Circadian Light Research Articles

Method to Calculate Melanopic Light Reaching the Retina Depending on the Optical Density of an Aging Crystalline Lens

Lighting studies that take into account the age of the inhabitants of an area and are related to circadian light are difficult to find. This study aims to simplify a method to approximately compute the circadian light reaching the retina based on photopic illuminance reaching the corneal plane and considering the optical density of an aging crystalline lens. As an example of this proposed method, calculations were performed with both the D65 and A standard illuminants, showing how the spectral power distribution is modified by the optical density of the crystalline lens, mainly at short wavelengths. Due to these selective wavelength absorptions of the aged lens, a significant variation in the level of daylight equivalent melanopic illuminance (EDI) is present in the retina. With levels of 200 lux at the corneal plane, these variations ranged from 204 EDI lux to 178 EDI lux for the D65 standard illuminant, and from 99 EDI lux to 101 EDI lux for the A standard illuminant for observers aged 10 and 90, respectively. In this work, we aimed to simplify the greatest possible level of calculation of melanopic light, while describing simple protocols that are easy to translate into practice. Our results will allow researchers to carry out optimized lighting designs from both the photometric and circadian perspectives considering the optical density of an aging lens.

Circadian rhythm entrainment of the jewel wasp, Nasonia vitripennis, by antagonistic interactions of multiple spectral inputs.

Circadian light entrainment in some insects is regulated by blue-light-sensitive cryptochrome (CRY) protein that is expressed in the clock neurons, but this is not the case in hymenopterans. The hymenopteran clock does contain CRY, but it appears to be light-insensitive. Therefore, we investigated the role of retinal photoreceptors in the photic entrainment of the jewel wasp Nasonia vitripennis. Application of monochromatic light stimuli at different light intensities caused phase shifts in the wasp's circadian activity from which an action spectrum with three distinct peaks was derived. Electrophysiological recordings from the compound eyes and ocelli revealed the presence of three photoreceptor classes, with peak sensitivities at 340 nm (ultraviolet), 450 nm (blue) and 530 nm (green). An additional photoreceptor class in the ocelli with sensitivity maximum at 560-580 nm (red) was found. Whereas a simple sum of photoreceptor spectral sensitivities could not explain the action spectrum of the circadian phase shifts, modelling of the action spectrum indicates antagonistic interactions between pairs of spectral photoreceptors, residing in the compound eyes and the ocelli. Our findings imply that the photic entrainment mechanism in N. vitripennis encompasses the neural pathways for measuring the absolute luminance as well as the circuits mediating colour opponency.

Light Pollution Disrupts Seasonal Differences in the Daily Activity and Metabolic Profiles of the Northern House Mosquito, Culex pipiens.

The Northern House mosquito, Culex pipiens, is an important disease vector, and females are capable of surviving the winter in a state of overwintering diapause. This species' diapause response has been extensively studied, and recent evidence suggests that the circadian clock is involved in measuring seasonal changes in daylength to initiate the diapause response. However, differences in the circadian activity of diapausing and non-diapausing Cx. pipiens have not been thoroughly investigated. Additionally, recent findings indicate that artificial light at night (ALAN) can disrupt mosquito diapause, potentially prolonging the mosquito biting season. We compared the circadian locomotor activity of mosquitoes reared in diapause-averting, long-day conditions and diapause-inducing, short-day conditions with and without ALAN to elucidate the interplay between circadian activity, diapause, and light pollution. We also uncovered metabolic differences between mosquitoes reared under diapausing and non-diapausing photoperiods with and without ALAN by measuring the concentration of protein, fructose, glycogen, water-soluble carbohydrates, and lipids. We found that ALAN exposure altered several diapause-associated phenotypes including slightly, but not significantly, increasing activity levels in short day-reared mosquitoes; and preventing some short day-reared mosquitoes from accumulating lipids. ALAN also significantly reduced glycogen and water-soluble carbohydrate levels in long day-reared mosquitoes. Based on our findings, light pollution may decrease insect fitness by perturbing metabolism, and may also impact several phenotypes associated with insect diapause, potentially extending the mosquito biting season and preventing insects in urban environments from overwintering successfully.

Genome-wide gene by environment study of time spent in daylight and chronotype identifies emerging genetic architecture underlying light sensitivity.

