Fig 5 - uploaded by Katharina C. Wollenberg Valero
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TS and SM alter the development and behavior of zebrafish embryos. A) Principal component analysis (PCA) of 1-dpf morphology. HPF, final stage in hours post fertilization; Stage, % in segmentation or pharyngula; EA, eye area; EL, eye length; DVL, dorsal-ventral length; HTA, head-trunk angle; LEL, longest embryo length; OVL, otic vesicle length; SEL, shortest embryo length; WBA, whole-body area; YBA/L, yolk ball area/length; YEA/L, yolk extension area/length; YE/YB, yolk extension/yolk ball ratio. Individual variables significantly altered by TS are shown by black arrows. B) Burst count per minute in active 1-dpf embryos. C) SEL (mm) increment from 1 to 4 dpf. D) Mean acceleration (m/s 2 ) of the first complete burst following the first and third touch stimuli. E) Density distribution of total burst count and total distance in cm F) after three touch stimuli. Effects of TS and SM across the two-way factorial design (C, SM, TS, and TS + SM) were computed using PERMANOVA A), ANOVA B-D, F), or negative binomial generalized linear model (E, confirmed by a density test) with significant predictors and covariates ("batch") reported on top-right corners. Pairwise tests with FDR P value correction compared CM to C and SM, or C to SM, TS, and TS + SM, with significant comparisons shown by horizontal bars. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001. Treatments were CM, control metabolites at 27°C; C, control in fresh medium at 27°C; SM, stress metabolites at 27°C; TS, fresh medium in thermal stress; and TS + SM, stress metabolites in thermal stress.
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Heat alters biology from molecular to ecological levels, but may also have unknown indirect effects. This includes the concept that animals exposed to abiotic stress can induce stress in naive receivers. Here, we provide a comprehensive picture of the molecular signatures of this process, by integrating multiomic and phenotypic data. In individual...
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... utilized laboratory inbred AB and endpoints 3/6 wilder PET embryos. the expression of genes expressed by SM and in lateral line hair cells. Remarkably, the DEGs in SM are also significantly upregulated in zebrafish lateral line hair cells in 5-dpf larvae, relative to their expression in neighboring epidermis cells (z-score = 8.71505; P < 0.0001, Fig. S5D). One interesting gene emerging from this comparison was xkr8.2 (ENSDARG00000076820), which is the highest up-regulated gene in both hair cells and SM (LFC = 1.68) compared with nonhair cells, and is a scramblase involved in the enriched GO term "membrane phospholipid transport" (GO:0015914, Dataset S1). Overall, our results show that ...
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... could cause changes in escape and startle behavioral responses, which are standard measurements to indicate anxiety and changes in sensory and motor functions (33)(34)(35). Heat stress (F = 30.1, R 2 = 0.25, P = 0.001), but neither SM (F = 0.16, P = 0.7610) nor the interaction term (F = 2.34, P = 0.1320), altered the phenotype of 1-dpf embryos ( Fig. 5A and Table S4). Post hoc tests confirmed that thermal stress significantly altered embryo growth, reaching the pharyngula stage earlier (z = −2.67, P = 0.0076, Figs. 5C and S10A and B). Fig. 4. Multiomic data evidence that stress metabolites (SM) differ in classes and functions from control metabolites (CM). A) Correlation plot showing ...
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... SM had no effect on growth at 1 dpf. However, an ANOVA of the two-way model terms showed that 4-dpf zebrafish embryos grew significantly longer in the presence of SM (t = 4.80, P = 0.0304, small effect size = 0.35) but not under thermal stress (t = 1.91, P = 0.1692) nor the interaction term (t = 0.11, P = 0.7353, Fig. 5D). An ANOVA revealed that both heat and SM significantly altered the increment in growth (ΔSEL) between 1 and 4 dpf, but in opposite directions (Fig. 5E and Tables S5 and S6). Heat-exposed embryos had a lower ΔSEL (t = 20.99, P < 0.0001, large effect size = 0.81), whereas those experiencing SM ...
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... grew significantly longer in the presence of SM (t = 4.80, P = 0.0304, small effect size = 0.35) but not under thermal stress (t = 1.91, P = 0.1692) nor the interaction term (t = 0.11, P = 0.7353, Fig. 5D). An ANOVA revealed that both heat and SM significantly altered the increment in growth (ΔSEL) between 1 and 4 dpf, but in opposite directions (Fig. 5E and Tables S5 and S6). Heat-exposed embryos had a lower ΔSEL (t = 20.99, P < 0.0001, large effect size = 0.81), whereas those experiencing SM ...
