Search

Broiler chickens reared under heat stress (HS) conditions have decreased growth performance and show metabolic and immunologic alterations. This study aimed to evaluate the effect of supplementation with a standardized blend of plant-derived isoquinoline alkaloids (IQ) on the growth performance, protein catabolism, intestinal barrier function, and inflammatory status of HS-treated chickens. Three hundred sixty 0-day-old Ross 308 male broiler chickens were randomly distributed into 2 treatment groups: control diet (no additives) or diet supplemented with 100 ppm IQ. At day 14, the chicks in each diet group were further divided into 2 groups, each of which was reared under thermoneutral (TN) (22.4°C) or constant HS (33.0°C) conditions until day 42. Each group consisted of 6 replicates with 15 birds per replicate, and chickens were provided ad libitum access to water and feed. During days 15–21, the body weight gain (BWG) and feed intake (FI) were significantly lower in the HS treatment group than in the TN group, and feed conversion ratio was higher (P

A study using pair-feeding technique was conducted to determine whether heat exposure directly or indirectly (via reduced feed intake) increases intestinal mucosal damage and permeability to endotoxin in broiler chickens. Male broiler chickens (Ross 308), 27-d-old, were subjected to one of the three treatments (n=8): 1) thermo-neutral conditions (24°C) with ad libitum feed intake, 2) heat stress conditions (33°C) with ad libitum feed intake, or 3) pair-feeding under thermo-neutral conditions, with the feed intake identical to that of heat-stressed chickens. Using these groups, two experiments were performed to evaluate temporal changes in the intestinal morphology in response to each treatment. In experiment 1, chickens were sacrificed after 24 h of exposure to the treatment conditions, while in experiment 2, chickens were sacrificed after 12 or 72 h of exposure to the treatment conditions. In experiment 1, exposure to heat stress conditions for 24 h significantly decreased both the villus height to crypt depth ratio and number of proliferating cell nuclear antigen (PCNA)-positive cells in the duodenum and increased the plasma endotoxin concentration. These findings were not observed in pair-fed chickens. In experiment 2, intestinal integrity and function were unaffected by 12 h of heat stress. On the other hand, chickens exposed to heat stress for 72 h exhibited significantly damaged intestinal morphology in the duodenum as well as increased plasma endotoxin concentration; these negative effects were not observed in pair-fed chickens. These findings suggest that the intestinal morphology and permeability changes observed in chickens that are heat-stressed for 24–72 h are due to the heat stress conditions and not due to reduced feed intake.

This study investigated the intracellular mechanism governing the effects of oleuropein (OLE), a phenolic compound of Olea europaea, on mRNA expression of avian uncoupling protein (avUCP) and mitochondrial biogenesis-related factors, and reactive oxygen species (mitROS) generation in a primary cultured chicken muscle cells. The OLE-treated cells exhibited increases in Avucp and ATP5a1z expression and a decrease in mitROS generation (p 0.05), while the effects was canceled by sirtuin-1 (SIRT1) or transient receptor potential vanilloid 1 (TRPV1) inhibitors, EX-527 or BCTC, respectively. Intracellular Ca

The aims of the present study were to identify the differences between two mouse lines (high (H)- and low (L)-oxygen consumption) in terms of mitochondrial respiratory activity when GMP (glutamate, malate, and pyruvate) and succinic acid are used as substrates and to examine the relationship between mitochondrial respiration activity and feed efficiency in both lines. The average daily feed intake, feed conversion ratio (FCR), and residual feed intake (RFI) were significantly higher in the H than the L line. The correlation between FCR and RFI was significant (r = 0.60, p < 0.05). RFI was effective as an indicator of feed efficiency. When succinic acid was used as a substrate, mitochondrial respiration states 2-4, ACR, and proton leak were significantly higher in the H than the L line. When GMP was used as a substrate, respiration states 3 and 4 in the H line were significantly higher than those in the L line, and there were significant positive correlations between FCR and RFI and mitochondrial respiration states 2-4. The results indicated that selection for high or low OC changed the basal metabolic rates estimated from liver mitochondrial respiration activity and feed efficiency.

Peptide transporter 1 (PepT1) is a transporter responsible for absorbing dipeptide and tripeptide in enterocytes and is upregulated by dipeptide in mammals. It has not been certain whether intestinal PepT1 expression is responsive to dipeptides in chickens because of the lack of in vitro study using the cultured enterocytes. This study established a primary culture model of chicken intestinal epithelial cells (IECs) in two-dimensional monolayer culture using collagen gel by which the response of chicken PepT1 gene expression to dipeptide stimuli was evaluated. The cultured chicken IECs showed the epithelial-like morphology attached in a patch-manner and exhibited positive expression of cytokeratin and epithelial cadherin, specific marker proteins of epithelial cells. Moreover, the chicken IECs exhibited the gene expression of intestinal cell type-specific marker, villin1, mucin 2, and chromogranin A, suggesting that the cultured IECs were composed of enterocytes as well as goblet and enteroendocrine cells. PepT1 gene expression was significantly upregulated by synthetic dipeptide, glycyl-l-glutamine, in the cultured IECs. From the results, we herein suggested that dipeptide is a factor upregulating PepT1 gene expression in chicken IECs.

