Subtle reproductive impairment through nitric oxide-mediated mechanisms in sea urchins from an area affected by harmful algal blooms
The health of the sea urchin Paracentrotus lividus, a key species in the Mediterranean Sea, is menaced by several pressures in coastal environments. Here, we aimed at assessing the reproductive ability of apparently healthy P. lividus population in a marine protected area affected by toxic blooms of Ostreospsis cf. ovata. Wide-ranging analyses were performed in animals collected prior to and during the bloom, as well as at several times thereafter, during the reproductive season. Adults showed a low fertilization rate, along with high nitric oxide (NO) levels in the gonads and the nitration of the major yolk protein toposome, which is an important player in sea urchin development. Serious developmental anomalies were observed in the progeny, which persist several months after the bloom. NO levels were high in the different developmental stages, which also showed variations in the transcription of several genes that were found to be directly or indirectly modulated by NO. These results highlight subtle but important reproductive flaws transmitted from the female gonads to the offspring with the NO involvement. Despite a recovery along time after the bloom, insidious damages can be envisaged in the local sea urchin population, with possible reverberation on the whole benthic system.
[…] For protein identification, sea urchins ovaries were collected and the protein extracts were digested by trypsin sequence grade upon precipitation in 10% TCA (trichloroacetic acid), reduction with 45 mM dithiothreitol and alkylation with 100 mM iodoacetamide. MS/MS analysis was carried out by a LTQ-Orbitrap Velos (Thermo Fisher Scientific), as previously described. Data Base searching was performed using the Sequest search engine of Proteome Discoverer 1.1 (Thermo Fisher Scientific). […]
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