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Blebbistatin potently inhibits actin-myosin interaction, preventing contractile activity of excitable cells including cardiac myocytes, despite electrical excitation of an action potential (AP). We collected intracellular microelectrode recordings of pacemaker cells located in the sinoatrial region (SAR) of the zebrafish heart at room temperature and during acute warming to investigate whether or not blebbistatin inhibition of contraction significantly alters pacemaker cell electrophysiology. Changes were evaluated based on 16 variables that characterized the AP waveform. None of these AP variables nor the spontaneous heart rate were significantly modified with the application of 10 μM blebbistatin when recordings were made at room temperature. Compared with the control group, the blebbistatin-treated group showed minor changes in the rate of spontaneous diastolic depolarization (P = 0.027) and the 50% and 80% repolarization (P = 0.008 and 0.010, respectively) in the 26°C–29°C temperature bin, but not at higher temperatures. These findings suggest that blebbistatin is an effective excitation-contraction uncoupler that does not appreciably affect APs generated in pacemaking cells of the SAR and can, therefore, be used in zebrafish cardiac studies.

We tested in six fish species [Pacific lamprey (Lampetra richardsoni), Pacific spiny dogfish (Squalus suckleyi), Asian swamp eel (Monopterus albus), white sturgeon (Acipenser transmontanus), zebrafish (Danio rerio), and starry flounder (Platichthys stellatus)] the hypothesis that elevated extracellular [HCO3-] protects spontaneous heart rate and cardiac force development from the known impairments that severe hypoxia and hypercapnic acidosis can induce. Hearts were exposed in vitro to either severe hypoxia (~ 3% of air saturation), or severe hypercapnic acidosis (either 7.5% CO2 or 15% CO2), which reduced heart rate (in six test species) and net force development (in three test species). During hypoxia, heart rate was restored by [HCO3-] in a dose-dependent fashion in lamprey, dogfish and eel (EC50 = 5, 25 and 30 mM, respectively), but not in sturgeon, zebrafish or flounder. During hypercapnia, elevated [HCO3-] completely restored heart rate in dogfish, eel and sturgeon (EC50 = 5, 25 and 30 mM, respectively), had a partial effect in lamprey and zebrafish, and had no effect in flounder. Elevated [HCO3-], however, had no significant effect on net force of electrically paced ventricular strips from dogfish, eel and flounder during hypoxia and hypercapnia. Only in lamprey hearts did a specific soluble adenylyl cyclase (sAC) inhibitor, KH7, block the HCO3--mediated rescue of heart rate during both hypoxia and hypercapnia, and was the only species where we conclusively demonstrated sAC activity was involved in the protective effects of HCO3- on cardiac function. Our results suggest a common HCO3--dependent, sAC-dependent transduction pathway for heart rate recovery exists in cyclostomes and a HCO3--dependent, sAC-independent pathway exists in other fish species.

Understanding how the current warming trends affect fish populations is crucial for effective conservation and management. To help define suitable thermal habitat for juvenile Chinook salmon, the thermal performance of juvenile Chinook salmon acclimated to either 15 or 19°C was tested across a range of environmentally relevant acute temperature changes (from 12 to 26°C). Swim tunnel respirometers were used to measure routine oxygen uptake as a measure of routine metabolic rate (RMR) and oxygen uptake when swimming maximally as a measure of maximal metabolic rate (MMR) at each test temperature. We estimated absolute aerobic scope (AAS = MMR - RMR), the capacity to supply oxygen beyond routine needs, as well as factorial aerobic scope (FAS = MMR/RMR). All fish swam at a test temperature of 23°C regardless of acclimation temperature, but some mortality occurred at 25°C during MMR measurements. Overall, RMR and MMR increased with acute warming, but aerobic capacity was unaffected by test temperatures up to 23°C in both acclimation groups. The mean AAS for fish acclimated and tested at 15°C (7.06 ± 1.76 mg O2 kg-1 h-1) was similar to that measured for fish acclimated and tested at 19°C (8.80 ± 1.42 mg O2 kg-1 h-1). Over the entire acute test temperature range, while MMR and AAS were similar for the two acclimation groups, RMR was significantly lower and FAS consequently higher at the lower test temperatures for the fish acclimated at 19°C. Thus, this stock of juvenile Chinook salmon shows an impressive aerobic capacity when acutely warmed to temperatures close to their upper thermal tolerance limit, regardless of the acclimation temperature. These results are compared with those for other salmonids, and the implications of our findings for informing management actions are discussed.

