RESUMEN
Vision enables both image-forming perception, driven by a contrast-based pathway, and unconscious non-image-forming circadian photoentrainment, driven by an irradiance-based pathway1,2. Although two distinct photoreceptor populations are specialized for each visual task3-6, image-forming photoreceptors can additionally contribute to photoentrainment of the circadian clock in different species7-15. However, it is unknown how the image-forming photoreceptor pathway can functionally implement the segregation of irradiance signals required for circadian photoentrainment from contrast signals required for image perception. Here we report that the Drosophila R8 photoreceptor separates image-forming and irradiance signals by co-transmitting two neurotransmitters, histamine and acetylcholine. This segregation is further established postsynaptically by histamine-receptor-expressing unicolumnar retinotopic neurons and acetylcholine-receptor-expressing multicolumnar integration neurons. The acetylcholine transmission from R8 photoreceptors is sustained by an autocrine negative feedback of the cotransmitted histamine during the light phase of light-dark cycles. At the behavioural level, elimination of histamine and acetylcholine transmission impairs R8-driven motion detection and circadian photoentrainment, respectively. Thus, a single type of photoreceptor can achieve the dichotomy of visual perception and circadian photoentrainment as early as the first visual synapses, revealing a simple yet robust mechanism to segregate and translate distinct sensory features into different animal behaviours.
Asunto(s)
Ritmo Circadiano , Drosophila melanogaster , Células Fotorreceptoras de Invertebrados , Percepción Visual , Animales , Acetilcolina/metabolismo , Relojes Biológicos/fisiología , Relojes Biológicos/efectos de la radiación , Ritmo Circadiano/fisiología , Ritmo Circadiano/efectos de la radiación , Drosophila melanogaster/citología , Drosophila melanogaster/fisiología , Drosophila melanogaster/efectos de la radiación , Retroalimentación Fisiológica , Histamina/metabolismo , Neurotransmisores/metabolismo , Células Fotorreceptoras de Invertebrados/metabolismo , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Receptores Colinérgicos/metabolismo , Receptores Histamínicos/metabolismo , Percepción Visual/fisiología , Percepción Visual/efectos de la radiaciónRESUMEN
Ciliary and rhabdomeric photoreceptor cells represent two main lines of photoreceptor-cell evolution in animals. The two cell types coexist in some animals, however how these cells functionally integrate is unknown. We used connectomics to map synaptic paths between ciliary and rhabdomeric photoreceptors in the planktonic larva of the annelid Platynereis and found that ciliary photoreceptors are presynaptic to the rhabdomeric circuit. The behaviors mediated by the ciliary and rhabdomeric cells also interact hierarchically. The ciliary photoreceptors are UV-sensitive and mediate downward swimming in non-directional UV light, a behavior absent in ciliary-opsin knockout larvae. UV avoidance overrides positive phototaxis mediated by the rhabdomeric eyes such that vertical swimming direction is determined by the ratio of blue/UV light. Since this ratio increases with depth, Platynereis larvae may use it as a depth gauge during vertical migration. Our results revealed a functional integration of ciliary and rhabdomeric photoreceptor cells in a zooplankton larva.
Asunto(s)
Cilios/fisiología , Opsinas/metabolismo , Células Fotorreceptoras de Invertebrados/fisiología , Zooplancton/fisiología , Animales , Cilios/efectos de la radiación , Larva/fisiología , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Natación , Rayos Ultravioleta , Zooplancton/efectos de la radiaciónRESUMEN
The purpose of this paper is to explore the firing rate of the caudal photoreceptors (CPRs) from the sixth abdominal ganglion of the crayfish Cherax quadricarinatus. We use simultaneous extracellular recordings on left and right CPR in the isolated ganglion (n = 10). The CPRs showed an asymmetry in the spontaneous activity and light-induced response. In darkness, we observed one subgroup (70%) in which the left CPR (CPR-L) and right CPR (CPR-R) had spontaneous firing rates with a median of 18 impulses/s and 6 impulses/s, respectively. In another subgroup (20%), the CPR-R had a median of 15 impulses/s and the CPR-L had 8 impulses/s. In both groups, the differences were significant. Furthermore, the CPRs showed an asymmetrical photoresponse induced by a pulse of white light (700 Lux, 4 s). In one subgroup (30%), the CPR-L showed light-induced activity with a median of 73%, (interquartile range, IQR = 51), while the CPR-R had a median of 41%, (IQR = 47). In another subgroup (70%), the CPR-R showed a median of 56%, (IQR = 51) and the CPR-L had a median of 42%, (IQR = 46). In both groups, the differences were significant. Moreover, we observed a differential effect of temperature on CPR activity. These results suggest a functional asymmetry in both activities from left and right CPRs. These CPR activity fluctuations may modulate the processing of information by the nervous system.