The skeletal muscle growth rate is a major feature differentiating meat- and laying-type chickens. A large amount of ATP is required during skeletal muscle synthesis, in which mitochondrial energy production capacities play a significant role. Additionally, mitochondria may participate in muscle protein degradation via reactive oxygen species generation. To investigate the differences in mitochondrial energetic characteristics between chickens exhibiting different growth rates, this study evaluated respiratory capacities in response to different types of respiratory substrate, protein abundances, assembly of individual respiratory complexes (I-V) and supercomplexes, and reactive oxygen species generation rates. These characteristics were compared between mitochondria from the breast muscle (M. pectoralis superficialis) of seven-week-old meat- and laying-type male chickens. Blue native polyacrylamide gel electrophoresis analysis revealed that meat-type chickens exhibited a significantly lower protein abundance of complex III (cytochrome bc 1 complex), complex V (F0F1 ATP synthase), and total amount of supercomplexes than did laying-type chickens. There were no differences between chicken types in the respiration rate of mitochondria incubated with either pyruvate/malate or succinate, each of which drives complex I- and complex II-linked respiration. Carnitine palmitoyltransferase-1-dependent and -independent respiration during ATP synthesis and carnitine palmitoyltransferase-2 enzymatic activity were significantly lower in meat-type chickens than in layingtype chickens. For mitochondria receiving pyruvate/malate plus succinate, the reactive oxygen species generation rate and its ratio to the oxygen consumed (the percentage of free radical leak) were also significantly lower in meat-type chickens than in laying-type chickens. These results suggested that the mitochondrial energetic capacities of the breast muscle of meat-type chickens could be lower than those of laying-type chickens at seven weeks of age. Furthermore, the lower reactive oxygen species generation rate in meat-type chickens might have implications for rapid muscle development, which is possibly related to their lower muscle protein degradation rates.

Excess glucocorticoid secretion induces oxidative damage and muscle proteolysis and modulates glucose and lipid metabolism. It is known that the high-temperature (HT) treatment enhances corticosterone (CORT) secretion, muscle proteolysis, and mitochondrial reactive oxygen species (mtROS) generation in chickens. The present study investigated the co-effects of CORT on proteolysis and mtROS production, together with glucose, fatty acid, and amino acid metabolism in HT-treated cells. Myoblast cells were isolated from the major pectoralis muscle of five 0- or 1-day-old neonatal chicks and were precultured at 37°C/CO2 conditions for 48 h to reach subconfluent (80%-90%) conditions. Cells were then reseeded onto a 6- or 24-well microplate for the subsequent measurement, followed by the culture under a control temperature (37°C, control) or HT (41°C) conditions for 1 or 6 h. The HT-treated cells were cocultured with physiologically relevant concentrations of CORT (20 ng/mL in dimethyl sulfoxide). The HT treatment decreased cellular protein content (P < 0.05) and increased atrogin-1 mRNA levels and mtROS generation levels compared to the control group (P < 0.05), whereas HT/CORT co-treatment did not induce changes in either parameter. The mRNA level of glucose transporter-1 was decreased in HT-treated cells compared to that in normal cells (P < 0.05), and the decrease was increased in the CORT co-treatment (P < 0.05). While HT treatment did not alter pyruvate dehydrogenase kinase-4 mRNA level, the level was increased in the CORT co-treatment compared to the control and HT-treated cells (P < 0.05). Neither HT nor HT/CORT treatments altered the mRNA levels of fatty acid oxidation-related factors, carnitine palmitoyl transferase-1, and cluster of differentiation 36. The study conducted a metabolic analysis using gas chromatography-mass spectrometry. The results showed that HT/CORT-treated cells had decreased intracellular citrate and α-ketoglutarate levels (P < 0.05) and increased extracellular alanine and amino acid that have gluconeogenic properties, as well as increased aspartate, isoleucine, serine, methionine, and threonine levels (P < 0.05) compared to HT-treated cells. These results suggest that CORT may not affect proteolysis and mtROS production but can suppress pyruvate oxidation and promote alanine production in HT-treated chickens.

Meat and bone meal (MBM) is an important protein source used in animal feeds. However, as the composition and availability of amino acids (AAs) in MBM fluctuate markedly, it is important to verify the magnitude of these parameters in different MBMs. In this study, the AA compositions of 19 MBMs were analyzed to confirm variations in lysine (Lys), threonine (Thr), and tryptophan (Trp), then which were compared with those of soybean meal (SBM) and fish meal (FM). Instability in Lys, Thr, and Trp availabilities in six MBMs were also considered after estimation using the slope-ratio growth assay technique in rats. Variations in AA composition were evaluated using the coefficient of variance (CV: Standard deviation/Mean). CVs for Lys, Thr, and Trp content were 9.40, 11.83, and 18.12 in MBM, 2.71, 2.48, and 3.19 in SBM, and 10.09, 10.44, and 13.47 in FM. Furthermore, means and SDs for Lys, Thr, and Trp availabilities in MBM were 53.3 ± 10.4% (CV: 19.5), 65.9 ± 17.6% (CV: 26.6), and 83.2 ± 11.2% (CV: 13.5), respectively. These results provide the first evidence that variations in MBM AA compositions were 3.5 to 5.7 times higher than those in SBM, but similar to FM, and that the large variation in availability substantially existed.

