Clock gene oscillation in the copepod Calanus finmarchicus in the Arctic: the effect of latitude and season

Life evolved under the permanent influence of environmental cycles, the most prominent being the daily light/dark cycle, caused by the earth’s rotation about its axis. As a consequence almost all organisms have developed biological clocks that allow them to anticipate cyclic changes in the environment and thus to adjust their behavior and physiology accordingly. A biological clock has also been identified in the copepod Calanus finmarchicus, where it is thought to underpin diel and seasonal rhythms in behavior and physiology. C. finmarchicus plays a central role in sustaining the food webs of the North Atlantic and Subarctic regions, however, climate change induced latitudinal range shifts have introduced C. finmarchicus into the Arctic region, where it experiences extreme light conditions, with almost constant light throughout the Summer months and constant darkness in Winter. Therefore, this thesis centers on the question whether the C. finmarchichus clock stays functional throughout the High Arctic Summer, when diel fluctuations in light reach a minimum. Net based 24 h samplings have been conducted at two stations along a latitudinal gradient from the southern Barents Sea (74.5 °N, 30 °E) to the Nansen Basin (82.56 °N, 30.85 °E) north of Svalbard, within 9 days of the Summer Solstice 2018. Further, temporal expression patterns of clock genes have been analyzed and the behavioral activity of individual C. finmarchicus has been assessed in onboard laboratory experiments. Results from gene expression analysis show significant rhythmic oscillations in a number of core clock genes in wild caught C. finmarchicus, suggesting a functional and synchronized endogenous clock during periods of minimal fluctuations in light intensity. Further, a period shortening could be observed in several clock genes at the northern station associated with lower diel oscillations in light properties. Results from behavioral experiments indicate overall low rhythmic behavioral activity during Summer in the High Arctic. The findings from this study are further discussed in the context of seasonal timing, concluding that the circadian clock likely stays functional throughout the whole active phase at high latitudes, including periods of Midnight Sun. This may further point out the importance of the circadian clock as a tool to track the progression of the season and help to time seasonal events, which is of fundamental importance for C. finmarchicus to survive in the extreme conditions of the Arctic

Timing requires the right amount and type of light

The quantity of UVA/deep violet light varies seasonally and affects locomotor activity in a marine annelid, providing cues for phenology in addition to those provided by change in photoperiod

Evidence for oscillating circadian clock genes in the copepod Calanus finmarchicus during the summer solstice in the high Arctic

The circadian clock provides a mechanism for anticipating environmental cycles and is synchronized by temporal cues such as daily light/dark cycle or photoperiod. However, the Arctic environment is characterized by several months of Midnight Sun when the sun is continuously above the horizon and where sea ice further attenuates photoperiod. To test if the oscillations of circadian clock genes remain in synchrony with subtle environmental changes, we sampled the copepod Calanus finmarchicus, a key zooplankter in the north Atlantic, to determine in situ daily circadian clock gene expression near the summer solstice at a southern (74.5° N) sea ice-free and a northern (82.5° N) sea ice-covered station. Results revealed significant oscillation of genes at both stations, indicating the persistence of the clock at this time. While copepods from the southern station showed oscillations in the daily range, those from the northern station exhibited an increase in ultradian oscillations. We suggest that in C. finmarchicus, even small daily changes of solar altitude seem to be sufficient to entrain the circadian clock and propose that at very high latitudes, in under-ice ecosystems, tidal cues may be used as an additional entrainment cue

Calanus finmarchicus seasonal cycle and diapause in relation to gene expression, physiology, and endogenous clocks: Calanus finmarchicus seasonal rhythmicity

The copepod Calanus finmarchicus plays a crucial role in the north Atlantic food web. Its seasonal life cycle involves reproduction and development in surface waters before overwintering in diapause at depth. Although diapause has been studied for more than a century, the factors responsible for the initiation and termination of it are still unclear. Endogenous clocks have been identified as potent tools for photoperiod measurement and seasonal rhythmicity in many terrestrial species, but knowledge of these remains scarce in the marine realm. Focusing on the dominant CV copepodid stage, we sampled a population of C. finmarchicus from a Scottish sea loch to characterize population dynamics, several physiological parameters, and diel and seasonal expression rhythms of 35 genes representing different metabolic pathways, including the circadian clock machinery. This generated a detailed overview of the seasonal cycle of C. finmarchicus including the most extensive field dataset on circadian clock gene expression in a marine species to date. Gene expression patterns revealed distinct gene clusters upregulated at different phases of the copepod's seasonal cycle. While diel clock cycling was restricted to the active spring/summer phase, many clock genes exhibited the highest expression during diapause. Our results provide new insights into diapause on physiological and genetic levels. We suggest that photoperiod, in interaction with internal and external factors (lipid content, temperature, food availability) and the endogenous clock mechanism, plays an important role in the timing of diapause in C. finmarchicus

