Thesis (Ph.D.) in Biological Sciences--University of Maine, 2009. / Includes vita. Includes bibliographical references (leaves 83-96).

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Anadromous pink salmon undergo several migratory movements between different habitats during their life history. These migrations are accurately timed on a seasonal basis. Annual rhythms or seasonally-timed events may result from interactions between daily rhythms and annual changes in environmental factors. Therefore, knowledge of daily behavioral rhythms in pink salmon may improve our current poor understanding of the seasonal timing of its migrations. Hence, the objective of this study was to investigate, in a seasonal context and mainly under laboratory conditions, diel rhythms of ecologically-relevant behavior in juvenile pink salmon, and their timing mechanisms. Fry emergence from a simulated gravel redd in fresh water was mainly nocturnal below 13°C. Diel emergence timing was synchronized with the onset of night, but was affected by temperature in a non-linear manner. Temperature affected negatively the duration of the intra-gravel alevin stage and the rate of emergence. Nocturnal emergence was considered an anti-predator adaptation. Fry exhibited mainly nocturnal rhythms of swimming activity and of vertical distribution during the first week after emergence. However, a gradual shift from a nocturnal to a diurnal swimming activity rhythm occurred 7 to 13 days after emergence, when wild fish are residing in estuaries and adjacent coastal waters. Coincident with this shift was an increasing tendency of the fry to swimnnear the water surface during the day. This suggested a weakening of their negative phototactic response during this period. Thereafter, the fish usually displayed diurnal rhythms of swimming activity and nocturnal rhythms of vertical distribution. The ontogenetic shift in the phase of the activity rhythm and in photobehavior was considered adaptive for schooling and feeding during the day. Wild fry fed mainly during daylight hours in littoral areas of two marine bays. However, their feeding rhythms varied among study sites. Laboratory experiments showed that hunger level affected fish feeding rate and ration consumed positively. Fish fed continuously on live copepods under idealized laboratory conditions. During a 12-h session they rapidly (< 30 min) filled their stomachs with prey; thereafter, they maintained their stomachs full by feeding at a rate that balanced the rate of evacuation of prey from the stomach. Hence, I concluded that pink salmon have flexible feeding activity rhythms, which may permit opportunistic exploitation of prey, and feed at a relatively low hunger threshold. This feeding strategy may explain in part their relatively high growth rates in nature. During the periods corresponding to their juvenile coastal and pelagic ocean phases, the fish exhibited generally diurnal rhythms of swimming activity and of aggression, and nocturnal rhythms of vertical distribution in response to simulated seasonal photoperiodic and temperature changes. These rhythms were synchronized with the artificial light-dark (LD) cycle throughout most of the year. Some parameters of these rhythms varied on a seasonal basis, but not according to the Aschoff-Wever model. Mean swimming speed, the degree of diurnalism of the swimming activity rhythm, and the timing of the daily peak of the rhythms were affected by daylength. Hence, photoperiod might be an important proximate factor that pink salmon use to time their oceanic migration on a seasonal basis. Some data suggested the existence of an endogenous, circadian activity rhythm, and thus a daily "clock", in pink salmon. However, this remains uncertain. The free-running period of their activity rhythm was not related negatively to constant light intensity, as predicted by the Circadian Rule. The LD cycle affected directly swimming activity (masking), rather than entraining an endogenous circadian system. Since the activity rhythm of pink salmon does not possess a strong endogenous component, it is doubtful that the seasonal timing of its migrations results from interactions between a circadian clock and seasonal changes in environmental factors. However, the flexibility and inter-individual variability of their behavioral rhythms may be adaptive responses to the instability and heterogeneity of the marine environment. / Science, Faculty of / Zoology, Department of / Graduate

