Intersexual differences of heat shock response between two amphipods ( Eulimnogammarus verrucosus and Eulimnogammarus cyaneus) in Lake Baikal

PeerJ. 2017 Feb 21;5:e2864. doi: 10.7717/peerj.2864. eCollection 2017.


Acute temperature fluctuations are common in surface waters, and aquatic organisms may manifest physiological responses to punctuated temperature spikes long before behavioral responses. Ectotherms, especially cryophilic stenotherms such as those endemic to Lake Baikal (Siberia), may demonstrate specialized physiological responses to acute temperature increases because their proteomes have evolved to function most efficiently at lower temperatures (e.g., <10 °C). Therefore, our study questioned the nature and degree of variation in physiological response to acute thermal stress in two congenerous, endemic Baikal amphipod species, Eulimnogammarus verrucosus and Eulimnogammarus cyaneus. We hypothesized that because interspecific and intersexual thermosensitivity varies significantly among ectotherms, there would be divergent intersexual and interspecific strategies to withstand acute thermal stress, manifested in different protein compositions and concentrations. We exposed individuals to the species' respective LT50 for one hour followed by a three-hour recovery period. We then performed 1D-PAGE, Western blotting, 2D-PAGE, and Mass Spectrometry techniques and assessed relative intersexual and interspecific changes in proteomic composition and heat shock protein 70 level. Our results demonstrate that females tend to be more sensitive to an acute thermal stimulus than males, most likely because females allocate significant energy to reproduction and less to heat shock response, evidenced by females' significantly lower LT50time. Lower level of Hsp70 was found in females of the thermosensitive E. verrucosus compared to males of this species. No intersexual differences were found in Hsp70 level in thermotolerant E. cyaneus. Higher levels of hemocyanin subunits and arginine kinase were found in E. cyaneus females after heat shock and recovery compared to males, which was not found for E. verrucosus, suggesting interspecific mechanisms for E. cyaneus's higher thermotolerance. These differing responses between species and sexes of Baikal amphipods may reflect more general strategies for maintaining homeostatic conditions during acute thermal stress. As mean surface water temperatures increase worldwide, the net efficiency and efficacy of these strategies could give rise to long term changes in physiology, behavior, and interactions with other species, potentially precipitating population and community level alterations.

Keywords: Amphipoda; Baikal; Eulimnogammarus cyaneus; Eulimnogammarus verrucosus; Freshwater benthic invertebrates; Hsp70; Proteome; Stress response; Thermotolerance; Two-dimensional gel electrophoresis.

Grant support

This work was supported by grants from the Russian Science Foundation (14-14-00400), RFBR (15-04-06685, 15-29-01003, 16-34-00687), CRDF Global (FSCX-15-61168-0); Ministry of Education of Russia (6.1387.2017/PCh, “Goszadanie”) and DAAD (6.731.2016.DAAD), in part by a Fulbright Fellowship to MFM and a Pushkin Grant from the Nanovic Institute of European Studies to REG. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.