Crickets in the spotlight: exploring the impact of light on circadian behavior.

Crickets serve as a well-established model organism in biological research spanning various fields, such as behavior, physiology, neurobiology, and ecology. Cricket circadian behavior was first reported over a century ago and prompted a wealth of studies delving into their chronobiology. Circadian rhythms have been described in relation to fundamental cricket behaviors, encompassing stridulation and locomotion, but also in hormonal secretion and gene expression. Here we review how changes in illumination patterns and light intensity differentially impact the different cricket behaviors as well as circadian gene expression. We further describe the cricket's circadian pacemaker. Ample anatomical manipulations support the location of a major circadian pacemaker in the cricket optic lobes and another in the central brain, possibly interconnected via signaling of the neuropeptide PDF. The cricket circadian machinery comprises a molecular cascade based on two major transcriptional/translational negative feedback loops, deviating somewhat from the canonical model of Drosophila and emphasizing the significance of exploring alternative models. Finally, the nocturnal nature of crickets has provided a unique avenue for investigating the repercussions of artificial light at night on cricket behavior and ecology, underscoring the critical role played by natural light cycles in synchronizing cricket behaviors and populations, further supporting the use of the cricket model in the study of the effects of light on insects. Some gaps in our knowledge and challenges for future studies are discussed.

Transcriptional Response of Circadian Clock Genes to an 'Artificial Light at Night' Pulse in the Cricket Gryllus bimaculatus.

Light is the major signal entraining the circadian clock that regulates physiological and behavioral rhythms in most organisms, including insects. Artificial light at night (ALAN) disrupts the natural light-dark cycle and negatively impacts animals at various levels. We simulated ALAN using dim light stimuli and tested their impact on gene expression in the cricket Gryllus bimaculatus, a model of insect physiology and chronobiology. At night, adult light-dark-regime-raised crickets were exposed for 30 min to a light pulse of 2-40 lx. The relative expression of five circadian-clock-associated genes was compared using qPCR. A dim ALAN pulse elicited tissue-dependent differential expression in some of these genes. The strongest effect was observed in the brain and in the optic lobe, the cricket's circadian pacemaker. The expression of opsin-Long Wave (opLW) was upregulated, as well as cryptochrome1-2 (cry) and period (per). Our findings demonstrate that even a dim ALAN exposure may affect insects at the molecular level, underscoring the impact of ALAN on the circadian clock system.

Lifelong exposure to artificial light at night impacts stridulation and locomotion activity patterns in the cricket Gryllus bimaculatus.

Living organisms experience a worldwide continuous increase in artificial light at night (ALAN), negatively affecting their behaviour. The field cricket, an established model in physiology and behaviour, can provide insights into the effect of ALAN on insect behaviour. The stridulation and locomotion patterns of adult male crickets reared under different lifelong ALAN intensities were monitored simultaneously for five consecutive days in custom-made anechoic chambers. Daily activity periods and acrophases were compared between the experimental groups. Control crickets exhibited a robust rhythm, stridulating at night and demonstrating locomotor activity during the day. By contrast, ALAN affected both the relative level and timing of the crickets' nocturnal and diurnal activity. ALAN induced free-running patterns, manifested in significant changes in the median and variance of the activity periods, and even arrhythmic behaviour. The magnitude of disruption was light intensity dependent, revealing an increase in the difference between the activity periods calculated for stridulation and locomotion in the same individual. This finding may indicate the existence of two peripheral clocks. Our results demonstrate that ecologically relevant ALAN intensities affect crickets' behavioural patterns, and may lead to decoupling of locomotion and stridulation behaviours at the individual level, and to loss of synchronization at the population level.

Artificial Light at Night Increases Recruitment of New Neurons and Differentially Affects Various Brain Regions in Female Zebra Finches.

Despite growing evidence that demonstrate adverse effects of artificial light at night (ALAN) on many species, relatively little is known regarding its effects on brain plasticity in birds. We recently showed that although ALAN increases cell proliferation in brains of birds, neuronal densities in two brain regions decreased, indicating neuronal death, which might be due to mortality of newly produced neurons or of existing ones. Therefore, in the present study we studied the effect of long-term ALAN on the recruitment of newborn neurons into their target regions in the brain. Accordingly, we exposed zebra finches (Taeniopygia guttata) to 5 lux ALAN, and analysed new neuronal recruitment and total neuronal densities in several brain regions. We found that ALAN increased neuronal recruitment, possibly as a compensatory response to ALAN-induced neuronal death, and/or due to increased nocturnal locomotor activity caused by sleep disruption. Moreover, ALAN also had a differential temporal effect on neuronal densities, because hippocampus was more sensitive to ALAN and its neuronal densities were more affected than in other brain regions. Nocturnal melatonin levels under ALAN were significantly lower compared to controls, indicating that very low ALAN intensities suppress melatonin not only in nocturnal, but also in diurnal species.