Asunto(s)
Potenciales de Acción/fisiología , Astacoidea/fisiología , Lateralidad Funcional/fisiología , Ganglios de Invertebrados/fisiología , Fototransducción/fisiología , Células Fotorreceptoras de Invertebrados/fisiología , Potenciales de Acción/efectos de la radiación , Animales , Astacoidea/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Lateralidad Funcional/efectos de la radiación , Ganglios de Invertebrados/efectos de la radiación , Luz , Fototransducción/efectos de la radiación , Estimulación Luminosa/métodos , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Dosis de RadiaciónRESUMEN
Noise is a prevalent and sometimes even dominant aspect of many biological processes. While many natural systems have adapted to attenuate or even usefully integrate noise, the variability it introduces often still delimits the achievable precision across biological functions. This is particularly so for visual phototransduction, the process responsible for converting photons of light into usable electrical signals (quantum bumps). Here, randomness of both the photon inputs (regarded as extrinsic noise) and the conversion process (intrinsic noise) are seen as two distinct, independent and significant limitations on visual reliability. Past research has attempted to quantify the relative effects of these noise sources by using approximate methods that do not fully account for the discrete, point process and time ordered nature of the problem. As a result the conclusions drawn from these different approaches have led to inconsistent expositions of phototransduction noise performance. This paper provides a fresh and complete analysis of the relative impact of intrinsic and extrinsic noise in invertebrate phototransduction using minimum mean squared error reconstruction techniques based on Bayesian point process (Snyder) filters. An integrate-fire based algorithm is developed to reliably estimate photon times from quantum bumps and Snyder filters are then used to causally estimate random light intensities both at the front and back end of the phototransduction cascade. Comparison of these estimates reveals that the dominant noise source transitions from extrinsic to intrinsic as light intensity increases. By extending the filtering techniques to account for delays, it is further found that among the intrinsic noise components, which include bump latency (mean delay and jitter) and shape (amplitude and width) variance, it is the mean delay that is critical to noise performance. As the timeliness of visual information is important for real-time action, this delay could potentially limit the speed at which invertebrates can respond to stimuli. Consequently, if one wants to increase visual fidelity, reducing the photoconversion lag is much more important than improving the regularity of the electrical signal.
Asunto(s)
Modelos Neurológicos , Modelos Estadísticos , Células Fotorreceptoras de Invertebrados/fisiología , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Visión Ocular/fisiología , Visión Ocular/efectos de la radiación , Animales , Teorema de Bayes , Simulación por Computador , Relación Dosis-Respuesta en la Radiación , Drosophila , Cadenas de Markov , Estimulación Luminosa/métodos , Dosis de Radiación , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Relación Señal-RuidoRESUMEN
Activation of a phytochrome photoreceptor triggers a program of Physarum polycephalum plasmodial cell differentiation through which a mitotic multinucleate protoplasmic mass synchronously develops into haploid spores formed by meiosis and rearrangement of cellular components. We have performed a transcriptome-wide RNAseq study of cellular reprogramming and developmental switching. RNAseq analysis revealed extensive remodeling of intracellular signaling and regulation in switching the expression of sets of genes encoding transcription factors, kinases, phosphatases, signal transduction proteins, RNA-binding proteins, ubiquitin ligases, regulators of the mitotic and meiotic cell cycle etc. in conjunction with the regulation of genes encoding metabolic enzymes and cytoskeletal proteins. About 15% of the differentially expressed genes shared similarity with members of the evolutionary conserved set of core developmental genes of social amoebae. Differential expression of genes encoding regulators that act at the transcriptional, translational, and post-translational level indicates the establishment of a new state of cellular function and reveals evolutionary deeply conserved molecular changes involved in cellular reprogramming and differentiation in a prototypical eukaryote.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica/fisiología , Redes Reguladoras de Genes/fisiología , Genes Protozoarios/fisiología , Physarum polycephalum/crecimiento & desarrollo , Proteínas Protozoarias/metabolismo , Animales , Diferenciación Celular/genética , Diferenciación Celular/efectos de la radiación , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica/efectos de la radiación , Redes Reguladoras de Genes/efectos de la radiación , Luz , Células Fotorreceptoras de Invertebrados/fisiología , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Physarum polycephalum/genética , Physarum polycephalum/efectos de la radiación , Fitocromo/genética , Fitocromo/metabolismo , Proteínas Protozoarias/genética , Transducción de Señal/genética , Transcriptoma/fisiología , Transcriptoma/efectos de la radiaciónRESUMEN
Ciliary opsins were classically thought to function only in vertebrates for vision, but they have also been identified recently in invertebrates for non-visual photoreception. Larvae of the annelid Platynereis dumerilii are used as a zooplankton model, and this zooplankton species possesses a "vertebrate-type" ciliary opsin (named c-opsin) in the brain. Platynereis c-opsin is suggested to relay light signals for melatonin production and circadian behaviors. Thus, the spectral and biochemical characteristics of this c-opsin would be directly related to non-visual photoreception in this zooplankton model. Here we demonstrate that the c-opsin can sense UV to activate intracellular signaling cascades and that it can directly bind exogenous all-trans-retinal. These results suggest that this c-opsin regulates circadian signaling in a UV-dependent manner and that it does not require a supply of 11-cis-retinal for photoreception. Avoidance of damaging UV irradiation is a major cause of large-scale daily zooplankton movement, and the observed capability of the c-opsin to transmit UV signals and bind all-trans-retinal is ideally suited for sensing UV radiation in the brain, which presumably lacks enzymes producing 11-cis-retinal. Mutagenesis analyses indicated that a unique amino acid residue (Lys-94) is responsible for c-opsin-mediated UV sensing in the Platynereis brain. We therefore propose that acquisition of the lysine residue in the c-opsin would be a critical event in the evolution of Platynereis to enable detection of ambient UV light. In summary, our findings indicate that the c-opsin possesses spectral and biochemical properties suitable for UV sensing by the zooplankton model.
