Scanning motion,ocellar morphology and orientation mechanisms in the phototaxis of the nematode Mermis nigrescens

Summary The putative ocellus of Mermis females consists of a hollow cylinder of dense hemoglobin pigmentation located in the anterior tip. The exact location of the photoreceptive nerve endings, however, is unknown. During phototaxis a continual bending or scanning motion of the head (anterior 2 mm) causes the orientation of the tip to swing about the direction of the source. By turning off (shuttering) the light source whenever the tip orientation was to one side of the source direction, the average orientation of the base of the head, and eventually the body orientation, was caused to be biased about 28° to the opposite side. Because the shuttering was synchronized with the scanning motion, the scanning motion must be involved in the maintenance of orientation to light. The direction of the bias rules out a two-signal comparison mechanism of orientation and demonstrates that a deviation of the tip from the source direction must decrease, rather than increase, the illumination of the photoreceptors. These findings, and the ocellar morphology, require that the photoreceptors be located inside the hollow tube of pigmentation where they can be shadowed by the pigment during deviations of the tip. Focusing by the curved anterior end should cause a similar modulation of the illumination at this location. The occasional episodes of transverse phototaxis can be explained by the leakiness of the pigment walls to transverse illumination. Analysis of the motion of the anterior in the presence and absence of shuttering indicates that the orientation of the base of the head, due to the motion of the neck, is controlled by the signals generated during one or more cycles of the scanning motion of the head. The orientation may be regulated by the phase relationship between the photoreceptor signal and putative proprioceptive signals that indicate the bending in the head.     

Photomovement in Dunaliella salina: Fluence rate-response curves and action spectra

We determined the action spectra of the photophobic responses as well as the phototactic response in Dunaliella salina (Volvocales) using both single cells and populations. The action spectra of the photophobic responses have maxima at 510 nm, the spectrum for phototaxis has a maximum at 450–460 nm. These action spectra are not compatible with the hypothesis that flavoproteins are the photoreceptor pigments, and we suggest that carotenoproteins or rhodopsins act as the photoreceptor pigments. We also conclude that the phototactic response in Dunaliella is an elementary response, quite independent of the step-up and step-down photophobic responses. We also determined the action spectra of the photoaccumulation response in populations of cells adapted to two different salt conditions. Both action spectra have a peak a 490 nm. The photoaccumulation response may be a complex response composed of the phototactic and photophobic responses. Blue or blue-green light does not elicit a photokinetic response in Dunaliella.Diagrams of the optical set-ups used for measuring the responses at the single-cell level and of the plans for building the phototaxometer described in this paper are available to the interested readerWe thank Mr. M. Kubota for a tremendous amount of technical assistance and Mr. R. Nagy for building the phototaxometer. We thank T. Kondo, Professor H. Imaseki and the members of the Laboratory of Biological Regulation, NIBB, for their help and support in various aspects of this research. This research was supported, in part, from grants from the Okazaki Large Spectrograph (Project Nos. 86-535, 87-518, 88-523), the Japanese Society for the Promotion of Science, and the College of Agriculture and Life Sciences at Cornell University to R. W.     

Effects of UV-B on motility and photobehavior in the green flagellate,Euglena gracilis

UV-B inhibits the motility of the green flagellate, Euglena gracilis, at fluences rates higher than those expected to occur in the natural sunlight even when the stratospheric ozone layer is partially reduced by manmade pollutants. The phototactic orientation of the cells, however, is drastically impaired by only slightly enhanced levels of UV-B irradiation. Since only negative phototaxis (movement away from a strong light source) is impaired while positive phototaxis (movement toward a weak light source) is not, the delicate balance by which the organisms adjust their position in their habitat is disturbed. Under these conditions the cells are unable to retreat from hazardous levels of radiation and are eventually killed not by the UV-B irradiation but by photobleaching of their photosynthetic pigments in the strong daylight at the surface.     

