Project description:Across evolutionary time, nearly all animal species have harnessed photoperiod to initiate processes that ultimately influence seasonal behavior and life history traits. In the freshwater cladoceran Daphnia magna, the effect of photoperiod on various life history traits has generally been investigated in conjunction with additional environmental stimuli. In the present study, we sought to untangling responses directly attributable to photoperiod in D. magna and identify the molecular processes underlying resultant behavioral and life history responses using functional analysis of global transcriptomic expression. D. magna were exposed to five different photoperiods immediately post-hatch for 21d where a standard long-day photoperiod of 16 hours light and 8 hours dark (16L:8D) served as the control relative to 4L:20D, 8L:16D, 12L:12L, and 20L:4D photoperiods. Entrainment to short-day photo-periods (4L:20D, 8L:16D, and 12L:12L) resulted in significantly increased light-avoidance behaviors relative to the control photoperiod where young Daphnia (7d old) displayed the most pronounced avoidance responses. Correspondingly, functional transcriptomics identified differential transcriptional expression of genes involved in glutamate signaling, which is critical in arthropod light-avoidance responses, as well as period circadian protein and proteins coding F-box/LRR-repeat domains, all of which contribute to establishing circadian rhythms in arthropods. Short-day photoperiods also induced increased metabolic rates which corresponded with broad-scale changes in transcriptional expression across multiple systems-level energy metabolism pathways. The most striking observation was increased male production across short-day photoperiods (4L:20D, 8L:16D, and 12L:12D). Transcriptional expression consistent with multiple putative mechanisms of male production were observed including expression suggestive of increased glutamate signaling; a response observed to induce male production in D. pulex via photo-period sensitive mechanisms. Overall, the results demonstrate the importance of photoperiod on behavior and life history trajectories in D. magna where we have now established multiple putative mechanistic pathways underlying several critical responses.

Project description:The US Department of Defense (DOD) is developing insensitive munitions (IMs) that are resistant to unintended detonation to protect warfighters. To enable evaluation of the material life-cycle for the IM, 1-methyl-3-nitro-1-nitroguanidine (MeNQ), ecotoxicological impacts assessment was required. A previous investigation of MeNQ exposures in Daphnia pulex revealed concentration-responsive decreases in reproduction relative to controls (0 mg/L) across a 174, 346, 709, 1385, and 2286 mg/L exposure range. The present study used those exposures to conduct global transcriptomic expression analyses to establish hypothetical mechanisms underlying inhibited reproduction. Changes in transcript expression tended to increase with increasing MeNQ concentration where hierarchical clustering analysis identified separation among the “low” (174 and 346 mg/L) and “high” (709, 1385, and 2286 mg/L) exposures. The MeNQ exposures significantly decreased transcriptional expression for a vitellogenin precursor and juvenile hormone-inducible protein, each having plausible mechanistic connections to impaired reproductive physiology and reduced offspring production in Daphnia. Further, MeNQ affected chorion peroxidase and high choriolytic enzyme transcriptional expression which are known to influence egg production / maturation processes in arthropods. Additionally, MeNQ exposure caused dose-responsive decreases in transcriptional expression for a diverse set of cuticle proteins which requires closer examination regarding Daphnia embryo development and brood release. Finally, canonical-pathway enrichment analysis identified significant effects on a broad array of energy metabolic processes in D. pulex which require consideration regarding potential shifts in homeostatic equilibrium that can have indirect costs on reproduction. Overall, plausible mechanisms of inhibited D. pulex reproduction have been identified and should be tested in targeted bioassays. see previous row.

Project description:This study measured the differential expression of genes from a single clonal Daphnia pulex isolate obtained from an eutrophic flooded opencast mine near Gräfenhain (Saxony, Germany) and has been kept in the laboratory since 2002. The daphniids (50 adult animals per batch) were raised in 3-L glass beakers under a 16 h:8 h L:D photoperiod. Three-quarter of the culture medium (M4) was renewed once a week, and the animals were equally fed ad libitum with algae (Desmodesmus subspicatus; SAG 53.80, Göttingen, Germany). Long-term-acclimated animals were kept at three different temperatures (i.e., 10 ± 0.5°C, 20 ± 0.3°C, and 24 ± 0.3°C) under normoxia (Po2 ≥ 20 kPa) for at least twelve weeks. Time-resolved experiments (acute heat stress) were carried out with long-term 20°C acclimated animals, which were exposed for three different time intervals (2, 4, and 8 h) to either 30 ± 0.2°C (test conditions) or 20 ± 0.3°C (control conditions). For this, 50 animals each were rapidly transferred with a sieve (mesh size: 1.2 mm) from 20°C-medium to a set of beakers containing 30°C-medium. As control, 50 animals each were transferred from 20°C-medium to another set of beakers with 20°C-medium. Only adult female animals with a body length of 2−2.5 mm (between the base of the apical spine and the anterior part of the head) and carrying parthenogenetic eggs and embryos were used for experiments. Animals were not fed during short-term exposures. During experiments, all animals were in good physical condition. After thermal acclimation or heat exposure, animals were transferred into 1.5-ml microcentrifuge tubes and shock-frozen in liquid nitrogen after removing adhering water with a tissue paper. Samples were short-term stored at -80°C. For all experimental conditions, four independent replicates (50 animals each) were analyzed. This GEO record is for the acute heat stress temperature contrasts between animals conditioned to 20 degrees Celsius (condition 1) versus heat shock to 30 degrees Celsius (condition 2) exposed for 2 hours (treatment 1), 4 hours (treatment 2), and 8 hours (treatment 3).

Dataset Information

Mechanistic Evaluation of Daphnia magna’s Behavioral and Life History Responses to Photoperiod [Experiment 1]

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