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Substitution of smooth muscle myosin's phosphorylation site Ser-19 and Thr-18 with aspartic acid or alanine results in a negative charge that is required at a specific spatial position for myosin activation. An analysis of the effects of Thr-18 phosphorylation on myosin function uses a regulatory light chain mutant that contains Ser-substituted with either Asp or Als. Asp incorporation increases phosphorylation and myosin configuration whereas Ala-substituted light chain has no effect on ATPase.

Previous studies in mammalian obesity models have suggested that central transforming growth factor- β (TGF- β ) controls the gene expression of appetite-regulating neuropeptides and peripheral energy metabolism. In the present study, we investigated the possible involvement of central TGF- β /Smad signaling in feeding regulation in chickens. Central administration of TGF- β 1 resulted in phosphorylation of Smad2 in the hypothalamus of chicks and suppressed feed intake without changing the gene expression of hypothalamic appetite-regulating neuropeptides (neuropeptide Y, agouti-related protein, proopiomelanocortin, and corticotropin-releasing factor). However, neither fasting nor refeeding induced the phosphorylation of hypothalamic Smad2. These findings suggest that the activation of hypothalamic TGF- β /Smad signaling suppresses feed intake in chicks but it might not occur in response to feeding status., Competing Interests: The authors declare no conflicts of interest that could inappropriately influence the work reported in this study., (2022, Japan Poultry Science Association.)

Atrogin-1 plays an important role in ubiquitin-proteasome proteolysis in vertebrate skeletal muscles. Recently, atrogin-1 has been shown to be involved in the autophagy-lysosome system, another proteolytic system, in the murine and fish hearts and skeletal muscles. With the aim to elucidate the effect of atrogin-1 on the autophagy-lysosome system in mammalian and avian skeletal muscles, this study has examined the effects of atrogin-1 knockdown on autophagy-lysosome-related proteins in C2C12 and chicken embryonic myotubes. Using the levels of microtubule-associated protein light chain 3 (LC3)-II protein, it was confirmed that atrogin-1 knockdown blocked the autophagic flux in both the myotubes. In addition, atrogin-1 knockdown in C2C12 myotubes significantly decreased the level of autophagy-related gene (ATG)12-ATG5 conjugate, which is supposedly necessary for the fusion of autophagosomes and lysosomes. Atrogin-1 knockdown also resulted in downregulation of forkhead box O3, a transcription factor for ATG12. These data suggest that atrogin-1 is essential for the normal autophagy-lysosome system in the striated muscles of vertebrates., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Objective: We examined whether auto/paracrine insulin-like growth factor-1 (IGF-1) contributes to the phosphorylation of Akt and ERK in chicken myotubes., Methods: Chicken myotubes were treated with IGF-1 siRNA, and then total RNA and protein were harvested for real-time PCR and western blot analysis., Results: Treatment with IGF-1 siRNA inhibited the phosphorylation of Akt and ERK, but not of ribosomal protein S6, in chicken myotubes. Interestingly, IGF-1 siRNA downregulated the expression of IGF-2., Conclusions: The results of this study suggest that auto/paracrine IGF-1 contributes to Akt and ERK phosphorylation in chicken myotubes., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Excessive accumulation of body fat in broiler chickens has become a serious problem in the poultry industry. However, the molecular mechanism of triglyceride accumulation in chicken white adipose tissue (WAT) has not been elucidated. In the present study, we investigated the physiological importance of the catabolic hormone corticosterone, the major glucocorticoid in chickens, in the regulation of chicken WAT lipid metabolism. We first examined the effects of fasting on the mRNA levels of lipid metabolism-related genes associated with WAT, plasma corticosterone, and non-esterified fatty acid (NEFA). We then examined the effects of corticosterone on the expression of these genes in vivo and in vitro . In 10-day-old chicks, 3 h of fasting significantly decreased mRNA levels of lipoprotein lipase ( LPL ) in WAT and significantly elevated plasma concentrations of NEFA. Six hours of fasting significantly increased mRNA levels of adipose triglyceride lipase ( ATGL ) in WAT and significantly elevated plasma concentrations of corticosterone. On the other hand, fasting significantly reduced mRNA levels of LPL in WAT and elevated plasma concentrations of NEFA in 29-day-old chicks without affecting mRNA levels of ATGL in WAT or plasma corticosterone concentrations. Oral administration of corticosterone significantly reduced mRNA levels of LPL and significantly increased the mRNA levels of ATGL in WAT in 29-day-old chicks without affecting plasma NEFA concentrations. The addition of corticosterone to primary chicken adipocytes significantly increased mRNA levels of ATGL , whereas mRNA levels of LPL tended to decrease. NEFA concentrations in the culture medium were not influenced by corticosterone levels. These results suggest that plasma corticosterone partly regulates the gene expression of lipid metabolism-related genes in chicken WAT and this regulation is different from the acute elevation of plasma NEFA due to short-term fasting., (2022, Japan Poultry Science Association.)