Photomovement in Cyanophora paradoxa

Photomovement has been studied in the symbiontic association of the colorless flagellate, Cyanophora paradoxa Korschikoff with the cyanelles, Cyanocyta korschikoffiana. There is no phototactic orientation in this organism, but a photokinetic effect. In addition, the cells show a pronounced step-up photophobic response (however no or only a weak step-down response). The phobic response is mediated by a subset of the photosynthetic pigments located in the symbiontic cyanelles. It is linked to the noncyclic photosynthetic electron transport chain but it is independent of the photosynthetic generation of a proton gradient and the ATP synthesis linked to it.Abbreviations CCCP carbonyl cyanide m-chlorophenyl hydrazone - DBMIB 2,5-dibromo-3-methyl-6-isopropylbenzo quinone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea     

Carotenoids in the eyespot apparatus of the flagellate green alga Spermatozopsis similis: Adaptation to the retinal-based photoreceptor

Isolated intact eyespot apparatuses, the photoreceptive organelles involved in blue-light-mediated photoresponses of flagellate green algae, were analyzed regarding their carotenoid composition. Carotenoids from the eyespot apparatuses of Spermatozopsis similis were identified by high-performance liquid chromatography, visible-light absorption spectra, mass spectroscopy and by ¹H-nuclear magnetic resonance spectroscopy (carotenes), and compared with those of whole-cell extracts. Both extracts contained ,-carotene, ,-carotene (formerly -carotene), lycopene, lutein, zeaxanthin, violaxanthin and all-E-and 9-Z-neoxanthin. The relative carotenoid compositions, however, differed significantly. A twofold relative increase in the total carotene level was evident in the fraction enriched in eyespot apparatuses. This was mainly due to an increase in the monocyclic ,-carotene and the aliphatic lycopene, whereas the relative content of ,-carotene remained unchanged. On the other hand a relative decrease in the total xanthophyll content, especially of lutein and the epoxidic carotenoid neoxanthin, was observed in the eyespot apparatuses compared with the whole-cell extracts. The decrease of the latter resulted almost solely from a reduction of the 9-Z-rather than the all-E-isomer. The bulk of the carotenes is thought to be localized in the highly organized eyespot lipid globules, which act as a combined quarter-wave interference reflector and absorption screen for the photoreceptor in green algae. The enrichment of ,-carotene and lycopene in the eyespot apparatuses, extending the range of visible light absorption to longer wavelengths, represents an adaptation of the screen to the retinal-based photoreceptor of flagellate green algae and is one of the prerequisites for maximal directional sensitivity of the eyespot apparatus.Abbreviations ¹H-NMR nuclear magnetic resonance - IUPAC International Union of Pure and Applied Chemistry - VIS visible absorption spectra This work was supported by the Deutsche Forschungsgemeinschaft (G.K. and M.M.). M.G. was supported by a fellowship from the Norwegian Research Council of Science and Humanities.     

Effects of inhibitors on photomovement in desmids

The effects of the inhibitors of the photosynthetic electron transport chain, 3-(3,4-dichlorophenyl) 1,1-dimethylurea (DCMU) and 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB), on the three phtoomovement responses known in the desmid Cosmarium cucumis have been studied. Both inhibitors block photokinesis very effectively in their respective specific concentration range. Most of the impairment of phototaxis and the photophobic response observed in population techniques seems to be due to a reduced motility of the cells, since microvideographic analysis of the cell movement indicated that the inhibitors do not affect the phobic response at all and that there is only partial inhibition of phototaxis. Both the fraction of motile cells and the duration of motility periods are affected by the inhibitors. The results demonstrate that, though all three photoresponses are mediated by chlorophyll acting as photoreceptor, at least the phobic response is independent of the photosynthetic electron transport chain.Abbreviations DCMU 3-(3,4 dichlorophenyl)-1,1-dimethylurea - DBMIB 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone     

Role of three basic light reactions in photomovement of desmids

Photoaccumulations in light trap experiments have been studied in the desmids, Cosmarium, Micrasterias and Euastrum. Dependence of accumulation density on exposure time follows saturation curves, while dose response curves show optima. Time-lapse microcinematography and population methods have revealed that all three basic light-induced motor responses known in microorganisms participate in producing photoaccumulations in desmids. During the initial phase the cells are phototactically attracted towards the trap by scattered light. In low light intensity traps photokinetic reactions may play only a minor role, since photokinesis could be evoked only by light intensities100 lx in Cosmarium cucumis. True photophobic reactions have been demonstrated for the first time in desmids. There are two types of phobic responses in desmids: either the cell reverses its movement or it swings sidewise into the new direction. Behaviour of partially shadowed cells suggests that perception of light direction is brough about by simultaneous intensity measurement at two or more sites within the cell.     

