White Muscle Research Articles

Swimming Performance of Schizothorax sp. and Planning of Fish Passages in Nepal

Swimming performance of migrating fish is crucial for the planning and design of effective fish passages. Commonly the fish swimming speeds are classified into five categories: 1) Optimum Swimming Speed, 2) Maximum Sustained Swimming Speed, 3) Critical Swimming Speed, 4) Maximum Domed Swimming Speed, and 5) Burst Swimming Speed. Swimming characteristics are related to the different species' adaptions to the environmental conditions. A variety of different factors affect the swimming capacity of fish which is directly related to fish size and species-specific morphology. A crucial abiotic factor is water temperature, which affects the fish physiology and thereby the swimming speed of the fish. This means that the fish's swimming speed can increase with increased water temperature. The swimming speed also increases with fish size. Fish often use burst speeds when passing a step in a fish ladder or entering a culvert. Burst speed is the highest speed attainable for fish, and it is the maximum energy output to be gained by using the white muscle (red muscle in salmonids). The burst speed can only be maintained for short periods of time (seconds). In the Himalayas, the main objective of many fisheries development plans is to secure future habitats and ensure up- and downstream migration of fish. Schizothorax richardsonii is often a targeted species in the area due to its importance as a food resource. If a fish passage is prepared for the upstream migration of fish, it is also crucial for the fish population development to ensure a safe downstream passage. Data on swimming performance for many of the native fish species in Nepal are scarce which complicates the design of suitable ecologically adapted fishways. In general, nature-like fish passes allow most of the species to migrate, followed by vertical slot fishways, meander fishways, and thereafter fish ladders of the pool-and-weir type. The key factor is to construct fish-passes that under all naturally required environmental circumstances can allow appropriate passage conditions of the targeted fish species.

Cloning, phylogenetic and expression analysis of two MyoDs in yellowtail kingfish (Seriola lalandi)

Yellowtail kingfish (Seriola lalandi) is a pelagic piscivore distributed circumglobally. Owing to its great market value, the growth mechanism of S. lalandi, including muscle development and growth, is a hot research topic. The myoblast determination protein (MyoD) gene has been shown to play an important role in formation of myoblasts and the function of somites in fish. The open reading frame (ORF) sequences of MyoD1 and MyoD2 in S. lalandi encoded 298 and 263 amino acids possessing three common characteristic domains, respectively, containing a myogenic basic domain, a bHLH domain, and a ser-rich region (helix III). S. lalandi MyoDs shared the highest identity with the MyoDs of S. dumerili. MyoDs are highly expressed in white muscle (P < 0.05) in S. lalandi. The expression level of MyoD1 mRNA was higher than that of MyoD2 mRNA during embryonic and early developmental stages, indicating that the two MyoD isoforms may have different roles in muscle formation. Moreover, the mRNA expression of MyoDs in the brain, pituitary, liver and muscle of endocrine growth axis were analyzed in the various sizes and ages stages. The expression levels of MyoDs in the different sizes and ages of S. lalandi showed that expression of both these genes was particularly high in 400-g fish and 2-year-old fish (P < 0.05). Moreover, the increases in the mRNA expression and plasma levels of growth hormone (GH) and insulin-like growth factor (IGF-I) were accompanied by an increase in mRNA expression of MyoDs, indicating the roles of GH and IGF-I in muscle development and growth of S. lalandi. Overall, the expression profiles of genes associated with muscle development are the first step taken towards deciphering fast growth mechanism in this important Seriola fish.

Non-lethal sampling method for the analysis of white muscle fatty acid profiles in European sea bass (Dicentrarchus labrax).

This study presents a novel non-lethal sampling method for assessing fatty acid (FA) composition in juvenile European sea bass (Dicentrarchus labrax) using subcutaneous white muscle biopsies. This research aimed to evaluate the suitability of the biopsy for FA analysis using two lipid extraction protocols and comparing them to a lethal routine method. The results showed that a mass of fresh tissue as low as 1.4 mg provided good quality FA chromatograms for both reserve and membrane lipids. Although the biopsy method displayed high variability in terms of FA quantity among intra-individual replicates, it showed good FA profile repeatability in both reserve and membrane lipids. The study highlights the potential of this non-lethal approach for studying FA dynamics in fish, with its application being particularly promising for ecological and experimental studies. However, careful biopsy implementation is recommended to account for potential lipid droplet and lipid distribution variability within the tissue.

