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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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All sourcesNASA NTRSDOE OSTI
NASA NTRS1999

Aging and space flight: findings from the University of Pittsburgh

For more than a decade, the Sleep and Chronobiology Center (SCC) at the University of Pittsburgh has received funding from the National Institute on Aging (NIA), the National Institute of Mental Health (NIMH) and the National Aeronautics and Space Administration (NASA) in order to study the sleep and circadian rhythms of healthy older people, as well as the sleep and circadian rhythms of astronauts and cosmonauts. We have always been struck by the strong synergism between the two endeavors. What happens to the sleep and circadian rhythms of people removed from the terrestrial time cues of Earth is in many ways similar to what happens to people who are advancing in years. Most obviously, sleep is shorter and sleep depth is reduced, but there are also more subtle similarities between the two situations, both in circadian rhythms and in sleep, and in the adaptive strategies needed to enhance 24h zeitgebers.

STS-78 Shuttle Project↗
NASA NTRS1996

Subjective alertness rhythms in elderly people

The aim of this study was to evaluate age-related changes in the circadian rhythm of subjective alertness and to explore the circadian mechanisms underlying such changes. Using a visual analogue scale (VAS) instrument, 25 older men and women (71 y and older; 15 female, 10 male) rated their subjective alertness about 7 times per day during 5 baseline days of temporal isolation during which habitual bedtimes and waketimes were enforced. Comparisons were made with 13 middle-aged men (37-52 y) experiencing the same protocol. Advancing age (particularly in the men) resulted in less rhythmic alertness patterns, as indicated by lower amplitudes and less reliability of fitted 24-h sinusoids. This appeared in spite of the absence of any reliable age-related diminution in circadian temperature rhythm amplitude, thus suggesting the effect was not due to SCN weakness per se, but to weakened transduction of SCN output. In a further experiment, involving 36 h of constant wakeful bedrest, differences in the amplitude of the alertness rhythm were observed between 9 older men (79 y+), 7 older women (79 y+), and 17 young controls (9 males, 8 females, 19-28 y) suggesting that with advancing age (particularly in men) there is less rhythmic input into subjective alertness from the endogenous circadian pacemaker. These results may explain some of the nocturnal insomnia and daytime hypersomnia that afflict many elderly people.

NASA Discipline Regulatory Physiology↗
DOE OSTI2022

Simulated Night- Shift Schedule Disrupts the Plasma Lipidome and Reveals Early Markers of Cardiovascular Disease Risk

Introduction: The circadian system coordinates daily rhythms in lipid metabolism, storage and utilization. Disruptions of internal circadian rhythms due to altered sleep/wake schedules, such as in night-shift work, have been implicated in increased risk of cardiovascular disease and metabolic disorders. To determine the impact of a night-shift schedule on the human blood plasma lipidome, an in-laboratory simulated shift work study was conducted. Methods: Fourteen healthy young adults were assigned to 3 days of either a simulated day or night-shift schedule, followed by a 24-h constant routine protocol with fixed environmental conditions, hourly isocaloric snacks, and constant wakefulness to investigate endogenous circadian rhythms. Blood plasma samples collected at 3-h intervals were subjected to untargeted lipidomics analysis. Results: More than 400 lipids were identified and quantified across 21 subclasses. Focusing on lipids with low between-subject variation per shift condition, alterations in the circulating plasma lipidome revealed generally increased mean triglyceride levels and decreased mean phospholipid levels after night-shift relative to day-shift. The circadian rhythms of triglycerides containing odd chain fatty acids peaked earlier during constant routine after night-shift. Regardless of shift condition, triglycerides tended to either peak or be depleted at 16:30 h, with chain-specific differences associated with the direction of change. Discussion: The simulated night-shift schedule was associated with altered temporal patterns in the lipidome. This may be premorbid to the elevated cardiovascular risk that has been found epidemiologically in night-shift workers.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1988