Asunto(s)
Proteínas del Tejido Nervioso/metabolismo , Opsinas/metabolismo , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Poliquetos/fisiología , Sistemas de Mensajero Secundario/efectos de la radiación , Zooplancton/fisiología , Sustitución de Aminoácidos , Animales , Células COS , Chlorocebus aethiops , Cilios/metabolismo , Cilios/efectos de la radiación , Canales de Potasio Rectificados Internamente Asociados a la Proteína G/genética , Canales de Potasio Rectificados Internamente Asociados a la Proteína G/metabolismo , Lisina/química , Mutación , Proteínas del Tejido Nervioso/química , Proteínas del Tejido Nervioso/genética , Oocitos/metabolismo , Oocitos/efectos de la radiación , Opsinas/química , Opsinas/genética , Técnicas de Placa-Clamp , Células Fotorreceptoras de Invertebrados/metabolismo , Filogenia , Poliquetos/efectos de la radiación , Estabilidad Proteica/efectos de la radiación , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/metabolismo , Retinaldehído/química , Retinaldehído/metabolismo , Estereoisomerismo , Rayos Ultravioleta , Xenopus , Zooplancton/efectos de la radiaciónRESUMEN
Dihydroceramide desaturases are evolutionarily conserved enzymes that convert dihydroceramide (dhCer) to ceramide (Cer). While elevated Cer levels cause neurodegenerative diseases, the neuronal activity of its direct precursor, dhCer, remains unclear. We show that knockout of the fly dhCer desaturase gene, infertile crescent (ifc), results in larval lethality with increased dhCer and decreased Cer levels. Light stimulation leads to ROS increase and apoptotic cell death in ifc-KO photoreceptors, resulting in activity-dependent neurodegeneration. Lipid-containing Atg8/LC3-positive puncta accumulate in ifc-KO photoreceptors, suggesting lipophagy activation. Further enhancing lipophagy reduces lipid droplet accumulation and rescues ifc-KO defects, indicating that lipophagy plays a protective role. Reducing dhCer synthesis prevents photoreceptor degeneration and rescues ifc-KO lethality, while supplementing downstream sphingolipids does not. These results pinpoint that dhCer accumulation is responsible for ifc-KO defects. Human dhCer desaturase rescues ifc-KO larval lethality, and rapamycin reverses defects caused by dhCer accumulation in human neuroblastoma cells, suggesting evolutionarily conserved functions. This study demonstrates a novel requirement for dhCer desaturase in neuronal maintenance in vivo and shows that lipophagy activation prevents activity-dependent degeneration caused by dhCer accumulation.