Effects of dapagliflozin monotherapy and combined aerobic exercise on skeletal muscle mitochondrial quality control and insulin resistance in type 2 diabetes mellitus rats

Type 2 diabetes mellitus (T2DM) is a prevalent, chronic metabolic disease. Sodium-glucose cotransporter-2 (SGLT2) inhibitors and aerobic exercise (AE) have shown promise in mitigating insulin resistance (IR) and T2DM. This study investigated the effects of dapagliflozin (Dapa) monotherapy and combined AE on mitochondrial quality control (MQC) in skeletal muscle and IR in T2DM rats. T2DM rats, induced by a high-fat diet/streptozotocin model, were randomly assigned to the following groups: T2DM+vehicle group (DMV), T2DM rats treated with Dapa (DMDa, 10 mg/kg/d), T2DM rats subjected to combined Dapa treatment and AE (DMDa+AE), and the standard control group (CON). Blood and skeletal muscle samples were collected after 6 weeks of intragastric administration and treadmill exercise. The results showed that DMDa monotherapy could reduce the accumulation of white adipose tissue and skeletal muscle lipid droplets and improve HOMA-IR. While the combined AE led to further reductions in subcutaneous white adipose tissue and fasting glucose levels, it did not confer additional benefits in terms of HOMA-IR. Furthermore, Dapa monotherapy enhanced skeletal muscle mitochondrial biogenesis (PGC-1α, NRF1, TFAM, and COX IV), mitochondrial dynamics (OPA1, DRP1, and MFN2), and mitophagy (PGAM5 and PINK1) related protein levels. Nevertheless, the combination of Dapa with AE treatment did not yield an additive effect. In conclusion, this study highlights the potential of SGLT2 inhibitors, specifically Dapa, in ameliorating IR and maintaining MQC in skeletal muscle in rats with T2DM. However, combined AE did not produce an additive effect, indicating the need for further research.

Effect on Muscle Cellularity of Diet Supplementation with Nannochloropsis gaditana Microalgae in the Final Fattening Phase of Gilthead Seabream Culture up to Commercial Size

Previous studies have shown that Nannochloropsis gaditana can partially replace fishmeal in the diet of gilthead seabream, Sparus aurata. However, its effect on muscle growth is hardly known. This experiment was carried out with gilthead seabream adults that were fed with N. gaditana at two inclusion levels (2.5 or 5%) either raw (R2.5 and R5 groups) or cellulose-hydrolyzed (H2.5 and H5 groups) for 45 days in the final fattening phase. The body length and body weight were measured in all fish at the beginning and end of the experiment. Also, the white muscle transverse area (WM), size, number and fibrillar density of the white fibers were measured in 9 fish group−1. After 45 days, the body parameters and the WM did not show significant differences among the groups. However, muscle cellularity did show significant differences, such that the hypertrophy values were higher in the H2.5 and H5 than in the R2.5 and R5 groups. On the contrary, R2.5 and R5 showed the highest fibrillar density and hyperplasia values, which are often positively correlated with the fillet firmness and therefore could improve the final quality of the fish. No significant differences attributable to the inclusion levels of N. gaditana were observed.

Transcriptomic analyses of juvenile Striped Bass (Morone saxatilis) exposed to chronic and acute temperature change.