Biological Rhythms, Sleep, and Wakefulness in Prolonged Confinement

The dysynchronization of human circadian rhythms during 7 long-term (2 to 6 months) confinement experiments in temporal isolation in caves was studied. Five subjects abandon the circadian period of sleep and wakefulness (S-W) and spontaneously reach a circabidian S-W cycle (34 to 36 hr waking, 14 to 12 hr sleep) they maintain during weeks. Some subjects reach the 48 hr cycle very quickly (8 to 15 days), others after months. Polygraphic analyses of sleep show that rapid eye movement state (REMS) duration is directly proportional to the total duration of sleep and that the ultradian periodicity of REMS remains constant when S-W cycle is circadian or circabidian. When S-W cycle desynchronizes from circadian to circabidian, REMS and S-4 increase at the expense of stages I-2 and remain in constant relationship with the duration of previous wakefulness period.

M Siffre↗
NASA NTRS1991

Crew factors in flight operations. 8: Factors influencing sleep timing and subjective sleep quality in commercial long-haul flight crews

How flight crews organize their sleep during layovers on long-haul trips is documented. Additionally, environmental and physiological constraints on sleep are examined. In the trips studied, duty periods averaging 10.3 hr alternated with layovers averaging 24.8 hr, which typically included two subject-defined sleep episodes. The circadian system had a greater influence on the timing and duration of first-sleeps than second-sleeps. There was also a preference for sleeping during the local night. The time of falling asleep for second-sleeps was related primarily to the amount of sleep already obtained in the layover, and their duration depended on the amount of time remaining in the layover. For both first- and second-sleeps, sleep durations were longer when subjects fell asleep earlier with respect to the minimum of the circadian temperature cycle. Naps reported during layovers and on the flight deck may be a useful strategy for reducing cumulative sleep loss. The circadian system was not able to synchronize with the rapid series of time-zone shifts. The sleep/wake cycle was forced to adopt a period different from that of the circadian system. Flight and duty time regulations are a means of ensuring that reasonable minimum rest periods are provided. This study clearly documents that there are physiologically and environmentally determined preferred sleep times within a layover. The actual time available for sleep is thus less than the scheduled rest period.

Gander, Philippa H.↗
NASA NTRS1991

Light as a chronobiologic countermeasure for long-duration space operations

Long-duration space missions require adaptation to work-rest schedules which are substantially shifted with respect to earth. Astronauts are expected to work in two-shift operations and the environmental synchronizers (zeitgebers) in a spacecraft differ significantly from those on earth. A study on circadian rhythms, sleep, and performance was conducted by exposing four subjects to 6 deg head-down tilt bedrest (to simulate the effects of the weightless condition) and imposing a 12-h shift (6 h delay per day for two days). Bright light was tested in a cross-over design as a countermeasure for achieving faster resynchronization and regaining stable conditions for sleep and circadian rhythmicity. Data collection included objective sleep recording, temperature, heart rate, and excretion of hormones and electrolytes as well as performance and responses to questionnaires. Even without a shift in the sleep-wake cycle, the sleep quantity, circadian amplitudes and 24 h means decreased in many functions under bedrest conditions. During the shift days, sleepiness and fatigue increased, and alertness decreased. However, sleep quantity was regained, and resynchronization was completed within seven days after the shift for almost all functions, irrespective of whether light was administered during day-time or night-time hours. The time of day of light exposure surprisingly appeared not to have a discriminatory effect on the resynchronization speed under shift and bedrest conditions. The results indicate that simulated weightlessness alters circadian rhythms and sleep, and that schedule changes induce additional physiological disruption with decreased subjective alertness and increased fatigue. Because of their operational implications, these phenomena deserve additional investigation.