Circadian Clock Involvement in Zooplankton Diel Vertical Migration

Biological clocks are a ubiquitous ancient and adaptive mechanism enabling organisms to anticipate environmental cycles and to regulate behavioral and physiological processes accordingly [1]. Although terrestrial circadian clocks are well understood, knowledge of clocks in marine organisms is still very limited [2, 3, 4, 5]. This is particularly true for abundant species displaying large-scale rhythms like diel vertical migration (DVM) that contribute significantly to shaping their respective ecosystems [6]. Here we describe exogenous cycles and endogenous rhythms associated with DVM of the ecologically important and highly abundant planktic copepod Calanus finmarchicus. In the laboratory, C. finmarchicus shows circadian rhythms of DVM, metabolism, and most core circadian clock genes (clock, period1, period2, timeless, cryptochrome2, and clockwork orange). Most of these genes also cycle in animals assessed in the wild, though expression is less rhythmic at depth (50–140 m) relative to shallow-caught animals (0–50 m). Further, peak expressions of clock genes generally occurred at either sunset or sunrise, coinciding with peak migration times. Including one of the first field investigations of clock genes in a marine species [5, 7], this study couples clock gene measurements with laboratory and field data on DVM. While the mechanistic connection remains elusive, our results imply a high degree of causality between clock gene expression and one of the planet’s largest daily migrations of biomass. We thus suggest that circadian clocks increase zooplankton fitness by optimizing the temporal trade-off between feeding and predator avoidance, especially when environmental drivers are weak or absent [8]

Calanus finmarchicus diel and seasonal rhythmicity in relation to endogenous timing under extreme polar photoperiods

Changing environmental conditions cause poleward distribution shifts in many marine organisms including the northern Atlantic key zooplankton species Calanus finmarchicus. The copepod has diel cycles of vertical migration and feeding, a seasonal life cycle with diapause in winter and a functioning circadian clock. Endogenous clock mechanisms control various aspects of rhythmic life and are heavily influenced by environmental light conditions. Here we explore how the extreme seasonal change in photoperiod (day length) in a high Arctic fjord affects circadian clock functioning as well as diel and seasonal cycles in C. finmarchicus. Expression of clock genes was measured in the active life phase at the end of midnight sun, in early diapause when photoperiod was ~12 h, and in late diapause during the polar night. While the clock maintained diel rhythmicity under extremely long photoperiods, it became arrhythmic during diapause. This was probably not due to a lack of light but was related to the physiological state of diapause. Seasonal expression analyses of 35 genes show distinct patterns for each investigated life phase. C. finmarchicus is able to maintain diel clock rhythmicity at photoperiods close to 24 h, and it is discussed how this may be related to the nature of the marine environment. The work also evaluates the potential negative consequences of rigid clock-based seasonal timing in a polar environment exposed to climate change and with high interannual variability

Residual volatile anesthetics after workstation preparation and activated charcoal filtration

Background Volatile anesthetics potentially trigger malignant hyperthermia crises in susceptible patients. We therefore aimed to identify preparation procedures for the Draeger Primus that minimize residual concentrations of desflurane and sevoflurane with and without activated charcoal filtration. Methods A Draeger Primus test workstation was primed with 7% desflurane or 2.5% sevoflurane for 2 hours. Residual anesthetic concentrations were evaluated with five preparation procedures, three fresh gas flow rates, and three distinct applications of activated charcoal filters. Finally, non‐exchangeable and autoclaved parts of the workstation were tested for residual emission of volatile anesthetics. Concentrations were measured by multicapillary column–ion mobility spectrometry with limits of detection/quantification being <1 part per billion (ppb) for desflurane and <2.5 ppb for sevoflurane. Results The best preparation procedure included a flushing period of 10 minutes between removal and replacement of all parts of the ventilator circuit which immediately produced residual concentrations <5 ppm. A fresh gas flow of 10 L/minute reduced residual concentration as effectively as 18 L/minute, whereas flows of 1 or 5 L/minute slowed washout. Use of activated charcoal filters immediately reduced and maintained residual concentrations <5 ppm for up to 24 hours irrespective of previous workstation preparation. The fresh gas hose, circle system, and ventilator diaphragm emitted traces of volatile anesthetics. Conclusion In elective cases, presumably safe concentrations can be obtained by a 10‐minute flush at ≥10 L/minute between removal and replacement all components of the airway circuit. For emergencies, we recommend using an activated charcoal filter