Glucagon-like peptide (GLP)-1 and GLP-2, proglucagon-derived brain-gut peptides, function as anorexigenic neuropeptides in mammals. We previously showed that central administration of GLP-1 and GLP-2 potently suppressed food intake in chicks. GLP-1 and GLP-2 specifically activate their receptors GLP-1 receptor (GLP1R) and GLP-2 receptor (GLP2R), respectively in chickens. In adult chickens, GLP1R and GLP2R are expressed in different brain regions. These findings raise the hypothesis that both GLP-1 and GLP-2 function as anorexigenic peptides in the chicken brain but the mechanisms underlying the anorexigenic effects are different between them. In the present study, we compared several aspects of GLP-1 and GLP-2 in chicks. GLP1R mRNA levels in the brain stem and optic lobes were significantly higher than in other parts of the brain, whereas GLP2R mRNA was densely expressed in the telencephalon. Intracerebroventricular administration of either GLP-1 or GLP-2 significantly reduced the mRNA levels of corticotrophin releasing factor and AMP-kinase (AMPK) α1. The mRNA level of proopiomelanocortin was significantly increased, and those of AMPKα2 and GLP2R were significantly decreased by GLP-2, whereas the mRNA level of pyruvate dehydrogenase kinase 4 was significantly increased, and that of GLP1R was significantly decreased by GLP-1. Intracerebroventricular administration of either GLP-1 or GLP-2 induced sleep-like behavior in chicks. Our findings suggest that the anorexigenic peptides GLP-1 and GLP-2 induce similar behavioral changes in chicks, but the mechanism may differ between them., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The purpose of this study was to investigate whether medullary cellular signaling pathways contribute to feeding regulation in chickens. Fasting inhibited the phosphorylated protein and its rates of ERK but not Akt in the chicken medulla, while refeeding promoted Akt and ERK. Intraperitoneal administration of sulfate cholecystokinin 8 did not affect medullary Akt and ERK phosphorylation in chickens. Intracerebroventricular administration of insulin significantly induced the phosphorylation of Akt and ERK in the chicken medulla. These findings suggest that the medullary Akt and ERK pathways are involved in the appetite-suppressive pathway of insulin in chickens., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Cholecystokinin (CCK) and peptide YY (PYY) have been investigated as gut hormones that send satiation signals to the brain in mammals. There is evidence that chicken PYY mRNA expression was the highest in the pancreas compared to other tissues. We recently suggested that insulin-like growth factor (IGF)-1 and its binding proteins (IGFBPs) may be involved in the appetite regulation system in chicks. In the present study, in order to evaluate the possible roles of CCK, PYY, and IGF-related proteins in the appetite regulation system in chicks, we analyzed changes in the mRNA levels of these genes in response to fasting and re-feeding in layer and hyperphagic broiler chicks. In layer chicks, 12 h of fasting reduced the mRNA levels of intestinal CCK, PYY, Y2 receptor, and pancreatic PYY, and these changes were reversed by 12 h of re-feeding. On the other hand, in broiler chicks 12 h of fasting reduced the mRNA levels of intestinal PYY and Y2 receptor, but not intestinal CCK and pancreatic PYY, and these changes were reversed by 12 h of re-feeding. Hypothalamic NPY mRNA significantly increased by 12 h of fasting in both chicks, and these changes were reversed by re-feeding. Also, 12 h of fasting significantly increased the mRNA levels of hypothalamic agouti-related protein and reduced the mRNA levels of hepatic IGF-1 only in broiler chicks, and 12 h of re-feeding did not change these. IGFBP-1 and -2 mRNA levels were markedly increased by 12 h of fasting in both chicks, and these changes were reversed by re-feeding. IGFBP-3 mRNA levels were increased by 12 h of fasting only in layer chicks, while re-feeding reduced the mRNA levels of IGFBP-3 in both types of chicks. These results suggest that several peripheral hormones, such as pancreatic PYY and intestinal CCK, may not play important roles in the regulation of food intake in broiler chicks., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Insulin-like growth factor (IGF)-2 is a multifunctional hormone with structural and functional similarity to IGF-1 in mammals and chickens. We previously showed that intracerebroventricular administration of IGF-1 suppresses food intake in chicks. Also, central administration of IGF-2 suppresses food intake in rats. In the present study, we evaluated whether IGF-2 is involved in the regulation of food intake in chicks. We also examined the effects of fasting on the mRNA levels of IGF binding proteins (IGFBPs) in the liver and hypothalamus, because IGFBPs bind IGF-1 and -2 in plasma and block their binding to the receptors, and locally expressed IGFBPs also influence IGFs binding to the receptors in mammals. Intracerebroventricular administration of IGF-2 significantly suppressed food intake in chicks. The mRNA levels of IGFBPs in the hypothalamus were not affected by six hours of fasting. On the other hand, six hours of fasting markedly increased the mRNA levels of hepatic IGFBP-1 and -2 (5.47- and 6.95-fold, respectively). The mRNA levels of IGFBP-3 were also significantly increased (1.36-fold) by six hours of fasting, whereas the mRNA levels of IGF-2, IGFBP-4, and -5 were unchanged. These findings suggest that circulating IGF-2 may be involved in satiety signals, but its physiological role may be regulated by IGFBPs production in the liver in chicks., (Copyright © 2021 Elsevier B.V. All rights reserved.)

