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Lake Winnipeg, Manitoba, is Canada’s second largest commercial inland freshwater fishery, and concern over collapse of the walleye fishery is growing. Molecular techniques have been increasingly used to study responses of wild organisms to environmental and anthropogenic stressors. The present study used transcriptomics to examine the physiology of wild-caught walleye (Sander vitreus(Mitchell, 1818)) across Lake Winnipeg using non-lethal techniques. Gill transcriptomes of walleye sampled from the north and south basins of Lake Winnipeg, and the channel connecting them, exhibited distinct profiles implicating regionally specific biological responses. North basin walleye exhibited transcriptomic responses indicative of exposure to environmental stressors. Transcriptomic patterns suggested a shift to increased reliance on anaerobic metabolism and up-regulation of hypoxia-sensitive genes in north basin fish, possibly representing exposure to low-oxygen conditions. Exposure to environmental stressors may also have driven increases in gene transcripts associated with proteasomal catabolism, DNA repair, molecular chaperones, and ion regulation. North basin fish also exhibited transcriptomic patterns indicative of gill remodeling via regulation of the mTOR pathway, cytoskeleton reorganization, and fatty acid synthesis. Our results highlight the complexity of examining wild fish across environmental gradients but also the potential use of molecular techniques in elucidating organismal sensitivity to local environmental factors.

Mitochondrial proton and electron leak have a major impact on mitochondrial coupling efficiency and production of reactive oxygen species. In the first part of this chapter, we address the molecular nature of the basal and inducible proton leak pathways, and their physiological importance. The basal leak is unregulated, and a major proportion can be attributed to mitochondrial anion carriers, whereas the proton leak through the lipid bilayer appears to be minor. The basal proton leak is cell-type specific and correlates with metabolic rate. The inducible leak through the ANT (adenine nucleotide translocase) and UCPs (uncoupling proteins) can be activated by fatty acids, superoxide or lipid peroxidation products. The physiological role of inducible leak through UCP1 in mammalian brown adipose tissue is heat production, whereas the roles of non-mammalian UCP1 and its paralogous proteins, in particular UCP2 and UCP3, are not yet resolved. The second part of the chapter focuses on the electron leak that occurs in the mitochondrial electron transport chain. Exit of electrons prior to the reduction of oxygen to water at cytochrome c oxidase causes superoxide production. As the mechanisms of electron leak are crucial to understanding their physiological relevance, we summarize the mechanisms and topology of electron leak from complexes I and III in studies using isolated mitochondria. We also highlight recent progress and challenges of assessing electron leak in the living cell. Finally, we emphasize the importance of proton and electron leak as therapeutic targets in body mass regulation and insulin secretion.

Rainbow smelt (Osmerus mordax) were maintained in a long term acclimation study to elucidate temperature effects on the accumulation of trimethylamine oxide (TMAO) and to determine if the activity of trimethylamine oxidase (TMAoxi) plays a role in modulating the seasonally variable levels of TMAO. In the same experiment, the TMAO content was determined for several tissues at varying plasma TMAO concentrations. TMAO accumulation begins at 5–7°C, well above that which might be normally associated with an antifreeze response. The plasma concentration reached a plateau of 20 mM as temperatures reached 0°C. Plasma TMAO concentration drops to pre‐accumulation levels, less than 5 mM, when fish are held at elevated temperature (8–11°C) and increases when fish are chilled below ambient seawater temperatures. However, despite temperatures near or below 0°C, plasma TMAO decreases after the winter season. Changes in TMAoxi activity do not correlate with TMAO levels, suggesting that the activity of this enzyme does not play a key role in regulating TMAO concentrations in smelt. For the first time in any teleost fish, tissue TMAO contents in liver, kidney, brain, and intestine were found to strongly correlate with plasma TMAO concentrations. For these tissues, the intracellular and extracellular concentration of TMAO appears to be approximately equal. Conversely, the heart and white muscle accumulate TMAO, and in the case of white muscle, intracellular concentration is maintained at a constant level of approximately 35 mmol/kg, despite fluctuating plasma concentrations over a range from 0 to over 25 mM. J. Exp. Zool. 303A:283–293, 2005. © 2005 Wiley‐Liss, Inc.

Tissue levels of trimethylamine oxide (TMAO) were compared for seven teleost and two elasmobranch species captured from three depth ranges: shallow (<150 m), moderate (500–700 m), and deep (1,000–1,500 m). Within the teleosts, the deep-caught species had significantly greater TMAO content than shallow- or moderate-caught species. In all teleosts, muscle had substantially more TMAO than all other tissues. Kidney or, in some cases, liver had elevated trimethylamine (TMA) content, 2.20–9.65 mmol/kg, along with appreciable trimethylamine oxidase (TMAoxi) activity, suggesting active TMAO synthesis. No correlation was found between TMAoxi activity and TMAO content. The elasmobranchs in this study, Squalus acanthias and Centroscyllium fabricii from shallow and deep water, respectively, were both squaliform sharks. The deep-caught species had significantly more TMAO in all tissues than the shallow species. Furthermore, urea was significantly less in the deep species in all tissues except liver, while the urea:TMAO ratio was significantly less in all tissues. As with teleosts, the TMAO content of muscle was substantially higher for both elasmobranchs than in all other tissues. TMAoxi was below levels of detection in both elasmobranch species, suggesting that TMAO is obtained solely from the diet. This study expands the trend of increased muscle TMAO in deep-sea fish to a variety of other tissues. The accumulation of TMAO in various tissues in deep-sea teleosts and the accumulation of TMAO and concurrent urea decrease in a deep-sea elasmobranch in comparison to a shallow water species strongly support the contention that TMAO is of physiological importance in deep-sea fish. © 2002 Wiley-Liss, Inc.