Photophobic Response Research Articles

Comparative analysis of light-stimulated motility responses in three diatom species

Diatoms have long been known to be photosensitive, altering the direction of their movement in response to changes in ambient light conditions detected at the tips of the cells. In order to better understand the light conditions responsible for triggering positive photophobic (out-of-light) and negative photophobic (into-light) responses of diatoms, cells from three species of diatoms, Craticula cuspidata (Kützing) D.G. Mann, Stauroneis phoenicenteron (Nitzsch) Ehrenberg, and Pinnularia viridis (Nitzsch) Ehrenberg, were irradiated at their leading or trailing ends during cell movement. The response times for direction changes when cells were irradiated at various irradiance levels and wavelengths were measured to determine the quality of light responsible for eliciting cell direction changes in each of the three species. All three species displayed strong out-of-light responses at the highest irradiances measured. Craticula cuspidata cells displayed negative photophobic sensitivity (into-light) responses in moderate level blue light, while S. phoenicenteron cells showed into-light responses with low-level red light. Pinnularia viridis cells showed less responsiveness to blue and green light than the other two species, and almost no sensitivity to red light. By re-irradiating cells a second time after a previous leading or trailing end irradiation, we observed a 2–3 fold (leading) or 3–8 fold (trailing) motility repression, caused by the initial light exposures, which lasted for approximately 30–60 sec. Irradiating the cells multiple times, upon each direction change, indicated some degree of habituation to irradiation over time. Multiple consecutive irradiations of the trailing end of diatoms resulted in strong repression of any direction change, with cells continuing to move in the same direction for up to 20 min; this repression became reduced as the interval time between trailing-end irradiations increased. These results suggest that diatoms display species-specific physiological responses to light irradiations that may help them to appropriately respond to ambient light conditions, and better organize and succeed within larger algal or multi-species diatom assemblages.

Tryptophan hydroxylase Is Required for Eye Melanogenesis in the Planarian Schmidtea mediterranea.

Melanins are ubiquitous and biologically important pigments, yet the molecular mechanisms that regulate their synthesis and biochemical composition are not fully understood. Here we present a study that supports a role for serotonin in melanin synthesis in the planarian Schmidtea mediterranea. We characterize the tryptophan hydroxylase (tph) gene, which encodes the rate-limiting enzyme in serotonin synthesis, and demonstrate by RNA interference that tph is essential for melanin production in the pigment cups of the planarian photoreceptors. We exploit this phenotype to investigate the biological function of pigment cups using a quantitative light-avoidance behavioral assay. Planarians lacking eye pigment remain phototactic, indicating that eye pigmentation is not essential for light avoidance in S. mediterranea, though it improves the efficiency of the photophobic response. Finally, we show that the eye pigmentation defect observed in tph knockdown animals can be rescued by injection of either the product of TPH, 5-hydroxytryptophan (5-HTP), or serotonin. Together, these results highlight a role for serotonin in melanogenesis, perhaps as a regulatory signal or as a pigment substrate. To our knowledge, this is the first example of this relationship to be reported outside of mammalian systems.