It is well known that dietary fiber stimulates the release of satiety hormones such as glucagon-like peptide-1 (GLP-1), which in turn suppresses appetite. In order to evaluate appetite regulating role of enzymatically synthesized glycogen (ESG, one of the resistant starch), we examined the effects of dietary supplementation of ESG on food intake and cecal proglucagon gene expression in normal and high fat diet-fed mice. Twenty four male ICR mice were weighed and assigned to four groups: normal diet group; normal diet containing 25% ESG group; high-fat diet (HFD) group; HFD containing 25% ESG group. Each group was fed the relevant diets for 3 wk. All data were analyzed by a two-way ANOVA with the main effects of HFD and ESG. ESG significantly decreased food intake and increased the weight of the cecum and cecal content. Plasma total short chain fatty acids concentration was significantly elevated by ESG. The mRNA levels of proglucagon in the cecum and plasma total GLP-1 concentration were significantly increased by ESG. The mRNA levels of appetite regulating neuropeptides such as neuropeptide Y, agouti-related protein, proopiomelanocortin, and cocain- and amphetamine-regulating transcript in the hypothalamus were not influenced by ESG. There is no significant interaction between diet and ESG in any parameters. These results suggest that ESG-induced upregulation of GLP-1 production in the cecum suppresses food intake in mice and that fecal fermentation may be involved in the anorexigenic effect.