Striped Bass are economically important, migratory fishes, which occur across a wide range of latitudes. Given their wide-ranging nature, Striped Bass can cope with a broad range of environmental temperatures, yet the mechanisms underlying this ability have not been thoroughly described. Heat shock proteins (HSPs) are inducible molecular chaperones, which help mitigate protein damage resulting from increased temperatures. The importance of HSPs has been demonstrated in a number of fish species, but their role in Striped Bass is poorly understood. This study characterizes changes in gene expression in juvenile Striped Bass, following acute and chronic temperature change. Fish were acclimated to one of three temperatures (15, 25 or 30°C) and sampled at one of two treatments (control or after CTmax), following which we assessed differential gene expression and gene ontology in muscle. It is clear from our differential expression analyses that acclimation to warm temperatures elicits more robust changes to gene expression, compared to acute temperature increases. Our differential expression analyses also revealed induction of many different heat shock proteins, including hsp70, hsp90, hsp40 and other small HSPs, after both acute and chronic temperature increase in white muscle. Furthermore, the most consistent gene ontology pattern that emerged following both acclimation and CTmax was upregulation of transcripts involved in "protein folding", which also include heat shock proteins. Gene ontology analyses also suggest changes to other processes after acclimation, including decreased growth pathways and changes to DNA methylation. Overall, these data suggest that HSPs likely play a major role in the Striped Bass's ability to tolerate warm waters.

Ocimum basilicum essential oil in pacu Piaractus mesopotamicus: anesthetic efficacy, distribution, and depletion in different tissues.

This study aimed to evaluate the anesthetic activity of Ocimum basilicum essential oil and the distribution and depletion of its major compounds in different tissues of the pacu, Piaractus mesopotamicus. Juveniles (319.08 ± 9.14g) were individually anesthetized with six concentrations of essential oil from O. basilicum (150, 180, 210, 240, 270, and 300mg L-1), while in a second experiment, fish (492.39 ± 51.51g) were subjected to a 10min immersion bath with essential oil from O. basilicum (300mg L-1). After anesthetic recovery, blood and tissue samples of the brain, gills, liver, spleen, and white muscle were collected at 0, 0.5, 1.0, 3.0, 6.0, 12.0, and 24h. A 300mg L-1 concentration induced anesthesia in the shortest time (193.11 ± 9.31), while at 270 and 300mg L-1 concentrations, the anesthetic recovery period was the longest (244.33 ± 12.44) Methyl chavicol and linalool were quantified in all tissue samples. The plasma concentrations of methyl chavicol differed (p < 0.05) at all evaluated times. Linalool decreased (p < 0.05) from 0 to 1h and decreased again only after 12h. Reduction percentages in 24h were 92.9% for methyl chavicol, and 97.2% for linalool. Elimination of the compounds methyl chavicol and linalool is slower in the gills, where lower elimination constants (0.03 and 0.15 per h) and longer half-lives (25.84 and 4.53h), respectively, are noted. In general, essential oil from O. basilicum compounds was readily eliminated, showing promising potential for use as an anesthetic in aquaculture.

Sarcolipin (sln) and Sarcoplasmic Reticulum calcium ATPase pump (serca1) expression increase in Japanese medaka (Oryzias latipes) skeletal muscle tissue following cold challenge

Endothermy is the process by which organisms maintain a constant body temperature despite dynamic environmental temperatures. There are two mechanisms organisms use to elevate body temperature: shivering thermogenesis (ST) and non-shivering thermogenesis (NST). Skeletal muscle NST is achieved through a futile Ca2+ cycling of sarcoplasmic reticulum Ca2+ ATPase (Serca1) in the presence of sarcolipin (Sln). Here we subjected Japanese medaka to a cold challenge to examine the expression of sln and serca1 transcripts from slow-twitch red and fast-twitch white muscle as environmental temperature decreased. We show a significant increase in relative sln and serca1 transcript expression in skeletal muscle tissues of cold-challenged Japanese medaka. The elevated transcripts support the role of Sln as a component of NST and support previous literature with the increase in serca1. To date, this is the first cold challenge on an ectothermic fish investigating sln transcripts. The ability of medaka to respond to a cold challenge with an increase in key calcium cycling components, specifically the calcium pump and sarcolipin suggest that teleost fish share a conserved transcriptional program in response to cold stimuli with fish species that possess the requisite anatomical adaptations to conserve metabolic heat.

Evolutionary Adaptation of Protein Turnover in White Muscle of Stenothermal Antarctic Fish: Elevated Cold Compensation at Reduced Thermal Responsiveness.