光谱和光强度对棕榈蓟马雌成虫行为反应的影响

PDF HTML阅读 XML下载 导出引用 引用提醒 光谱和光强度对棕榈蓟马雌成虫行为反应的影响 DOI: 10.5846/stxb201308112058 作者: 作者单位: 山东省农业科学院植物保护研究所/山东省植物病毒学重点实验室,山东省农业科学院植物保护研究所/山东省植物病毒学重点实验室,山东省农业科学院植物保护研究所/山东省植物病毒学重点实验室,山东省农业科学院植物保护研究所/山东省植物病毒学重点实验室,山东省农业科学院植物保护研究所/山东省植物病毒学重点实验室,山东省农业科学院植物保护研究所/山东省植物病毒学重点实验室,山东省农业科学院植物保护研究所/山东省植物病毒学重点实验室,山东省农业科学院植物保护研究所/山东省植物病毒学重点实验室 作者简介: 通讯作者: 中图分类号: 基金项目: 国家"十二五"农村领域科技计划课题(2012BAD19B06);山东省科技发展计划项目(2012GNC11112) Effect of spectral sensitivity and intensity on the behavioral response of the Thrips palmi female adult Author: Affiliation: Institute of Plant Protection, Shangdong Academy of Agricultural Sciences,,,,,,, Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:室内研究了光谱、光强度对棕榈蓟马雌成虫的趋、避光行为的影响。结果显示:在340-605 nm 波谱内棕榈蓟马雌成虫对14个单色光刺激的趋光行为反应为多峰型。其中蓝光483 nm处峰最高,趋光反应率达34.96%,其次为绿光498-524 nm、562-582 nm、紫外光340 nm处;其避光行为反应共有3个峰,其中紫外光 380 nm 处最高,避光率18.08%,另外2个峰分别在橙光605 nm、紫光420 nm处。在趋光率较高的单色光(340、483、524、582 nm)和避光率较高的单色光(380、605 nm)以及白光刺激下,棕榈蓟马雌成虫的趋光率随光强增强的增强而提高,而避光率则随着光强的增强而降低;光强最弱时仍均有一定趋光率,最强时均未出现高端平台。因此:棕榈蓟马雌成虫对不同单色光具有明显的选择性,光谱和光强度对其趋光行为和避光行为有较大影响,光强度的影响作用与波长因素有关。 Abstract:Thrips palmi is one of the most serious insect pests, causing adverse effects on vegetable production. In China, the insect pest was first discovered in Guangdong Province in the 1970s and has gradually become a major insect pest of vegetables, especially solar greenhouse vegetables. For example, in 1993 solanaceous fruit vegetables such as eggplant, green pepper and potato were damaged seriously by T. palmi, and the damage rate was about 90% in the Hangzhou district. In recent years, with the development of solar greenhouse vegetables in northern China, T. palmi has survived through the winter in solar greenhouses and damaged vegetables throughout the year, so controlling these insect pests has become more difficult. Physical control is an important control technique within integrated pest management (IPM), and the color plate trap is one of the most important methods of physical control. The color plate trap can not only reduce the number of T. palmi insects effectively, but can also forecast the occurrence of the insect pests in the field, so this method plays an important role in the IPM of T. palmi. The selectivity of T. palmi to colors in the field has been reported to show significant variation. To discover the photosensitivity of the insect pest, we studied the phototactic and photophobic behavior of T. palmi female adults in response to spectral sensitivity and light intensity by behavior method in the laboratory. The results showed that the phototactic response of T. palmi female adults to monochromatic light at 14 different wavelengths within a 340-605 nm range occurred in a curve with multiple peaks. The primary peak, representing the highest phototactic response (34.96%), occurred at a wavelength of 483 nm (blue), the secondary peak at 498-524 nm (green), and the remainder at 562-582 nm (green) and 340 nm (ultra-violet). The photophobic response occurred in a curve with three peaks: one peak at 380 nm (ultra-violet) represented the highest photophobic response rate (18.08%), and the other two peaks at 605 nm (orange) and 420 nm (violet). Under the stimulus of monochromatic light-at which the T. palmi showed high phototactic response rates (340 nm, 483 nm, 524 nm, 582 nm, 380 nm, 605 nm) and high photophobic response rates (380 nm, 605 nm)-and white light, the phototactic response rate of T. palmi female adults increased with increasing light intensity, while the photophobic response rate decreased. The response rate of T. palmi female adults could be detected at the lowest intensity but not at the height of the strongest intensity. Therefore, in T. palmi female adults, both the spectrum and intensity of light stimulus resulted in phototactic and photophobic responses under monochromatic light conditions, and the level of response was proportional to the wavelength. The results obtained provided a theoretical basis for further investigation and applications as well as for the evaluation of scientific light traps used to monitor and control T. palmi. 参考文献 相似文献 引证文献

Clustering and dynamics of phototransducer signaling domains revealed by site-directed spin labeling electron paramagnetic resonance on SRII/HtrII in membranes and nanodiscs.