In mammalian skeletal muscles, protein synthesis rates vary according to fiber types. We herein demonstrated differences in the regulatory mechanism underlying the protein synthesis in the pectoralis major (a glycolytic twitch muscle), adductor superficialis (an oxidative twitch muscle), and adductor profound (a tonic muscle) muscles of 14-day-old chickens. Under ad libitum feeding conditions, protein synthesis is significantly higher in the adductor superficialis muscle than in the pectoralis major muscle, suggesting that protein synthesis is upregulated in oxidative muscles in chickens, similar to that in mammals. In the pectoralis major muscle, fasting significantly inhibited the Akt/S6 pathway and protein synthesis with a corresponding decrease in plasma insulin concentration. Conversely, the insulin like growth factor-1 (IGF-1) mRNA levels significantly increased. These findings suggest that the insulin/Akt/S6 pathway plays an important role in the regulation of protein synthesis in the pectoralis major muscle. Interestingly, protein synthesis in the adductor superficialis muscle appears to be regulated in an Akt-independent manner, because fasting significantly decreased S6 phosphorylation and protein synthesis without affecting Akt phosphorylation. In the adductor profound muscle, IGF-1 expression, phosphorylation of Akt and S6, and protein synthesis were decreased by fasting, suggesting that insulin and/or skeletal IGF-1 appear contribute to protein synthesis via the Akt/S6 pathway. These findings revealed the differential regulation of protein synthesis depending on skeletal muscle types in chickens., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Insulin-like growth factor-1 (IGF-1) is a key regulator of muscle development and metabolism in chickens. Recently, we have demonstrated that intracerebroventricular administration of IGF-1 significantly decreased food intake in broiler chicks. However, the molecular mechanisms underlying the IGF-1-induced anorexia and the anorexigenic effect of IGF-1 in different strains of commercial chicks have not been investigated. Neuropeptide Y (NPY, a hypothalamic orexigenic neuropeptide), agouti-related protein (AgRP, a hypothalamic orexigenic neuropeptide), and proopiomelanocortin (POMC, the precursor of hypothalamic anorexigenic neuropeptides) play important roles in the regulation of food intake in both mammals and chickens. Evidence shows that several cell signaling pathways in the hypothalamus are involved in regulating the feeding behavior of mammals. In the present study, we first investigated the effects of IGF-1 on the expression of appetite-regulating neuropeptides and phosphorylation of signaling molecules in the hypothalamus of broiler chicks. Intracerebroventricular administration of IGF-1 significantly increased the mRNA levels of POMC, whereas the mRNA levels of NPY and AgRP were not significantly altered. IGF-1 also significantly induced the phosphorylation of v-Akt murine thymoma viral oncogene homolog 1 (AKT) in the hypothalamus of chicks, but did not influence the phosphorylation of forkhead box O1, S6 protein, AMP-activated protein kinase, and extracellular signal-regulated kinase 1/2. We also compared the effect of IGF-1 on food intake in broiler chicks (a hyperphagic strain of chickens) and layer chicks. Results demonstrated that the threshold of IGF-1-induced anorexia in broiler chicks was higher than that in layer chicks. Our observations suggest that hypothalamic POMC and AKT may be involved in the IGF-1-induced anorexigenic pathway and that high threshold of IGF-1-induced anorexia in broiler chicks might be one of the causes of hyperphagia in broiler chicks. Overall, it appears that IGF-1 plays important roles in the central regulation of feeding behavior in chicks., Competing Interests: The authors declare no conflict of interest., (2019, Japan Poultry Science Association.)

Several studies in rodents and layer chickens have demonstrated that insulin upregulates hypothalamic AKT-mediated signaling and expression of proopiomelanocortin (POMC, the precursor of alpha-melanocyte stimulating hormone, an anorexigenic peptide) and suppresses appetite in these animals. However, a previous study has also reported that insulin fails to suppress food intake in broiler chicks. In the present study, no significant differences were observed in hypothalamic AKT and forkhead box O1 (FOXO1) phosphorylation levels between broiler and layer chicks. The phosphorylation rate of AKT, but not that of FOXO1, increased in the hypothalami of broilers refed for 1 h after a 24-h fast, with a corresponding increase in plasma insulin concentration. Intracerebroventricular (ICV) administration of 50 pmol insulin, which could decrease food intake in broiler chicks, significantly increased the AKT phosphorylation rate, whereas no significant change was observed in FOXO1 phosphorylation or POMC expression after ICV insulin administration. These findings suggest that hypothalamic AKT responds to insulin in broiler chicks, but FOXO1-mediated regulation of POMC expression is not induced by insulin, which may be one of the causes of excessive food intake in broiler chickens., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Skeletal muscle mass is an important trait in poultry meat production. In mammals, myostatin, a negative regulator of skeletal muscle growth, activates Smad transcription factors and induces the expression of atrogin-1 by regulating the Akt/FOXO pathway. Although the amino acid sequence of chicken myostatin is known to be completely identical to its mammalian counterpart, previous studies in chicken skeletal muscles have implied that the physiological roles of chicken myostatin are different from those of mammals. Furthermore, it remains to be elucidated whether myostatin affects cellular signaling factors and atrogin-1 expression. In this study, using chick embryonic myotubes, we found that myostatin significantly increased the phosphorylation rate of Smad2 and mRNA levels of atrogin-1. No significant change was observed in the phosphorylation of Akt and FOXO1. These in vitro results suggest that the molecular mechanisms underlying myostatin-induced expression of atrogin-1 might be different between chickens and mammals., (2019, Japan Poultry Science Association.)

