investigate whether in the crab Neohelice granulata melatonin was produced in the optic lobes of these animals and underwent rhythmic fluctuations related to the daily light/dark cycle. Our experimental animals were divided into three groups exposed to different photoperiods: normal photoperiod (12L:12D), constant dark (DD), and constant light (LL). The optic lobes were collected every 4 hours over a 24-h period for melatonin quantification by radioimmunoassay (RIA). N. granulata kept under 12 L:12D and DD conditions, showed daily melatonin variations with two peaks of abundance (p<0.05), one during the day and another, more extensive one, at night. Under LL-conditions no significant daily variations were noticeable (p>0.05). These results demonstrate the presence of a daily biphasic fall and rise of melatonin in the eyestalk of N. granulata and suggest that continuous exposure to light inhibits the production of melatonin synthesis.

Ultrastructural study of first and second order neurons in the visual system of the crab Ucides cordatus following exposure to ultraviolet radiation.

The visual system as an interface between the environment and the living organism can serve as a sensitive indicator especially in studies that deal with effects of radiation. The crab retina as the seat of the photoreceptors and the lamina ganglionaris as the place of second order neurons were the targets of our study. Conventional specimen preparation techniques for transmission electron microscopy (TEM) are adequate to preserve any modifications that may occur as a consequence of the experimental treatment. In this study we analyzed by TEM how retinal and lamina ganglionaris cells of the crab Ucides cordatus responded to a 30 min exposure to ultraviolet C (UV-C) and ultraviolet B (UV-B) radiation with doses of 7200 and 590J/cm(2), respectively. The results show that damaged cells occurred in both retina and lamina ganglionaris, but that the retinal cells were affected to a greater extent. Morphological alterations of the pigment granules and an increase in the quantity of lipid droplets of the retinal cells were also observed. Additional changes include an increase in heterochromatin, nuclear karyolyses and karyorrhexes, distention of rough endoplasmic reticulum and mitochondrial disruptions. The observed morphological changes are indicative of apoptotic processes and show that an exposure to light of wavelengths of 254 and 312 nm may be injurious to the visual system of invertebrates.

Biological Monitoring of Solar UV Radiation at 17 Sites in Asia, Europe and South America from 1999 to 2004

Abstract A small and robust dosimeter for determining the biologically effective dose of ambient UV radiation has been developed using UV-sensitive mutant spores of Bacillus subtilis strain TKJ6312. A membrane filter with four spots of the spores was snapped to a slide mount. The slide was wrapped and covered with two or more layers of polyethylene sheet to protect the sample from rain and snow and to reduce monthly-cumulative doses within the measurable range. From 1999, monthly data were collected at 17 sites for more than 1 year, and data for 4 to 6 consecutive years were obtained from 12 sites. Yearly total values of the spore inactivation dose (SID) ranged from 3200 at subarctic Oulu to 96 000 at tropical Denpasar, and the mean yearly values of SID exhibited an exponential dependence on latitude in both hemispheres with a doubling for about every 14 degrees of change. During the observation period, increasing trends of UV doses have been observed at all sites with more than 5 years of data available. Year-to-year variations at high and middle latitude sites are considered due mostly to climatic variation. At three tropical sites, negative correlations between the yearly doses and the column ozone amounts were observed. The results verified the applicability of spore dosimetry for global and long-time monitoring of solar UV radiation, in particular at tropical sites where no monitoring is taking place.

Polarotaxis in egg-laying yellow fever mosquitoes Aedes (Stegomyia) aegypti is masked due to infochemicals

Aquatic and water-associated insects need to locate suitable bodies of water to lay their eggs in and allow their aquatic larvae to develop. More than 300 species are known to solve this task by positive polarotaxis, relying primarily on the horizontally polarized light reflected from the water surface. The yellow fever mosquito Aedes (Stegomyia) aegypti has been thought to be an exception, locating its breeding habitats by chemical cues like odour of conspecifics, their eggs, or water vapour. We now demonstrate through dual-choice experiments that horizontally polarized light can also attract ovipositing Ae. aegypti females when the latter are deprived of chemical cues: water-filled transparent egg-trays illuminated by horizontally polarized light from below gained a 94.2% higher total number of eggs than trays exposed to unpolarized light, but only when no chemical substances capable of functioning as cues were present. Ae. aegypti is the first known water-associated insect in which polarotaxis exists, but does not play a dominant role in locating water bodies and can be constrained in the presence of chemical cues.

A structural study of the retinal photoreceptor, plexiform and ganglion cell layers following exposure to UV-B and UV-C radiation in the albino rat

Over the last two decades, ultraviolet radiation levels (UV), reaching the Earths surface, have been increasing at a rate of 1.5% per each 1% loss of the ozone layer. Moreover, artificial UV-sources have also proliferated and contributed to the rising UV-stress that many organisms have to face. To assess how the vertebrate retina responds to an exposure of short wavelength UV, we focused our attention on the rat retina, observing photoreceptor (containing outer and inner segments of rods and cones), inner plexiform, and ganglion cell layers by light and transmission electron microscopy using conventional and cytochemical techniques. We analyzed how cells of the layers in question responded to a 30 min exposure to UV-C and UV-B radiation with doses of 7200 and 590 J/cm(2), respectively. The results show that there are significant changes in the nuclei and cytoplasmic organelles of the exposed retinae when compared with those of the unexposed controls. The changes include an increase in heterochromatin, distension of rough endoplasmic reticulum, mitochondrial disruptions, and increases in the number of myelin bodies. The recorded morphological changes, especially those of the ganglion cells, are suggestive of apoptotic processes and show that the exposure of vertebrate retina to wavelengths ranging from 254 to 312 nm can produce alterations that are likely to impact negatively on the retinas proper functioning.