Daily transcriptomes of the copepod Calanus finmarchicus during the summer solstice at high Arctic latitudes

The zooplankter Calanus finmarchicus is a member of the so-called “Calanus Complex”, a group of copepods that constitutes a key element of the Arctic polar marine ecosystem, providing a crucial link between primary production and higher trophic levels. Climate change induces the shift of C. finmarchicus to higher latitudes with currently unknown impacts on its endogenous timing. Here we generated a daily transcriptome of C. finmarchicus at two high Arctic stations, during the more extreme time of Midnight Sun, the summer solstice. While the southern station (74.5 °N) was sea ice-free, the northern one (82.5 °N) was sea ice-covered. The mRNAs of the 42 samples have been sequenced with an average of 126 ± 5 million reads (mean ± SE) per sample, and aligned to the reference transcriptome. We detail the quality assessment of the datasets and the complete annotation procedure, providing the possibility to investigate daily gene expression of this ecologically important species at high Arctic latitudes, and to compare gene expression according to latitude and sea ice-coverage

Widely rhythmic transcriptome in Calanus finmarchicus during the high Arctic summer solstice period

Solar light/dark cycles and seasonal photoperiods underpin daily and annual rhythms of life on Earth. Yet, the Arctic is characterized by severalmonths of permanent illumination (‘‘midnight sun’’). To determine the persistence of 24h rhythms during the midnight sun, we investigated transcriptomic dynamics in the copepod Calanus finmarchicus during the summer solstice period in the Arctic, with the lowest diel oscillation and the highest altitude of the sun’s position. Here we reveal that in these extreme photic conditions, a widely rhythmic daily transcriptome exists, showing that very weak solar cues are sufficient to entrain organisms. Furthermore, at extremely high latitudes and under sea-ice, gene oscillations become re-organized to include <24h rhythms. Environmental synchronization may therefore be modulated to include non-photic signals (i.e. tidal cycles). The ability of zooplankton to be synchronized by extremely weak diel and potentially tidal cycles, may confer an adaptive temporal reorganization of biological processes at high latitudes

Quantification of Volatile Aldehydes Deriving from In Vitro Lipid Peroxidation in the Breath of Ventilated Patients

Exhaled aliphatic aldehydes were proposed as non-invasive biomarkers to detect increased lipid peroxidation in various diseases. As a prelude to clinical application of the multicapillary column–ion mobility spectrometry for the evaluation of aldehyde exhalation, we, therefore: (1) identified the most abundant volatile aliphatic aldehydes originating from in vitro oxidation of various polyunsaturated fatty acids; (2) evaluated emittance of aldehydes from plastic parts of the breathing circuit; (3) conducted a pilot study for in vivo quantification of exhaled aldehydes in mechanically ventilated patients. Pentanal, hexanal, heptanal, and nonanal were quantifiable in the headspace of oxidizing polyunsaturated fatty acids, with pentanal and hexanal predominating. Plastic parts of the breathing circuit emitted hexanal, octanal, nonanal, and decanal, whereby nonanal and decanal were ubiquitous and pentanal or heptanal not being detected. Only pentanal was quantifiable in breath of mechanically ventilated surgical patients with a mean exhaled concentration of 13 ± 5 ppb. An explorative analysis suggested that pentanal exhalation is associated with mechanical power—a measure for the invasiveness of mechanical ventilation. In conclusion, exhaled pentanal is a promising non-invasive biomarker for lipid peroxidation inducing pathologies, and should be evaluated in future clinical studies, particularly for detection of lung injury