In halophilic archaea the photophobic response is mediated by the membrane-embedded 2:2 photoreceptor/-transducer complex SRII/HtrII, the latter being homologous to the bacterial chemoreceptors. Both systems bias the rotation direction of the flagellar motor via a two-component system coupled to an extended cytoplasmic signaling domain formed by a four helical antiparallel coiled-coil structure. For signal propagation by the HAMP domains connecting the transmembrane and cytoplasmic domains, it was suggested that a two-state thermodynamic equilibrium found for the first HAMP domain in NpSRII/NpHtrII is shifted upon activation, yet signal propagation along the coiled-coil transducer remains largely elusive, including the activation mechanism of the coupled kinase CheA. We investigated the dynamic and structural properties of the cytoplasmic tip domain of NpHtrII in terms of signal transduction and putative oligomerization using site-directed spin labeling electron paramagnetic resonance spectroscopy. We show that the cytoplasmic tip domain of NpHtrII is engaged in a two-state equilibrium between a dynamic and a compact conformation like what was found for the first HAMP domain, thus strengthening the assumption that dynamics are the language of signal transfer. Interspin distance measurements in membranes and on isolated 2:2 photoreceptor/transducer complexes in nanolipoprotein particles provide evidence that archaeal photoreceptor/-transducer complexes analogous to chemoreceptors form trimers-of-dimers or higher-order assemblies even in the absence of the cytoplasmic components CheA and CheW, underlining conservation of the overall mechanistic principles underlying archaeal phototaxis and bacterial chemotaxis systems. Furthermore, our results revealed a significant influence of the NpHtrII signaling domain on the NpSRII photocycle kinetics, providing evidence for a conformational coupling of SRII and HtrII in these complexes.

Planarian Phototactic Assay Reveals Differential Behavioral Responses Based on Wavelength.

Planarians are free-living aquatic flatworms that possess a well-documented photophobic response to light. With a true central nervous system and simple cerebral eyes (ocelli), planarians are an emerging model for regenerative eye research. However, comparatively little is known about the physiology of their photoreception or how their behavior is affected by various wavelengths. Most phototactic studies have examined planarian behavior using white light. Here, we describe a novel planarian behavioral assay to test responses to small ranges of visible wavelengths (red, blue, green), as well as ultraviolet (UV) and infrared (IR) which have not previously been examined. Our data show that planarians display behavioral responses across a range of wavelengths. These responses occur in a hierarchy, with the shortest wavelengths (UV) causing the most intense photophobic responses while longer wavelengths produce no effect (red) or an apparent attraction (IR). In addition, our data reveals that planarian photophobia is comprised of both a general photophobic response (that drives planarians to escape the light source regardless of wavelength) and wavelength-specific responses that encompass specific behavioral reactions to individual wavelengths. Our results serve to improve the understanding of planarian phototaxis and suggest that behavioral studies performed with white light mask a complex behavioral interaction with the environment.

Transient freezing behavior in photophobic responses of Euglena gracilis investigated in a microfluidic device.

We found that the transient freezing behavior in photophobic responses of Euglena gracilis is a good indicator of the metabolic status of the cells. The transient blue light photophobic responses of E. gracilis cells were investigated on-chip using a new measurement, 'trace momentum' (TM), to evaluate their swimming activity quantitatively in real time. When blue light of intensity >30 mW cm(-2) was repeatedly switched on and off, a large negative spike in the TM was observed at the onset of the 'blue-light-off' phase. Single-cell trace analysis at a blue light intensity of 40 mW cm(-2) showed that 48% (on average, n = 15) of tumbling Euglena cells ceased activity ('freezing') for 2-30 s at the onset of blue-light-off before commencing forward motion in a straight line (termed 'straightforward swimming'), while 45% smoothly commenced straightforward swimming without delay. The proportion of freezing Euglena cells depended on the blue light intensity (only 20% at 20 mW cm(-2)). When the cells were stimulated by four blue light pulses at the higher intensity, without pre-exposure, the transient freezing behavior was more prominent but, on repeating the stimuli after an 80 min interval in red light, the same cells did not freeze. This shows that the metabolism of the cells had changed to anti-freezing during the interval. The relationship between the interval time with/without light irradiation and the blue light adaptation was elucidated experimentally. The origin of the freezing behavior is considered to be a shortage of a metabolic substance that promotes smooth switching of flagellum movement from in situ rotation mode to a straightforward swimming mode.

