Expression of the human hepatocyte growth factor cDNA in primary cultures of rat hepatocytes

Hepatocyte growth factor (HGF) and epidermal growth factor (EGF) are primary mitogens for hepatocytes in culture. hepatocytes express the HGF-receptor MET but not HGF itself. To investigate the influence of autocrine HGF expression on the proliferative potential of hepatocytes, primary cultures were submitted to retrovirus-mediated transduction of the human hgf (huHGF) cDNA. Expression of the transduced cDNA revealed a minimum 2-fold increase in HGF-mRNA, whereas expression of the Escherichia coli beta-galactosidase gene remained even. Estimation of huHGF copy numbers showed there was a minimum 4-fold increase, suggesting an increase in the population of transduced cells. Immunoprecipitation of excreted huHGF and growth bioassays proofed that HGF was present and functional. HGF is excreted into the medium and therefore, by diffusion, available to transduced and non-transduced cells. The increase in huHGF-transduced cells suggests that the autocrine pathway as opposed to the paracrine pathway, which are both present at the same time, confers a growth advantage to these cells.     

Assessment of largemouth bass (<Emphasis Type="Italic">Micropterus salmoides</Emphasis>) behaviour and activity at multiple spatial and temporal scales utilizing a whole-lake telemetry array

A whole-lake acoustic telemetry array was utilized to monitor the three-dimensional position of 20 largemouth bass (Micropterus salmoides). Code division multiple access (CDMA) technology enabled the simultaneous monitoring of the 20 transmitters (equipped with pressure and temperature sensors) at 15 s intervals with sub-meter accuracy. Fish were monitored between November 2003 and April 2004 to evaluate the behaviour of fish across different temporal and spatial scales. The distance moved by largemouth bass, assessed both on a daily and hourly basis, varied by season and was positively correlated with water temperature. For example, daily movement rates were 2.69 ± 1.45 km/day in mid November (average daily water temperature 5.9°C), 2.24 ± 0.73 km/day in early January (5.1°C), and 7.28 ± 2.62 km/day in mid April (7.7°C). Interestingly, daily movement rates varied by as much as 25 fold among individual fish. Visualization of fish swimming paths revealed that whereas some fish occupied discrete areas and made only localized movements, other individuals made lengthier journeys covering much of the lake in periods of as little as one day. Analysis of fish behaviour at a finer temporal scale revealed that during the winter, fish spend more than 95% of their time swimming at speeds less than 0.1 m/s (0.07 ± 0.24 m/s). During late fall, and especially in spring, swimming speeds were higher with mean swimming speeds of 0.11 ± 0.27 m/s and 0.19 ± 0.29 m/s, respectively. When the telemetry dataset was queried to simulate 24 h manual tracking intervals, it was clear that manual tracking data would not have been representative of actual daily movement rates, underestimating daily movement and swimming speeds by at least 75 fold. This study identifies the importance of evaluating fish activity at multiple spatial (whole lake to sub-meter position) and temporal (seasonal to seconds) scales and illustrates the potential of CDMA telemetry to yield such data.     

Facing the River Gauntlet: Understanding the Effects of Fisheries Capture and Water Temperature on the Physiology of Coho Salmon

An improved understanding of bycatch mortality can be achieved by complementing field studies with laboratory experiments that use physiological assessments. This study examined the effects of water temperature and the duration of net entanglement on physiological disturbance and recovery in coho salmon (Oncorhynchus kisutch) after release from a simulated beach seine capture. Heart rate was monitored using implanted electrocardiogram biologgers that allowed fish to swim freely before and after release. A subset of fish was recovered in respirometers to monitor metabolic recovery, and separate groups of fish were sacrificed at different times to assess blood and white muscle biochemistry. One hour after release, fish had elevated lactate in muscle and blood plasma, depleted tissue energy stores, and altered osmoregulatory status, particularly in warmer (15 vs. 10°C) and longer (15 vs. 2 min) capture treatments. A significant effect of entanglement duration on blood and muscle metabolites remained after 4 h. Oxygen consumption rate recovered to baseline within 7–10 h. However, recovery of heart rate to routine levels was longer and more variable, with most fish taking over 10 h, and 33% of fish failing to recover within 24 h. There were no significant treatment effects on either oxygen consumption or heart rate recovery. Our results indicate that fishers should minimize handling time for bycatch and maximize oxygen supply during crowding, especially when temperatures are elevated. Physiological data, such as those presented here, can be used to understand mechanisms that underlie bycatch impairment and mortality, and thus inform best practices that ensure the welfare and conservation of affected species.     