Light is the primary stimulus for synchronizing the circadian clock in humans. There are very large interindividual differences in the sensitivity of the circadian clock to light. Little is currently known about the genetic basis for these interindividual differences. We performed a genome-wide gene-by-environment interaction study (GWIS) in 280 897 individuals from the UK Biobank cohort to identify genetic variants that moderate the effect of daytime light exposure on chronotype (individual time of day preference), acting as "light sensitivity" variants for the impact of daylight on the circadian system. We identified a genome-wide significant SNP mapped to the ARL14EP gene (rs3847634; p < 5 × 10-8), where additional minor alleles were found to enhance the morningness effect of daytime light exposure (βGxE = -.03, SE = 0.005) and were associated with increased gene ARL14EP expression in brain and retinal tissues. Gene-property analysis showed light sensitivity loci were enriched for genes in the G protein-coupled glutamate receptor signaling pathway and genes expressed in Per2+ hypothalamic neurons. Linkage disequilibrium score regression identified Bonferroni significant genetic correlations of greater light sensitivity GWIS with later chronotype and shorter sleep duration. Greater light sensitivity was nominally genetically correlated with insomnia symptoms and risk for post-traumatic stress disorder (PTSD). This study is the first to assess light as an important exposure in the genomics of chronotype and is a critical first step in uncovering the genetic architecture of human circadian light sensitivity and its links to sleep and mental health.

O024 Modifying light environments according to individual circadian light sensitivity

Abstract Introduction Large inter-individual differences exist in how sensitive the circadian system is to light. Circadian light sensitivity can be affected by medications, such as antidepressants, and varies as a function of age and some mood disorders. Using a computational model, we investigated how differences in an individual’s light sensitivity can be offset by changes in their light environment to maintain stable circadian timing. Methods A previously validated computational model was used to simulate sleep and circadian timing under realistic assumptions about light and sleep schedules in day workers across two weeks. The model predicted circadian phase (dim light melatonin onset) and sleep onset/offset times for each day. Simulations were repeated varying two parameters: (i) light sensitivity, representing changes in the dose-response curve to light, and (ii) evening illuminance, representing home lighting levels after sunset. Results Higher light sensitivity and higher evening illuminance levels resulted in systematically later predicted sleep and circadian timing. The effects of increasing light sensitivity could be offset by reducing the level of evening illuminance, with this relationship holding over a wide range of light sensitivity values. Low light sensitivity combined with high evening illuminance produced a non-entrained (non-24) phenotype; decreasing evening illuminance from this state resulted in stable entrainment. Discussion We are only beginning to understand the influence of medications and health conditions on circadian light sensitivity. Our results show how modifications to evening light levels can be used to offset impacts of variable light sensitivity on sleep and circadian timing.

Dimming the Lights to Support the Overall Human Wellbeing In An Urban Context

In modern times we are exposing ourselves to artificial light sources more than our ancestors were and it is conflicting with learned behaviours developed during the evolutionary process over thousands of years. In this review paper, the effect of this rapid change and possible effects on human wellbeing is questioned. Publications detailing, the health consequences of circadian disruptions and the current stage of global artificial light at night publications have been reviewed to see the level of intrusive lighting at night and to understand the possible connection. The current level of light pollution is mainly questioned as it is reported that the human circadian system and melatonin suppression is more sensitive to a lower level of light than has been previously reported [1]. During the time of review “Light at Night and Circadian” terms used in at Google Chrome search engine and, 24,557 publications were found on Science Direct [2], 17,500 publications were found on Google Scholar [3], and 1,240 free publications were found on PubMed [4] sites that are directly related to the circadian rhythm and light at night topics. Among those publications, recently published research papers (2010-2020) have been selected for review to narrow down the reviewed light sources to light emitting diodes (LEDs) as it is noted that the circadian system is susceptible to the blue wavelengths and LEDs are showing a peak in their spectrum at the same sensitivity area. At the end of the review, possible direct connections between the use of artificial light and adverse physiological and psychological effects with more severe health problems have been found. In addition, the current light pollution studies showed that vast majority of the global population is living under light polluted skies and possibly experiencing the negative effects of disrupted circadian system daily and therefore experiencing deteriorating effects of circadian disruptions and personal wellbeing. As the use of artificial light rapidly increases with global population, exposure to artificial light will increase exponentially in the near future and not only human beings but also the rest of the ecosystem will suffer from its negative effects. Because of this rapid change we should start implementing more stringent guidelines written with consideration for the health implications of artificial light while controlling the excessive use of it. Also, as the LED based lighting systems increase their popularity worldwide, we should start researching a possible modification to reduce the impact of the LED lighting on circadian system and its negative effects on human wellbeing.