Protein turnover is highly energy consuming and overall relates to an organism's growth performance varying largely between species, e.g., due to pre-adaptation to environmental characteristics such as temperature. Here, we determined protein synthesis rates and capacity of protein degradation in white muscle of the cold stenothermal Antarctic eelpout (Pachycara brachycephalum) and its closely related temperate counterpart, the eurythermal common eelpout (Zoarces viviparus). Both species were exposed to acute warming (P. brachycephalum, 0 °C + 2 °C day-1; Z. viviparus, 4 °C + 3 °C day-1). The in vivo protein synthesis rate (Ks) was monitored after injection of 13C-phenylalanine, and protein degradation capacity was quantified by measuring the activity of cathepsin D in vitro. Untargeted metabolic profiling by nuclear magnetic resonance (NMR) spectroscopy was used to identify the metabolic processes involved. Independent of temperature, the protein synthesis rate was higher in P. brachycephalum (Ks = 0.38-0.614 % day-1) than in Z. viviparus (Ks= 0.148-0.379% day-1). Whereas protein synthesis remained unaffected by temperature in the Antarctic species, protein synthesis in Z. viviparus increased to near the thermal optimum (16 °C) and tended to fall at higher temperatures. Most strikingly, capacities for protein degradation were about ten times higher in the Antarctic compared to the temperate species. These differences are mirrored in the metabolic profiles, with significantly higher levels of complex and essential amino acids in the free cytosolic pool of the Antarctic congener. Together, the results clearly indicate a highly cold-compensated protein turnover in the Antarctic eelpout compared to its temperate confamilial. Constant versus variable environments are mirrored in rigid versus plastic functional responses of the protein synthesis machinery.

Daily thermal variability does not modify long-term gene expression relative to stable thermal environments: A case study of a tropical fish

Global warming is leading to an increase in the frequency and intensity of extreme weather events, magnifying the breadth of temperatures faced by ectotherms across days and seasons. Despite the importance and ecological relevance of diurnal thermal variability, the vast majority of knowledge on gene expression patterns and physiology stems from animals acclimated to constant temperatures or in the early stages of exposure to a new temperature regime. If heterothermal environments modulate responses differently from constant thermal environments, our existing capacity to forecast impacts of climate warming may be compromised. To address this knowledge gap, we acclimated barramundi (Lates calcarifer) to 23 °C, 29 °C (optimal), 35 °C and to thermal cycling conditions (23–35 °C daily with a mean of 29 °C) and sampled liver and white muscle tissue before acclimation and after 2 and 17 weeks of acclimation. NanoString nCounter technologies were used to measure expression of 20 genes related to metabolism, growth and maintenance of cellular homeostasis. Acclimation to cool and warm conditions caused predictable changes in whole-animal performance (metabolism and growth) and the underlying gene expression patterns. Acclimation to a cycling temperature regime did not change the molecular regulation of metabolism or growth compared with barramundi acclimated to constant 29 °C, nor did it cause any discernible effects on whole-animal performance. However, the heat shock response was higher in the former group, suggesting that barramundi under a daily temperature cycle have an increased need for cellular chaperoning to minimise detrimental effects of temperature on proteins. We conclude that the genetic regulation of metabolism and growth may be more dependent on the mean daily temperature than on the daily temperature range.

THU507 Tumor Activin A Mediates Cachexia Primarily Through White Adipose Tissue Hydrolysis In Pancreatic Ductal Adenocarcinoma