The central anorexigenic mechanism seems to be similar in mammals and chicks, because the appetite-suppressive action of a number of peptide hormones is similar in both species. Accumulating evidence in mammals has revealed that hypothalamic Akt-mediated signaling factors (for instance, mTOR and FOXO1) are significantly involved in the regulation of food intake. However, the role of hypothalamic Akt in feeding regulation is yet to be determined in chickens. In this study, we showed that pAkt (Thr308)/Akt, pFOXO1/FOXO1, and pS6 levels were significantly increased in the hypothalami of chicks refed 1 h after a 24 h-fast in correlation to increases in the plasma concentrations of insulin, one of the activators of the Akt-mediated signaling pathways. In addition, central administration of insulin increased the phosphorylation of Akt, FOXO1, and S6 in chicken hypothalami. Furthermore, intracerebroventricular injections of both phosphoinositide 3-kinase inhibitor LY294002 and mTOR inhibitor rapamyacin enhanced the food intake of chicks. These findings suggest that hypothalamic Akt-mediated signaling pathways contribute to the regulation of food intake in chicks., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The physiological functions of insulin-like growth factor-binding proteins (IGFBPs) in mammals have been evaluated in several studies. However, the physiological roles of IGFBPs in chickens have not yet been elucidated. In this study, we examined the effects of short-term (6 h) fasting and refeeding on the mRNA levels of IGFBPs in chick liver and brain. Eighteen 8-day-old chicks were weighed and allocated to three groups on the basis of body weight, and subjected to ad libitum feeding, 6 h of fasting, or 6 h of fasting followed by 6 h of refeeding. After the chicks were euthanized by decapitation, the liver and brain were excised, and the brain was dissected into six segments (telencephalon, optic lobes, cerebellum, rostral part of the brainstem, middle part of the brainstem, and caudal part of the brainstem). IGFBP mRNA levels were determined by qRT-PCR. Fasting significantly increased the mRNA levels of IGFBP-1 and -2 in the chick liver, and these changes were reversed by 6 h of refeeding. The mRNA levels of IGFBP-3 in the middle part of the brainstem and IGFBP-5 in the optic lobes were decreased by 6 h of fasting and were not reversed after 6 h of refeeding. These findings suggest that IGFBP-1 and -2 in the liver, IGFBP-3 in the middle part of the brainstem, and IGFBP-5 in the optic lobes may play physiological roles in response to short-term changes in the nutritional status of chicks., (2018 by Japan Poultry Science Association.)

Skeletal muscle is composed of four types of fibers in mammals; oxidative slow-twitch type I, oxidative fast-twitch IIA, and glycolytic fast-twitch IIB and IIX/D. In this study using C2C12 myotubes, an extract of soybean protein significantly upregulated mRNA level of myosin heavy chain 7 (Myh7), the predominant isoform expressed in oxidative slow-twitch type I and downregulated mRNA levels of Myh4, the predominant isoform expressed in glycolytic fast-twitch IIB. Similarly, its hydrolysate prepared using digestive enzyme also significantly increased Myh7 expression. In contrast, no significant change was observed in Myh4 mRNA level after the hydrolysate treatment. These findings suggest that dietary intake of the soybean protein extract may increase oxidative slow-twitch fiber in skeletal muscle.

Skeletal muscle mass is an important trait in the animal industry. We previously reported an age-dependent downregulation of the insulin-like growth factor 1 (IGF-1)/Akt/S6 pathway, major protein synthesis pathway, in chicken breast muscle after 1 week of age, despite a continuous increase of breast muscle weight. Myosin heavy chain (HC), a major protein in muscle fiber, has several isoforms depending on chicken skeletal muscle types. HC I (fast-twitch glycolytic type) is known to be expressed in adult chicken breast muscle. However, little is known about the changes in the expression levels of protein synthesis-related factors and HC isoforms in perihatching chicken muscle. In the present study, protein synthesis-related factors, such as IGF-1 messenger RNA (mRNA) levels, phosphorylation of Akt, and phosphorylated S6 content, increased in an age-dependent manner after post-hatch day (D) 0. The mRNA levels of HC I, III and V (fast-twitch glycolytic type) dramatically increased after D0. The increase ratio of breast muscle weight was approximately 1100% from D0 to D7. To our knowledge, these findings provide the first evidence that upregulation of protein synthesis pathway and transcription of fast twitch glycolytic HC isoforms play critical roles in the increase of chicken breast muscle weight during the first week after hatching., (© 2017 Japanese Society of Animal Science.)