GSK-3ß Inhibition Affects Singing Behavior and Neurogenesis in Adult Songbirds.

GSK-3 (glycogen synthase kinase-3) is a serine/threonine kinase which is a critical regulator in neuronal signaling, cognition, and behavior. We have previously shown that unlike other vertebrates that harbor both α and ß GSK-3 genes, the α gene is missing in birds. Therefore, birds can be used as a new animal model to study the roles of GSK-3ß in behavior and in regulating adult neurogenesis. In the present study, we inhibited GSK-3ß in brains of adult male zebra finches (Taeniopygia guttata) and accordingly investigated how this inhibition affects behavior and cell proliferation. Our results show that GSK-3 inhibition: (1) affects specific aspects of singing behavior, which might be related to social interactions in birds, and (2) differentially affects cell proliferation in various parts of the ventricular zone. Taken together, our study demonstrates a role of GSK-3ß in regulating singing behavior and neuronal proliferation in birds and highlights the importance of GSK-3ß in modulating cognitive abilities as well as social behavior.

When night becomes day: Artificial light at night alters insect behavior under semi-natural conditions.

Light is the most important Zeitgeber for temporal synchronization in nature. Artificial light at night (ALAN) disrupts the natural light-dark rhythmicity and thus negatively affects animal behavior. However, to date, ALAN research has been mostly conducted under laboratory conditions in this context. Here, we used the field cricket, Gryllus bimaculatus, to investigate the effect of ALAN on insect behavior under semi-natural conditions, i.e., under shaded natural lighting conditions, natural temperature and soundscape. Male crickets were placed individually in outdoor enclosures and exposed to ALAN conditions ranging from <0.01 to 1500 lx intensity. The crickets' stridulation behavior was recorded for 14 consecutive days and nights and their daily activity patterns were analysed. ALAN impaired the crickets' stridulation rhythm, evoking a change in the crickets' naturally synchronized daily activity period. This was manifested by a light-intensity-dependent increase in the proportion of insects demonstrating an intrinsic circadian rhythm (free-run behavior). This also resulted in a change in the population's median activity cycle period. These ALAN-induced effects occurred despite the crickets' exposure to almost natural conditions. Our findings provide further validity to our previous studies on ALAN conducted under lab conditions and establish the deleterious impacts of ALAN on animal behavioral patterns. TEASER: Artificial light at night alters cricket behavior and desynchronizes their stridulation even under near-natural conditions.

Exposure to a nocturnal light pulse simultaneously and differentially affects stridulation and locomotion behaviors in crickets.

It is crucial for living organisms to be in synchrony with their environment and to anticipate circadian and annual changes. The circadian clock is responsible for entraining organisms' activity to the day-night rhythmicity. Artificial light at night (ALAN) was shown to obstruct the natural light cycle, leading to desynchronized behavioral patterns. Our knowledge of the mechanisms behind these adverse effects of ALAN, however, is far from complete. Here we monitored the stridulation and locomotion behavior of male field crickets (Gryllus bimaculatus), raised under light:dark conditions, before, during, and after exposure to a nocturnal 3-h pulse of different ALAN intensities. The experimental insects were then placed under a constant light regime (of different intensities); their behavior was continuously monitored; and the period of their daily activity rhythms was calculated. The light pulse treatment induced a simultaneous negative (suppressing stridulation) and positive (inducing locomotion) effect, manifested in significant changes in the average level of the specific activity on the night of the pulse compared to the preceding and the following nights. The transition to constant light conditions led to significant changes in the period of the circadian rhythms. Both effects were light-intensity-dependent, indicating the importance of dark nights for both individual and population synchronization.

GSK-3ß Inhibition in Birds Affects Social Behavior and Increases Motor Activity.