Asunto(s)
Autofagia , Ceramidas/metabolismo , Metabolismo de los Lípidos , Animales , Apoptosis , Línea Celular Tumoral , Ceramidas/análisis , Drosophila , Proteínas de Drosophila/deficiencia , Proteínas de Drosophila/genética , Ácido Graso Desaturasas/genética , Técnicas de Inactivación de Genes , Humanos , Luz/efectos adversos , Lipólisis , Proteínas de la Membrana/deficiencia , Proteínas de la Membrana/genética , Enfermedades Neurodegenerativas/prevención & control , Células Fotorreceptoras de Invertebrados/patología , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Esfingolípidos/metabolismoRESUMEN
The nervous system has the remarkable ability to adapt and respond to various stimuli. This neural adjustment is largely achieved through plasticity at the synaptic level. The Active Zone (AZ) is the region at the presynaptic membrane that mediates neurotransmitter release and is composed of a dense collection of scaffold proteins. AZs of Drosophila melanogaster (Drosophila) photoreceptors undergo molecular remodeling after prolonged exposure to natural ambient light. Thus the level of neuronal activity can rearrange the molecular composition of the AZ and contribute to the regulation of the functional output. Starting from the light exposure set-up preparation to the immunohistochemistry, this protocol details how to quantify the number, the spatial distribution, and the delocalization level of synaptic molecules at AZs in Drosophila photoreceptors. Using image analysis software, clusters of the GFP-fused AZ component Bruchpilot were identified for each R8 photoreceptor (R8) axon terminal. Detected Bruchpilot spots were automatically assigned to individual R8 axons. To calculate the distribution of spot frequency along the axon, we implemented a customized software plugin. Each axon's start-point and end-point were manually defined and the position of each Bruchpilot spot was projected onto the connecting line between start and end-point. Besides the number of Bruchpilot clusters, we also quantified the delocalization level of Bruchpilot-GFP within the clusters. These measurements reflect in detail the spatially resolved synaptic dynamics in a single neuron under different environmental conditions to stimuli.
Asunto(s)
Luz , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Transmisión Sináptica/fisiología , Animales , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Proteínas Fluorescentes Verdes/metabolismo , Sustancias Luminiscentes/metabolismo , Células Fotorreceptoras de Invertebrados/metabolismo , Terminales Presinápticos , Unión Proteica , Transporte de Proteínas , Sinapsis/metabolismoRESUMEN
Homoeostatic regulation of the light sensor, rhodopsin, is critical for the maintenance of light sensitivity and survival of photoreceptors. The major fly rhodopsin, Rh1, undergoes light-induced endocytosis and degradation, but its protein and mRNA levels remain constant during light/dark cycles. It is not clear how translation of Rh1 is regulated. Here, we show that adult photoreceptors maintain a constant, abundant quantity of ninaE mRNA, which encodes Rh1. We demonstrate that the Fmr1 protein associates with ninaE mRNA and represses its translation. Further, light exposure triggers a calcium-dependent dephosphorylation of Fmr1, which relieves suppression of Rh1 translation. We demonstrate that Mts, the catalytic subunit of protein phosphatase 2A (PP2A), mediates light-induced Fmr1 dephosphorylation in a regulatory B subunit of PP2A (CKa)-dependent manner. Finally, we show that blocking light-induced Rh1 translation results in reduced light sensitivity. Our results reveal the molecular mechanism of Rh1 homoeostasis and physiological consequence of Rh1 dysregulation.
Asunto(s)
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Proteína de la Discapacidad Intelectual del Síndrome del Cromosoma X Frágil/metabolismo , Homeostasis , Rodopsina/metabolismo , Envejecimiento/metabolismo , Animales , Proteínas de Drosophila/genética , Drosophila melanogaster/efectos de la radiación , Homeostasis/efectos de la radiación , Luz , Fototransducción/efectos de la radiación , Fosforilación/efectos de la radiación , Células Fotorreceptoras de Invertebrados/metabolismo , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Biosíntesis de Proteínas/efectos de la radiación , ARN Mensajero/genética , ARN Mensajero/metabolismo , Rodopsina/genéticaRESUMEN
During illumination, the light-sensitive plasma membrane (rhabdomere) of Drosophila photoreceptors undergoes turnover with consequent changes in size and composition. However, the mechanism by which illumination is coupled to rhabdomere turnover remains unclear. We find that photoreceptors contain a light-dependent phospholipase D (PLD) activity. During illumination, loss of PLD resulted in an enhanced reduction in rhabdomere size, accumulation of Rab7 positive, rhodopsin1-containing vesicles (RLVs) in the cell body and reduced rhodopsin protein. These phenotypes were associated with reduced levels of phosphatidic acid, the product of PLD activity and were rescued by reconstitution with catalytically active PLD. In wild-type photoreceptors, during illumination, enhanced PLD activity was sufficient to clear RLVs from the cell body by a process dependent on Arf1-GTP levels and retromer complex function. Thus, during illumination, PLD activity couples endocytosis of RLVs with their recycling to the plasma membrane thus maintaining plasma membrane size and composition.