Samel, Alexander↗
NASA NTRS1997

Light-Induced Alterations in Striatal Neurochemical Profiles

Much of our present knowledge regarding circadian rhythms and biological activity during space flight has been derived from those missions orbiting the Earth. During space missions, astronauts can become exposed to bright/dark cycles that vary considerably from those that entrain the mammalian biological timing system to the 24-hour cycle found on Earth. As a spacecraft orbits the Earth, the duration of the light/dark period experienced becomes a function of the time it takes to circumnavigate the planet which in turn depends upon the altitude of the craft. Orbiting the Earth at an altitude of 200-800 km provides a light/dark cycle lasting between 80 and 140 minutes, whereas a voyage to the moon or even another planet would provide a light condition of constant light. Currently, little is known regarding the effects of altered light/dark cycles on neurochemical levels within the central nervous system (CNS). Many biochemical, physiological and behavioral phenomena are under circadian control, governed primarily by the hypothalamic suprachiasmatic nucleus. As such, these phenomena are subject to influence by the environmental light/dark cycle. Circadian variations in locomotor and behavioral activities have been correlated to both the environmental light/dark cycle and to dopamine (DA) levels within the CNS. It has been postulated by Martin-Iverson et al. that DA's role in the control of motor activity is subject to modulation by circadian rhythms (CR), environmental lighting and excitatory amino acids (EAAs). In addition, DA and EAA receptor regulated pathways are involved in both the photic entrainment of CR and the control of motor activity. The cellular mechanisms by which DA and EAA-receptor ligands execute these functions, is still unclear. In order to help elucidate these mechanisms, we set out to determine the effects of altered environmental light/dark cycles on CNS neurotransmitter levels. In this study, we focused on the striatum, a region of the brain that receives a number of dopaminergic and glutamatergic input and is known to be involved in the modulation of locomotor and behavioral responses.

Sroufe, Angela E.↗
NASA NTRS1994

The Influence of Sound Cues on the Maintenance of Temporal Organization in the Sprague-Dawley Rat

Temporal organization is a fundamental property of living matter. From single cells to complex animals including man, most physiological systems undergo daily periodic changes in concert with environmental cues (e.g., light, temperature etc.). It is known that pulsed Environmental synchronizers, zeitgebers, (e.g. light) can modify rhythm parameters. Rhythm stability is a necessary requirement for most animal experiments. The extent to which sound can influence the circadian system of laboratory rats is poorly understood. This has implications to animal habitats in the novel environments of the Space-Laboratory or Space Station. A series of three white noise (88+/-0.82 db) zeitgeber experiments were conducted (n=6/experiment).The sound cue was introduced in the circadian free-running phase (DD-NQ) and in one additional case sound was added to the usual photoperiod (12L:12D) to determine masking effects. Circadian rhythm parameters of drinking frequency, feeding frequency, and gross locomotor activity were continuously monitored. Data analysis for these studies included macroscopic and microscopic methods. Raster plots to visually detect entrainment versus free-running period, were plotted for each animal, for all three parameters, during all sound perturbation tested. These data were processed through a series of detrending (robust locally weighted regression analyses) and complex demodulation analyses. In summary, these findings show that periodic "white" noise "influences" the rats circadian system but does not "entrain" the feeding, drinking or locomotor activity rhythms.

Winget, C. M.↗
NASA NTRS1996

Light-Emitting Diodes (LED) for Primary Animal Habitat Lighting in Highly Controlled Environments

Significant alterations in Biological Clock responses have been reported following sidereal time changes (e.g., Jet-lag), and exposure to microgravity (e.g., daytime sleepiness). Additionally, light reduces circulating melatonin (spectral specificity greatest between 450-500 nm). It was hypothesized that LEDs can replace the current light sources used in zero gravity and terrestrial research laboratories because of their small size, low mass, low energy consumption and long functional life. This report evaluates the capacity of LEDs to entrain the circadian system of rats as judged by measurement of overt behavioral circadian rhythms (activity, feeding, drinking). These data were collected in highly controlled environments similar to the shuttle Animal Enclosure Modules. Two groups were compared: control - animals exposed to standard cool-white fluorescent lights, and test - animals exposed to LEDs with a spectral power distribution matching the fluorescent lights. Gross locomotor activity, feeding and drinking frequencies were continuously monitored and stored at 10 minute intervals. Animals were exposed to the following photoperiods: 28 days of 12L:12D, 19 days of 24L:0D and 16 days of 12L:12D. Light intensities tested varied between 0.1 to 100 lux. Rats received food and water ad libitum, and temperature and humidity were controlled throughout the study. The general health status of all rats was acceptable for each day of this study. No incidents of aggressive behavior were observed. Growth, locomotor activity, food and water consumption were comparable for all groups of animals, i.e, the circadian characteristics of the animals under these conditions were comparable. These results indicate that LED arrays are as effective in maintaining circadian rhythm stability as the commonly used cool-white fluorescent light sources. LEDs with their flexible spectrum, low energy requirements and minimal heat production have advantages for some chronopharmacology studies and for microgravity animal habitats.