The excessive accumulation of body fat has become a serious problem in the broiler industry. However, the molecular mechanisms underlying the regulation of lipid metabolism-related genes in broiler chickens are not fully understood. In the present study, we investigated the role of glucagon on the expression of lipid metabolism-related genes in chicken white adipose tissue (WAT). Four hours of fasting significantly increased plasma levels of free fatty acid in broiler chickens. The mRNA levels of adipose triglyceride lipase (ATGL) and pyruvate dehydrogenase kinase 4 (PDK4) in abdominal WAT significantly increased by fasting, whereas the mRNA levels of diacylglycerol O-acyl-transferase homolog 2 (DGAT2) and peroxisome proliferator-activated receptor- γ (PPAR γ ) significantly decreased. The results suggest that fasting stimulates lipolysis and suppresses adipogenesis and re-esterification of TG in chicken WAT. Glucagon significantly increased the mRNA levels of PDK4 in chicken primary adipocytes, whereas there were no significant changes in the mRNA levels of ATGL, DGAT2, and PPAR γ . Our findings suggest that glucagon upregulates PDK4 expression and may stimulate lipolysis without affecting the expression of ATGL in chicken WAT., (2017, Japan Poultry Science Association.)

A number of studies have been made on the physiological actions of insulin-like growth factor-1 (IGF-1) in mammals and birds. In mammals, the effects of central administration of IGF-1 on food intake have been examined. For example, intracerebroventricular administration of IGF-1 significantly decreased food intake in diabetic rats, but not in sheep and nondiabetic rats. The chicken is known to be a hyperglycemic animal. Like satiety hormones, plasma IGF-1 levels are elevated postprandially in chickens. In this study, we hypothesized that IGF-1 is involved in the regulation of food intake in chickens. Intracerebroventricular administration of IGF-1 significantly suppressed food intake in chicks in a dose dependent manner. Both the mRNAs of IGF-1 and its receptor were expressed throughout the brain. However, the mRNA levels of IGF-1 were not influenced by fasting and refeeding in all regions of the brain. On the other hand, 6h of fasting significantly suppressed mRNA expression of hepatic IGF-1, and this effect was significantly reversed by 6h of refeeding. Furthermore, intravascular administration of IGF-1 significantly suppressed food intake in chicks. These findings suggest that IGF-1 may function as a satiety hormone in chickens., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Animal by-products can be recycled and used as sources of essential nutrients. Water-soluble heme iron (WSHI), a functional food additive for supplementing iron, is produced by processing animal blood. In this study, we investigated the effects of dietary supplementation of 3% WSHI and exercise training for 4 weeks on the accumulation of abdominal fat and lipid metabolism in mice fed high-fat diet. Exercise-trained mice had significantly less perirenal adipose tissue, whereas WSHI-fed mice tended to have less epididymal adipose tissue. In addition, total weight of abdominal adipose tissues was significantly decreased in the Exercise + WSHI group. Dietary WSHI significantly increased the messenger RNA (mRNA) levels of lipoprotein lipase and hormone-sensitive lipase. WSHI-fed mice also tended to show increased mRNA levels of adipose triglyceride lipase in their epididymal adipose tissue. Dietary WSHI also significantly decreased the mRNA levels of fatty acid oxidation-related enzymes in the liver, but did not influence levels in the Gastrocnemius muscle. Exercise training did not influence the mRNA levels of lipid metabolism-related enzymes in the epididymal adipose tissue, liver or the Gastrocnemius muscle. These findings suggest that the accumulation of abdominal fat can be efficiently decreased by the combination of dietary WSHI and exercise training in mice fed high-fat diet., (© 2016 Japanese Society of Animal Science.)

