Comparative Biochemistry and Physiology A-Molecular & Integrative Physiology最新文献

The creatine kinase system is crucial for maintaining cellular energy homeostasis and plays a role in regulating locomotor behavior in organisms, but its significance in the regulating the motionless behavior in olive flounder is limited. In the first experiment of this study, elevated levels of creatine kinase (CK) activity in the spinal cord were detected in the juvenile group (JG) flounder compared to the adult group (AG) flounder. In the second experiment, to further confirm the involvement of CK in the locomotor behavior, the adult flounder was given an intraperitoneal injection of creatine (150 mg/kg), while the flounder in the control group received a saline solution. After one week post-injection, the behavioral analysis revealed that the flounder in the creatine-treated group displayed higher levels of locomotor activity and a greater number of escape attempts in response to external stimuli when compared to the control group. However, the acute stress response, induced by intraperitoneal injection and characterized by tail beating, was significantly alleviated in the flounder in the creatine-treated group. Additionally, there was an upregulation of the UII and AchR genes in the spinal cord, as well as increased levels of UII and AchR in the muscle tissues of the creatine-treated flounder. However, a reduction in UI mRNA levels was observed in the brain of the flounder. Collectively, our data provide the evidence that the elevated enzyme activity and gene expression of creatine kinase play important roles in off-bottom swimming behavior in the JG flounder. Furthermore, administration of creatine improved the locomotor activity and alleviated the stress response in flounder, which is associated with regulation of the locomotor- and stress-related gene in the brain, spinal cord, and muscle.

Low-temperature stress poses a significant risk to the survival of both cultivated and wild fish populations. Existing studies have found that the pre-acclimation of fishes to moderate cold stress can stimulate the activation of acclimation pathways, thereby enhancing their tolerance to cold stress. The fitness of fish relies heavily on appropriately controlled transcriptional reactions to environmental changes. Despite previous characterization of gene expression profiles in various fish species during cold acclimation, the specific genes responsible for essential functions in this process remain largely unknown, particularly the down-regulated genes induced by cold acclimation. To investigate the genes involved in cold acclimation, this study employed real-time quantitative PCR (RT-qPCR), molecular cloning, microinjection techniques, and cold stress experiments to determine the genes that play an essential part in cold acclimation. Consequently, 18 genes were discovered to be down-regulated in larval zebrafish experiencing cold stress. All 18 genes successfully detected overexpression in zebrafish at 96 and 126 hpf (fold change ≥3), which declined with the growth of zebrafish. Following microinjection, it was observed that her8a, cyp51, lss, txnipb, and bhlha9 had an adverse impact on the survival rate of zebrafish larvae under cold stress. These genes have been identified to play significant roles in various biological processes. For instance, bhlha9 has been found to be involved in both limb development and temperature sensing and her8a has been implicated in neural development. Additionally, cyp51 and lss have been identified as participants in the cholesterol synthesis pathway. Txnipb has been reported to induce cell apoptosis, thereby potentially influencing the survival rate of zebrafish larvae under cold stress. These findings offered crucial data for the analysis of molecular processes related to cold tolerance and the development of cold-resistant fish breeding.

Quantifying physiological stress in wild animals is essential for understanding their health, reproductive success, and survival in a variable environment. The yellow-bellied marmot (Marmota flaviventer) study at the Rocky Mountain Biological Laboratory near Crested Butte, Colorado, USA is the world's second longest study of free-living mammals. Historically, we used a validated corticosterone radioimmunoassay (RIA) to measure fecal glucocorticoid metabolites (FGMs) as a proxy for physiological stress. However, the costs and risks associated with working with radioisotopes drove us to consider a more sustainable method. Here we evaluate the suitability of two competitive corticosterone enzyme assays (EIA), one from Cayman Chemical Company (CCC) and one from Arbor Assays (AA), to measure marmot FGMs via their cross-reaction. The findings revealed that the AA EIA better matched the RIA in terms of accuracy across high and low FGM concentrations, had superior assay parameters, showed the highest correlations with RIA results and effectively captured the annual variations in FGM concentrations, thus demonstrating its reliability for use in longitudinal studies. We further analytically validated the AA EIA for FGMs and confirmed its efficacy and lack of matrix effects, thus establishing its suitability for ongoing and future studies of FGMs in marmots. The transition to the AA EIA from the RIA ensures continued data integrity while enhancing safety and environmental sustainability.