Although neural modulation of heart rate is well established among chordate animals, the Pacific hagfish (Eptatretus stoutii) lacks any cardiac innervation, yet it can increase its heart rate from the steady, depressed heart rate seen in prolonged anoxia to almost double its normal normoxic heart rate, an almost fourfold overall change during the 1-h recovery from anoxia. The present study sought mechanistic explanations for these regulatory changes in heart rate. We provide evidence for a bicarbonate-activated, soluble adenylyl cyclase (sAC)-dependent mechanism to control heart rate, a mechanism never previously implicated in chordate cardiac control.

Transformation of earth's ecosystems by anthropogenic climate change is predicted for the 21st century. In many regions, the associated increase in environmental temperatures and reduced precipitation will have direct effects on the physiological performance of terrestrial and aquatic ectotherms and have already threatened fish biodiversity and important fisheries. The threat of elevated environmental temperatures is particularly salient for members of the Oncorhynchus genus living in California, which is the southern limit of their range. Here, we report the first assessments of the aerobic capacity of a Californian population of wild Oncorhynchus mykiss Walbaum in relationship to water temperature. Our field measurements revealed that wild O. mykiss from the lower Tuolumne River, California maintained 95% of their peak aerobic scope across an impressive temperature range (17.8-24.6°C). The thermal range for peak performance corresponds to local high river temperatures, but represents an unusually high temperature tolerance compared with conspecifics and congeneric species from northern latitudes. This high thermal tolerance suggests that O. mykiss at the southern limit of their indigenous distribution may be locally adjusted relative to more northern populations. From fisheries management and conservation perspectives, these findings challenge the use of a single thermal criterion to regulate the habitat of the O. mykiss species along the entirety of its distribution range.

This comprehensive review synthesizes findings from 43 studies in which students with disabilities utilized behavioral self-management (BSM) techniques in general education settings. Findings suggest that the long-standing promise of BSM as an inclusive technique has been partially fulfilled. The review identifies strengths and limitations of BSM studies and BSM techniques, provides recommendations for future research and practice, and identifies BSM training materials.

Abstract There is a paucity of information on the physiological changes that occur over the course of salmon early marine migration. Here we aim to provide insight on juvenile Coho salmon (Oncorhynchus kisutch) physiology using the changes in gene expression (cGRASP 44K microarray) of four tissues (brain, gill, muscle, and liver) across the parr to smolt transition in freshwater and through the first eight months of ocean residence. We also examined transcriptome changes with body size as a covariate. The strongest shift in the transcriptome for brain, gill, and muscle occurred between summer and fall in the ocean, representing physiological changes that we speculate may be associated with migration preparation to feeding areas. Metabolic processes in the liver were positively associated with body length, generally consistent with enhanced feeding opportunities. However, a notable exception to this metabolic pattern was for spring post-smolts sampled soon after entry into the ocean, which showed a pattern of gene expression more likely associated with depressed feeding or recent fasting. Overall, this study has revealed life stages that may be the most critical developmentally (fall post-smolt) and for survival (spring post-smolt) in the early marine environment. These life stages may warrant further investigation. [ABSTRACT FROM AUTHOR]