Mitochondrial content is regarded a useful feature to distinguish muscle-fiber types in terms of energy metabolism in skeletal muscles. Increasing evidence suggests that specific mitochondrial bioenergetic phenotypes exist in metabolically different muscle fibers. A few studies have examined the energetic properties of skeletal muscle in domestic fowls; however, no information on muscle bioenergetics in broiler chickens selectively bred for faster growth is available. In this study, we aimed to characterize the mitochondrial contents and functions of chicken skeletal muscle consisting entirely of type I (oxidative) (M. pubo-ischio-femoralis pars medialis), type IIA (glycolytic/oxidative) (M. pubo-ischio-femoralis pars lateralis), and type IIB (glycolytic) (M. pectoralis) muscle fibers. Citrate synthase (CS) activity was the highest in type IIA muscle tissues and isolated mitochondria, among the muscle tissues tested. Although no difference was registered in mitochondrial CS activity between type IIB and type I muscles, tissue CS activity was significantly higher in the latter. Histochemical staining for NADH tetrazolium reductase and the ratio of muscle-tissue to mitochondrial CS activity indicated that type I, type IIA, and type IIB muscle-fiber types showed decreasing mitochondrial content. Mitochondria from type I muscle exhibited a higher coupled respiration rate induced by pyruvate/malate, palmitoyl-CoA/malate, and palmitoyl-carnitine, as respiratory substrates, than type IIB-muscle mitochondria, while the response of mitochondria from type IIA muscle to those substrates was comparable to that of mitochondria from type I muscle. Type IIA-muscle mitochondria exhibited the highest carnitine palmitoyltransferase-2 level among all tissues tested, which may contribute to the higher fatty acid oxidation in these mitochondria. The results suggest that mitochondrial abundance is one of the features differentiating metabolic characteristics of different chicken skeletal muscle types. Moreover, the study demonstrated that type IIA-muscle mitochondria may have distinct metabolic capacities.

Mitochondrial dysfunction increases oxidative stress and depletes ATP in a variety of disorders. Several antioxidant therapies and drugs affecting mitochondrial biogenesis are undergoing investigation, although not all of them have demonstrated favorable effects in the clinic. We recently reported a therapeutic mitochondrial drug mitochonic acid MA-5 (Tohoku J. Exp. Med., 2015). MA-5 increased ATP, rescued mitochondrial disease fibroblasts and prolonged the life span of the disease model “Mitomouse” (JASN, 2016). To investigate the potential of MA-5 on various mitochondrial diseases, we collected 25 cases of fibroblasts from various genetic mutations and cell protective effect of MA-5 and the ATP producing mechanism was examined. 24 out of the 25 patient fibroblasts (96%) were responded to MA-5. Under oxidative stress condition, the GDF-15 was increased and this increase was significantly abrogated by MA-5. The serum GDF-15 elevated in Mitomouse was likewise reduced by MA-5. MA-5 facilitates mitochondrial ATP production and reduces ROS independent of ETC by facilitating ATP synthase oligomerization and supercomplex formation with mitofilin/Mic60. MA-5 reduced mitochondria fragmentation, restores crista shape and dynamics. MA-5 has potential as a drug for the treatment of various mitochondrial diseases. The diagnostic use of GDF-15 will be also useful in a forthcoming MA-5 clinical trial., Graphical Abstract Image 1, Highlights • MA-5 rescues patient's cell survival independent of genetic mutation backgrounds. • MA-5 promotes ATP synthase dimer formation followed by the precipitation of supercomplex. • The cell protective effect of MA-5 will be predicted by GDF-15 in vitro and in vivo. Mitochondrial diseases are life-threatening and progressive, yet largely untreatable because of the lack of adequate drug. We found a mitochondria-homing drug Mitochonic acid-5 (MA-5) that facilitates ATP production and restoring mitochondrial dynamics, rescuing a wide variety of human mitochondrial diseases cell survival. MA-5 binds to mitochondrial protein mitofilin/Mic60 and facilitating ATP synthase oligomerization and supercomplex formation that increase local ATP production, even under the genetically proton-limited condition in mitochondrial diseases. Therefore, MA-5 will be a candidate chemical for modulating mitochondrial function and remedy for not only mitochondrial diseases but also mitochondrial-related diseases (diabetes, diabetic nephropathy, cardiomyopathy, longevity etc.).

1. Oleuropein (Ole) is a major phenolic compound in Olea europaea, with anti-oxidative, anti-obesity, and anti-inflammatory properties. To explore the effect of Ole on the physiology and metabolism...