An organism's oxygen supply capacity, measured as a ratio of a metabolic rate to its critical oxygen partial pressure, describes the efficacy of oxygen uptake and transport. This metric is sensitive to errors in oxygen measurement, especially near anoxia where the magnitude of instrument error as a proportion of total signal is magnified. Here, we present a conceptual and mathematical method that uses this sensitivity to identify, quantify, and therefore correct oxygen measurements collected using inaccurately calibrated sensors. When appropriate, adding a small correction value to each oxygen measurement counteracts the effects of this error and provides results that are comparable to data from accurately calibrated oxygen probes. We demonstrate, using simulated, laboratory, and literature datasets, how this method can be used post hoc to diagnose error in, correct the magnitude of, and reduce the variability in repeat measures of traits relevant to oxygen tolerance.

The embryonic chicken is a valuable model for studying the maturation of cardiovascular physiology and the responses of this organ system to environmental manipulations such as acute hypoxia. Hypoxia determines not only the general cardiovascular response but also is a tool to determine the system's maturation of reflexive control. Several studies suggest embryonic chicken's regulation of the cardiovascular response to hypoxia, but no studies have measured the blood chemistry changes that accompany these responses. To clarify the changes in blood parameters accompanying cardiovascular function changes during acute hypoxia, we designed a study to investigate the blood chemistry (pO₂, pCO₂, pH, lactate, glucose, and blood ions) in developing embryos during acute hypoxia (O₂ = 10 %). Embryos ranging from day 13 to 21 of incubation were sampled during a control period and at the end of a 5-min of hypoxia. Hypoxia caused bradycardia on all days of incubation. The maximal blood hypoxic response occurred on day 15, with lactate increasing 7-fold (2.5 to 16.6 mmol/l) while glucose levels decreased by 50 % (136 to 63 mg/dl). Furthermore, hypoxia reduced pH (7.40 to 7.26), which peaked on day 15. These data indicate that a 5-min exposure to 10 % O₂ is sufficient to induce dramatic changes in blood chemistry however chorioallantoic arterial blood pO₂ was unchanged on most days of the study. Therefore, given the cardiovascular response to hypoxia and the increase in blood lactate prior to airbreathing in the chicken embryo, the embryonic tissues experienced an acute stress that may be the basis for the change in cardiovascular function during the exposure.

The honey bee (Apis mellifera L.), as an eusocial insect species, is an important model organism in research focusing on ageing and longevity, due to prominent seasonal lifespan plasticity within the worker caste (summer and winter worker bees). In this study, we employed a screening approach to evaluate several molecular parameters, providing comprehensive insights into the antioxidative (superoxide dismutase and catalase activity, reduced glutathione and sulfhydryl group content, total antioxidative capacity), detoxifying (glutathione S-transferase and acetylcholinesterase activity), and immune (phenol oxidase and glucose oxidase activity) status, as well as vitellogenin content, in the summer and winter generation of honey bees, across ageing stages and in two body compartments: the whole abdomen and the head. Summer worker bees were collected weekly for six weeks, while winter bees were collected monthly for five months. The results of our study clearly indicate a reduced overall antioxidative capacity of older groups of worker bees from both generations, while the parameters of immune responsiveness mostly contributed to the separation between the two generations based on season rather than age categories. Detoxification ability appeared to be more susceptible to environmental factors. An age-dependent increase in vitellogenin content was recorded in the abdomen, but without seasonal differences. These findings provide an excellent starting point for further investigations into age-related changes, particularly within the context of honey bee sociality.

Red drum, Sciaenops ocellatus, are a marine teleost native to the Gulf of Mexico that routinely experiences periods of low oxygen (hypoxia). Recent work has demonstrated this species has the capacity to improve aerobic performance in hypoxia through respiratory acclimation. However, it remains unknown how hypoxia acclimation impacts anaerobic metabolism in red drum, and the consequences of exhaustive exercise and recovery. Juvenile fish were acclimated to normoxia (n = 15, DO 90.4 ± 6.42 %) or hypoxia (n = 15, DO 33.6 ± 7.2 %) for 8 days then sampled at three time points: at rest, after exercise, and after a 3 h recovery period. The resting time point was used to characterize the acclimated phenotype, while the remaining time points demonstrate how this phenotype responds to exhaustive exercise. Whole blood, red muscle, white muscle, and heart tissues were sampled for metabolites and enzyme activity. The resting phenotype was characterized by lower pH_e and changes to skeletal muscle ATP. Exhaustive exercise increased muscle lactate, and decreased phosphocreatine and ATP with no effect of acclimation. Interestingly, hypoxia-acclimated fish had higher pH_e and pH_i than control in all exercise time points. Red muscle ATP was lower in hypoxia-acclimated fish versus control at each sample period. Moreover, acclimated fish increased lactate dehydrogenase activity in the red muscle. Hypoxia acclimation increased white muscle ATP and hexokinase activity, a glycolytic enzyme. In a gait-transition swim test, hypoxia-acclimated fish recruited anaerobic-powered burst swimming at lower speeds in normoxia compared to control fish. These data suggest that acclimation increases reliance on anaerobic metabolism, and does not benefit recovery from exhaustive exercise.