Glycogen synthase kinase-3 (GSK-3) is a highly conserved serine/threonine protein kinase that plays a central role in a wide variety of cellular processes, cognition and behaviour. In a previous study we showed that its α and ß isozymes are highly conserved in vertebrates, however the α gene is missing in birds. This selective loss offers a unique opportunity to study the role of GSK-3ß independently. Accordingly, in the present study we aimed to investigate the role of GSK-3ß in social behaviour, motivation, and motor activity in zebra finches (Taeniopygia guttata). We did that by selective inhibition of GSK-3ß and by using tests that were specifically designed in our laboratory. Our results show that GSK-3ß inhibition: 1) Affected social recognition, because the treated birds tended to move closer towards a stranger, unlike the control birds that stood closer to a familiar bird. 2) Caused the treated birds to spend more time in the more middle parts of the cage compared to controls, a behaviour that might indicate anxiety. 3) As the experiment progressed, the treated birds took less time to make a decision where to stand in the cage compared to controls, suggesting an effect on decision-making. 4) Increased in the motor activity of the treated birds compared to the controls, which can be regarded as hyperactivity. 5) Caused the treated birds to pass through a barrier in order to join their flock members faster compared to controls, and regardless of the increase in the level of difficulty, possibly suggesting increased motivation. Our study calls for further investigation, because GSK-3 is well acknowledged as a central player in regulating mood behaviour, cognitive functions, and neuronal viability. Therefore, studying its impact on normal behaviour as we did in the current study, unlike most studies that were done in diseases models, can advance our understanding regarding GSK-3 various roles and can contribute to the discovery and development of effective treatments to repair cognition and behaviour.

Birds as a model to study adult neurogenesis: bridging evolutionary, comparative and neuroethological approaches.

During the last few decades, evidence has demonstrated that adult neurogenesis is a well-preserved feature throughout the animal kingdom. In birds, ongoing neuronal addition occurs rather broadly, to a number of brain regions. This review describes adult avian neurogenesis and neuronal recruitment, discusses factors that regulate these processes, and touches upon the question of their genetic control. Several attributes make birds an extremely advantageous model to study neurogenesis. First, song learning exhibits seasonal variation that is associated with seasonal variation in neuronal turnover in some song control brain nuclei, which seems to be regulated via adult neurogenesis. Second, food-caching birds naturally use memory-dependent behavior in learning the locations of thousands of food caches scattered over their home ranges. In comparison with other birds, food-caching species have relatively enlarged hippocampi with more neurons and intense neurogenesis, which appears to be related to spatial learning. Finally, migratory behavior and naturally occurring social systems in birds also provide opportunities to investigate neurogenesis. This diversity of naturally occurring memory-based behaviors, combined with the fact that birds can be studied both in the wild and in the laboratory, make them ideal for investigation of neural processes underlying learning. This can be done by using various approaches, from evolutionary and comparative to neuroethological and molecular. Finally, we connect the avian arena to a broader view by providing a brief comparative and evolutionary overview of adult neurogenesis and by discussing the possible functional role of the new neurons. We conclude by indicating future directions and possible medical applications.

Evidence That Artificial Light at Night Induces Structure-Specific Changes in Brain Plasticity in a Diurnal Bird.

We recently reported that artificial light at night (ALAN), at ecologically relevant intensities (1.5, 5 lux), increases cell proliferation in the ventricular zone and recruitment of new neurons in several forebrain regions of female zebra finches (Taeniopygia guttata), along with a decrease of total neuronal densities in some of these regions (indicating possible neuronal death). In the present study, we exposed male zebra finches to the same ALAN intensities, treated them with 5'-bromo-2'-deoxyuridine, quantified cell proliferation and neuronal recruitment in several forebrain regions, and compared them to controls that were kept under dark nights. ALAN increased cell proliferation in the ventricular zone, similar to our previous findings in females. We also found, for the first time, that ALAN increased new neuronal recruitment in HVC and Area X, which are part of the song system in the brain and are male-specific. In other brain regions, such as the medial striatum, nidopallium caudale, and hippocampus, we recorded an increased neuronal recruitment only in the medial striatum (unlike our previous findings in females), and relative to the controls this increase was less prominent than in females. Moreover, the effect of ALAN duration on total neuronal densities in the studied regions varied between the sexes, supporting the suggestion that males are more resilient to ALAN than females. Suppression of nocturnal melatonin levels after ALAN exhibited a light intensity-dependent decrease in males in contrast to females, another indication that males might be less affected by ALAN. Taken together, our study emphasizes the importance of studying both sexes when considering ALAN effects on brain plasticity.

Artificial light at night affects brain plasticity and melatonin in birds.