The day-to-day physiologies are largely influenced by circadian rhythms. Disruptions of such rhythms are associated with many diseases. Adjusting them to a healthy one can be promising to treat different circadian rhythms disruption associated diseases such as sleep disorders, cardiovascular disorders, metabolic disorders. The regulations underlying the circadian rhythms are much complicated and systematic which may involve thousands of genes. In mammals, these robust circadian rhythms are primarily intended by the concerted molecular interplays, knowingly, transcriptional-translational feedback loops (TTFLs). The collaborative interactions among a large number of genes intend to sustain the TTFLs. It facilitates to generate the primary transcriptional oscillations among the clock genes and genome-wide rhythmic oscillations. The collaborative transcriptional events act as dominant driving forces underpinning such rhythmic expressions. The mode of the transcriptional regulations depends on the concentration of the transcriptional factors (TFs) at the promoter region at a particular time point. The inclusive mechanisms of their regulations and the associated circadian rhythmic outputs across the physiology are not well defined yet. However, temporal recruitment of core-clock proteins, different transcriptional and translational regulators and chromatin modifications are imperative towards a comprehensive understanding of the spatio-temporal regulation of such complex rhythms. Despite many experimental signs of progress about the circadian transcriptional controls, there is still an interesting question remains unexplored that how do these few components belonging to the same molecular architecture are capable to govern such divergent gene expressions? Nevertheless, how they are being regulated and the landscape of their combinatorial regulatory controls have not gained any inclusive attention yet. Thus, a systematic understanding considering all-encompassing circadian TFs and their relational interplay could help us to decode the intricated transcriptional regulatory logics composed by different TFs. Such comprehensive understanding may lead to unleashing their potential to therapeutically modulate the circadian rhythms. Experiments alone are indeed quite challenging to achieve this. Decoding the inclusive transcriptional insights along with multifaceted molecular regulations remained out of reach with prevailing approaches. They are limited by the complexities of more integrative algorithms that accommodate different layers of molecular information quantitatively into a single framework. Studies indicated the knockout of the circadian TFs results in changing the rhythms. And, rescuing them helps to regain the circadian functionality substantially. However, knocking out all possible combinations of circadian TF-genes experimentally is merely very tedious, time-consuming and expensive. And, some essential genes cannot be knocked out. The magnitude of disruption of the circadian rhythmic fluctuations may also vary in disease conditions and even from individual to individual. These are serious concerns which were weakly understood. Due to the lack of advanced quantitative approach, these have remained a great challenge with traditional practices for reversing the disrupted circadian rhythms. Another level of challenge is not only aligning the rhythms but also, having a strong understanding of the directionality of the alignment varying in different clinical contexts is the most crucial. Consequently, a thorough quantitative understanding at the molecular level of the clock control mechanisms is essential. To address these ambiguities, a quantitative understanding of the circadian gene regulation and the molecular interplay among the key regulators are quite important. An alternative yet the operative approach is the reconstruction of transcriptional network with those genes having circadian fluctuations by computational simulations. It may capture a systematic snapshot of such gene regulation network at a dynamic scale. Inferring them is again a complicated task as the large numbers of variables are unknown in the systems. There is also a lack of tools to capture and integrate the dynamic view which is biologically relevant. Virtual knockout experiments leverage in inferring such dynamic transcriptional regulatory networks iteratively and effectively. The molecular machinery underpinning the circadian rhythms possess high-temporal resolutions. Thus, it is also quite challenging to construct the network of those genes under the influence of TTFLs at dynamic scale using existing methods. Most of the prevailing approaches are quite limited by the quantitative understanding of the transcriptional landscape thoroughly. Recently, one of our computational approaches, LogicTRN was proposed for modelling the transcriptional regulatory networks quantitatively. Deploying the high-resolution temporal gene expression data and the TF-DNA binding data, it calculates the TF-DNA binding occupancy, which is a quantitative estimation. It also predicts the all possible combinatorial TF-logics influencing those target genes' regulations. Here, we introduced an extended computational approach based on LogicTRN to decode the quantitative transcriptional regulatory landscapes of circadian genes. We introduced the reconstruction of quantitative transcriptional regulatory networks (qTRNs) for circadian gene regulations using LogicTRN framework. The qTRNs facilitated to discover a wide range of genes exhibiting circadian fluctuations. Their dynamic behaviours and the cis-regulatory logics in the networks were also estimated precisely. Based on quantitative knowledge from qTRNs, we have further developed a method for virtually knocking out the core clock component TFs to estimate the influence to perturb the circadian rhythmic fluctuations at a dynamic scale. Consecutively, the method of single/multiple genes virtual knockout was developed and used to screen the best TF/TFs combination that effectively modulates the circadian rhythmic output at a dynamic scale. They were also ordered by their influence to perturb the circadian fluctuations in the qTRNs. In future, it may indicate a way to target the molecular regulators to therapeutically modulate the circadian period lengths in a specific direction based on an individual's clinical conditions. Our results indicate the reconstruction of highly accurate quantitative regulatory networks for the transcriptional controls of the circadian gene regulation at a dynamic scale. We have also identified the best plausible transcription factors or their combinations those can effectively modulate the circadian rhythms. Of them, the CLOCK and CRY1 double knockout preserve the highest capacity to modulate the circadian rhythm dynamics. Besides, all possible TFs/TF-combinations were ordered in terms of their capacity to influence the qTRNs at dynamic scales. Finally, our quantitative framework offers a quick, robust, and physiologically relevant way to screen and identify the most effective TFs/TF-combinations to modulate circadian rhythms. This foundation may potentially enable us to engineer the molecular regulators underpinning the circadian rhythms. This potentially indicates a clue towards adjusting the circadian rhythmic phases in desired directions depending on clinical requirements