A Role for Thalamic Projection GABAergic Neurons in Circadian Responses to Light.

The thalamus is an important hub for sensory information and participates in sensory perception, regulation of attention, arousal and sleep. These functions are executed primarily by glutamatergic thalamocortical neurons that extend axons to the cortex and initiate cortico-thalamocortical connectional loops. However, the thalamus also contains projection GABAergic neurons that do not extend axons toward the cortex. Here, we have harnessed recent insight into the development of the intergeniculate leaflet (IGL) and the ventral lateral geniculate nucleus (LGv) to specifically target and manipulate thalamic projection GABAergic neurons in female and male mice. Our results show that thalamic GABAergic neurons of the IGL and LGv receive retinal input from diverse classes of retinal ganglion cells (RGCs) but not from the M1 intrinsically photosensitive retinal ganglion cell (ipRGC) type. We describe the synergistic role of the photoreceptor melanopsin and the thalamic neurons of the IGL/LGv in circadian entrainment to dim light. We identify a requirement for the thalamic IGL/LGv neurons in the rapid changes in vigilance states associated with circadian light transitions.SIGNIFICANCE STATEMENT The intergeniculate leaflet (IGL) and ventral lateral geniculate nucleus (LGv) are part of the extended circadian system and mediate some nonimage-forming visual functions. Here, we show that each of these structures has a thalamic (dorsal) as well as prethalamic (ventral) developmental origin. We map the retinal input to thalamus-derived cells in the IGL/LGv complex and discover that while RGC input is dominant, this is not likely to originate from M1ipRGCs. We implicate thalamic cells in the IGL/LGv in vigilance state transitions at circadian light changes and in overt behavioral entrainment to dim light, the latter exacerbated by concomitant loss of melanopsin expression.

Effect of Circadian Rhythm Control Light on Sleep State and Mental Health of Students

Effect of Circadian Rhythm Control Light on Sleep State and Mental Health of Students

Circadian rhythm promotes the biomass and amylose hyperaccumulation by mixotrophic cultivation of marine microalga Platymonas helgolandica

BackgroundMicroalgal starch can be exploited for bioenergy, food, and bioplastics. Production of starch by green algae has been concerned for many years. Currently commonly used methods such as nutrient stress will affect cell growth, thereby inhibiting the production efficiency and quality of starch production. Simpler and more efficient control strategies need to be developed.ResultWe proposed a novel regulation method to promote the growth and starch accumulation by a newly isolated Chlorophyta Platymonas helgolandica. By adding exogenous glucose and controlling the appropriate circadian light and dark time, the highest dry weight accumulation 6.53 g L−1 (Light:Dark = 12:12) can be achieved, and the highest starch concentration could reach 3.88 g L−1 (Light:Dark = 6:18). The highest production rate was 0.40 g L−1 d−1 after 9 days of production. And this method helps to improve the ability to produce amylose, with the highest accumulation of 39.79% DW amylose. We also discussed the possible mechanism of this phenomenon through revealing changes in the mRNA levels of key genes.ConclusionThis study provides a new idea to regulate the production of amylose by green algae. For the first time, it is proposed to combine organic carbon source addition and circadian rhythm regulation to increase the starch production from marine green alga. A new starch-producing microalga has been isolated that can efficiently utilize organic matter and grow with or without photosynthesis.

Improving the accuracy of circadian lighting simulation with field measurement

For improving the simulation accuracy of circadian lighting, a workflow was established to first calibrate the simulation model under electric lighting followed by validation with integrated daylighting and electric lighting. A case study was conducted in a daylit office, with the evident impact of varying lighting parameters (light level, intensity, SPD, reflectance) on the simulation accuracy. In the fully calibrated model, the simulated and field-measured SPDs are highly correlated (rSPD ≥ 0.9), providing the acceptable accuracy of the simulated melanopic/photopic(M/P) ratio with the error rate (MBErel) less than 4% and residual deviation (RMSErel) within 7%. The MBErel and RMSErel of corneal illuminance (EV), EML, and CS are within 10% and 20%, respectively. The ratio of data points with the residual rate (e) less than 20% is over 75% in the calibration and validation phases. Conclusively, the proposed procedure could help prepare for further analysis of circadian light exposure.