Artificial light at night (ALAN), which disrupts the daily cycle of light, has vast biological impacts on all organisms, and is also associated with several health problems. The few existing studies on neuronal plasticity and cognitive functions in mammals indicate that a disruption of the circadian cycle impairs learning and memory and suppresses neurogenesis. However, nothing is known about the effect of ALAN on neuronal plasticity in birds. To this end, zebra finches (Taeniopygia guttata) were exposed to ecologically relevant ALAN intensities (0.5, 1.5 and 5 lx), treated with BrdU to quantify cell proliferation in their ventricular zone (VZ), and compared to controls that were kept under dark nights. We found, in our diurnal birds, that ALAN significantly increased cell proliferation in the VZ. However, neuronal densities in two brain regions decreased under ALAN, suggesting neuronal death. In addition, ALAN suppressed nocturnal melatonin production in a dose-dependent manner, and might also increase body mass. Taken together, our findings add to the notion of the deleterious effect of ALAN.

The relationship between nature of social change, age, and position of new neurons and their survival in adult zebra finch brain.

Some kinds of neurons are spontaneously recruited in the intact, healthy adult brain, but the variables that affect their survival are not always clear. We show that in caudal nidopallium of adult male zebra finches, the rostrocaudal position of newly recruited neurons, their age (1 vs 3 months), and the nature of social change (complex vs simple) after the neurons were born affect their survival. Greater social complexity promoted the survival of younger new neurons, and the demise of older ones; a less marked social change promoted the survival of older new neurons. These effects were position dependent. We suggest that functional correlations between new neuron recruitment/survival and its inferred benefit to the animal might be better perceived when taking into account the position of cells, their age at the time of life style changes, and the nature and magnitude of the life style change.

Frequent summer nuptial flights of ants provide a primary food source for bats.

In many ant species, nuptial flight tends to be short in time and assumed to be synchronous across a large area. Here, we report that, in the upper Jordan Valley, northern Israel, massive nuptial flights of Carpenter ants (Camponotus sp.) occur frequently throughout the summer, and their alates form up to 90% of the diet of the greater mouse-tailed bat (Rhinopoma microphyllum) during this period. This fat and protein-rich diet enables female bats to lactate during summer, and the large amount of fat that both sexes accumulate may serve as an energy source for their following winter hibernation and posthibernation mating in early spring (March-April). We suggest that the annual movement of these bats to the Mediterranean region of Israel may have evolved in order to enable them to exploit the extremely nutritious forms of ant alates when the bats' energetic demands are highest.

Interactions between environmental changes and brain plasticity in birds.

Neurogenesis and neuronal recruitment occur in many vertebrates, including humans. Most of the new neurons die before reaching their destination. Those which survive migrate to various brain regions, replace older ones and connect to existing circuits. Evidence suggests that this replacement is related to acquisition of new information. Therefore, neuronal replacement can be seen as a form of brain plasticity that enables organisms to adjust to environmental changes. However, direct evidence of a causal link between replacement and learning remains elusive. Our hypothesis is that increased neuronal recruitment is associated with increase in memory load. Moreover, since neuronal recruitment is part of a turnover process, we assume that the same conditions that favor survival of some neurons induce the death of others. I present studies that investigated the effect of various behaviors and environmental conditions (food-hoarding, social change, reproductive cycle) on neuronal recruitment and survival in adult avian brains, and discuss how these phenomena relate to the life of animals. I offer a frame and rationale for comparing neuronal replacement in the adult brain, in order to uncover the pressures, rules, and mechanisms that govern its constant rejuvenation. The review emphasizes the importance of using various approaches (behavioral, anatomical, cellular and hormonal) in neuroethological research, and the need to study natural populations, in order to fully understand how neurogenesis and neuronal replacement contribute to life of animals. Finally, the review indicates to future directions and ends with the hope that a better understanding of adult neuronal replacement will lead to medical applications.

The effect of social environment on singing behavior in the zebra finch (Taeniopygia guttata) and its implication for neuronal recruitment.

Previous studies found that complex social environment increases new neuronal recruitment in brains of adult male zebra finches, in comparison with exposure to a simple social environment. These experiments could not determine, however, whether this increase was due to greater amounts of auditory input (amount of auditory information the male is exposed to), or auditory output (amount of song it produces). To answer this question, we raised male zebra finches to adulthood in a controlled environment, and were then exposed them to either a single unfamiliar female (simple social environment) or to 45 unfamiliar zebra finches of both sexes (complex social environment). Their singing behavior was monitored in these new social environments. Birds which were exposed to a simple social environment sang significantly more than birds which were exposed to a complex social environment. This supports the hypothesis that increased neuronal recruitment in birds exposed to a complex social environment correlates with processing and storing of auditory input, and not with song produced by the bird.

Questioning Seasonality of Neuronal Plasticity in the Adult Avian Brain.

