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Long-Term Toxicity and Reactive Nitrogen Species (RNS) Production by the Mediterranean Sponge Gemmula foliacea Growing in Nitrogen-Limited Conditions.
To examine whether low nitrogen (N) concentrations promote diverse physiological strategies to alleviate N shortage, we evaluated the effects of N limitation on the reactive nitrogen species (RNS) production and the induction of a N-stress response by the Mediterranean sponge Gemmula foliacea. Cultured G. foliacea with or without N limitation were fed with a N-free diet for up to 3 months and N-ammonium (NH4+ ), -nitrate (NO3-), or -nitrite (NO2- ) treatments for up to 60 days. Sponge growth was controlled by the intensity of RNS-mediated signaling without adverse effects on the N-storage proteins (histones and actin) in the spongin organic matrix. Samples collected at day 0, 7, 14, 21, and 30, post-N-treatment, were used to assess the effect of N limitation and RNS on the sponge’s transcriptome. Both NO3- and NO2- enhanced sponge growth at 7 and 14 days (but not beyond) of the N-supply experiment, whereas ammonium treatments did not promote significant growth. The transcriptome responses of N- and RNS-treated G. foliacea were similar to those observed in the sponge G. williamsoni. All N- and RNS-responsive genes presented a time-dependent regulation. Transcripts related to the N-stress response, such as homologs to the NKA and GNA2 proteins, glutathione synthetase, and chaperone-like genes, were up-regulated at day 7, 14, and 21 in the N- and RNS-treated G. foliacea

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