Maintenance energy requirements (MER) of mice selected for high (H) or low (L) oxygen consumption (OC) were compared. Forty-four mice from H and L OC lines were weaned at 3 weeks and divided into four experimental groups: group A were sacrificed at 4 weeks; group B were fed ad libitum, and groups C and D were fed 2.8 and 2.4 g/day, respectively, from 4 to 8 weeks of age. Groups B-D were sacrificed at 8 weeks. Chemical components were estimated for all groups. MER was estimated using a model that partitioned metabolizable energy intake into that used for maintenance, and protein and fat deposition. The feed conversion ratio for the B group was significantly higher in the H than in the L line. Feed intake for metabolic energy content per metabolic body size was significantly also higher in the H line, whereas accumulated energy content per metabolic body size was significantly higher in the L line. MER of the H line was greater than that of the L line (P

Heat stress (HS) induces muscle protein degradation as well as production of mitochondrial reactive oxygen species (ROS). In the present study, to improve our understanding of how protein degradation is induced by HS treatment in birds, a time course analysis of changes in the circulating levels of glucocorticoid and N(τ)-methylhistidine, muscle proteolysis-related gene expression, and mitochondrial ROS generation, was conducted. At 25 days of age, chickens were exposed to HS conditions (33 °C) for 0, 0.5, 1 or 3 days. While no alteration in plasma N(τ)-methylhistidine concentration relative to that of the control group was observed in the 0.5 day HS group, the concentration was significantly higher in the 3-d HS treatment group. Plasma corticosterone concentrations increased in response to 0.5-d HS treatment, but subsequently returned to near-normal values. HS treatment for 0.5 days did not change the levels of μ-calpain, cathepsin B, or proteasome C2 subunit mRNA, but increased the levels of mRNA encoding atrogin-1 (P

It has been shown that oleuropein, a phenolic compound in the fruit and leaves of the olive tree (Olea europaea) induces mammalian uncoupling protein 1 (UCP1) expression via an increased secretion of noradrenaline and adrenaline. This study investigated the effects of oleuropein on avian UCP (avUCP) expression as well as genes related to mitochondrial oxidative phosphorylation and biogenesis in cultured avian muscle cells, together with reactive oxygen species generation. Oleuropein induced avUCP as well as peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor-1 (NRF1), mitochondrial transcription factor A (TFAM) and ATP5a1 (a component of mitochondrial adenosine triphosphate synthase) gene expression and cytochrome c oxidase activity, indicating the induction of mitochondrial biogenesis. Sirtuin-1 (SIRT1) gene expression was also up-regulated by this compound, which could contribute to an increase in PGC-1α activity. Oleuropein suppressed the level of superoxide generation per mitochondrion, possibly via the up-regulation of avUCP and manganese superoxide dismutase (MnSOD) expression. Based on these findings, this study is the first to show that oleuropein may induce avUCP expression in avian muscle cells independent of the catecholamines, in which PGC-1α may be involved.

This study was conducted to determine if dietary supplementation with coenzyme Q10 (CoQ10 ), which can act as a potent antioxidant and is an obligatory cofactor of mitochondrial uncoupling protein, suppresses the heat stress (HS)-induced overproduction of mitochondrial reactive oxygen species (ROS) and oxidative damage in the skeletal muscle of birds. The carbonyl protein content of skeletal muscle was significantly higher in birds exposed to HS treatment (34°C, 12 h) than in thermoneutral birds (25°C). This increase was suppressed by CoQ10 supplementation (40 mg/kg diet). Succinate-supported mitochondrial ROS production was increased by HS treatment, and this increase was also suppressed by CoQ10 supplementation. In contrast, CoQ10 supplementation did not affect the HS-induced decrease in mitochondrial proton leak. The mitochondrial membrane potential (ΔΨ), to which HS-induced ROS production was previously shown to be sensitive, tended to be increased by HS treatment, but this rise in ΔΨ was not affected by CoQ10 supplementation. Taken together, these results suggest that dietary CoQ10 supplementation attenuates HS-induced oxidative damage to skeletal muscle, by preventing the overproduction of succinate-supported mitochondrial ROS in a manner that is independent of ΔΨ. © 2015 Japanese Society of Animal Science.

Heat stress is a major factor inducing oxidative disturbance in cells. In the present study, we investigated the mechanism of overproduction of reactive oxygen species (ROS) in cultured avian muscle cells in response to heat stress, and also focused attention on the interaction of mitochondrial superoxide anions with altered NADPH oxidase (NOX), superoxide dismutase (SOD) and heme oxygenase-1 (HO-1) mRNA levels in heat-stressed cells. Exposure of cells to heat stress conditions (41°C, 6h) resulted in increased mitochondrial superoxide and intracellular ROS levels, and increased carbonyl protein content as compared with that of normal cells (37°C). The mitochondrial uncoupler 2,4-dinitrophenol lowered intracellular ROS levels in heat-stressed cells. Heat stress increased NOX4 mRNA and decreased HO-1 mRNA levels, while SOD1 and SOD2 mRNA levels remained relatively stable in heat-stressed cells. Addition of the superoxide scavenger 4-hydroxy TEMPO to the culture medium of heat-stressed cells restored mitochondrial superoxide and intracellular ROS levels as well as NOX4 and HO-1 mRNA levels to near-normal values. We suggest that mitochondrial superoxide production could play an influential role in augmenting oxidative damage to avian muscle cells, possibly via the up-regulation of NOX4 and down-regulation of HO-1 in heat-stressed avian muscle cells.