Abstract Disclosure: S. Yu: None. Y. Luan: None. R. Dong: None. A. Abazarikia: None. S. Kim: None. Pancreatic ductal adenocarcinoma (PDAC) commonly harbors mutations in KRAS and TP53 genes and its overall 5-year survival rate is only 10% in the US. Among risk factors associated with the dismal PDAC prognosis, cachexia accounts for about 30% of PDAC-related death and is present in nearly 80% of PDAC patients. Significant adipose tissue loss followed by skeletal muscle loss characterizes PDAC-related cachexia in humans and mice. Accumulated evidence indicates that activin A, a homodimer of inhibin βA that is encoded by the INHBA gene, is a recognized key cachexia mediator as supported by mitigation of weight loss by activin A antagonists in PDAC-bearing mice. However, much remains unclear regarding the primary source cells of activin A and the mechanisms by which activin A mediates cachexia in PDAC. To address the unknowns, we employed CRISPR/Cas9 system for Inhba gene knock-down (KD) in mouse KPC (KrasG12D and Trp53R172H) PDAC cells and generated orthotopic mice using Inhba KD or control KPC cells. Sham group which received the same operation except for cell injection was included. Body weight was measured on a daily basis, and blood and white adipose tissues (WATs) and skeletal muscle tissues were harvested on post-surgery day 12. We found that, when compared to the Sham group, injection of control KPC cells significantly elevated serum activin A levels up to 6 ng/mL and temporally decreased the average body weight in the orthotopic mice (Ctrl group), whereas Inhba gene KD significantly reduced serum activin A levels by ∼ 2 ng/mL and prevented weight loss in the orthotopic mice (KD group). When compared to the Sham and KD groups, notable reductions in organ weights of epididymal, mesenteric or subcutaneous WATs were found in the Ctrl group, however, weights for gastrocnemius and plantaris or soleus muscle tissues were statistically similar between the Ctrl and KD groups at post-surgery day 12. Correspondingly, the average serum free fatty acids levels were higher in the Ctrl group than the KD group. Histological observations support that dramatic adipocyte size reductions in WATs were prevented by serum activin A suppression. Immunofluorescence analysis showed that WATs from the Ctrl group had considerably higher protein expression of hormone-sensitive or adipose triglyceride lipase than the KD and Sham groups. Furthermore, phosphorylation of SMAD3 and phosphorylated PKA substrate which regulate adipocyte lipolysis were intensely stained in WATs from the Ctrl group when compared to the KD and Sham groups. Collectively, our findings identify PDAC cancer cells as the major source of activin A in PDAC and suggest systemic activin A as a potential mediator of PDAC-related adipose tissue loss. The current findings warrant further investigation to address clinical relevance. Presentation: Thursday, June 15, 2023

Acute thermal stress increased enzyme activity and muscle energy distribution of yellowfin tuna.

Heat is a powerful stressor for fish living in natural and artificial environments. Understanding the effects of heat stress on the physiological processes of fish is essential for better aquaculture and fisheries management. In this experiment, a heating rod was used to increase the temperature at 2°C/h to study the changes of energy allocation (CEA) and energy metabolity-related enzyme activities, including pepsin, trypsin, amylase, lipase, acid phosphatase, lactate dehydrogenase, alanine aminotransferase, glutamic oxalic aminotransferase and energy reserve (Ea), energy expenditure (ETS), in juvenile yellowfin tuna cells under acute temperature stress. The results showed that the Ea of juvenile yellowfin tuna muscles in response to high temperature (34°C) was significantly lower than that of the control (28°C), and it also increased ETS. At 6 h, CEA decreased slightly in the high-temperature group, but, the difference in CEA between 24 h and 0 h decreased. After heat stress for 6 h, the activities of acid phosphatase (ACP), lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and glutamic oxalacetic transaminase (AST) increased, indicating that the metabolic rate was accelerated. After heat stress for 24 h, the activity of ALT decreased, indicating that with time elapsed, the activities of some protein metabolizing enzymes increased, and some decreased. In this study, digestive enzymes, trypsin and lipase increased gradually. After heat stress, Ea and Ec change significantly. Yellowfin tuna muscles use lipids in response to sharp temperature increases at high temperatures, red muscles respond to temperature changes by increasing energy in the early stages, but not nearly as much, and white muscles reduce lipids.

Physiological responses to acute warming at the agitation temperature in a temperate shark.

