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Abstract: Heat shock proteins are important components in the cellular defense against proteotoxic stress. This work aimed to reveal HSP70 (hsc70 plus hsp70) expression patterns in several marine species (fish, crabs and shrimps) within a community along a temperature gradient and at the upper thermal limit. The organisms were collected in the Tagus estuary and adjacent shore (in Cabo Raso), Portugal. Exposure trials were performed using the critical thermal maximum (CTMax) method in order to recreate a stress gradient of ecological relevance. Protein analysis was performed using an enzyme linked immunosorbent assay (ELISA). Organisms within each community (estuary, coast; subtidal, intertidal, supratidal) responded in several different ways: no change in HSP70 levels, an increase in HSP70 levels, or increases and decreases in HSP70 levels. These patterns of response occurred independently of taxa, CTMax and habitat type. Magnitude of expression relates to the habitat''s thermal conditions. Species from highly variable and hot habitats i.e. intertidal/supratidal zone, and living in greater shore heights produce higher amounts of HSP70. Demersal and subtidal species inhabit colder and more stable waters thus they seem to have a slower heat shock response. No clear pattern was observed for species of the same group (fish, crabs and shrimps) or congeneric species. HSP70 expression showed high intraspecific variability potentially due to genetic traits, environmental traits and condition status. [Copyright &y& Elsevier]

• Mussels exhibited intraspecific differences depending on each environmental scenario. • Harbour mussels exhibited differences in lethal and sublethal responses. • Female mussels showed more sensitivity after chemical and warming insults. • Lethal responses could determine the non-reversibility imprint of biomarker responses. The intertidal mussel B. rodriguezii is a representative species from hard bottom substrates where both anthropogenic and natural stressors are present. Pre-exposure to these different stressors can modify the tolerance to additional stressors such as warming. Moreover, this tolerance can vary depending on intraspecific variables such as the organism's sex. The effects of warming and its intraspecific variability in representative coastal species are crucial to understanding the tolerance to future environmental scenarios. The mussels were collected in different environmental scenarios, including low (Control), chemical (Harbour) and natural stressed (Estuary) sites, and then exposed to different water temperatures (10–30 °C) for 14 days. Lethal and sublethal responses were evaluated in different mussel populations. Thus, cumulative death rate, air survival time, heat shock proteins (HSC70/HSP70), total ubiquitin, catalase (CAT), glutathione-s-transferase (GST) and lipid peroxidation (TBARS) were assessed in mussels from different areas and different sexes. The results revealed diminished air survival time and high cumulative mortality rate in mussels collected at the harbour and those exposed to higher temperatures, respectively. The sublethal responses of the field animals showed different patterns according to the different areas investigated. Besides, the results revealed that these differences were also observed between sexes. Regarding the sublethal responses in mussels exposed to warming, the interactive effects of temperature and sites showed a strong influence on all biochemical parameters analyzed (p < 0.001). Therefore, harbour mussels showed a distinct pattern compared to other locations and reflecting the most damaging effects of warming. The influence of sex and its interactions with warming were also crucial in most of the sublethal responses (p < 0.05). Multivariate analysis was performed with all sublethal responses, and the different warming scenarios showed different groups according to the sites. In the predicted warming scenarios, males showed no differences between sites. In contrast to males, females showed differences between sites in the predicted and the worse-case warming scenarios. Our results highlight the importance of compensatory mechanisms in the mussel warming tolerance like HSP70. The influence of sex is also crucial in understanding warming tolerance in mussels chronically exposed to pollutants in their natural environment. Also, lethal endpoints are essential for understanding the non-reversibility signature of the observed biochemical responses. [ABSTRACT FROM AUTHOR]