SUMMARYButoxybutyl alcohol (BBA) is a possible growth promoter contained in the fermentation and distillation by-products of a traditional Japanese spirit, shochu. In the present study, BBA was synthesized and its chemical structure was confirmed by gas chromatography mass spectrometry and nuclear magnetic resonance. Then, two studies were conducted to investigate the effects of feeding the synthesized BBA on the growth and skeletal muscle proteolysis of broiler chickens. Ross male broiler chickens were divided into two groups, control (basal diet: 219 g crude protein/kg and 12·66 MJ metabolizable energy/kg) and BBA diet (30 mg BBA/kg basal diet), with the experimental diets being provided from 15 to 27 days and 0 to 27 days of age, for Studies 1 and 2, respectively. Butoxybutyl alcohol supplementation increased final body weight in both studies, whereas feed intake was unchanged, thereby indicating significantly increased feed efficiency. Furthermore, the synthesized BBA increased the weights of the pectoralis superficialis and profundus muscles, and the leg. The BBA decreased theNτ-methylhistidine concentration in the excrement and plasma, which are indices of the rate of skeletal muscle protein degradation. It also decreased the mRNA levels of μ-calpain large subunit, atrogin-1/muscle atrophy F-box (MAFbx), ubiquitin and 20S proteasome C2 subunit. These suggest that growth promotion due to the feeding of synthesized BBA is caused by the suppression of skeletal muscle protein degradation, which is related to a decrease in gene expression in the calpain and ubiquitin–proteasome systems.

Mitochondria are key organelles implicated in a variety of processes related to energy and free radical generation, the regulation of apoptosis, and various signaling pathways. Mitochondrial dysfunction increases cellular oxidative stress and depletes ATP in a variety of inherited mitochondrial diseases and also in many other metabolic and neurodegenerative diseases. Mitochondrial diseases are characterized by the dysfunction of the mitochondrial respiratory chain, caused by mutations in the genes encoded by either nuclear DNA or mitochondrial DNA. We have hypothesized that chemicals that increase the cellular ATP levels may ameliorate the mitochondrial dysfunction seen in mitochondrial diseases. To search for the potential drugs for mitochondrial diseases, we screened an in-house chemical library of indole-3-acetic-acid analogs by measuring the cellular ATP levels in Hep3B human hepatocellular carcinoma cells. We have thus identified mitochonic acid 5 (MA-5), 4-(2,4-difluorophenyl)-2-(1H-indol-3-yl)-4-oxobutanoic acid, as a potential drug for enhancing ATP production. MA-5 is a newly synthesized derivative of the plant hormone, indole-3-acetic acid. Importantly, MA-5 improved the survival of fibroblasts established from patients with mitochondrial diseases under the stress-induced condition, including Leigh syndrome, MELAS (myopathy encephalopathy lactic acidosis and stroke-like episodes), Leber's hereditary optic neuropathy, and Kearns-Sayre syndrome. The improved survival was associated with the increased cellular ATP levels. Moreover, MA-5 increased the survival of mitochondrial disease fibroblasts even under the inhibition of the oxidative phosphorylation or the electron transport chain. These data suggest that MA-5 could be a therapeutic drug for mitochondrial diseases that exerts its effect in a manner different from anti-oxidant therapy.

1. The present study was designed to achieve a reduction of reactive oxygen species (ROS)-induced oxidative damage to skeletal muscle and to improve the performance of broiler chickens exposed to chronic heat stress. 2. Chickens were given a control diet with normal drinking water, or diets supplemented with cashew nut shell liquid (CNSL) or grape seed extract (GSE), or a control diet with electrolysed reduced water (ERW) for 19 d after hatch. Thereafter, chickens were exposed to a temperature of either 34°C continuously for a period of 5 d, or maintained at 24°C, on the same diets. 3. The control broilers exposed to 34°C showed decreased weight gain and feed consumption and slightly increased ROS production and malondialdehyde (MDA) concentrations in skeletal muscle. The chickens exposed to 34°C and supplemented with ERW showed significantly improved growth performance and lower ROS production and MDA contents in tissues than control broilers exposed to 34°C. Following heat exposure, CNSL chickens performed better with respect to weight gain and feed consumption, but still showed elevated ROS production and skeletal muscle oxidative damage. GSE chickens did not exhibit improved performance or reduced skeletal muscle oxidative damage. 4. In conclusion, this study suggests that ERW could partially inhibit ROS-induced oxidative damage to skeletal muscle and improve growth performance in broiler chickens under medium-term chronic heat treatment.