Pipelines carrying diluted bitumen (dilbit) from Canada's oil sands traverse North America, including the freshwater habitat of Pacific salmon, posing a risk of environmental release and aquatic exposure. Swimming performance is impacted in juvenile sockeye ( Oncorhynchus nerka ) exposed to dilbit; therefore biomarkers of dilbit exposure will be valuable for monitoring at-risk salmon stocks. This study characterized changes in the serum proteome of sockeye exposed to a sub-lethal and environmentally relevant concentration of dilbit using isobaric tags for relative and absolute quantitation (iTRAQ), and included a range of experimental conditions to permit identification of biomarkers that are robust across time (1 and 4 wk) and exercise level (at rest and following a swim test). Over 500 proteins were identified and quantified in sockeye serum, with dilbit exposure significantly altering the abundance of 24 proteins irrespective of time and exercise, including proteins associated with immune and inflammatory responses, coagulation, and iron homeostasis. An increase in creatine kinase (CK) activity in serum of dilbit-exposed salmon confirmed the higher CK protein abundance measured using iTRAQ. The combination of 4 wk dilbit exposure and a swim test had a greater effect on the serum proteome than either treatment alone, including a marked increase in tissue leakage proteins, suggesting that aerobic exercise exacerbates the serum proteome response to dilbit, and the increased cellular damage could impede exercise recovery. This study provides a foundation for the development of bio-monitoring tools for salmon stock assessments, and offers new insights into the sub-lethal toxicity of crude oil exposure in fish. [ABSTRACT FROM AUTHOR]

Few estimates of migration rates or descriptions of behavior or survival exist for wild populations of out-migrating Pacific salmon smolts from natal freshwater rearing areas to the ocean. Using acoustic transmitters and fixed receiver arrays across four years (2010-2013), we tracked the migration of >1850 wild sockeye salmon ( Oncorhynchus nerka) smolts from Chilko Lake, British Columbia, to the coastal Pacific Ocean (>1000 km distance). Cumulative survival to the ocean ranged 3-10% among years, although this may be slightly underestimated due to technical limitations at the final receiver array. Distinct spatial patterns in both behavior and survival were observed through all years. In small, clear, upper-river reaches, downstream migration largely occurred at night at speeds up to 50 km/d and coincided with poor survival. Among years, only 57-78% of smolts survived the first 80 km. Parallel laboratory experiments revealed excellent short-term survival and unhindered swimming performance of dummy-tagged smolts, suggesting that predators rather than tagging effects were responsible for the initial high mortality of acoustic-tagged smolts. Migration speeds increased in the Fraser River mainstem (~220 km/d in some years), diel movement patterns ceased, and smolt survival generally exceeded 90% in this segment. Marine movement rates and survival were variable across years, with among-year segment-specific survival being the most variable and lowest (19-61%) during the final (and longest, 240 km) marine migration segment. Osmoregulatory preparedness was not expected to influence marine survival, as smolts could maintain normal levels of plasma chloride when experimentally exposed to saltwater (30 ppt) immediately upon commencing their migration from Chilko Lake. Transportation of smolts downstream generally increased survival to the farthest marine array. The act of tagging may have affected smolts in the marine environment in some years as dummy-tagged fish had poorer survival than control fish when transitioned to saltwater in laboratory-based experiments. Current fisheries models for forecasting the number of adult sockeye returning to spawn have been inaccurate in recent years and generally do not incorporate juvenile or smolt survival information. Our results highlight significant potential for early migration conditions to influence adult recruitment. [ABSTRACT FROM AUTHOR]

We compared exhaustion-related physiological stress and physical injury as contributors to fish condition, longevity, and egg retention in two Pacific salmon species after their arrival at spawning areas. Adult female Pink SalmonOncorhynchus gorbuschaand Chum SalmonO. ketawere exposed to six experimental capture treatments that represented different levels of exhaustive exercise, air exposure, and injury. After we evaluated its reflex impairment and obtained a blood sample, each fish was released into its natal spawning channel with an external tag and later retrieved postmortem to evaluate spawning success via examining egg retention. Reflex impairment, plasma lactate, chloride, potassium, and osmolality varied among treatments, with differences generally driven by the length of exposure to capture stress, which included exhaustive exercise and air exposure. However, overall prespawn mortality was negligible (about 5%) and consistent across treatments for both species. We hypothesize that Pink and Chum Salmon are resilient to capture-related exhaustion upon reaching spawning areas because of a combination of low water temperature (about 12°C in this study) and a physiological shift towards increased use of anaerobic pathways during their final weeks of life. The capture and release of fish arriving at the spawning ground does not appear to influence survival, in contradiction to the results of other studies, which focused on earlier components of Pacific salmon spawning migrations. Fisheries adjacent to spawning sites represent the end of the continuum of salmon fisheries that begin with the high seas fishery and extend through the coastal and riverine environments. The mortality rates in this study should be interpreted cautiously by management until research efforts are broadened to provide a better understanding of how postrelease outcomes at different life stages compare in natural systems and under conditions more representative of real fisheries. Received May 20, 2012; accepted October 25, 2012 [ABSTRACT FROM AUTHOR]