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... accelerated growth from 1 to 4 dpf (t = 6.65, P = 0.0112, small effect size = 0.43) and slightly surpassed the TS treatment in average final length at 4 dpf ( Fig. 5D and E). Heat stress (t = 18.15, P < 0.0001), but neither SM (t = 0.29, P = 0.5891) nor their interaction (t = 0.07, P = 0.7872), significantly reduced the startle response of 1-dpf zebrafish embryos (Fig. 5B and Tables S7 and S8). Heat also slowed mean acceleration (t = 4.26, P = 0.0412, small effect size = 0.34), and TS marginally reduced ...
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... S8). Heat also slowed mean acceleration (t = 4.26, P = 0.0412, small effect size = 0.34), and TS marginally reduced mean speed compared with C (t = −2.74, P = 0.0212) in the touch-evoked swimming response of 4-dpf larvae (Fig. S11B and C). Larvae incubated in SM, however, swam fewer burst counts compared with fresh medium (t = −2.1, P = 0.0381, Fig. 5F). Since the metabolomic analysis supported our hypothesis that SM from stressed animals are different compared with CM from undisturbed controls, we explored the consequences of SM vs. CM at the phenotypic level. We found that SM tended to induce less hatching (Fig. S10D) and tended to have altered startle responses at 1 dpf (Figs. 5B ...
Citations
... Climate adaptation at the molecular scale is often acknowledged, but rarely adequately addressed (Gienapp et al. 2008;Bodensteiner, Agudelo-Cantero, et al. 2021), although modern technologies have now begun to explore the role of epigenomics (McGuigan et al. 2021;McCaw et al. 2020), transcriptomics (Wang et al. 2021;Hamann et al. 2021;Rosso et al. 2024;Feugere et al. 2022), proteomics (Sun et al. 2022;Luo et al. 2024), and genomics (Saravanan et al. 2024;Wollenberg Valero et al. 2022;Turbek et al. 2023) in climate adaptation. The increasing availability of sequencing data has allowed scans for selection associated with traits or environmental parameters across genomes, a step change from studying a narrow range of predefined genes for adaptive evolution (Biswas & Akey 2006). ...
Climate adaptation is caused by multiple mechanisms, including evolution of regulatory promoter sequences under broad environmental pressures, however, little is known about these processes in response to climate pressures in animals. Here, we examined spatial and temporal evolution of climate-related gene promoters in an insular species, the Western-Canaries lizard Gallotia galloti, across diverse environmental conditions over nine years. Outlier SNPs linked to elevation-sensitive environmental factors identified adaptively significant loci, including in aquaporin and Electron Transfer Flavoprotein Subunit Alpha (ETFA) gene promoters, demonstrating roles in local climate adaptation. Adaptive indexing mapped climate-driven genetic differentiation across the landscape, revealing local adaptation along an elevational gradient. Signatures of climate-related selection at the temporal scale showed specific promoters shaped by balancing selection supporting survival across environments, while others experienced positive selection, revealing a selective sweep of beneficial alleles. In addition, elevation-based ancestry groups of climate loci differed from the pattern observed in neutral loci, and temporal shifts in ancestry composition responded to preceding extreme weather events over the span of a single generation. Selection on specific promoter regions indicated adaptive evolution in transcription factor binding sites with the potential to enhance transcriptional flexibility, resulting in increased cellular stress tolerance. Our findings open new avenues into genomic mechanisms of resilience across spatially heterogeneous environments and how rapid evolution correlates with extreme weather events associated with climate change.
... Notably, animals exposed to 1 and 10 ppm 44 nm NanoPS at 32 • C showed the most pronounced increases in antioxidant response. Thermal fluctuations can induce oxidative stress in zebrafish by increasing the production of mitochondrial reactive oxygen species, or either by altering redox-related genes' expression or activity or affecting cellular metabolism (Feugere et al., 2023;Rollwitz and Jastroch, 2021;Fig. 5. Overall antioxidant responses of zebrafish embryos and larvae exposed to polystyrene nanoplastics (NanoPS) at 28 • C or 32 • C. Tg(EPRE:EGFP) at 6 h postfertilization (hpf) were exposed to increasing concentrations of NanoPS at 28 • C or 32 • C and activation of electrophile responsive element (EPRE)-mediated responses were estimated by EGFP quantification through fluorescence microscopy in 96 hpf larvae. ...
... Loughland et al., 2022). For instance, zebrafish embryos exposed to cadmium, copper, or lead at warmer temperatures (26 • C vs 34 • C) have increased expression of the antioxidant genes catalase, superoxide dismutase, and glutathione S-transferase (Park et al., 2020a;Park et al., 2020b;Park and Kwak, 2022), while heat stress alone can increase the activity of the pentose phosphate pathway to produce reducing equivalents to the antioxidant system (Feugere et al., 2023). Increasing temperatures stimulate the synthesis of glutathione, specifically the expression of glutathione synthetase, in zebrafish larvae (Long et al., 2012). ...