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 flavo-proteins 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 at 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.

Site-directed spin labeling EPR spectroscopy in protein research

Site-directed spin labeling (SDSL) in combination with electron paramagnetic resonance (EPR) spectroscopy has emerged as an efficient tool to elucidate the structure and the conformational dynamics of proteins under conditions close to the native state. This review article summarizes the basics as well as the recent progress in SDSL and EPR methods, especially for investigations on protein structure, protein function, and interaction of proteins with other proteins or nucleic acids. Labeling techniques as well as EPR methods are introduced and exemplified with applications to systems that have been studied in the author's laboratory in the past 15 years, headmost the sensory rhodopsin-transducer complex mediating the photophobic response of the halophilic archaeum Natronomonas pharaonis. Further examples underline the application of SDSL EPR spectroscopy to answer specific questions about the system under investigation, such as the nature and influence of interactions of proteins with other proteins or nucleic acids. Finally, it is discussed how SDSL EPR can be combined with other biophysical techniques to combine the strengths of the different methodologies.

A Genetic Dissection of the Photophobic Response of Paramecium tetraurelia

Paramecium tetraurelia displayed two behavioral responses upon the initiation of a light stimulus at 7 x 10(4) lux. The cells exhibited a photophobic response in the form of behavioral avoiding reactions, followed by an increase in forward swimming velocity that was significantly higher than prior to the light stimulus activation. It was determined that an intensity of approximately 6.5 x 10(3) lux was required to initiate a moderate avoidance behavioral response. Following the avoiding response, a gradual increase in speed occurred as the intensity increased, indicating that increased swimming speeds are dependent on the light intensity. Two mutants, pawnA and Dancer, were utilized since they affect known Ca(2+)-currents of the cell. The use of pawnA cells, which lack voltage-dependent Ca(2+) channel activity, showed that the two responses to light could be genetically separated, in that the cells showed no avoiding reactions, but did increase their swimming speed. The Dancer cells, which display exaggerated Ca(2+) channel activity, exhibited similar initial avoiding responses as the wild type cells, however did not increase their swimming speed as the intensity of the light was increased. This phenotype as replicated in wildtype cells that had been placed in 25 μM 8-Br-cGMP. These data demonstrate that the photophobic light response of Paramecium tetraurelia can be genetically dissected as a means of elucidating the molecular mechanisms of the light response.

Encephalic photoreception and phototactic response in the troglobiont Somalian blind cavefish Phreatichthys andruzzii.

Many physiological and behavioural responses to changes in environmental lighting conditions are mediated by extraocular photoreceptors. Here we investigate encephalic photoreception in Phreatichthys andruzzii, a typical cave-dwelling fish showing an extreme phenotype with complete anophthalmy and a reduction in size of associated brain structures. We firstly identified two P. andruzzii photopigments, orthologues of rod opsin and exo-rod opsin. In vitro, both opsins serve as light-absorbing photopigments with λ(max) around 500 nm when reconstituted with an A(1) chromophore. When corrected for the summed absorption from the skin and skull, the spectral sensitivity profiles shifted to longer wavelengths (rod opsin: 521 nm; exo-rod opsin: 520 nm). We next explored the involvement of both opsins in the negative phototaxis reported for this species. A comparison of the spectral sensitivity of the photophobic response with the putative A(2) absorbance spectra corrected for skin/skull absorbance indicates that the A(2) versions of either or both of these pigments could explain the observed behavioural spectral sensitivity.

CALCIUM RELEASE FROM INTRACELLULAR STORES IS NECESSARY FOR THE PHOTOPHOBIC RESPONSE IN THE BENTHIC DIATOM NAVICULA PERMINUTA (BACILLARIOPHYCEAE)(1).