Abstract: A large-scale functional genomics study revealed shifting metabolic processes in white muscle during the final 1300 km migration of wild sockeye salmon to their spawning grounds in the Fraser River, British Columbia. In 2006, Lower Adams stock sockeye salmon ceased feeding after passing the Queen Charlotte Islands, 850 km from the Fraser River. Enhanced protein turnover and reduced transcription of actin, muscle contractile and heme-related proteins were early starvation responses in saltwater. Arrival to the estuarine environment triggered massive protein turnover through induction of proteasomal and lysosomal proteolysis and protein biosynthesis, and a shift from anaerobic glycolysis to oxidative phosphorylation. Response to entry into freshwater was modest, with up-regulation of heat shock proteins and nitric oxide biosynthesis. High river temperatures resulted in a strong defense/immune response and high mortalities in 50% of fish. Arrival to the spawning grounds triggered further up-regulation of oxidative phosphorylation and proteolysis, down-regulation of protein biosynthesis and helicase activity, and continued down-regulation of muscle proteins and most glycolytic enzymes. However, sharp up-regulation of PFK-I indicated induction of glycolytic potential at the spawning grounds. The identification of potential environmental cues triggering genome-wide transcriptional shifts in white muscle associated with migration and the strong activation of proteasomal proteolysis were both novel findings. [Copyright &y& Elsevier]

Abstract: The effects of acute and subchronic aqueous hydrocarbon exposures in the ppb range (0.2–127μg/L total PAH) on the immune system in Pacific herring (Clupea pallasi) were examined through specific immunocompetency assays and a host resistance model using Listonella anguillarum. Short-term hydrocarbon exposure at the highest concentration significantly enhanced respiratory burst activity (RBA) in macrophages and decreased plasma lysozyme concentrations, however, subchronic exposure (4–57d) reduced RBA. Fish in the high exposure group were also less susceptible to the pathogen L. anguillarum following acute hydrocarbon exposure; however, this group was the most susceptible following subchronic exposures. These results are explained by a measured transient physiological stress response and long-term effects on ionoregulation. This study illustrates that hydrocarbon-elicited effects are dynamic and that toxic outcomes with respect to the teleost immune system depend on chemical concentrations and composition, exposure durations and the specific pathogen challenge. [Copyright &y& Elsevier]

A comprehensive data set for the acute toxicity of monochloramine to juvenile chinook salmon, Oncorhynchus tshawytscha, and the aquatic invertebrate Ceriodaphnia dubia are presented. For exposures up to 10 days, the equation LC50 = 7244t[sup -0.4525] (where LC50 = monochloramine concentration in mg/L for 50% lethality and t = time in minutes) can predict the LC50 value for monochlofamine in juvenile chinook salmon and accounts for 94.4% of the variability in the experimental data. Predictions of acute toxicity were less reliable for concentrations in the range 0.5 mg/L to 0.8 mg/L since the LT50 value varied substantially in this range. Post-exposure mortality occurred only with exposures of >0.67 mg/L monochloramine. Feeding fish immediately before and during acute monochloramine exposure created a large chloramine demand in test chambers. In all tests where fish were fed, measured monochloramine concentrations were below detectable levels (<10 mg/L) and no fish mortality occurred. Monochloramine toxicity to the freshwater invertebrate Ceriodaphnia dubia was studied with time-to-lethality tests at 26 different concentrations. The equation LC[sub 50] = 61600t[sup -1.0748] predicted the LC50 value for chloramine in C. dubia and accounted for 94.8% of the variability in the experimental data. The mathematical equations that describe the LC50 for juvenile chinook salmon and C. dubia were used as reference lines for the evaluation of relative monochloramine toxicity. Both juvenile chinook salmon and C. dubia were among the most sensitive fish and aquatic invertebrate species, respectively, C. dubia being more sensitive than juvenile chinook salmon for exposure durations >30 minutes. [ABSTRACT FROM AUTHOR]