Circadian clocks are vital to the proper functioning of organisms’ internal processes and behavioral outputs and typically have endogenous periods that approximate (within 1-2 hours) the 24-hour solar day. Clocks that deviate significantly from about 24 hours are often associated with metabolic syndromes or other disease states. For instance, organisms with near-24-hour clocks have higher survivorship under 24-h light:dark (LD) cycles than with 22- or 26-hour cycles. Likewise, mutant organisms with 22-hour clocks survive better under 22-h cycles but fare poorly under 24- and 26-h cycles. In other words, organisms suffer if their circadian clocks do not “resonate” with environmental cycles. Organisms fail to synchronize (entrain) their activity with non-resonant LD cycles and this failure typically leads to a number of physiological disruptions. Interestingly, several spider species have endogenous circadian periods that deviate by several hours from the period of the Earth’s solar day. The object of the present study is to investigate whether the phenomenon of circadian resonance also pertains to these atypical spider circadian rhythms. We investigated three spider species, two of which have internal periods (τ) significantly different from 24 hours. Approximately 50 individuals of each species of spider (Frontinella communis: τ=29.05±0.62 hours; Metazygia wittfeldae: τ=22.74±0.24h; and Cyclosa turbinata: τ=18.54±0.28h) were placed into chambers with periods of 19 (9.5:9.5h L:D), 24 (12:12h L:D), or 29 hours (14.5:14.5h L:D). If resonance is pertinent for spiders, we would expect survivorship to decrease in non-resonant LD cycles. Instead, no spider species exhibited decreased longevity in non-resonant L:D cycles. These findings contradict all previous research into circadian resonance and suggest that spiders do not suffer the costs of extreme desynchronization. In a second experiment, 10-11 spiders from each species were placed into infrared activity monitors to determine if their locomotor activity could entrain to (synchronize with) the three different LD cycles. Individuals from all three spider species entrained to all LD period lengths, again in contrast with prior research in other species. These results indicate that spider circadian clocks have highly unusual limits of entrainment and suggest a remarkable level of plasticity in their release from the selective pressure to maintain an internal period of approximately 24 hours.

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Shi, Yujian. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 83-101). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts also in Chinese.

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