Complex photoreceptor pathways exist in algae to exploit light as a sensory stimulus. Previous studies have implicated calcium in blue-light signaling in plants and algae. A photophobic response to high-intensity blue light was characterized in the marine benthic diatom Navicula perminuta (Grunow) in van Heurck. Calcium modulators were used to determine the involvement of calcium in the signaling of this response, and the fluorescent calcium indicator Calcium Crimson was used to image changes in intracellular [Ca(2+) ] during a response. A localized, transient elevation of Calcium Crimson fluorescence was seen at the cell tip at the time of cell reversal. Intracellular calcium release inhibitors produced a significant decrease in the population photophobic response. Treatments known to decrease influx of extracellular calcium had no effect on the population photophobic response but did cause a significant decrease in average cell speed. As the increase in intracellular [Ca(2+) ] at the cell tip corresponded to the time of direction change rather than the onset of the light stimulus, it would appear that Ca(2+) constitutes a component of the switching mechanism that leads to reversal of the locomotion machinery. Our current evidence suggests that the source of this Ca(2+) is intracellular.

Comprehensive Determination of Protein Tyrosine pKa Values for Photoactive Yellow Protein Using Indirect 13C NMR Spectroscopy

Upon blue-light irradiation, the bacterium Halorhodospira halophila is able to modulate the activity of its flagellar motor and thereby evade potentially harmful UV radiation. The 14 kDa soluble cytosolic photoactive yellow protein (PYP) is believed to be the primary mediator of this photophobic response, and yields a UV/Vis absorption spectrum that closely matches the bacterium's motility spectrum. In the electronic ground state, the para-coumaric acid (pCA) chromophore of PYP is negatively charged and forms two short hydrogen bonds to the side chains of Glu-46 and Tyr-42. The resulting acid triad is central to the marked pH dependence of the optical-absorption relaxation kinetics of PYP. Here, we describe an NMR approach to sequence-specifically follow all tyrosine side-chain protonation states in PYP from pH 3.41 to 11.24. The indirect observation of the nonprotonated 13Cγ resonances in sensitive and well-resolved two-dimensional 13C-1H spectra proved to be pivotal in this effort, as observation of other ring-system resonances was hampered by spectral congestion and line-broadening due to ring flips. We observe three classes of tyrosine residues in PYP that exhibit very different pKa values depending on whether the phenolic side chain is solvent-exposed, buried, or hydrogen-bonded. In particular, our data show that Tyr-42 remains fully protonated in the pH range of 3.41–11.24, and that pH-induced changes observed in the photocycle kinetics of PYP cannot be caused by changes in the charge state of Tyr-42. It is therefore very unlikely that the pCA chromophore undergoes changes in its electrostatic interactions in the electronic ground state.

Modeling Filamentous Cyanobacteria Reveals the Advantages of Long and Fast Trichomes for Optimizing Light Exposure

Cyanobacteria form a very large and diverse phylum of prokaryotes that perform oxygenic photosynthesis. Many species of cyanobacteria live colonially in long trichomes of hundreds to thousands of cells. Of the filamentous species, many are also motile, gliding along their long axis, and display photomovement, by which a trichome modulates its gliding according to the incident light. The latter has been found to play an important role in guiding the trichomes to optimal lighting conditions, which can either inhibit the cells if the incident light is too weak, or damage the cells if too strong. We have developed a computational model for gliding filamentous photophobic cyanobacteria that allows us to perform simulations on the scale of a Petri dish using over 105 individual trichomes. Using the model, we quantify the effectiveness of one commonly observed photomovement strategy—photophobic responses—in distributing large populations of trichomes optimally over a light field. The model predicts that the typical observed length and gliding speeds of filamentous cyanobacteria are optimal for the photophobic strategy. Therefore, our results suggest that not just photomovement but also the trichome shape itself improves the ability of the cyanobacteria to optimize their light exposure.