Sensory rhodopsin I: Receptor activation and signal relay

Recent progress is summarized on the mechanism of phototransduction by sensory rhodopsin I (SR-I), a phototaxis receptor inHalobacterium halobium. Two aspects are emphasized: (i)The coupling of retinal isomerization to protein conformational changes. Retinal analogs have been used to probe chromophore-apoprotein interactions during the receptor activation process. One of the most important results is the finding of a steric trigger deriving from the interaction of residues on the protein with a methyl group near the isomerizing bond of the retinal (at carbon 13). Recent work on molecular genetic methods to further probe structure/function includes the synthesis and expression of an SR-I apoprotein gene designed for residue replacements by cassette mutagenesis, and transformation of anH. halobium mutant lacking all retinylidene proteins known in this species to SR-I⁺ and bacteriorhodopsin (BR)⁺. (ii)The relay of the SR-I signal to a post-receptor component. A carboxylmethylated protein (MPP-I) associated with SR-I and found in theH. halobium membrane exhibits homology with the signaling domain of eubacterial chemotaxis transducers (e.g.,Escherichia coli Tar, Tsr, and Trg proteins), suggesting a model based on SR-I MPP-I signal relay.     

Photoresponses of late instar <Emphasis Type="Italic">Chaoborus punctipennis</Emphasis> larvae to UVR

With a few clear exceptions (e.g., Daphnia) it is uncertain if most aquatic invertebrates can detect and respond to ultraviolet radiation (UVR). It is known that many aquatic invertebrates are vulnerable to UVR and that anthropogenically-induced increases in surface UVR have occurred in recent decades. We examined the photoresponses of late larval instars of Chaoborus punctipennis to different combinations of UVA (320–400 nm), UVB (300–320 nm) and visible light (400–700 nm) to determine whether the larvae can detect and/or avoid UVR. To accomplish this, we exposed late instar C. punctipennis larvae to a directional light source of UVR only (peak wavelength at 360 nm), visible light only or visible plus various wavebands of UVR. We examined negative phototaxis for 10 min at a quantum flux of 2.62 x 10¹³ quanta s^–1 cm^–2 (S.D. = 3.63 x 10¹² quanta s^–1 cm^–2). In the dark, larvae stayed close to the surface of the experimental vessels. Under all treatments containing visible light the larvae exhibited negative phototaxis and occupied the bottom of the vessels. Under UVR only, the larvae occupied the middle of the water column. Our results suggest that late instar C. punctipennis larvae are unable to detect and avoid UVB and short UVA wavelengths but they can detect long UVA wavelengths.     

Survival and behavioral characteristics of amphidromous goby larvae of Sicyopterus japonicus (Tanaka, 1909) during their downstream migration

To understand the ecology and environmental tolerances of newly hatched larvae of the amphidromous fish Sicyopterus japonicus during their downstream migration, the salinity tolerance of eggs, 0-15 day old larvae, and adults, and the temperature tolerance, specific gravity and phototaxis of hatched larvae were examined. Tolerances of adults were measured as survival after a 24 h challenge in freshwater (FW), brackish water (1/3 SW) and seawater (SW). The survival rate of adult S. japonicus was 100% in FW and 1/3 SW, while none survived in SW. Hatching success of eggs (30 eggs each) was significantly higher in FW (mean: 73%) and 1/3 SW (73%) than in SW (19%). Tolerance of newly hatched larvae to salinity and temperature was investigated in different combinations of salinities (FW, 1/3 SW and SW) and temperatures (18, 23 and 28 °C). Larval survival was significantly different in each salinity and temperature. Survival rate was significantly higher in 1/3 SW than in FW and higher in SW than in FW at 23 °C and 28 °C. At the latter part of the experiment, there was no survival in FW and at 28 °C. Survival was higher in lower temperatures, but larval development did not occur in FW. Specific gravity of newly hatched larvae was 1.036 at 28 °C and 1.034 at 23 °C. When exposed to a light source on one side of an aquarium, larval distribution was not affected. Our results indicated larval S. japonicus are more adapted to brackish water and seawater than freshwater, while the adults and eggs are more adapted to freshwater and brackish water than seawater. This is consistent with their amphidromous life history with growth and spawning occurring in freshwater and the larval stage utilizing marine habitats.