Core circadian clock and light signaling genes brought into genetic linkage across the green lineage.

The circadian clock is conserved at both the level of transcriptional networks as well as core genes in plants, ensuring that biological processes are phased to the correct time of day. In the model plant Arabidopsis (Arabidopsis thaliana), the core circadian SHAQKYF-type-MYB (sMYB) genes CIRCADIAN CLOCK ASSOCIATED 1 (CCA1) and REVEILLE (RVE4) show genetic linkage with PSEUDO-RESPONSE REGULATOR 9 (PRR9) and PRR7, respectively. Leveraging chromosome-resolved plant genomes and syntenic ortholog analysis enabled tracing this genetic linkage back to Amborella trichopoda, a sister lineage to the angiosperm, and identifying an additional evolutionarily conserved genetic linkage in light signaling genes. The LHY/CCA1–PRR5/9, RVE4/8–PRR3/7, and PIF3–PHYA genetic linkages emerged in the bryophyte lineage and progressively moved within several genes of each other across an array of angiosperm families representing distinct whole-genome duplication and fractionation events. Soybean (Glycine max) maintained all but two genetic linkages, and expression analysis revealed the PIF3–PHYA linkage overlapping with the E4 maturity group locus was the only pair to robustly cycle with an evening phase, in contrast to the sMYB–PRR morning and midday phase. While most monocots maintain the genetic linkages, they have been lost in the economically important grasses (Poaceae), such as maize (Zea mays), where the genes have been fractionated to separate chromosomes and presence/absence variation results in the segregation of PRR7 paralogs across heterotic groups. The environmental robustness model is put forward, suggesting that evolutionarily conserved genetic linkages ensure superior microhabitat pollinator synchrony, while wide-hybrids or unlinking the genes, as seen in the grasses, result in heterosis, adaptation, and colonization of new ecological niches.

The CLASS Study (Circadian Light in Adolescence, Sleep and School): protocol for a prospective, longitudinal cohort to assess sleep, light, circadian timing and academic performance in adolescence

BackgroundDuring adolescence, sleep and circadian timing shift later, contributing to restricted sleep duration and irregular sleep-wake patterns. The association of these developmental changes in sleep and circadian timing with cognitive...

Rest-Activity Circadian Rhythm and Light Exposure Using Wrist Actigraphy in ICU Patients

Actigraphy (ACTG) is easy to use and has proved reliable in non-critical populations of patients. It has been used to track circadian rest-activity cycles and to identify states of wakefulness and sleep [1]. A more recent study has found that ACTG have a moderate level of agreement with PSG in distinguishing between sleep and wakeful states which is more profound in non-ventilated patients [2].

GIGANTEA regulates PAD4 transcription to promote pathogen defense against Hyaloperonospora arabidopsidis in Arabidopsis thaliana

ABSTRACT Plants have evolved a network of complex signaling pathways that allow them to cope with the fluctuations of internal and external environmental cues. GIGANTEA (GI), a well-known, highly conserved plant nuclear protein, has been shown to regulate multiple biological functions in plants such as circadian rhythm, light signaling, cold tolerance, hormone signaling, and photoperiodic flowering. Recently, the role of GI in disease tolerance against different pathogens has come to light; however, a detailed mechanism to understand the role of GI in pathogen defense remains largely unexplained. Here, we report that GIGANTEA is upregulated upon infection with a virulent oomycete pathogen, Hyaloperonospora arabidopsidis (Hpa), in Arabidopsis thaliana accession Col-0. To investigate the role of GI in Arabidopsis defense, we examined the pathogen infection phenotype of gi mutant plants and found that gi-100 mutant was highly susceptible to Hpa Noco2 infection. Notably, the quantitative real-time PCR showed that PHYTOALEXIN DEFICIENT4 (PAD4) and several PAD4-regulated downstream genes were downregulated upon Noco2 infection in gi-100 mutant as compared to Col-0 plants. Furthermore, the chromatin immunoprecipitation results show that GI can directly bind to the intronic region of the PAD4 gene, which might explain the mechanism of GI function in regulating disease resistance in plants. Taken together, our results suggest that GI expression is induced upon Hpa pathogen infection and GI can regulate the expression of PAD4 to promote resistance against the oomycete pathogen Hyaloperonospora arabidopsidis in Arabidopsis thaliana.