Meat-type chickens show high feed efficiency and have a very rapid growth rate compared with laying-type chickens. To clarify whether the type-specific difference in feed conversion efficiency is involved in mitochondrial bioenergetics, modular kinetic analysis was applied to oxidative phosphorylation in skeletal muscle mitochondria of both type chickens. Mitochondria from skeletal muscle of meat-type chickens showed greater substrate oxidation and phosphorylating activities, and less proton leak than those of the laying-type, resulting in a higher efficiency of oxidative phosphorylation. Gene expression and protein content of uncoupling protein (avUCP) but not adenine nucleotide translocase (avANT) gene expression were lower in skeletal muscle mitochondria of meat-type chickens than the laying-type. The current results regarding a higher efficiency of oxidative phosphorylation and UCP content may partially support the high feed efficiency of meat-type chickens.

Heat stress (HS) causes poor growth, immunosupression and high mortality, through physiological dysfunction. We have reported that ‘acute’ HS increases mitochondrial reactive oxygen species (ROS) and oxidative damage in the skeletal muscle of broiler chickens (Cobb), but that this is not for the case for males of the laying-type white leghorn (WLH) strain of chickens (Mujahid et al., 2005b). We have now studied the effects of ‘chronic’ HS on performance and oxidative damage to skeletal muscle in different strains of chickens. Meat-type male chickens (Ross and Cobb) and male laying-type chickens (WLH) of 14d of age were arranged according to a factorial design (3 strains×4 conditions): control (24°C), cyclic (32-24-32°C: 32°C for 8h/d, 32-24-32HS), constant (32 and 34°C, 32HS and 34HS, respectively) for 14d. Feed consumption and body weight gain were measured. Thiobarbituric acid reactive substances (TBARS) content in skeletal muscle was measured colorimetrically with BHT (butylated hydroxytoluene) and expressed as malondialdehyde (MDA) equivalent. Body weight gain and feed consumption for the 3 strains (Ross, Cobb and WLH) were decreased by 32HS and 34HS, but not by 32-24-32HS. The decrease in weight gain of WLH chickens for the 32HS and 34HS conditions was smaller than for broiler chickens (Ross and Cobb). The 32HS and 34HS groups showed slightly enhanced MDA levels (P

Maintenance energy requirements (MER) of mice selected for high (H) or low (L) oxygen consumption (OC) were compared. Forty-four mice from H and L OC lines were weaned at 3 weeks and divided into four experimental groups: group A were sacrificed at 4 weeks; group B were fed ad libitum, and groups C and D were fed 2.8 and 2.4 g/day, respectively, from 4 to 8 weeks of age. Groups B-D were sacrificed at 8 weeks. Chemical components were estimated for all groups. MER was estimated using a model that partitioned metabolizable energy intake into that used for maintenance, and protein and fat deposition. The feed conversion ratio for the B group was significantly higher in the H than in the L line. Feed intake for metabolic energy content per metabolic body size was significantly also higher in the H line, whereas accumulated energy content per metabolic body size was significantly higher in the L line. MER of the H line was greater than that of the L line (P 0.10). These results suggest that selection for H or L OC produced differences in chemical components, feed efficiency, and MER between the H and L lines.

Mitochondria are a major source of reactive oxygen species production in cells, and the production level is sensitive to the magnitude of the membrane potential (ΔΨ). The present study investigated the level of superoxide production in mitochondria oxidizing glycerol 3-phosphate (GP) and its dependence on ΔΨ in isolated avian muscle mitochondria. The levels of superoxide produced in mitochondria oxidizing GP were lower than those obtained with succinate and were similar to those obtained with NADH-linked substrates (glutamate/malate/pyruvate). The dependence of superoxide production on ΔΨ in mitochondria oxidizing GP was lower than that of mitochondria oxidizing succinate, and a weak dependence of GP-supported superoxide production on ΔΨ was observed in the presence of NADH-linked substrates or succinate. These results suggest that the levels of superoxide generated in response to GP are quantitatively low, but they are unsusceptible to changes in ΔΨ in avian muscle mitochondria.

It has been shown that oleuropein, a phenolic compound in the fruit and leaves of the olive tree (Olea europaea) induces mammalian uncoupling protein 1 (UCP1) expression via an increased secretion of noradrenaline and adrenaline. This study investigated the effects of oleuropein on avian UCP (avUCP) expression as well as genes related to mitochondrial oxidative phosphorylation and biogenesis in cultured avian muscle cells, together with reactive oxygen species generation. Oleuropein induced avUCP as well as peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor-1 (NRF1), mitochondrial transcription factor A (TFAM) and ATP5a1 (a component of mitochondrial adenosine triphosphate synthase) gene expression and cytochrome c oxidase activity, indicating the induction of mitochondrial biogenesis. Sirtuin-1 (SIRT1) gene expression was also up-regulated by this compound, which could contribute to an increase in PGC-1α activity. Oleuropein suppressed the level of superoxide generation per mitochondrion, possibly via the up-regulation of avUCP and manganese superoxide dismutase (MnSOD) expression. Based on these findings, this study is the first to show that oleuropein may induce avUCP expression in avian muscle cells independent of the catecholamines, in which PGC-1α may be involved.