Winget, C. M.↗
NASA NTRS2003

Retinal ganglion cell projections to the hamster suprachiasmatic nucleus, intergeniculate leaflet, and visual midbrain: bifurcation and melanopsin immunoreactivity

The circadian clock in the suprachiasmatic nucleus (SCN) receives direct retinal input via the retinohypothalamic tract (RHT), and the retinal ganglion cells contributing to this projection may be specialized with respect to direct regulation of the circadian clock. However, some ganglion cells forming the RHT bifurcate, sending axon collaterals to the intergeniculate leaflet (IGL) through which light has secondary access to the circadian clock. The present studies provide a more extensive examination of ganglion cell bifurcation and evaluate whether ganglion cells projecting to several subcortical visual nuclei contain melanopsin, a putative ganglion cell photopigment. The results showed that retinal ganglion cells projecting to the SCN send collaterals to the IGL, olivary pretectal nucleus, and superior colliculus, among other places. Melanopsin-immunoreactive (IR) ganglion cells are present in the hamster retina, and some of these cells project to the SCN, IGL, olivary pretectal nucleus, or superior colliculus. Triple-label analysis showed that melanopsin-IR cells bifurcate and project bilaterally to each SCN, but not to the other visual nuclei evaluated. The melanopsin-IR cells have photoreceptive characteristics optimal for circadian rhythm regulation. However, the presence of moderately widespread bifurcation among ganglion cells projecting to the SCN, and projection by melanopsin-IR cells to locations distinct from the SCN and without known rhythm function, suggest that this ganglion cell type is generalized, rather than specialized, with respect to the conveyance of photic information to the brain. Copyright 2003 Wiley-Liss, Inc.

NASA Discipline Neuroscience↗
NASA NTRS2014

Fatigue‐Related Countermeasures for Long‐Duration Exploration Missions

The NASA Human Research Program's (HRP) Behavioral Health and Performance Element (BHP) supports and conducts research to mitigate deleterious outcomes related to fatigue, sleep loss, circadian desynchronization, and work overload. Objective evidence indicates that within the context of the International Space Station (ISS), sleep is reduced and there is circadian misalignment. Despite chronic sleep loss and high workloads; however, astronauts successfully complete their missions. Contributing to their success is not only the tremendous skills and capabilities of each astronaut, but also the collaborative team efforts amongst the crew, between flight and ground crews, and through real‐time care provided by medical personnel. It is anticipated that risks to human health and performance will increase in the context of exploration missions, where crewmembers will venture to deep space for extended durations and in small vehicles with limited communication with home. Hence, fatigue‐related countermeasures are being developed and/or validated that include unobtrusive monitoring technologies to detect fatigue‐related performance decrements, environmental countermeasures, and sleep education and training for flight and ground crews. Given that fatigue is an issue in current ISS missions, the BHP works collaboratively with Space Medicine operations to collect data in the operational environment, to validate fatigue-related countermeasures, and provide evidence‐based mitigations. Our presentation will summarize fatigue‐related operational research that is underway through NASA's BHP in partnership with its operational counterparts. Efforts include studies evaluating the effects of hypnotics, lighting protocols as countermeasures for circadian entrainment, and investigations involving education and training. This presentation will further identify, based on flight and terrestrial evidence, additional sleep and circadian countermeasures that may still be needed to support exploration missions. Lessons learned from transitioning research deliverables into ISS operations will also be discussed.

