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The Journal of Experimental Biology 205, 1831-1841 (2002)
© 2002 The Company of Biologists Limited

Temperature-dependence of mitochondrial function and production of reactive oxygen species in the intertidal mud clam Mya arenaria

D. Abele^1,*, K. Heise¹, H. O. Pörtner¹ and S. Puntarulo²

¹ Alfred Wegener Institut for Polar and Marine Research, Columbusstraße, 27568 Bremerhaven, Germany
² Physical Chemistry, School of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina

^* Author for correspondence (e-mail: abele{at}awi-bremerhaven.de )

Accepted 3 April 2002

Mitochondrial respiration, energetic coupling to phosphorylation and the production of reactive oxygen species (ROS) were studied in mitochondria isolated from the eurythermal bivalve Mya arenaria (Myoidea) from a low-shore intertidal population of the German Wadden Sea. Measurements were conducted both within the range of the habitat temperatures (5-15 °C) and when subjected to heat exposure at 20 °C and 25 °C. Experimental warming resulted in an increase in the rate of state 3 and state 4 respiration in isolated mitochondria. The highest respiratory coupling ratios (RCR) were found at 15 °C; at higher temperatures mitochondrial coupling decreased, and release of ROS doubled between 15 and 25 °C. ROS production was 2-3% of total oxygen consumption in state 3 (0.3-0.5 nmol ROS mg^-1 protein min^-1) at the habitat temperature, reaching a maximum of 4.3 % of state 3 respiration and 7 % of oligomycin-induced state 4+ respiration under heat stress. Thus, state 4 respiration, previously interpreted exclusively as a measure of proton leakage, included a significant contribution from ROS formation in this animal, especially under conditions of heat stress. Oxygen radical formation was directly dependent on temperature-controlled respiration rates in states 3 and 4 and inversely related to mitochondrial coupling (RCR+) in state 4. Mitochondrial ROS formation is therefore involved in cellular heat stress in this eurythermal marine ectotherm.

Key words: mitochondrial function, reactive oxygen species, heat stress, antioxidant, mud clam, Mya arenaria

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