The Effects of Light and the Circadian System on Rhythmic Brain Function

Life on earth has evolved under the influence of regularly recurring changes in the environment, such as the 24 h light/dark cycle. Consequently, organisms have developed endogenous clocks, generating 24 h (circadian) rhythms that serve to anticipate these rhythmic changes. In addition to these circadian rhythms, which persist in constant conditions and can be entrained to environmental rhythms, light drives rhythmic behavior and brain function, especially in nocturnal laboratory rodents. In recent decades, research has made great advances in the elucidation of the molecular circadian clockwork and circadian light perception. This review summarizes the role of light and the circadian clock in rhythmic brain function, with a focus on the complex interaction between the different components of the mammalian circadian system. Furthermore, chronodisruption as a consequence of light at night, genetic manipulation, and neurodegenerative diseases is briefly discussed.

The potential anti-cancer effects of melatonin on breast cancer

Melatonin is the primary hormone of the pineal gland that is secreted at night. It regulates many physiological functions, including the sleep-wake cycle, gonadal activity, free radical scavenging, immunomodulation, neuro-protection, and cancer progression. The precise functions of melatonin are mediated by guanosine triphosphate (GTP)-binding protein (G-protein) coupled melatonin receptor 1 (MT1) and MT2 receptors. However, nuclear receptors are also associated with melatonin activity. Circadian rhythm disruption, shift work, and light exposure at night hamper melatonin production. Impaired melatonin level promotes various pathophysiological changes, including cancer. In our modern society, breast cancer is a serious problem throughout the world. Several studies have been indicated the link between low levels of melatonin and breast cancer development. Melatonin has oncostatic properties in breast cancer cells. This indolamine advances apoptosis, which arrests the cell cycle and regulates metabolic activity. Moreover, melatonin increases the treatment efficacy of cancer and can be used as an adjuvant with chemotherapeutic agents.

Circadian Strain, Light Exposure, and Depressive Symptoms in Rural Communities of Southern Brazil.

Irregular light-dark cycles and circadian/sleep disturbances have been suggested as risk or co-occurring factors in depression. Among a set of metrics developed to quantify strain on the circadian system, social jetlag (SJL) has been put forward as a measure of the discrepancy between biological and social clocks. Here, we approached the question on whether light exposure and SJL would also be associated with depressive symptoms in Quilombola communities in Southern Brazil. These rural communities are void of potential confounders of modern lifestyles and show low levels of SJL. 210 Quilombolas (age range 16-92; 56% women) were asked about their sleep times and light exposure using the Munich ChronoType Questionnaire (MCTQ). The Beck Depression Inventory (BDI) was used to assess depressive symptoms. Additionally, we analyzed 7-day actimetry recordings in 124 subjects. BDI scores higher than 10 (having clinically significant depressive symptoms; controlled for age and sex in the multivariate analysis) were positively associated with SJL >1h and negatively associated with median light exposure during the day, especially in the morning from 8:00 to 10:00. Our results suggest that low light exposure during the day, and higher levels of SJL are associated with depressive symptoms; longitudinal and experimental studies are needed to understand the underlying mechanisms. Nevertheless, we highlight the potential of treatment strategies aimed at decreasing circadian strain and insufficient light exposure, which are suggested as areas of further research in Psychiatry.

Relationship Between Circadian Strain, Light Exposure, and Body Mass Index in Rural and Urban Quilombola Communities.

Industrialization has greatly changed human lifestyle; work and leisure activities have been moved indoors, and artificial light has been used to illuminate the night. As cyclic environmental cues such as light and feeding become weak and/or irregular, endogenous circadian systems are increasingly being disrupted. These disruptions are associated with metabolic dysfunction, possibly contributing to increased rates of overweight and obesity worldwide. Here, we aimed to investigate how activity-rest rhythms, patterns of light exposure, and levels of urbanization may be associated with body mass index (BMI) in a sample of rural and urban Quilombola communities in southern Brazil. These are characterized as remaining social groups who resisted the slavery regime that prevailed in Brazil. Quilombola communities were classified into five groups according to their stage of urbanization: from rural areas with no access to electricity to highly urbanized communities. We collected anthropometric data to calculate BMI, which was categorized as follows: from ≥ 18.5 kg/m2 to < 25 kg/m2 = normal weight; from ≥ 25 kg/m2 to < 30 kg/m2 = overweight; and ≥ 30 kg/m2 = obese. Subjects were asked about their sleep routines and light exposure on workdays and work-free days using the Munich Chronotype Questionnaire (N = 244 included). In addition, we analyzed actimetry data from 121 participants with seven consecutive days of recordings. Living in more urbanized areas and higher intradaily variability (IV) of activity-rest rhythms were associated with an increased risk of belonging to the overweight or obese group, when controlling for age and sex. These findings are consistent with preclinical data and point to potential strategies in obesity prevention and promotion of healthy metabolic profiles.