Whitmire, A.↗
NASA NTRS2016

Variations in Sleep and Performance by Duty Start Time in Short Haul Operations

Prior studies have confirmed that commercial airline pilots experience circadian phase shifts and short sleep duration following travel with layovers in different time zones. Few studies have examined the impact of early and late starts on the sleep and circadian phase of airline pilots who return to their domicile after each duty period. We recruited 44 pilots (4 female) from a short-haul commercial airline to participate in a study examining sleep and circadian phase over four duty schedules (baseline, early starts, mid-day starts, late starts). Each duty schedule was five days long, separated by three rest days. Participants completed the rosters in the same order. Sleep outcomes were estimated using wrist-borne actigraphy (Actiware Software, Respironics, Bend, OR) and daily sleep diaries. Thirteen participants volunteered to collect urine samples for the assessment of 6-sulfatoxymelatonin (aMT6s). Urine samples were collected in four-hourly bins during the day and eight-hourly bins during sleep episodes, for 24 hours immediately following each experimental duty schedule. The aMT6s results were fit to a cosine in order to obtain the acrophase to estimate circadian phase. Univariate statistics were calculated for acrophase changes, schedule start times and sleep times. All statistical analyses were computed using SAS software (Cary, IN).

fatigue↗
NASA NTRSDate not supplied

Sweating the Small Stuff: A Sensor for Real-Time Neuro-Immune Axis Monitoring

Although manageable with certain precautions, circadian rhythm misalignment and sleep disturbances pose a potential threat to astronaut health. It is well-known that sleep is essential for proper immunological and neurological functioning, while impairments in these functions result in risks for spaceflight success. Neuropeptides and hormones are involved in regulating the circadian clock and are used as biomarkers for circadian alignment. However, in-flight monitoring of these biomarkers is limited due to the lack of real-time sensor systems and sample collection/processing confines. Therefore, real-time measurements of biomarkers in-flight are necessary for mission success and crew health. For this, a robust biomarker involved in the neuro-immune axis of circadian rhythm cycling, physiological stress responses, and inflammation would be well-received as a viable biomarker for assessing physiological health for crew during long-duration missions, as these are currently NASA Human Research Program defined risks. Therefore, this paper describes the development of a wearables we at biosensor to measure the biological clock neuropeptide, orexin/hypocretin. Additionally, potential for implementation of a user-friendly sensor of orexin/hypocretin to be telemetrically reported in real-time, is proposed. In brief, the proposed system has the possibility to be used as a biomarker monitor to support preventive and personalized medicine.

Amber M Paul↗
NASA NTRSDate not supplied

Changes in Oculomotor Behavior and PVT Reaction Time During One Night of Sleep Deprivation

INTRODUCTION: The Psychomotor Vigilance Test (PVT) is a widely used objective measure of sustained attention, alertness, and fatigue. Previous studies consistently show that prolonged wakefulness and disrupted sleep patterns lead to increased lapses of attention and slower reaction times. These findings have critical implications for various professions, including healthcare, aviation, transportation safety, and spaceflight. In this study, we employed linear mixed models (LMM) to investigate hourly-measured 5-minute PVT reaction times during one night of total sleep deprivation. Our goal was to characterize the contribution of homeostatic sleep pressure (time awake) and circadian phase (salivary melatonin levels) on the dynamics of PVT reaction time. MATERIALS & METHODS: Data from twelve human participants were used in the analysis. Participants were healthy non-smokers, aged 18 and 40, with normal sleep habits defined as Pittsburg Sleep Quality Index scores < 5, and Morningness – Eveningness Questionnaire scores > 42 and < 58, respectively. Participants followed a constant-routine protocol and arrived at the NASA sleep laboratory approximately 1 to 2 hours after their habitual waking time. Tympanic temperature was measured every 30 min to provide a real-time estimate of circadian phase. Participants were required to stay awake throughout the laboratory experiment, which continued until their tympanic temperature had returned to baseline levels. This approach ensured that we could capture each participant's circadian trough and recovery, typically occurring between 24 and 26 hours after waking. The PVT and saliva melatonin were measured every hour, starting approximately from 3 hours after awakening. More information about the measurement protocol can be found in the publication by Stone et al. We used PVT reaction times between 100 and 10000 ms to calculate Reciprocal Reaction Times (RRTs). LMM models were employed to assess the contribution of time awake and saliva melatonin levels on changes in RRTs. Two models were created: the first model treated time awake as a fixed effect and the second model considered both the salivary melatonin level and time awake as fixed effects. Participants were included as a random effect in both models. We compared the models using an Analysis of Variance (ANOVA). The models were computed using the 'lmer' function from the 'lme4' package in R, and LMM p-values were determined using Satterthwaite's method. RESULTS: We found that time awake had a statistically significant effect on PVT RRTs during one night of sleep deprivation [χ2 = 2132.4, df = 1, p < 0.001]. In addition, the model including both saliva melatonin levels and time awake was statistically significantly better than one that uses just time awake [χ2 = 17.4, df = 1, p < 0.001]. DISCUSSION & CONCLUSION: As expected, these preliminary results suggest that the time awake and circadian phase each contribute to performance reductions as measured by the PVT. Future analyses will explore the contributions of other factors to the model.

