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Sci Rep
2019 Jan 24;91:517. doi: 10.1038/s41598-018-37546-y.
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Toxicity of diatom-derived polyunsaturated aldehyde mixtures on sea urchin Paracentrotus lividus development.
Ruocco N
,
Annunziata C
,
Ianora A
,
Libralato G
,
Manfra L
,
Costantini S
,
Costantini M
.
Abstract
Diatom-derived polyunsaturated aldehydes (PUAs), decadienal, heptadienal and octadienal, derive from the oxidation of fatty acids and have cytotoxic and anticancer effects. PUAs, tested separately, induce malformations in sea urchin Paracentrotus lividus embryos. Decadienal induces the worst malformations and lowest survival rates. Interestingly, decadienal, heptadienal and octadienal place in motion several genes to counteract their negative effects. To date, no studies are available reporting on the effects of PUA mixtures on marine invertebrates. Here we test binary and ternary mixtures on embryonic development of P. lividus. Our findings demonstrate that mixtures of PUAs act (i) at morphological level in synergistic way, being much more severe compared to individual PUAs; (ii) at molecular level also reveal an additive effect, affecting almost all fifty genes, previously tested using individual PUAs. This study is relevant from an ecological point of view since diatoms are a major food source for both pelagic and benthic organisms. This work opens new perspectives for understanding the molecular mechanisms that marine organisms use in reacting to environmental natural toxin mixtures such as diatom PUAs.
Figure 1. Examples of malformations induced in (b–d) P. lividus plutei at 48 hpf after incubation with binary (decadienal 0.5 µM + heptadienal 1.0 µM; decadienal 0.5 µM + octadienal 1.5 µM; heptadienal 1.0 µM + octadienal 1.5 µM) and ternary (decadienal 0.5 µM + heptadienal 1.0 µM + octadienal 1.5 µM) mixtures in comparison with (a) the control (embryos in sea water without PUA mixtures); (e) embryos still at the gastrula stage. Bar, 50 µm.
Figure 2. Real-Time qPCR at blastula (5hpf), gastrula (21 hpf) and pluteus (48 hpf) stages. Histograms show the differences in expression levels of fifty genes involved in different embryonic processes, divided in four classes: stress, skeletogenesis, development/differentiation and detoxification. P. lividus embryos were grown in the presence of ternary PUA mixture, consisting of decadienal 0.5 µM, heptadienal 1.0 µM and octadienal 1.5 µM. Data are reported as a fold difference compared with control (mean ± SD) embryos in sea water without PUA mixture. Fold differences greater than ±2 (see red dotted horizontal guidelines at values of +2 and −2) were considered significant (see Supplementary Table S2 for the values).
Figure 3. Heat map (using Heatmapper available at www.heatmappear.ca; https://creativecommons.org/licenses/by-sa/2.0/) of differentially expressed genes versus the three developmental stages (early blastula, late gastrula and pluteus), at which developing sea urchin P. lividus embryos have been collected after PUA mixture at 0.5 µM decadienal, 1.0 µM heptadienal and 1.5 µM octadienal for Real Time qPCR. Color code: red, negative values of gene expression (down-regulated genes respect to the control, embryos developed in sea water without PUA mixture); green, positive values of gene expression (up-regulated genes respect to the control); black, genes for which there was no variation of expression respect to the control.
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