The effects of temperature change on the hatching success and larval survival of largemouth bass Micropterus salmoides and smallmouth bass Micropterus dolomieu

In this study, the effects of abrupt temperature change on the hatching success and larval survival of eggs, yolk-sac larvae (YSL) and larvae above nest (LAN), for both largemouth bass Micropterus salmoides and smallmouth bass Micropterus dolomieu were quantified. Temperature had a significant effect on hatching success and time to 50% mortality, with large heat shocks causing accelerated mortality. The temperature changes shown to influence survival of all life stages, however, were beyond what is typically experienced in the wild. Micropterus salmoides had greater egg hatching success rates and increased survival rates at YSL and LAN stages, relative to M. dolomieu. Additionally, egg hatching success and survival of LAN varied across nests within the study. These findings suggest that temperature alone may not account for variations in year-class strength and may emphasize the need for protection of the nest-guarding male Micropterus spp. to ensure recruitment.     

Calcium signaling around Mitochondria Associated Membranes (MAMs)

Calcium (Ca²⁺) homeostasis is fundamental for cell metabolism, proliferation, differentiation, and cell death. Elevation in intracellular Ca²⁺ concentration is dependent either on Ca²⁺ influx from the extracellular space through the plasma membrane, or on Ca²⁺ release from intracellular Ca²⁺ stores, such as the endoplasmic/sarcoplasmic reticulum (ER/SR). Mitochondria are also major components of calcium signalling, capable of modulating both the amplitude and the spatio-temporal patterns of Ca²⁺ signals. Recent studies revealed zones of close contact between the ER and mitochondria called MAMs (Mitochondria Associated Membranes) crucial for a correct communication between the two organelles, including the selective transmission of physiological and pathological Ca²⁺ signals from the ER to mitochondria. In this review, we summarize the most up-to-date findings on the modulation of intracellular Ca²⁺ release and Ca²⁺ uptake mechanisms. We also explore the tight interplay between ER- and mitochondria-mediated Ca²⁺ signalling, covering the structural and molecular properties of the zones of close contact between these two networks.     

Effects of an experimental short‐term cortisol challenge on the behaviour of wild creek chub Semotilus atromaculatus in mesocosm and stream environments

The consequences of stress on the behaviour of wild creek chub Semotilus atromaculatus outside the reproductive period were studied using a single intra‐coelomic injection of cortisol, suspended in coconut butter, to experimentally raise plasma cortisol levels. Behaviour between cortisol‐treated, sham‐treated (injected with coconut butter) and control S. atromaculatus was compared in a mesocosm system, using a passive integrated transponder array, and in a natural stream system (excluding shams), using surgically implanted radio transmitters. While laboratory time‐course studies revealed that the cortisol injection provided a physiologically relevant challenge, causing prolonged (c. 3 days) elevations of plasma cortisol similar to that achieved with a standardized chasing protocol, no differences in fine‐scale movements were observed between cortisol‐treated, sham‐treated and control S. atromaculatus nor in the large‐scale movements of cortisol‐treated and control S. atromaculatus. Moreover, no differences were observed in diel activity patterns among treatments. Differential mortality, however, occurred starting 10 days after treatment where cortisol‐treated S. atromaculatus exhibited nearly twice as many mortalities as shams and controls. These results suggest that, although the experimental manipulation of cortisol titres was sufficient to cause mortality in some individuals, there were compensatory mechanisms that maintained behaviours (i.e. including activity and movement) prior to death. This study is one of the first to use experimental cortisol implants outside a laboratory environment and during the non‐reproductive period and yields insight into how wild animals respond to additional challenges (in this case elevated cortisol) using ecologically meaningful endpoints.     

Molecular population genetics of the alcohol dehydrogenase locus in the Hawaiian drosophilid D. mimica

Sequence variation among 10 alleles of the alcohol dehydrogenase (Adh) gene of the Hawaiian drosophilid D. mimica was analyzed with reference to the evolutionary history of the Hawaiian subgroup as well as to levels and patterns of polymorphism of the Adh gene in continental drosophilid species. The Adh gene of D. mimica is less polymorphic than that of other drosophilid species, and no replacement substitutions were found. Statistical analyses of the Adh alleles suggested the action of balancing selection and revealed significant linkage disequilibrium among three of the variable sites. The effective population size was estimated to be only slightly smaller than that of continental species and, surprisingly, on the same order of magnitude as the actual size.      

Exposure to elevated pCO2 alters post‐treatment diel movement patterns of largemouth bass over short time scales

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