Imaging polarimetry of forest canopies: how the azimuth direction of the sun, occluded by vegetation, can be assessed from the polarization pattern of the sunlit foliage

Radiance, color, and polarization of the light in forests combine to create complex optical patterns. Earlier sporadic polarimetric studies in forests were limited by the narrow fields of view of the polarimeters used in such studies. Since polarization patterns in the entire upper hemisphere of the visual environment of forests could be important for forest-inhabiting animals that make use of linearly polarized light for orientation, we measured 180 degrees field-of-view polarization distributions in Finnish forests. From a hot air balloon we also measured the polarization patterns of Hungarian grasslands lit by the rising sun. We found that the pattern of the angle of polarization alpha of sunlit grasslands and sunlit tree canopies was qualitatively the same as that of the sky. We show here that contrary to an earlier assumption, the alpha-pattern characteristic of the sky always remains visible underneath overhead vegetation, independently of the solar elevation and the sky conditions (clear or partly cloudy with visible suns disc), provided the foliage is sunlit and not only when large patches of the clear sky are visible through the vegetation. Since the mirror symmetry axis of the alpha-pattern of the sunlit foliage is the solar-antisolar meridian, the azimuth direction of the sun, occluded by vegetation, can be assessed in forests from this polarization pattern. Possible consequences of this robust polarization feature of the optical environment in forests are briefly discussed with regard to polarization-based animal navigation.

Polarized light and oviposition site selection in the yellow fever mosquito: no evidence for positive polarotaxis in Aedes aegypti.

Aquatic insects and insects associated with water use horizontally polarized light (i.e., positive polarotaxis) to detect potential aquatic or moist oviposition sites. Mosquitoes lay their eggs onto wet substrata, in water, water-filled tree/rock holes, or man-made small containers/bottles/old tyres containing water. Until now it has remained unknown whether mosquitoes are polarotactic or not. The knowledge how mosquitoes locate water would be important to develop new control measures against them. Thus, we studied in dual-choice laboratory experiments the role of horizontally polarized light in the selection of oviposition sites in blood-fed, gravid females of the yellow fever mosquito, Aedes aegypti. On the basis of our results we propose that Ae. aegypti is not polarotactic. Thus the yellow fever mosquito is the first known water-associated insect species that does not detect water by means of the horizontally polarized water-reflected light. This can be explained by the reflection-polarization characteristics of small-volume water-filled cavities/containers preferred by Ae. aegypti as oviposition sites.

Understanding the Cephalic Eyes of Pulmonate Gastropods: A Review*

Abstract: This review showcases one group of gastropods ability to perceive light through the eyes. The central question is simple: what are the visual performances and tasks of cephalic eyes in gastropods? That topic in itself is rather broad and is here applied to pulmonate gastropods, coming from terrestrial and aquatic biomes as well as different habitats and microhabitats, exhibiting different life-styles and light-tolerances. Therefore, the main objectives have been to analyze (1) anatomical and ultrastructural eye characteristics, (2) optical systems, (3) image-forming capabilities and possible functional consequences of eye size and design, (4) interactions between gastropods and their environment mediated by the visual information obtained through the eye, and (5) the specific visual tasks that the eyes serve. During the course of this study, a range of variations (= adaptations) in both optical and retinal design parameters, including eye size, aperture size, quality of optical image, retinal shape, sampling density, and optical sensitivity were discovered. All species of pulmonate gastropods studied have paired simple camera-type eyes that operate with advanced fixed focal-length optics. However, in terrestrial snails and slugs as well as freshwater limpets, the optics cannot produce a focused image on their shallow retinas. This seems to indicate that eyes in these species are not designed to receive a focused image and are likely to measure only the average light intensity or quality over large angles rather than resolve fine image details. The aquatic snails examined are able to focus a sharp image on the photoreceptive layer of the retina due to the deepenings of the latter (at least in a localized region). Although there is a significant correlation between specialization of the eye (e.g., quality of optical image, sensitivity, and resolution) for a particular visual task in a specific habitat that the species encounters, there is no correlation between cellular composition of the retina and light/dark preferences. Their high optical sensitivity allows terrestrial snails to perform the necessary visual tasks in both bright and dim light, whereas the eye in aquatic species functions preferentially under bright light conditions. In conclusion, pulmonate gastropods use their eyes primarily for the following two kinds of visual tasks: (1) discriminating objects and possible enemies in their environment and (2) monitoring the environmental brightness level to orient towards dark places. The first type of visual task is characteristic of the aquatic snails and is served by image-forming eyes; the second is typical of terrestrial snails and slugs and is best served by a blurred image. Attention is given to visual ecological adaptations, specific visual needs, and the evolutionary history of gastropods.