Numerous studies report on the influence of temperature on blood gases in ectothermic vertebrates, but there is merely a cursory understanding of these effects in developing animals. Animals that develop in eggs are at the mercy of environmental temperature and are expected to lack the capacity to regulate gas exchange and may regulate blood gases by means of altered conductance for gas exchange. We, therefore, devised a series of studies to characterize the developmental changes in blood gases when embryonic alligators were exposed to 25, 30 and 35 °C. To determine how blood parameters were impacted by changes in embryonic temperature, blood was sampled from the chorioallantoic membrane artery. The blood in the chorioallantoic membrane artery is a mixture of oxygen-poor and oxygen-rich blood, which based on the embryonic vascular anatomy may reflect blood that perfuses the chemoreceptors of the developing animal. Our findings indicate that following a 48 h exposure to 25 °C or 35 °C, there was a positive relationship between CAM artery blood PO₂, PCO₂ and glucose. However, blood pH suggests embryonic alligators lack an acute regulatory mechanism for adjusting blood pH.

Climate change alters multiple abiotic environmental factors in aquatic environments but relatively little is known about their interacting impacts, particularly in developing organisms where these exposures have the potential to cause long-lasting effects. To explore these issues, we exposed developing killifish embryos (Fundulus heteroclitus) to 26 °C or 20 °C and 20 ppt or 3 ppt salinity in a fully-factorial design. After hatching, fish were transferred to common conditions of 20 °C and 20 ppt to assess the potential for persistent developmental plasticity. Warm temperature increased hatching success and decreased hatch time, whereas low salinity negatively affected hatching success, but this was only significant in fish developed at 20 °C. Temperature, salinity, or their interaction affected mRNA levels of genes typically associated with thermal and hypoxia tolerance (hif1a, hsp90b, hsp90a, hsc70, and hsp70.2) across multiple developmental timepoints. These differences were persistent into the juvenile stage, where the fish that developed at 26 °C had higher expression of hif1a, hsp90b, hsp90a, and hsp70.2 than fish developed at 20 °C, and this was particularly evident for the group developed at both high temperature and salinity. There were also long-lasting effects of developmental treatments on body size after four months of rearing under common conditions. Fish developed at low salinity or temperature were larger than fish developed at high temperature or salinity, but there was no interaction between the two factors. These data highlight the complex nature of the developmental effects of interacting stressors which has important implications for predicting the resilience of fishes in the context of climate change.

The classic melatonin biosynthesis pathway (Mel; N-acetyl-5-methoxytryptamine) involves two consecutive enzymatic steps that are decisive in hormone production: conversion of serotonin (5-hydroxytryptamine; 5-HT) to N-acetylserotonin (NAS) and the methylation of the last compound to Mel. This pathway requires the activity of the enzymes: the first is of the category of N-acetyltransferases (AANAT, SNAT, or NAT) and the second is N-acetylserotonin O-methyltransferase (ASMT; also known as HIOMT). However, quite recently, new information has been provided on the possibility of an alternative Mel synthesis pathway; it would include a two-step action by these enzymes, but in reverse order, where ASMT (or ASMTL, the enzyme related to ASMT) methylates 5-HT to 5-methoxytryptamine (5-MT), and then the last compound is acetylated by an enzyme of the category of N-acetyltransferases to Mel. In our study on the activity of enzymes in the Mel biosynthesis pathway in flounder skin, we have found an increase in 5-MT level, as a result of the increase in 5-HT concentration, which is followed by a growing concentration of Mel. However, we have not found any increase in Mel concentration, despite an increase in NAS in the samples. Our data strongly suggest an alternative way of Mel production in flounder skin in which 5-HT is first methylated to 5-MT, which is then acetylated to Mel.