The core circadian clock mechanism relies on a feedback loop comprised of clock genes, such as the brain and muscle Arnt-like 1 (Bmal1), chriptochrome 1 (Cry1), and period 3 (Per3). Exposure to the light-dark cycle synchronizes the master circadian clock in the brain, and which then synchronizes circadian clocks in peripheral tissues. Birds have long been used as a model for the investigation of circadian rhythm in human neurobiology. In the present study, we examined the effects of continuous light and the combination of white and blue light on the expression of clock genes (Bmal1, Cry1, and Per3) in the central and peripheral tissues in chicks. Seventy two day-old male chicks were weighed, allocated to three groups and maintained under three light schedules: 12h white light-12h dark-cycles group (control); 24h white light group (WW group); 12h white light-12h blue light-cycles group (WB group). The mRNA levels of clock genes in the diencephalon were significantly different between the control and WW groups. On the other hand, the alteration in the mRNA levels of clock genes was similar between the control and WB groups. Similar phenomena were observed in the liver and skeletal muscle (biceps femoris). These results suggest that 12h white-12h blue light-cycles did not disrupt the circadian rhythm of clock gene expression in chicks., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Gut hormones act as appetite regulatory hormones in mammals. For example, the hunger hormone ghrelin, which is released from the stomach before food intake, stimulates appetite. In contrast, satiety hormones such as cholecystokinin, glucagon-like peptide-1, and peptide YY, which are released from the intestines after food intake, suppress appetite. The effects of these peptides on food intake have been shown to be similar in both mammals and fishes. However, evidence suggests that the physiological roles of these gut hormones may be different between birds and other vertebrates. This review summarizes the current information on the roles of gut hormones in the regulation of food intake in birds, especially in chickens., (2017, Japan Poultry Science Association.)

Peptide YY (PYY) functions as a postprandial satiety signal in mammals. However, the genomic information and physiological roles of chicken PYY have not yet been clarified, although PYY peptide was isolated from chicken intestines in 1992. In this study, we identified a full-length complementary DNA (cDNA) sequence encoding the chicken PYY precursor. The deduced amino acid sequence of chicken PYY was completely consistent with the previously identified peptide sequence. PYY mRNA was abundantly expressed in the small intestine compared with the large intestine. PYY mRNA levels in the jejunum were significantly higher during ad libitum feeding compared with fasting, suggesting that intestinal PYY expression is altered in response to nutritional status in chicks. Intravenous administration of PYY significantly suppressed food intake in chicks. Furthermore, neuropeptide Y receptor Y2, a possible target of PYY, was expressed in various brain regions including the appetite-regulating centers in chicks. This is the first evidence that the intestinal hormone PYY may function as an anorexigenic hormone in chicks., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The combination of diet and exercise is the first choice for the treatment of obesity and metabolic syndrome. We previously reported that enzymatically synthesized glycogen (ESG) suppresses abdominal fat accumulation in obese rats. However, the effect of the combination of ESG and exercise on abdominal fat accumulation has not yet been investigated. Our goal in this study was therefore to evaluate the effects of dietary ESG and its combination with exercise on abdominal fat accumulation in high-fat diet (HFD)-fed mice. Male ICR mice were assigned to four groups: HFD, HFD containing 20% ESG, HFD with exercise, HFD containing 20% ESG with exercise. Treadmill exercise was performed for 3 wk (25 m/min, 30 min/d, 3 d/wk) after 5-d adaption to running at that speed. Both ESG and exercise significantly reduced the weights of abdominal adipose tissues. In addition, the combination of ESG and exercise significantly suppressed abdominal fat accumulation, suggesting that ESG and exercise showed an additive effect. Exercise significantly increased the mRNA levels of lipid metabolism-related genes such as lipoprotein lipase, peroxisome proliferator-activated receptor delta; factor-delta (PPARδ), carnitin palmitoyltransferase b, adipose triglyceride lipase (ATGL), and uncoupling protein-3 in the gastrocnemius muscle. On the other hand, dietary ESG significantly decreased the mRNA levels of PPARδ and ATGL in the gastrocnemius muscle. These results suggest that the combined treatment of ESG and exercise effectively suppresses abdominal fat accumulation in HFD-fed mice by different mechanisms.

