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1. Diapause vs. reproductive programs: transcriptional phenotypes in a keystone copepod

   https://doi.org/10.1038/s42003-021-01946-0  

   Lenz, Petra H.; Roncalli, Vittoria; Cieslak, Matthew C.; Tarrant, Ann M.; Castelfranco, Ann M.; Hartline, Daniel K. (December 2021, Communications Biology)

   Abstract Many arthropods undergo a seasonal dormancy termed “diapause” to optimize timing of reproduction in highly seasonal environments. In the North Atlantic, the copepod Calanus finmarchicus completes one to three generations annually with some individuals maturing into adults, while others interrupt their development to enter diapause. It is unknown which, why and when individuals enter the diapause program. Transcriptomic data from copepods on known programs were analyzed using dimensionality reduction of gene expression and functional analyses to identify program-specific genes and biological processes. These analyses elucidated physiological differences and established protocols that distinguish between programs. Differences in gene expression were associated with maturation of individuals on the reproductive program, while those on the diapause program showed little change over time. Only two of six filters effectively separated copepods by developmental program. The first one included all genes annotated to RNA metabolism and this was confirmed using differential gene expression analysis. The second filter identified 54 differentially expressed genes that were consistently up-regulated in individuals on the diapause program in comparison with those on the reproductive program. Annotated to oogenesis, RNA metabolism and fatty acid biosynthesis, these genes are both indicators for diapause preparation and good candidates for functional studies. 

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2. Post-diapause transcriptomic restarts: insight from a high-latitude copepod

   https://doi.org/10.1186/s12864-021-07557-7  

   Roncalli, Vittoria; Cieslak, Matthew C; Castelfranco, Ann M; Hopcroft, Russell R; Hartline, Daniel K; Lenz, Petra H (December 2021, BMC Genomics)

   Abstract BackgroundDiapause is a seasonal dormancy that allows organisms to survive unfavorable conditions and optimizes the timing of reproduction and growth. Emergence from diapause reverses the state of arrested development and metabolic suppression returning the organism to an active state. The physiological mechanisms that regulate the transition from diapause to post-diapause are still unknown. In this study, this transition has been characterized for the sub-arctic calanoid copepod Neocalanus flemingeri, a key crustacean zooplankter that supports the highly productive North Pacific fisheries. Transcriptional profiling of females, determined over a two-week time series starting with diapausing females collected from > 400 m depth, characterized the molecular mechanisms that regulate the post-diapause trajectory. ResultsA complex set of transitions in relative gene expression defined the transcriptomic changes from diapause to post-diapause. Despite low temperatures (5–6 °C), the switch from a “diapause” to a “post-diapause” transcriptional profile occurred within 12 h of the termination stimulus. Transcriptional changes signaling the end of diapause were activated within one-hour post collection and included the up-regulation of genes involved in the 20E cascade pathway, the TCA cycle and RNA metabolism in combination with the down-regulation of genes associated with chromatin silencing. By 12 h, females exhibited a post-diapause phenotype characterized by the up-regulation of genes involved in cell division, cell differentiation and multiple developmental processes. By seven days post collection, the reproductive program was fully activated as indicated by up-regulation of genes involved in oogenesis and energy metabolism, processes that were enriched among the differentially expressed genes. ConclusionsThe analysis revealed a finely structured, precisely orchestrated sequence of transcriptional changes that led to rapid changes in the activation of biological processes paving the way to the successful completion of the reproductive program. Our findings lead to new hypotheses related to potentially universal mechanisms that terminate diapause before an organism can resume its developmental program. 

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3. Post-diapause transcriptomic restarts: insight from a high-latitude copepod

   https://doi.org/10.1186/s12864-021-07868-9  

   Roncalli, Vittoria; Cieslak, Matthew C.; Castelfranco, Ann M.; Hopcroft, Russell R.; Hartline, Daniel K.; Lenz, Petra H. (January 2021, BMC genomics)