To date, studies that reported seasonal patterns of adult neurogenesis and neuronal recruitment have correlated them to seasonal behaviors as the cause or as a consequence of neuronal changes. The aim of our study was to test this correlation, and to investigate whether there is a seasonal pattern of new neuronal recruitment that is not correlated to behavior. To do this, we used adult female zebra finches (songbirds that are not seasonal breeders), kept them under constant social, behavioral, and spatial environments, and compared neuronal recruitment in their brains during two seasons, under natural and laboratory conditions. Under natural conditions, no significant differences were found in the pattern of new neuronal recruitment across seasons. However, under artificial indoor conditions that imitated the natural conditions, higher neuronal recruitment occurred in late summer (August) compared to early spring (February). Moreover, our data indicate that "mixing" temperature and day length significantly reduces new neuronal recruitment, demonstrating the importance of the natural combination of temperature and day length. Taken together, our findings show, for the first time, that neuroplasticity changes under natural vs. artificial conditions, and demonstrate the importance of both laboratory and field experiments when looking at complex biological systems.

Exploring the Relationship between Brain Plasticity, Migratory Lifestyle, and Social Structure in Birds.

Studies in Passerines have found that migrating species recruit more new neurons into brain regions that process spatial information, compared with resident species. This was explained by the greater exposure of migrants to spatial information, indicating that this phenomenon enables enhanced navigational abilities. The aim of the current study was to test this hypothesis in another order-the Columbiformes - using two closely-related dove species-the migrant turtle-dove (Streptopelia turtur) and the resident laughing dove (S. senegalensis), during spring, summer, and autumn. Wild birds were caught, treated with BrdU, and sacrificed 5 weeks later. New neurons were recorded in the hyperpallium apicale, hippocampus and nidopallium caudolaterale regions. We found that in doves, unlike passerines, neuronal recruitment was lower in brains of the migratory species compared with the resident one. This might be due to the high sociality of doves, which forage and migrate in flocks, and therefore can rely on communal spatial knowledge that might enable a reduction in individual navigation efforts. This, in turn, might enable reduced levels of neuronal recruitment. Additionally, we found that unlike in passerines, seasonality does not affect neuronal recruitment in doves. This might be due to their non-territorial and explorative behavior, which exposes them to substantial spatial information all year round. Finally, we discuss the differences in neuronal recruitment between Columbiformes and Passeriformes and their possible evolutionary explanations. Our study emphasizes the need to further investigate this phenomenon in other avian orders and in additional species.

Social and spatial changes induce multiple survival regimes for new neurons in two regions of the adult brain: An anatomical representation of time?

Male zebra finches reared in family groups were housed initially in small indoors cages with three other companions. At 4-5 months of age these birds were treated with [(3)H]-thymidine and then placed in large outdoors aviaries by themselves or with other zebra finches. Counts of new neurons were made 40, 60 and 150 days after the change in housing. Recruitment of new neurons in nidopallium caudale (NC) was higher than in the hippocampal complex (HC); but in both brain regions it was higher in communally housed birds than in birds housed singly, suggesting that the complexity of the social setting affects new neuron survival. In addition, the new neurons lived longer in rostral NC than in its caudal counterpart, and neuronal turnover was faster and more significant in NC than in HC. Albeit indirect, this may be the first suggestion that different parts of the brain upgrade memories at different time intervals, yielding an anatomical representation of time.

Theta EEG neurofeedback benefits early consolidation of motor sequence learning.

Procedural learning is subject to consolidation processes believed to depend on the modulation of functional connections involved in representing the acquired skill. While sleep provides the most commonly studied framework for such consolidation processes, posttraining modulation of oscillatory brain activity may also impact on plasticity processes. Under the hypothesis that consolidation of motor learning is associated with theta band activity, we used EEG neurofeedback (NFB) to enable participants to selectively increase either theta or beta power in their EEG spectra following the acquisition phase of motor sequence learning. We tested performance on a motor task before and after training, right after the NFB session to assess immediate NFB effects, 1 day after NFB to assess interaction between NFB effects and overnight sleep-dependent stabilization, and 1 week after the initial session, to assess the effects of NFB on long-term stabilization of motor training. We also explored the extent of the influence of single-electrode NFB on EEG recorded across the scalp. Results revealed a significantly greater improvement in performance immediately after NFB in the theta group than in the beta group. This effect continued for testing up to 1 week following training. Across participants, post-NFB improvement correlated positively with theta/beta ratio change achieved during NFB. Additionally, NFB was found to cause widespread band-power modulation beyond the electrode used for feedback. Thus, upregulating postlearning theta power may yield contributions to the immediate performance and subsequent consolidation of an acquired motor skill.