sleep deprivation↗
DOE OSTI2025

Gene network centrality analysis identifies key regulators coordinating day-night metabolic transitions in Synechococcus elongatus PCC 7942 despite limited accuracy in predicting direct regulator-gene interactions

Synechococcus elongatus PCC 7942 is a model organism for studying circadian regulation and bioproduction, where precise temporal control of metabolism significantly impacts photosynthetic efficiency and CO 2 -to-bioproduct conversion. Despite extensive research on core clock components, our understanding of the broader regulatory network orchestrating genome-wide metabolic transitions remains incomplete. We address this gap by applying machine learning tools and network analysis to investigate the transcriptional architecture governing circadian-controlled gene expression. While our approach showed moderate accuracy in predicting individual transcription factor-gene interactions - a common challenge with real expression data - network-level topological analysis successfully revealed the organizational principles of circadian regulation. Our analysis identified distinct regulatory modules coordinating day-night metabolic transitions, with photosynthesis and carbon/nitrogen metabolism controlled by day-phase regulators, while nighttime modules orchestrate glycogen mobilization and redox metabolism. Through network centrality analysis, we identified potentially significant but previously understudied transcriptional regulators: HimA as a putative DNA architecture regulator, and TetR and SrrB as potential coordinators of nighttime metabolism, working alongside established global regulators RpaA and RpaB. This work demonstrates how network-level analysis can extract biologically meaningful insights despite limitations in predicting direct regulatory interactions. The regulatory principles uncovered here advance our understanding of how cyanobacteria coordinate complex metabolic transitions and may inform metabolic engineering strategies for enhanced photosynthetic bioproduction from CO 2 .

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Energy impact of human health and wellness lighting recommendations for office and classroom applications

The goal of this investigation was to evaluate potential energy impacts of circadian lighting design recommendations gaining attention in a variety of common applications such as offices and classrooms. The renewed focus on health along with advances in SSL technology capabilities has underscored that there is still much to learn regarding the relationship between light and human physiology. The energy implications of designing to address these possible physiological effects are not yet fully understood. Beyond the fact that the basic metric of luminous efficacy (lumens per watt) does not cover these other effects, the emerging science seems to indicate that addressing a holistic view of the human needs in most applications may mean a need for increased light and associated energy use by electric lighting systems. Within the two applications considered, lumen output, spectral characteristics, surface reflectance distribution and desk orientation were varied to explore the magnitude of potential effects. Meeting current IES illuminance recommendations did not satisfy existing EML and CS recommendations for any of the simulations. In some cases, satisfying circadian metric recommendations required average illuminance that was more than double IES recommendations, which may negatively impact lighting quality as well. Using results from 45 unique simulation conditions, it was estimated that lighting energy use may increase between 10% and 100% due to increased luminaire light levels used to meet circadian lighting design recommendations listed in current building standards such as WELL v2 Q2 2019, UL Design Guideline 24480, and CHPS Core Criteria 3.0.