Asunto(s)
Membrana Celular/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/enzimología , Endocitosis/efectos de la radiación , Fosfolipasa D/metabolismo , Células Fotorreceptoras de Invertebrados/metabolismo , Factor 1 de Ribosilacion-ADP/genética , Factor 1 de Ribosilacion-ADP/metabolismo , Animales , Membrana Celular/efectos de la radiación , Membrana Celular/ultraestructura , Vesículas Citoplasmáticas/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster/citología , Drosophila melanogaster/genética , Drosophila melanogaster/efectos de la radiación , Expresión Génica , Prueba de Complementación Genética , Guanosina Trifosfato/metabolismo , Luz , Ácidos Fosfatidicos/metabolismo , Fosfolipasa D/genética , Estimulación Luminosa , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Células Fotorreceptoras de Invertebrados/ultraestructura , Rodopsina/genética , Rodopsina/metabolismo , Visión Ocular/fisiología , Proteínas de Unión al GTP rab/genética , Proteínas de Unión al GTP rab/metabolismo , Proteínas de Unión a GTP rab7RESUMEN
Many invertebrates carry out a daily cycle of shedding and rebuilding of the photoreceptor's photosensitive rhabdomeric membranes. The mosquito Aedes aegypti shows a robust response, losing nearly all Aaop1 rhodopsin from the rhabdomeric membranes during the shedding process at dawn. Here, we made use of Aaop1 antibodies capable of distinguishing newly synthesized, glycosylated rhodopsin from mature nonglycosylated rhodopsin to characterize the fate of Aaop1 during the shedding and rebuilding processes. The rhabdomeric rhodopsin is moved into large cytoplasmic vesicles at dawn and is subsequently degraded during the standard 12 h daytime period. The endocytosed rhodopsin is trafficked back to the photosensitive membranes if animals are shifted back to dark conditions during the morning hours. During the daytime period, small vesicles containing newly synthesized and glycosylated Aaop1 rhodopsin accumulate within the cytoplasm. At dusk, these vesicles are lost as the newly synthesized Aaop1 is converted to the nonglycosylated form and deposited in the rhabdomeres. We demonstrate that light acts though a novel signaling pathway to block rhodopsin maturation, thus inhibiting the deglycosylation and rhabdomeric targeting of newly synthesized Aaop1 rhodopsin. Therefore, light controls two cellular processes responsible for the daily renewal of rhodopsin: rhodopsin endocytosis at dawn and inhibition of rhodopsin maturation until dusk. SIGNIFICANCE STATEMENT: Organisms use multiple strategies to maximize visual capabilities in different light conditions. Many invertebrates show a daily cycle of shedding the photoreceptor's rhabdomeric membranes at dawn and rebuilding these during the following night. We show here that the Aedes aegypti mosquito possesses two distinct light-driven cellular signaling processes for modulating rhodopsin content during this cycle. One of these, endocytosis of rhabdomeric rhodopsin, has been described previously. The second, a light-activated cellular pathway acting to inhibit the anterograde movement of newly synthesized rhodopsin, is revealed here for the first time. The discovery of this cellular signaling pathway controlling a G-protein-coupled receptor is of broad interest due to the prominent role of this receptor family across all areas of neuroscience.
Asunto(s)
Ritmo Circadiano/fisiología , Culicidae/fisiología , Culicidae/efectos de la radiación , Estimulación Luminosa/métodos , Células Fotorreceptoras de Invertebrados/fisiología , Rodopsina/metabolismo , Animales , Ritmo Circadiano/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Luz , Tasa de Depuración Metabólica/fisiología , Tasa de Depuración Metabólica/efectos de la radiación , Fotoperiodo , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Dosis de RadiaciónRESUMEN
The biflagellate green alga Chlamydomonas reinhardtii exhibits both positive and negative phototaxis to inhabit areas with proper light conditions. It has been shown that treatment of cells with reactive oxygen species (ROS) reagents biases the phototactic sign to positive, whereas that with ROS scavengers biases it to negative. Taking advantage of this property, we isolated a mutant, lts1-211, which displays a reduction-oxidation (redox) dependent phototactic sign opposite to that of the wild type. This mutant has a single amino acid substitution in phytoene synthase, an enzyme that functions in the carotenoid-biosynthesis pathway. The eyespot contains large amounts of carotenoids and is crucial for phototaxis. Most lts1-211 cells have no detectable eyespot and reduced carotenoid levels. Interestingly, the reversed phototactic-sign phenotype of lts1-211 is shared by other eyespot-less mutants. In addition, we directly showed that the cell body acts as a convex lens. The lens effect of the cell body condenses the light coming from the rear onto the photoreceptor in the absence of carotenoid layers, which can account for the reversed-phototactic-sign phenotype of the mutants. These results suggest that light-shielding property of the eyespot is essential for determination of phototactic sign.