Thermal tolerance and associated mechanisms are often tested via the critical thermal maximum (CTMax). The agitation temperature is a recently described thermal limit in fishes that has received little mechanistic evaluation. The present study used a temperate elasmobranch fish to test the hypothesis that this thermal tolerance trait is partially set by the onset of declining cardiorespiratory performance and the cellular stress response. Pacific spiny dogfish (Squalus suckleyi) were screened for cardiorespiratory and whole-organism thermal limits to test for associations between thermal performance and tolerance. Then, biochemical markers of secondary stress, aerobic and anaerobic enzyme activities, and molecular markers of cellular stress were determined for various tissues at the agitation temperature, and secondary stress markers were determined at CTMax. In dogfish, the agitation temperature was characterised by increased turning activity within experimental chambers and was equal to the temperature where dogfish exhibited maximum heart rate. Citrate synthase activity increased at the agitation temperature in white muscle relative to unmanipulated dogfish. Further, lactate dehydrogenase activity and accumulated lactate in the plasma and muscle were not affected by acute warming. Cellular stress was apparent in hypothalamus, gill filament, and ventricle, denoted by elevated transcript abundance of hsp70, but not hif1α. Conversely, CTMax was characterised by metabolic acidosis driven by anaerobic lactate production, signifying an increased reliance on anaerobic metabolism between the agitation temperature and CTMax. Together, these data provide partial support for our hypothesis, in that cellular stress, but not declining thermal performance, occurred at the agitation temperature.

Seasonal Dynamics of Protein Synthesis in the Tissues Of &lt;i&gt;Channa punctatus&lt;/i&gt; [Bloch], an Indian Air-Breathing Teleost

The rates of initial synthesis and maximal net synthesis of total protein at the prevailing water-temperatures were determined by measuring the 14 levels of incorporation at 1h and 6h after administration of C-leucine respectively into total protein of liver, gill, brain, white muscle and red muscle of Chana punctatus in different months in an annual cycle. A clear biphasic pattern of seasonal variation of protein synthesis ( particularly of the net synthesis) and break-down i s evident in liver, gill and brain. This is much less discernible in muscles (white and red) . Five distinct periods of the seasonal dynamics of protein synthesis and degradation have been described as "the rising phase-I" (January-April/May-June), "the declining phase" (April/May-June-July), "the rising phase-II"(July-August) , "the maintenance phase-I" (August-October/November),"the maintenance phase-II" (October/November-December/January), and their significance discussed.

Genetic Deletion of DNAJB3 Using CRISPR-Cas9, Produced Discordant Phenotypes.

Several pathways and/or genes have been shown to be dysregulated in obesity-induced insulin resistance (IR) and type 2 diabetes (T2D). We previously showed, for the first time, impaired expression of DNAJB3 mRNA and protein in subjects with obesity, which was concomitant with increased metabolic stress. Restoring the normal expression of DNAJB3 attenuated metabolic stress and improved insulin signaling both in vivo and in vitro, suggesting a protective role of DNAJB3 against obesity and T2D. The precise underlying mechanisms remained, however, unclear. This study was designed to confirm the human studies in a mouse model of dietary obesity-induced insulin resistance, and, if validated, to understand the underlying mechanisms. We hypothesized that mice lacking DNAJB3 would be more prone to high-fat (HF)-diet-induced increase in body weight and body fat, inflammation, glucose intolerance and insulin resistance as compared with wild-type (WT) littermates. Three DNAJB3 knockout (KO) lines were generated (KO 30, 44 and 47), using CRISPR-Cas9. Male and female KO and WT mice were fed a HF diet (45% kcal fat) for 16 weeks. Body weight was measured biweekly, and a glucose tolerance test (GTT) and insulin tolerance test (ITT) were conducted at week 13 and 14, respectively. Body composition was determined monthly by nuclear magnetic resonance (NMR). Following euthanasia, white adipose tissue (WAT) and skeletal muscle were harvested for further analyses. Compared with WT mice, male and female KO 47 mice demonstrated higher body weight and fat mass. Similarly, KO 47 mice also showed a slower rate of glucose clearance in GTT that was consistent with decreased mRNA expression of the GLUT4 gene in WAT but not in the muscle. Both male and female KO 47 mice exhibited higher mRNA levels of the pro-inflammatory marker TNF-a in WAT only, whereas increased mRNA levels of MCP1 chemokine and the ER stress marker BiP/Grp78 were observed in male but not in female KO 47 mice. However, we did not observe the same changes in the other KO lines. Taken together, the phenotype of the DNAJB3 KO 47 mice was consistent with the metabolic changes and low levels of DNAJB3 reported in human subjects. These findings suggest that DNAJB3 may play an important role in metabolic functions and glucose homeostasis, which warrants further phenotyping and intervention studies in other KO 47 and other KO mice, as well as investigating this protein as a potential therapeutic target for obesity and T2D.