Plastic pollution is a growing environmental concern due to its ubiquitous impact on aquatic ecosystems. Nanoplastics can be generated from the breakdown of plastic waste and interact with organisms at the cellular level, potentially disrupting cellular physiology. We investigated the effects of 44 nm polystyrene nanoparticles (44 nm NanoPS) on the development and physiology of zebrafish (Danio rerio) in the presence of sublethal heat stress (32 °C vs control, 28 °C). We hypothesized that the simultaneous exposure to nanoplastics and rising temperatures seriously threaten developing fish. This combination could create a critical imbalance: rising temperatures may lead to heightened energy demands, while nanoplastic exposure reduces energy production, threatening animal survival. As expected, 32 °C increased markers associated with animal metabolism and developmental timing, such as growth, hatching, heart rate, and feeding. Changes in apoptosis dynamics, oxygen consumption rates, and a decrease in mitochondrial content were detected as adaptive processes to temperature. 44 nm NanoPS alone did not alter development but decreased mitochondrial efficiency in ATP production and increased apoptosis in the heart. Surprisingly, exposure to 44 nm NanoPS at 32 °C did not cause major implications to survival, developmental success, or morphology. Still, 44 nm NanoPS mitigated the temperature-driven change in heart rate, increased oxidative stress, and decreased the coupling efficiency of the less abundant and highly active mitochondria under heat stress. We highlight the interplay between temperature and nanoplastics exposure and suggest that the combined impact of nanoplastics and temperature stress results in a scenario where physiological adaptations are strained, potentially leading to compromised development. This research underscores the need for further investigation into the metabolic costs of plastic pollution, particularly in the context of global warming, to better understand its long-term implications for aquatic life.
... Expression of TLR18 during embryogenesis has been observed in P. fulvidraco, where TLR18 gene expression could be seen earliest at the early blastula stage with a continued downregulation from gastrula till hatching but was then again abundantly expressed in 1-day-old spawns . TLR18 gene activation has also been reported in D. rerio embryos collected 1-day post-fertilization in response to heat stress (Feugere et al. 2023). In rohu, during the embryogenesis, TLR18 was highly expressed in the late blastula and gastrula stage; however, low expression continued from then onwards even in the ~ 2-day-old spawns. ...
Toll-like receptors (TLRs) are key pattern recognition receptors (PRRs) that detect pathogen-associated molecular patterns (PAMPs) and elicit broadly acting innate immune responses. This study accomplished the identification, cloning, and sequencing of Labeo rohita TLR18 (LrTLR18). Structurally, LrTLR18 possesses a signal peptide, six leucine-rich repeats (LRRs), an LRR-C-terminal (LRR-CT) domain, a transmembrane (TM) region, and a Toll/IL-1 receptor (TIR) domain. Phylogenetically, LrTLR18 is evolutionarily closely related to the Schizothorax prenanti and Cyprinus carpio TLR18. The quantitative real-time PCR analysis revealed that LrTLR18 gene was expressed in all tested tissues, and among the tissues, the highest expression was observed in the eye followed by spleen, intestine, and gill in the descending order. During the ontogenic developments, the highest expression of LrTLR18 gene has been detected in the late blastula and gastrula stages. In response to the Aeromonas hydrophila and Edwardsiella tarda infections, LrTLR18 gene was differentially expressed in the blood, kidney, liver, and gills. Labeo rohita gills (LRG) cells in vitro infected with A. hydrophila and E. tarda, and red blood cells (RBCs) and peripheral blood leucocytes (PBLs) isolated from the rohu fingerlings infected with these pathogens revealed significantly (p < 0.05) enhanced expression of LrTLR18 gene. Following in vivo and in vitro stimulations with lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I: C), LrTLR18 gene expression was also significantly (p < 0.05) enhanced in various tissues, RBCs, PBLs, and macrophages. Together, these results highlight the important roles of the TLR18 against pathogenic invasions in fish.
Our viewpoint is that ethanol could act as a molecular messenger in animal and plant organisms under conditions of hypoxia or other stresses and could elicit physiological responses to such conditions. There is evidence that both animal and plant organisms have endogenous levels of ethanol, but reports on the changes induced by this alcohol at physiological levels are sparse. Studies have shown that ethanol has different effects on cell metabolism at low and high concentrations, resembling a hormetic response. Further studies have addressed the potential cellular and molecular mechanisms used by organisms to sense changes in physiological concentrations of ethanol. This article summarizes the possible mechanisms by which ethanol may be sensed, particularly at the cell membrane level. Our analysis shows that current knowledge on this subject is limited. More research is required on the effects of ethanol at very low doses, in plants and animals at both molecular and physiological levels. We believe that further research on this topic could lead to new discoveries in physiology and may even help us understand metabolic adjustments related to climate change. As temperatures rise more frequently, dissolved oxygen levels drop, leading to hypoxic conditions and consequently, an increase in cellular ethanol levels.