Asunto(s)
Carotenoides/fisiología , Movimiento Celular/fisiología , Chlamydomonas reinhardtii/fisiología , Células Fotorreceptoras de Invertebrados/fisiología , Fototaxis/fisiología , Animales , Carotenoides/efectos de la radiación , Movimiento Celular/efectos de la radiación , Chlamydomonas reinhardtii/citología , Chlamydomonas reinhardtii/efectos de la radiación , Luz , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Pigmentación/fisiología , Pigmentación/efectos de la radiación , Dosis de RadiaciónRESUMEN
The ability to perceive geomagnetic fields (GMFs) represents a fascinating biological phenomenon. Studies on transgenic flies have provided evidence that photosensitive Cryptochromes (Cry) are involved in the response to magnetic fields (MFs). However, none of the studies tackled the problem of whether the Cry-dependent magnetosensitivity is coupled to the sole MF presence or to the direction of MF vector. In this study, we used gene silencing and a directional MF to show that mammalian-like Cry2 is necessary for a genuine directional response to periodic rotations of the GMF vector in two insect species. Longer wavelengths of light required higher photon fluxes for a detectable behavioral response, and a sharp detection border was present in the cyan/green spectral region. Both observations are consistent with involvement of the FADox, FAD(â¢-) and FADH(-) redox forms of flavin. The response was lost upon covering the eyes, demonstrating that the signal is perceived in the eye region. Immunohistochemical staining detected Cry2 in the hemispherical layer of laminal glia cells underneath the retina. Together, these findings identified the eye-localized Cry2 as an indispensable component and a likely photoreceptor of the directional GMF response. Our study is thus a clear step forward in deciphering the in vivo effects of GMF and supports the interaction of underlying mechanism with the visual system.
Asunto(s)
Cucarachas/metabolismo , Criptocromos/metabolismo , Campos Magnéticos , Células Fotorreceptoras de Invertebrados/metabolismo , Animales , Cucarachas/efectos de la radiación , Ojo Compuesto de los Artrópodos/efectos de la radiación , Fenotipo , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Rayos UltravioletaRESUMEN
Two insults often underlie a variety of eye diseases including glaucoma, optic atrophy, and retinal degeneration--defects in mitochondrial function and aberrant Rhodopsin trafficking. Although mitochondrial defects are often associated with oxidative stress, they have not been linked to Rhodopsin trafficking. In an unbiased forward genetic screen designed to isolate mutations that cause photoreceptor degeneration, we identified mutations in a nuclear-encoded mitochondrial gene, ppr, a homolog of human LRPPRC. We found that ppr is required for protection against light-induced degeneration. Its function is essential to maintain membrane depolarization of the photoreceptors upon repetitive light exposure, and an impaired phototransduction cascade in ppr mutants results in excessive Rhodopsin1 endocytosis. Moreover, loss of ppr results in a reduction in mitochondrial RNAs, reduced electron transport chain activity, and reduced ATP levels. Oxidative stress, however, is not induced. We propose that the reduced ATP level in ppr mutants underlies the phototransduction defect, leading to increased Rhodopsin1 endocytosis during light exposure, causing photoreceptor degeneration independent of oxidative stress. This hypothesis is bolstered by characterization of two other genes isolated in the screen, pyruvate dehydrogenase and citrate synthase. Their loss also causes a light-induced degeneration, excessive Rhodopsin1 endocytosis and reduced ATP without concurrent oxidative stress, unlike many other mutations in mitochondrial genes that are associated with elevated oxidative stress and light-independent photoreceptor demise.
Asunto(s)
Proteínas de Drosophila/genética , Mitocondrias/metabolismo , Proteínas Mitocondriales/genética , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Enfermedades de la Retina/genética , Adenosina Trifosfato/biosíntesis , Animales , Citrato (si)-Sintasa/genética , Drosophila , Proteínas de Drosophila/metabolismo , Electrorretinografía , Endocitosis , Mitocondrias/genética , Proteínas Mitocondriales/metabolismo , Estrés Oxidativo , Complejo Piruvato Deshidrogenasa/genética , Complejo Piruvato Deshidrogenasa/metabolismo , Proteínas de Unión al ARN/genética , Proteínas de Unión al ARN/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Rodopsina/metabolismo , Visión OcularRESUMEN
The cranchiid Teuthowenia pellucida, like many deep-sea squid species, possesses large eyes that maximise light sensitivity in a nearly aphotic environment. To assess ontogenetic changes in the visual system, we conducted morphometric and histological analyses of the eyes using specimens from New Zealand collections. While the ratio between eye diameter and mantle length maintained a linear relationship throughout development, histological sections of the retina revealed that the outer photoreceptor layer became proportionally longer as the animal aged, coincident with a habitat shift into deeper, darker ocean strata. Other retinal layers maintained the same absolute thickness as was observed in paralarvae. Granules of the pigment ommin, normally located in the screening layer positioned at the base of the photoreceptors, were also observed at the outer end of the photoreceptor segments throughout the retina in young and mid-sized specimens. Early developmental stages of this species, dwelling in shallow waters, may therefore rely on migratory ommin to help shield photoreceptors from excess light and prevent over-stimulation. The oldest, deeper-dwelling specimens of T. pellucida examined had longer photoreceptors, and little or no migrated ommin was observed; we suggest therefore that short-term adaptive mechanisms for bright light conditions may be used primarily during epipelagic, early life stages in this species.