New Channelrhodopsin with a Red-Shifted Spectrum and Rapid Kinetics from Mesostigma viride

ABSTRACTLight control of motility behavior (phototaxis and photophobic responses) in green flagellate algae is mediated by sensory rhodopsins homologous to phototaxis receptors and light-driven ion transporters in prokaryotic organisms. In the phototaxis process, excitation of the algal sensory rhodopsins leads to generation of transmembrane photoreceptor currents. When expressed in animal cells, the algal phototaxis receptors function as light-gated cation channels, which has earned them the name “channelrhodopsins.” Channelrhodopsins have become useful molecular tools for light control of cellular activity. Only four channelrhodopsins, identified in Chlamydomonas reinhardtii and Volvox carteri, have been reported so far. By screening light-induced currents among algal species, we identified that the phylogenetically distant flagellate Mesostigma viride showed photoelectrical responses in vivo with properties suggesting a channelrhodopsin especially promising for optogenetic use. We cloned an M. viride channelrhodopsin, MChR1, and studied its channel activity upon heterologous expression. Action spectra in HEK293 cells match those of the photocurrents observed in M. viride cells. Comparison of the more divergent MChR1 sequence to the previously studied phylogenetically clustered homologs and study of several MChR1 mutants refine our understanding of the sequence determinants of channelrhodopsin function. We found that MChR1 has the most red-shifted and pH-independent spectral sensitivity so far reported, matches or surpasses known channelrhodopsins’ channel kinetics features, and undergoes minimal inactivation upon sustained illumination. This combination of properties makes MChR1 a promising candidate for optogenetic applications.

Picosecond transient circular dichroism of the photoreceptor protein of the light-adapted form of Blepharisma japonicum

We present a picosecond transient circular dichroism study of OBIP, the putative photoreceptor protein involved in the photophobic response of Blepharisma japonicum. The probe wavelength was chosen at 230 nm. The results are compared to those of the isolated chromophore, OxyBP, in solution. The CD changes in OBIP and OxyBP do not show the same dynamics: OBIP’s signal relaxes in a few ps whereas no such decay is obtained for OxyBP. This observation brings support to the formerly evoked existence of a fast photoinduced reaction in the chromoprotein, and demonstrates the implication of local geometrical changes that accompany this process.

Spin labeling EPR

Site-directed spin labeling in combination with electron paramagnetic resonance spectroscopy has emerged as an efficient tool to elucidate the structure and conformational dynamics of biomolecules under native-like conditions. This article summarizes the basics as well as recent progress of site-directed spin labeling. Continuous wave EPR spectra analyses and pulse EPR techniques are reviewed with special emphasis on applications to the sensory rhodopsin-transducer complex mediating the photophobic response of the halophilic archaeum Natronomonas pharaonis and the photosynthetic reaction center from Rhodobacter sphaeroides R26.

Single-Molecule Force Spectroscopy Measures Structural Changes Induced by Light Activation and Transducer Binding in Sensory Rhodopsin II

Microbial rhodopsins are a family of seven-helical transmembrane proteins containing retinal as chromophore. Sensory rhodopsin II (SRII) triggers two very different responses upon light excitation, depending on the presence or the absence of its cognate transducer HtrII: Whereas light activation of the NpSRII/ NpHtrII complex activates a signalling cascade that initiates the photophobic response, NpSRII alone acts as a proton pump. Using single-molecule force spectroscopy, we analysed the stability of NpSRII and its complex with the transducer in the dark and under illumination. By improving force spectroscopic data analysis, we were able to reveal the localisation of occurring forces within the protein chain with a resolution of about six amino acids. Distinct regions in helices G and F were affected differently, depending on the experimental conditions. The results are generally in line with previous data on the molecular stability of NpSRII. Interestingly, new interaction sites were identified upon light activation, whose functional importance is discussed in detail.