Genome-wide identification of CCT genes in wheat (Triticum aestivum L.) and their expression analysis during vernalization

Numerous CCT genes are known to regulate various biological processes, such as circadian rhythm regulation, flowering, light signaling, plant development, and stress resistance. The CCT gene family has been characterized in many plants but remains unknown in the major cereal wheat (Triticum aestivum L.). Extended exposure to low temperature (vernalization) is necessary for winter wheat to flower successfully. VERNALIZATION2 (VRN2), a specific CCT-containing gene, has been proved to be strongly associated with vernalization in winter wheat. Mutation of all VRN2 copies in three subgenomes results in the eliminated demands of low temperature in flowering. However, no other CCT genes have been reported to be associated with vernalization to date. The present study screened CCT genes in the whole wheat genome, and preliminarily identified the vernalization related CCT genes through expression analysis. 127 CCT genes were identified in three subgenomes of common wheat through a hidden Markov model-based method. Based on multiple alignment, these genes were grouped into 40 gene clusters, including the duplicated gene clusters TaCMF6 and TaCMF8, each tandemly arranged near the telomere. The phylogenetic analysis classified these genes into eight groups. The transcriptome analysis using leaf tissues collected before, during, and after vernalization revealed 49 upregulated and 31 downregulated CCT genes during vernalization, further validated by quantitative real-time PCR. Among the differentially expressed and well-investigated CCT gene clusters analyzed in this study, TaCMF11, TaCO18, TaPRR95, TaCMF6, and TaCO16 were induced during vernalization but decreased immediately after vernalization, while TaCO1, TaCO15, TaCO2, TaCMF8, and TaPPD1 were stably suppressed during and after vernalization. These data imply that some vernalization related CCT genes other than VRN2 may exist in wheat. This study improves our understanding of CCT genes and provides a foundation for further research on CCT genes related to vernalization in wheat.

Molecular determinants of response kinetics of mouse M1 intrinsically-photosensitive retinal ganglion cells

Intrinsically-photosensitive retinal ganglion cells (ipRGCs) are non-rod/non-cone retinal photoreceptors expressing the visual pigment, melanopsin, to detect ambient irradiance for various non-image-forming visual functions. The M1-subtype, amongst the best studied, mediates primarily circadian photoentrainment and pupillary light reflex. Their intrinsic light responses are more prolonged than those of rods and cones even at the single-photon level, in accordance with the typically slower time course of non-image-forming vision. The short (OPN4S) and long (OPN4L) alternatively-spliced forms of melanopsin proteins are both present in M1-ipRGCs, but their functional difference is unclear. We have examined this point by genetically removing the Opn4 gene (Opn4−/−) in mouse and re-expressing either OPN4S or OPN4L singly in Opn4−/− mice by using adeno-associated virus, but found no obvious difference in their intrinsic dim-flash responses. Previous studies have indicated that two dominant slow steps in M1-ipRGC phototransduction dictate these cells’ intrinsic dim-flash-response kinetics, with time constants (τ1 and τ2) at room temperature of ~ 2 s and ~ 20 s, respectively. Here we found that melanopsin inactivation by phosphorylation or by β-arrestins may not be one of these two steps, because their genetic disruptions did not prolong the two time constants or affect the response waveform. Disruption of GAP (GTPase-Activating-Protein) activity on the effector enzyme, PLCβ4, in M1-ipRGC phototransduction to slow down G-protein deactivation also did not prolong the response decay, but caused its rising phase to become slightly sigmoidal by giving rise to a third time constant, τ3, of ~ 2 s (room temperature). This last observation suggests that GAP-mediated G-protein deactivation does partake in the flash-response termination, although normally with a time constant too short to be visible in the response waveform.