   Background: Diapause is a seasonal dormancy that allows organisms to survive unfavorable conditions and optimizes the timing of reproduction and growth. Emergence from diapause reverses the state of arrested development and metabolic suppression returning the organism to an active state. The physiological mechanisms that regulate the transition from diapause to post-diapause are still unknown. In this study, this transition has been characterized for the sub-arctic calanoid copepod Neocalanus flemingeri, a key crustacean zooplankter that supports the highly productive North Pacific fisheries. Transcriptional profiling of females, determined over a two-week time series starting with diapausing females collected from > 400m depth, characterized the molecular mechanisms that regulate the post-diapause trajectory. Results: A complex set of transitions in relative gene expression defined the transcriptomic changes from diapause to post-diapause. Despite low temperatures (5–6 °C), the switch from a “diapause” to a “post-diapause” transcriptional profile occurred within 12 h of the termination stimulus. Transcriptional changes signaling the end of diapause were activated within one-hour post collection and included the up-regulation of genes involved in the 20E cascade pathway, the TCA cycle and RNA metabolism in combination with the down-regulation of genes associated with chromatin silencing. By 12 h, females exhibited a post-diapause phenotype characterized by the up-regulation of genes involved in cell division, cell differentiation and multiple developmental processes. By seven days post collection, the reproductive program was fully activated as indicated by up-regulation of genes involved in oogenesis and energy metabolism, processes that were enriched among the differentially expressed genes. Conclusions: The analysis revealed a finely structured, precisely orchestrated sequence of transcriptional changes that led to rapid changes in the activation of biological processes paving the way to the successful completion of the reproductive program. Our findings lead to new hypotheses related to potentially universal mechanisms that terminate diapause before an organism can resume its developmental program. 

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4. Lipid metabolism in Calanus finmarchicus is sensitive to variations in predation risk and food availability

   https://doi.org/10.1038/s41598-020-79165-6  

   Skottene, Elise; Tarrant, Ann M.; Altin, Dag; Olsen, Rolf Erik; Choquet, Marvin; Kvile, Kristina Ø. (December 2020, Scientific Reports)

   Abstract Late developmental stages of the marine copepods in the genus Calanus can spend extended periods in a dormant stage (diapause) that is preceded by the accumulation of large lipid stores. We assessed how lipid metabolism during development from the C4 stage to adult is altered in response to predation risk and varying food availability, to ultimately understand more of the metabolic processes during development in Calanus copepods. We used RNA sequencing to assess if perceived predation risk in combination with varied food availability affects expression of genes associated with lipid metabolism and diapause preparation in C. finmarchicus . The lipid metabolism response to predation risk differed depending on food availability, time and life stage. Predation risk caused upregulation of lipid catabolism with high food, and downregulation with low food. Under low food conditions, predation risk disrupted lipid accumulation. The copepods showed no clear signs of diapause preparation, supporting earlier observations of the importance of multiple environmental cues in inducing diapause in C. finmarchicus . This study demonstrates that lipid metabolism is a sensitive endpoint for the interacting environmental effects of predation pressure and food availability. As diapause may be controlled by lipid accumulation, our findings may contribute towards understanding processes that can ultimately influence diapause timing. 

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5. Diapause and Anoxia-Induced Quiescence Are Unique States in Embryos of the Annual Killifish, Austrofundulus limnaeus

   https://doi.org/10.3390/biom15040515  

   Clouser, Patrick R; Riggs, Claire L; Romney, Amie_L T; Podrabsky, Jason E (April 2025, Biomolecules)

   Diapause is a state of developmental and metabolic dormancy that precedes exposure to environmental stresses. Yet, diapausing embryos are typically stress-tolerant. Evidence suggests that diapausing embryos “prepare” for stress as part of a gene expression program as they enter dormancy. Here, we investigate if diapause II embryos of the annual killifish Austrofundulus limnaeus, which can survive for hundreds of days of anoxia, can mount a transcriptomic response to anoxic insult. Bulk RNAseq was used to characterize the transcriptomes of diapause II embryos exposed to normoxia, 4 h and 24 h anoxia, and 2 h and 24 h normoxic recovery from anoxia. Differential expression and gene ontology analyses were used to probe for pathways that may mitigate survival. Transcriptional factor analysis was used to predict potential mediators of this response. Diapausing embryos exhibited a robust transcriptomic response to anoxia and recovery that returns to near baseline conditions after 24 h. Anoxia induced an upregulation of genes involved in the integrated stress response, lipid metabolism, p38mapk kinase signaling, and apoptosis. Developmental and mitochondrial genes decreased. We conclude that diapause II embryos mount a robust transcriptomic stress response when faced with anoxic insult. This response is consistent with mediating expected challenges to cellular homeostasis in anoxia. 

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