Mitochondrial dysfunction causes increased oxidative stress and depletion of ATP, which are involved in the etiology of a variety of renal diseases, such as CKD, AKI, and steroid–resistant nephrotic syndrome. Antioxidant therapies are being investigated, but clinical outcomes have yet to be determined. Recently, we reported that a newly synthesized indole derivative, mitochonic acid 5 (MA-5), increases cellular ATP level and survival of fibroblasts from patients with mitochondrial disease. MA-5 modulates mitochondrial ATP synthesis independently of oxidative phosphorylation and the electron transport chain. Here, we further investigated the mechanism of action for MA-5. Administration of MA-5 to an ischemia-reperfusion injury model and a cisplatin–induced nephropathy model improved renal function. In in vitro bioenergetic studies, MA-5 facilitated ATP production and reduced the level of mitochondrial reactive oxygen species (ROS) without affecting activity of mitochondrial complexes I–IV. Additional assays revealed that MA-5 targets the mitochondrial protein mitofilin at the crista junction of the inner membrane. In Hep3B cells, overexpression of mitofilin increased the basal ATP level, and treatment with MA-5 amplified this effect. In a unique mitochondrial disease model (Mitomice with mitochondrial DNA deletion that mimics typical human mitochondrial disease phenotype), MA-5 improved the reduced cardiac and renal mitochondrial respiration and seemed to prolong survival, although statistical analysis of survival times could not be conducted. These results suggest that MA-5 functions in a manner differing from that of antioxidant therapy and could be a novel therapeutic drug for the treatment of cardiac and renal diseases associated with mitochondrial dysfunction.

This study was conducted to determine if dietary supplementation with coenzyme Q

Selection for high (H) and low (L) oxygen consumption (OC) as an indirect estimation of maintenance energy requirement was determined. Feed intake and body weight were measured and feed conversion ratio (FCR) of 4-8-week-old mice was calculated. Respiratory activity of hepatic mitochondria was measured at 12 weeks. Total feed intake (H: 103.74 g, L: 97.92 g, P

1. To clarify the underlying mechanism of hepatic fat accretion due to methionine (Met) deficiency in broiler chickens, the present study investigated the effect of Met deficiency on the hepatic carnitine palmitoyltransferase (CPT) system, which imports fatty acids into mitochondria. 2. Fifteen-d-old male meat-type chickens were fed on either a control diet (containing 0.52 g/100 g Met) or a Met-deficient diet (containing 0.27 g Met/100 g). After a 10-d feeding period, the birds were killed by decapitation and their livers excised to determine hepatic CPT1 and CPT2 mRNA levels and for the related hepatic fatty acid-supported mitochondrial respiration to be measured. 3. Met deficiency decreased body weight gain and feed efficiency and increased hepatic lipid content compared to the control group. Whereas the hepatic CPT2 mRNA level in the Met-deficient group remained unchanged compared to that of the control group, the CPT1 mRNA level was decreased in the Met-deficient group and CPT1-dependent hepatic mitochondrial respiration was impaired. 4. Our results suggest that the hepatic lipid accretion that occurs in response to Met deficiency might be attributable to the impairment of CPT1-mediated fatty acid import into mitochondria.

Selection for high (H) and low (L) oxygen consumption (OC) as an indirect estimation of maintenance energy requirement was determined. Feed intake and body weight were measured and feed conversion ratio (FCR) of 4-8-week-old mice was calculated. Respiratory activity of hepatic mitochondria was measured at 12 weeks. Total feed intake (H: 103.74 g, L: 97.92 g, P 0.01), daily feed intake (H: 3.70 g/day, L: 3.50 g/day, P 0.01) and FCR (H: 18.79, L: 15.50, P 0.01) were significantly different between lines. The line by sex interaction was significant for FCR. No line differences were observed in males; and the FCR of the H line was greater than in the L line in females. H line mice had the highest hepatic mitochondrial respiratory activity in state 2 (P 0.01), the highest uncoupled respiratory rate of mitochondria in the presence of an uncoupling agent (P 0.001), and the mitochondrial proton leak. The adenosine diphosphate/ O ratio was highest in the L line (P 0.05). This suggests that the selection for high and low OC induced differences in basal mitochondrial respiration and basal metabolism, resulting in difference in FCR between H and L lines.