Light‐induced and apoptosis‐like cell death in the unicellular eukaryote, Blepharisma japonicum

The unicellular eukaryote, Blepharisma japonicum, is a light-sensitive ciliated protozoa. It possesses a photoreceptor pigment called blepharismin that plays critical roles in defensive behavior against predators and step-up photophobic response. In addition, the pigment generates reactive oxygen species such as singlet oxygen and hydroxyl radicals which contribute to photodynamic action. Previous studies reported that intense light (>300W m(-2)) induced rapid photodynamic killing (necrosis) characterized by cell swelling and plasma efflux, while moderate light (3-30W m(-2)) only induced pigment extrusion and photooxidation. We have found that moderate light (5W m(-2)) induced apoptosis-like cell death. Microscopically it was found that >3h of moderate light irradiation induced macronuclear condensation and plasma efflux without cell swelling. Single cell gel electrophoresis assay showed that DNA fragmentation occurred between 1 and 3h of irradiation, and the condensed macronuclei contained quite fragmented DNA. Macronuclear DNA extracted from light-irradiated cells contained DNA fragments of 180-200 and 360-400bp, which were seen as apoptosis ladders.

Role of serotonergic neurons in the Drosophila larval response to light

BackgroundDrosophila larval locomotion consists of forward peristalsis interrupted by episodes of pausing, turning and exploratory behavior (head swinging). This behavior can be regulated by visual input as seen by light-induced increase in pausing, head swinging and direction change as well as reduction of linear speed that characterizes the larval photophobic response. During 3rd instar stage, Drosophila larvae gradually cease to be repelled by light and are photoneutral by the time they wander in search for a place to undergo metamorphosis. Thus, Drosophila larval photobehavior can be used to study control of locomotion.ResultsWe used targeted neuronal silencing to assess the role of candidate neurons in the regulation of larval photobehavior. Inactivation of DOPA decarboxylase (Ddc) neurons increases the response to light throughout larval development, including during the later stages of the 3rd instar characterized by photoneutral response. Increased response to light is characterized by increase in light-induced direction change and associated pause, and reduction of linear movement. Amongst Ddc neurons, suppression of the activity of corazonergic and serotonergic but not dopaminergic neurons increases the photophobic response observed during 3rd instar stage. Silencing of serotonergic neurons does not disrupt larval locomotion or the response to mechanical stimuli. Reduced serotonin (5-hydroxytryptamine, 5-HT) signaling within serotonergic neurons recapitulates the results obtained with targeted neuronal silencing. Ablation of serotonergic cells in the ventral nerve cord (VNC) does not affect the larval response to light. Similarly, disruption of serotonergic projections that contact the photoreceptor termini in the brain hemispheres does not impact the larval response to light. Finally, pan-neural over-expression of 5-HT1ADro receptors, but not of any other 5-HT receptor subtype, causes a significant decrease in the response to light of 3rd instar larvae.ConclusionOur data demonstrate that activity of serotonergic and corazonergic neurons contribute to the control of larval locomotion by light. We conclude that this control is carried out by 5-HT neurons located in the brain hemispheres, but does not appear to occur at the photoreceptor level and may be mediated by 5-HT1ADro receptors. These findings provide new insights into the function of 5-HT neurons in Drosophila larval behavior as well as into the mechanisms underlying regulation of larval response to light.

Excited states of photobiological receptors. II. Chlorophylls, phytochrome, and stentorin

In this series on the application of configuration analysis (CA) to the electronic excited states of photobiological receptors, we have extended CA to two important photobiological receptors, phytochrome of photomorphogenic and biological clock systems and stentorin of the photophobic response system of stentors. In addition, the effect of substituents in chlorophylls on the electronic excitation of the chlorin and bacteriochlorin ring systems has been examined. The open-shell calculations of chlorophylls have also been performed. In the singlet excited state of phytochrome, there is a significant charge transfer from ring A to the violin moiety of the phytochrome molecule, with a tendency for the localized excitation in the A ring. The charge transfer contribution to the singlet-singlet excitation is also significant. These results are correlated with the photomorphogenic transformation of phytochrome which involves deprotonation and/or tautomerism of the ring A in the excited state. In the case of stentorin, the excitation resides largely in the aromatic system, with some charge transfer to the carbonyl groups. The photobiological significance of these results are discussed.