Exhaustive exercise alters native and site-specific H2O2 emission in red and white skeletal muscle mitochondria

Mitochondrial reactive oxygen species (ROS) homeostasis is intricately linked to energy conversion reactions and entails regulation of the mechanisms of ROS production and removal. However, there is limited understanding of how energy demand modulates ROS balance. Skeletal muscle experiences a wide range of energy requirements depending on the intensity and duration of exercise and therefore is an excellent model to probe the effect of altered energy demand on mitochondrial ROS production. Because in most fish skeletal muscle exists essentially as pure spatially distinct slow-twitch red oxidative and fast-twitch white glycolytic fibers, it provides a natural system for investigating how functional specialization affects ROS homeostasis. We tested the hypothesis that acute increase in energy demand imposed by exhaustive exercise will increase mitochondrial H2O2 emission to a greater extent in red muscle mitochondria (RMM) compared with white muscle mitochondria (WMM). We found that native H2O2 emission rates varied by up to 6-fold depending on the substrate being oxidized and muscle fiber type, with RMM emitting at higher rates with glutamate-malate and palmitoylcarnitine while WMM emitted at higher rates with succinate and glyceral-3-phosphate. Exhaustive exercise increased the native and site-specific H2O2 emission rates; however, the maximal emission rates depended on the substrate, fiber type and redox site. The H2O2 consumption capacity and activities of individual antioxidant enzymes including the glutathione- and thioredoxin-dependent peroxidases as well as catalase were higher in RMM compared with WMM indicating that the activity of antioxidant defense system does not explain the differences in H2O2 emission rates in RMM and WMM. Overall, our study suggests that substrate selection and oxidation may be the key factors determining the rates of ROS production in RMM and WMM following exhaustive exercise.

Lactate signaling and fuel selection in rainbow trout: mobilization of energy reserves.

Lactate is now recognized as a regulator of fuel selection in mammals because it inhibits lipolysis by binding to the hydroxycarboxylic acid receptor 1 (HCAR1). The goals of this study were to quantify the effects of exogenous lactate on: 1) lipolytic rate or rate of appearance of glycerol in the circulation (Ra glycerol) and hepatic glucose production (Ra glucose), and 2) key tissue proteins involved in lactate signaling, glucose transport, glycolysis, gluconeogenesis, lipolysis, and β-oxidation in rainbow trout. Measurements of fuel mobilization kinetics show that lactate does not affect lipolysis as it does in mammals (Ra glycerol remains at 7.3 ± 0.5 µmol·kg-1·min-1), but strongly reduces hepatic glucose production (16.4 ± 2.0 to 8.9 ± 1.2 µmol·kg-1·min-1). This reduction is likely induced by decreasing gluconeogenic flux through the inhibition of cytosolic phosphoenolpyruvate carboxykinase (Pck1, alternatively called Pepck1; 60% and 24% declines in gene expression and protein level, respectively). It is also caused by lactate substituting for glucose as a fuel in all tissues except white muscle that increases glut4a expression and has limited capacity for monocarboxylate transporter (Mct)-mediated lactate import. We conclude that lipolysis is not affected by hyperlactatemia because trout show no activation of autocrine Hcar1 signaling (gene expression of the receptor is unchanged or even repressed in red muscle). Lactate regulates fuel mobilization via Pck1-mediated suppression of gluconeogenesis and by replacing glucose as a fuel. This study highlights important functional differences in the Hcar1 signaling system between fish and mammals for the regulation of fuel selection.