Asunto(s)
Decapodiformes/crecimiento & desarrollo , Estadios del Ciclo de Vida/fisiología , Células Fotorreceptoras de Invertebrados/ultraestructura , Pigmentos Retinianos/química , Animales , Decapodiformes/anatomía & histología , Decapodiformes/efectos de la radiación , Estadios del Ciclo de Vida/efectos de la radiación , Luz , Nueva Zelanda , Células Fotorreceptoras de Invertebrados/fisiología , Células Fotorreceptoras de Invertebrados/efectos de la radiaciónRESUMEN
Drosophila phototransduction is a model system for the ubiquitous phosphoinositide signaling. In complete darkness, spontaneous unitary current events (dark bumps) are produced by spontaneous single Gqα activation, while single-photon responses (quantum bumps) arise from synchronous activation of several Gqα molecules. We have recently shown that most of the spontaneous single Gqα activations do not produce dark bumps, because of a critical phospholipase Cß (PLCß) activity level required for bump generation. Surpassing the threshold of channel activation depends on both PLCß activity and cellular [Ca(2+)], which participates in light excitation via a still unclear mechanism. We show here that in IP3 receptor (IP3R)-deficient photoreceptors, both light-activated Ca(2+) release from internal stores and light sensitivity were strongly attenuated. This was further verified by Ca(2+) store depletion, linking Ca(2+) release to light excitation. In IP3R-deficient photoreceptors, dark bumps were virtually absent and the quantum-bump rate was reduced, indicating that Ca(2+) release from internal stores is necessary to reach the critical level of PLCß catalytic activity and the cellular [Ca(2+)] required for excitation. Combination of IP3R knockdown with reduced PLCß catalytic activity resulted in highly suppressed light responses that were partially rescued by cellular Ca(2+) elevation, showing a functional cooperation between IP3R and PLCß via released Ca(2+). These findings suggest that in contrast to the current dogma that Ca(2+) release via IP3R does not participate in light excitation, we show that released Ca(2+) plays a critical role in light excitation. The positive feedback between PLCß and IP3R found here may represent a common feature of the inositol-lipid signaling.
Asunto(s)
Drosophila/fisiología , Receptores de Inositol 1,4,5-Trifosfato/fisiología , Fosfolipasa C beta/fisiología , Células Fotorreceptoras de Invertebrados/fisiología , Animales , Animales Modificados Genéticamente , Señalización del Calcio/fisiología , Electrorretinografía , Hipoxia/metabolismo , Receptores de Inositol 1,4,5-Trifosfato/genética , Luz , Masculino , Técnicas de Placa-Clamp , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Interferencia de ARNRESUMEN
In a previous study of the phototaxis of green rice leafhoppers, Nephotettix cincticeps (Hemiptera, Cicadellidae), we found positive responses to 735 nm light. Here, we investigated the mechanism underlying this sensitivity to near-infrared light. We first measured the action spectrum using a Y-maze with monochromatic lights from 480 to 740 nm. We thus found that the action spectrum peaks at 520 nm in the tested wavelength range, but that a significant effect is still observed at 740 nm, albeit with a sensitivity 5 log units lower than the peak. Second, we measured the spectral sensitivity of the eye, and found that the sensitivity in the long-wavelength region parallels the behaviorally determined action spectrum. We further identified mRNAs encoding opsins of ultraviolet, blue, and green-absorbing visual pigments, and localized the mRNAs in the ommatidia by in situ hybridization. The electrophysiology, molecular biology and the anatomy of the eye together indicate that the eyes of N. cincticeps do not contain true "red" receptors, but rather that the behavioral response to near-infrared light is mediated by the tail sensitivity of the green receptors in the long-wavelength region of the spectrum.