The global demand for corn to be used in the production of feed and fuel is increasing at a rapid rate. Many types of grain have been proposed as substitutes for corn in broiler chicken diets, thereby serving as alternative sources of dietary carbohydrates. We have already demonstrated that paddy rice show some potential for use as a substitute for corn in poultry feed under thermoneutral conditions (Nanto et al., 2012). To the authors’ knowledge, virtually no information is available to confirm a direct effect of the rice feeding on performance on chickens exposed to heat stress. Heat stress is a major issue for the poultry industry because of the growth retardation and high mortality. We have previously found that heat stress induces oxidative damage, resulting in growth performance in birds (Azad et al., 2010). Furthermore, it has been reported that intestinal morphology was altered by heat stress (Quinteiro-Filho et al., 2010). From these findings, we could hypothesize that oxidative stress and intestinal morphology might be factor responsible for growth retardation under heat stress conditions. Therefore, the present study was conducted to not only determine the effects of whole-grain paddy rice-based diets on growth performance in birds under heat stress conditions, but also to clarify the involvement of intestinal morphological alterations and oxidative stress in the performance under chronic heat stress conditions.

Heat stress is a major factor affecting meat production in the poultry industry, especially in hotter regions of the world. A previous in vivo study by our group highlighted that acute heat stress stimulates mitochondrial reactive oxygen species (ROS) production in the skeletal muscle of broiler chickens. Recently, we further clarified that the overproduction of mitochondrial ROS resulted from an increase in the mitochondrial membrane potential, probably due to an increase in substrate oxidation (Kikusato et al., 2010) and a decrease in avian uncoupling protein (avUCP) content (Mujahid et al., 2006). Furthermore, it was reported that heat stress increases protein oxidation and myofibrillar proteolysis in chick cultured skeletal muscle cells (Nakashima et al., 2004). Considering the fact that ROS play a pivotal role in muscle proteolysis in sepsis (Supinski and Callahan, 2007), it could be postulated that heat treatment stimulates mitochondrial ROS production, thereby leading to an increase in skeletal muscle proteolysis. To verify this possibility, we have investigated the effect of heat stress on ROS production, protein content, and the ROS-related expression of genes for avUCP, heme oxygenase-1 (HO-1) and NADPH oxidase 4 (NOX4) in chick skeletal muscle cells in primary culture. We also examine the reducing effects of 4-hydroxy-TEMPO (Tempol), which is a strong antioxidant, on these parameters.

This study was designed to elucidate physiological changes of skeletal muscle mitochondria from broiler chickens (Gallus gallus) during chronic heat exposure. Chickens (19-day-old) were exposed to either constant heat stress (34 degrees C) or kept at control temperature (24 degrees C) for 14days. Mitochondrial ROS production for control group showed little changes during the experimental periods, whereas that for the heat-stressed group was increased after 3, 5 and 9days of heat exposure and returned to original levels at day 14. Mitochondrial membrane potential in state 4 for heat-stressed birds was higher than those of control birds after 3 and 5days, but was not at day 14. Mitochondrial oxygen consumption rate in state 3 was increased after 3 and 5days, and also returned to original levels by day 14. These results suggest that chronic heat stress induces increased ROS production in skeletal muscle mitochondria, probably via elevation of the membrane potential in state 4, resulting from enhanced oxygen consumption in the initial stage of heat exposure. These physiological changes were no longer observed at day 14, possibly because the animals had acclimatized to environmental heat stress.

We evaluated the cryosurvival of rat epididymal sperm preserved in raffinose-modified Krebs-Ringer bicarbonate-egg yolk extender supplemented with various energy-yielding substrates (glucose, pyruvate, lactate, and ATP) and assessed the effect on sperm oxygen consumption. The incubation of sperm at 37 degrees C for 10 min in lactate-free extender decreased sperm motility and oxygen consumption before and after thawing compared with those of sperm in glucose- and pyruvate-free mediums. We then focused on the effect of supplementing the extender with lactate (0, 10.79, 21.58, 32.37, and 43.16 mM) and found that sperm frozen and thawed in extender supplemented with 32.37 mM lactate exhibited the highest motility. When we supplemented extender containing 32.37 mM lactate with ATP (0, 0.92, 1.85, 3.70, and 5.55 mM), sperm frozen and thawed in the extender supplemented with 1.85 mM ATP exhibited considerably higher motility and viability than those of sperm frozen and thawed in ATP-free extender. These results provide the first evidence that supplementation of the raffinose-modified Krebs-Ringer bicarbonate-egg yolk extender with 32.37 mM lactate and 1.85 mM ATP increases of number of motile sperm before freezing and enhances the cryosurvival of rat sperm. These supplements to the extender may enhance sperm cryosurvival by improving the metabolic capacity of sperm before freezing.

Emerging evidence has shown that acute heat exposure affects metabolic characteristics and causes oxidative damage to skeletal muscle in birds. Little is known, however, about such phenomena under chronic heat stress conditions. To address this, we designed the present study to determine the influence of cyclic (32 to 24 to 32 degrees C: 32 degrees C for 8 h/d, 32-24-32HS ), and constant (32 and 34 degrees C, 32HS and 34HS, respectively) heat exposure on the metabolic and peroxide status in skeletal muscle of 4-wk-old male broiler chickens. Heat stress, particularly in the 32HS and 34HS groups, depressed feed intake and growth, while cyclic high temperature gave rise to a less severe stress response in performance terms. Malondialdehyde (MDA) levels in skeletal muscle were enhanced (P