circadian lighting, lighting simulation, solid sta↗
DOE OSTI2026

Rhythmic Mechanisms Governing CAM Photosynthesis in Kalanchoe fedtschenkoi : High-Resolution Temporal Transcriptomics

Crassulacean acid metabolism (CAM) is a specialized photosynthetic pathway that enhances water-use efficiency by temporally separating nocturnal CO 2 uptake from daytime decarboxylation and carbon fixation. To uncover the regulatory mechanisms coordinating these temporal dynamics, we generated high-resolution, 48 h time-course transcriptomes for the CAM model Kalanchoe fedtschenkoi under both 12 h/12 h light/dark (LD) cycles and continuous light (LL). A rhythmicity analysis revealed that diel light cues are the dominant driver of transcript oscillations: 16,810 genes (54.3% of annotated genes) exhibited rhythmic expression only under LD, whereas just 399 genes (1.3%) remained rhythmic under LL. A smaller set of 3009 genes (9.7%) oscillated in both conditions, indicating that the intrinsic circadian clock sustains rhythmicity for a limited subset of the transcriptome. A gene co-expression network analysis revealed extensive integration between circadian clock components, core CAM pathway enzymes, and stomatal regulators, defining regulatory modules that coordinate metabolic and physiological timing. Notably, key hub genes associated with post-translational and post-transcriptional regulation, including the E3 ubiquitin ligase HUB2 and several pentatricopeptide repeat (PPR) proteins, act as central nodes in CAM-associated networks. This discovery implicates epigenetic and organellar regulation as previously unrecognized critical tiers of control in CAM. Together, our results support a regulatory model in which CAM rhythmicity is governed by both external light/dark cues and the endogenous circadian clock through multi-level control spanning transcriptional and protein-level regulation. To support community exploration, we also provide an interactive eFP (electronic Fluorescent Pictograph) browser for visualizing time-resolved gene expression profiles.

09 BIOMASS FUELS↗
DOE OSTI2022

Comparative Phenotyping of Two Commonly Used Chlamydomonas reinhardtii Background Strains: CC-1690 (21gr) and CC-5325 (The CLiP Mutant Library Background)

The unicellular green alga Chlamydomonas reinhardtii is an excellent model organism to investigate many essential cellular processes in photosynthetic eukaryotes. Two commonly used background strains of Chlamydomonas are CC-1690 and CC-5325. CC-1690, also called 21gr, has been used for the Chlamydomonas genome project and several transcriptome analyses. CC-5325 is the background strain for the Chlamydomonas Library Project (CLiP). Photosynthetic performance in CC-5325 has not been evaluated in comparison with CC-1690. Additionally, CC-5325 is often considered to be cell-wall deficient, although detailed analysis is missing. The circadian rhythms in CC-5325 are also unclear. To fill these knowledge gaps and facilitate the use of the CLiP mutant library for various screens, we performed phenotypic comparisons between CC-1690 and CC-5325. Our results showed that CC-5325 grew faster heterotrophically in dark and equally well in mixotrophic liquid medium as compared to CC-1690. CC-5325 had lower photosynthetic efficiency and was more heat-sensitive than CC-1690. Furthermore, CC-5325 had an intact cell wall which had comparable integrity to that in CC-1690 but appeared to have reduced thickness. Additionally, CC-5325 could perform phototaxis, but could not maintain a sustained circadian rhythm of phototaxis as CC1690 did. Finally, in comparison to CC-1690, CC-5325 had longer cilia in the medium with acetate but slower swimming speed in the medium without nitrogen and acetate. Our results will be useful for researchers in the Chlamydomonas community to choose suitable background strains for mutant analysis and employ the CLiP mutant library for genome-wide mutant screens under appropriate conditions, especially in the areas of photosynthesis, thermotolerance, cell wall, and circadian rhythms.

59 BASIC BIOLOGICAL SCIENCES↗