Asunto(s)
Mariposas Diurnas/fisiología , Percepción de Color/fisiología , Opsinas/metabolismo , Células Fotorreceptoras de Invertebrados/fisiología , Animales , Color , Percepción de Color/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Masculino , Aprendizaje por Laberinto/efectos de la radiación , Opsinas/clasificación , Opsinas/genética , Estimulación Luminosa/métodos , Células Fotorreceptoras de Invertebrados/metabolismo , Células Fotorreceptoras de Invertebrados/efectos de la radiación , ARN Mensajero , Rayos UltravioletaRESUMEN
Medicinal leeches are aquatic predators that inhabit surface waters during daylight and also leave the water where they might be exposed to less screened light. Whereas the leech visual system has been shown to respond to visible light, leeches in the genus Hirudo do not appear to be as negatively phototactic as one might expect in order to avoid potential ultraviolet radiation (UVR)-induced damage. I used high intensity light emitting diodes to test the hypothesis that leeches could detect and specifically avoid near UVR (395-405 nm). Groups of unfed juvenile leeches exhibited a robust negative phototaxis to UVR, but had no behavioral response to blue or red and only a slight negative phototaxis to green and white light. Individual leeches also exhibited a vigorous negative phototaxis to UVR; responding in 100% of trials compared with modest negative responses to visible light (responding in ~8% of the trials). The responses in fed and unfed leeches were comparable for UVR stimuli. The responses depended upon the stimulus site: leeches shortened away from UV light to the head, and extended away from UV light to the tail. Electrophysiological nerve recordings showed that the cephalic eyes responded vigorously to UVR. Additionally, individual leech photoreceptors also showed strong responses to UVR, and a higher-order neuron associated with shortening and rapid behavioral responses, the S-cell, was activated by UVR, on both the head and tail. These results demonstrate that the leech can detect UVR and is able to discriminate behaviorally between UVR and visible light.
Asunto(s)
Hirudo medicinalis/efectos de la radiación , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Rayos Ultravioleta , Animales , Reacción de Prevención/efectos de la radiación , Hirudo medicinalis/fisiología , Locomoción/efectos de la radiación , Células Fotorreceptoras de Invertebrados/fisiología , Vías Visuales/fisiología , Vías Visuales/efectos de la radiaciónRESUMEN
Most living organisms possess a circadian clock (24 h period) which allows them to adapt to environmental conditions. Numerous studies in Drosophila allowed to discover various key clock genes, such as period and timeless. The powerful tools of drosophila genetics have shown that the molecular clock relies on negative feedback loops that generate oscillations of the clock genes mRNA. A delay between the accumulation of mRNAs and proteins is required for the feedback loop. It is generated by post-translational modifications as phosphorylations and ubiquitinations, which control protein stability and determine the period of their oscillations. Clock cells are present in brain as well as in multiple peripheric tissues where they run autonomously. The synchronisation of clock cells by light relies on cryptochrome in both brain and peripheral tissues. In the brain, synchronisation also involves the eye photoreceptors. The clock that drives sleep-wake rhythms is controlled by different groups of neurons in the brain. Each group has a distinct function in the generation of the behavioral rhythm and this function is modulated by environmental conditions.
Asunto(s)
Péptidos y Proteínas de Señalización del Ritmo Circadiano/genética , Ritmo Circadiano/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Genes de Insecto , Animales , Conducta Animal/fisiología , Conducta Animal/efectos de la radiación , Encéfalo/fisiología , Ritmo Circadiano/efectos de la radiación , Péptidos y Proteínas de Señalización del Ritmo Circadiano/fisiología , Criptocromos/fisiología , Criptocromos/efectos de la radiación , Proteínas de Drosophila/fisiología , Proteínas de Drosophila/efectos de la radiación , Drosophila melanogaster/fisiología , Drosophila melanogaster/efectos de la radiación , Proteínas del Ojo/fisiología , Proteínas del Ojo/efectos de la radiación , Luz , Mamíferos/genética , Mamíferos/fisiología , Neuronas/clasificación , Neuronas/fisiología , Células Fotorreceptoras de Invertebrados/fisiología , Células Fotorreceptoras de Invertebrados/efectos de la radiación , Procesamiento Proteico-Postraduccional , ARN Mensajero/genética , ARN Mensajero/fisiología , Sueño/genética , Sueño/fisiología , Especificidad de la Especie , Transcripción Genética , Vigilia/genética , Vigilia/fisiologíaRESUMEN
The compound eyes of the eastern pale clouded yellow butterfly, Colias erate, contain three types of ommatidia (I, II and III), identifiable by the differing arrangements of pigment clusters around the rhabdoms. The pigment color is red in all ommatidial types except for type II ommatidia of females, where the pigment is orange. Intracellular recordings demonstrated that the spectral sensitivities of the proximal photoreceptors (R5-8) of all ommatidia in both sexes are strongly tuned by the perirhabdomal pigments. These pigments act as long-pass filters, shifting the peak sensitivities into the wavelength range above 600 nm. Due to the sex-specific pigments in type II ommatidia, the spectral sensitivities of the R5-8 photoreceptors of females peaked at 620 nm while those in males peaked at 660 nm. The measured spectral sensitivities could be well reproduced by an optical model assuming a long-wavelength-absorbing visual pigment with peak absorbance at 565 nm. Whereas the sexual dimorphism was unequivocally demonstrated for the ventral eye region, dimorphism in the dorsal region was not found. Presumably the ventral region is adapted for sexual behaviors such as courtship and oviposition.