Osmoregulatory responses in the neotropical fish species Astyanax lacustris, exposed to single and combined microplastics, polycyclic aromatic hydrocarbons, and their mixture

Microplastic (MP) pollution poses a significant environmental threat. These MPs can adsorb toxic compounds such as polycyclic aromatic hydrocarbons (PAH), which are highly lipophilic and carcinogenic. To assess the potential effects of virgin MP, PAH, and MP+PAH in association with osmoregulation and energetic substrate, we conducted experiments with the tetra cardinal Astyanax lacustris. The environmentally relevant concentration of MP (10 mg L−1) and 20 % of the LC50–96 h of crude oil for A. lacustris (2.28 µg L−1) were used during the 96-h exposure. Fish were exposed to virgin MP, PAH, MPC (MP loaded with PAH), PAH+MP (PAH and MP in association), and the control without (CT) and with handling (CH). After 96 h, blood was collected for osmoregulatory parameters (plasma osmolality; Na+, K+, Cl−, Mg2+; glycose and lactate); gills for osmoregulatory enzyme activities (Na+, K+ ATPase, H+ ATPase, and carbonic anhydrase); and white muscle samples were used to determine glycogen as an energetic substrate. The low molecular weight PAH was not detected in PAH-loaded MP (MPC) and PAH in combination with MP (PAH+MP). The PAH concentration of the MPC and PAH+MP was similar and low compared to other works. Virgin MP, PAH, MPC, and PAH+MP were able to cause muscle glycogen depletion. The activity of v-type H+ ATPase and plasma Na+ concentrations were lower in PAH with MP (MPC). However, the hydromineral balance (K+, Mg2+, Cl−, and osmolality) was not affected by any treatment. In this sense, we can conclude that the MPC caused osmoregulatory disturbances not seen in the MP associated with PAH (MP+PAH). However, this seems unrelated to the PAH leaking from the MPC or the PAH absorption to the virgin MP once the PAH concentrations from the MPC and PAH+MP were similar.

Red and melanized focal changes in white skeletal muscle in Atlantic salmon (Salmo salar): Comparative analysis of farmed, wild and hybrid fish reared under identical conditions.

Selective breeding plays a vital role in the production of farmed Atlantic salmon and has shown success in many aspects. Still, challenges related to fish health and welfare continue to result in significant economic losses. One such challenge is red and melanized focal changes (RFC/MFC), which result from acute and chronic inflammation, respectively, in the skeletal muscle. Importantly, RFC/MFC has not been observed in wild Atlantic salmon, suggesting that both external and genetic factors may contribute to the development of inflammation. To investigate the underlying cause of RFC/MFC, we conducted a study involving 1854 Atlantic salmon of farmed, wild and hybrid origin. All fish were reared under identical conditions to minimize the influence of external factors. Throughout the production cycle, the fish was monitored for growth parameters and examined for RFC/MFC using macroscopic and histological analysis. We found no association between the experimental groups and the presence of RFC/MFC. Histological investigations revealed melano-macrophages in the soft tissue in freshwater smolt, although no macroscopic discoloration was observed. MFC showed granulomas in various stages, suggesting a complex progression of the condition. In summary, we conclude that RFC/MFC is primarily caused by external factors found in the rearing facilities of farmed Atlantic salmon.

Daily Rhythmicity of Muscle-Related and Rhythm Genes Expression in Mackerel Tuna (Euthynnus affinis).

The aim of this study was to investigate the circadian rhythm of muscle-related gene expression in mackerel tuna under different weather conditions. The experiment was carried out under two weather conditions at four sampling times (6:00, 12:00, 18:00, and 24:00) to determine the expression of growth, function, and rhythm genes: white muscle rhythm genes were rhythmic on sunny and cloudy days, except for PER3 and RORA; all functional genes had daily rhythmicity. Red muscle had daily rhythmicity on both sunny and cloudy days; functional genes had daily rhythmicity except for MBNL. The expression levels of the rhythm gene PER1 were determined to be significantly different by independent t-test samples in white muscle at 6:00, 12:00, 18:00, and 24:00 under different weather conditions; the expression levels of the functional genes MBNL and MSTN were both significantly different. In the red muscle, the expression of the rhythm genes PER3, REVERBA, and BMAL1 was determined by independent t-test samples at 6:00, 12:00, 18:00, and 24:00 on cloudy and sunny days; the functional gene MBNL was significantly different. The present study showed that mackerel tuna muscle rhythm genes and functional genes varied significantly in expression levels depending on weather, time of day, and light intensity and that the expression levels of myogenic genes were closely related to clock gene expression. The fish were also able to adapt to changes in light intensity in different weather conditions through positive physiological regulation.