The skeletal muscle mass is known to be controlled by the balance between protein synthesis and degradation. The fractional rate of protein synthesis has been reported to decrease age-dependently from 1 to 4 weeks of age in the chicken breast muscle (pectoralis major muscle). On the other hand, age-dependent change of the fractional protein degradation rate was reported to be less in the skeletal muscle of chickens. These findings suggest that protein synthesis is age-dependently downregulated in chicken muscle. We herein investigated the age-dependent changes in protein synthesis or proteolysis-related factors in the breast muscle of 7, 14, 28, and 49-day old broiler chickens. IGF-1 mRNA level, phosphorylation rate of Akt, and phospho-S6 content were coordinately decreased in an age-dependent manner, suggesting that IGF-1-stimulated protein synthesis is downregulated with age in chicken breast muscle. In contrast, atrogin-1, one of the proteolysis-related factors, gradually increased with age at mRNA levels. However, plasma N τ -methylhistidine concentration, an indicator of skeletal muscle proteolysis, did not coordinately change with atrogin-1 mRNA levels. Taken together, our results suggest that the IGF-1/Akt/S6 signaling pathway is age-dependently downregulated in the chicken breast muscle., (2016, Japan Poultry Science Association.)

Liver is the most important target organ for investigation of lipid metabolism in domestic fowls. However, little is known about the regulatory mechanism of fatty acid oxidation in chicken liver. In mammals, proliferator-activated receptor alpha (PPARα), a transcription factor, plays an essential role in the regulation of hepatic fatty acid oxidation. The aim of the present study was to investigate the regulatory mechanisms of PPARα-induced gene expression involved in hepatic fatty acid oxidation in chickens in vivo and in vitro. WY14643, a PPARα agonist, significantly increased the messenger RNA (mRNA) levels of carnitine palmitoyltransferase 1a (CPT1a) and acyl-coenzyme A oxidase (ACO), but not long-, middle- and short-chain acyl-coenzyme A dehydrogenase (LCAD, MCAD and SCAD, respectively), hydroxyacyl-coenzyme A dehydrogenase (HAD), and PPARα itself in chicken hepatoma cells. In contrast, WY14643 significantly increased the mRNA levels of CPT1a, ACO, MCAD, SCAD, HAD and PPARα in human hepatoma cells. The mRNA levels of CPT1a and ACO in the liver were significantly increased by 6 h of fasting in chickens, whereas the mRNA levels of LCAD, MCAD, SCAD and HAD were unchanged. These results suggest that, unlike in mammals, CPT1a and ACO might play an important role in PPARα-induced fatty acid oxidation in the liver of chickens., (© 2015 Japanese Society of Animal Science.)

The regulatory mechanisms of carbohydrate and lipid metabolism are known to differ among skeletal muscle types in mammals. For example, glycolytic muscles prefer glucose as an energy source, whereas oxidative muscles prefer fatty acids (FA). We herein demonstrated differences in the expression of genes involved in carbohydrate and lipid metabolism in the pectoralis major (a glycolytic twitch muscle), adductor superficialis (an oxidative twitch muscle), and adductor profound (a tonic muscle) of 14-day-old chicks. Under ad libitum feeding conditions, the mRNA levels of muscle type phosphofructokinase-1 were markedly lower in the adductor superficialis muscle, suggesting that basal glycolytic activity is very low in this type of muscle. In contrast, high mRNA levels of lipoprotein lipase (LPL) and fatty acid translocase/cluster of differentiation 36 (FAT/CD36) in the adductor superficialis muscle suggest that FA uptake is high in this type of muscle. The mRNA levels of adipose triglyceride lipase (ATGL) and carnitine palmitoyltransferase 1b (CPT1b) were significantly higher in the adductor profound muscle than in other muscles, suggesting that basal lipolytic activity is high in this type of muscle. Furthermore, the mRNA levels of peroxisome proliferator activated receptor δ and CPT1b were significantly increased in the adductor superficialis muscle, but not in other muscles, after 24h of fasting. Therefore, the availability of FA in the oxidative twitch muscles in growing chickens appears to be upregulated by fasting. Our results suggest that lipid metabolism-related genes are upregulated under both basal and fasting conditions in the adductor superficialis in growing chickens., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Genetic selection results in a higher growth rate and meat yield in broiler chickens than in layer chickens. We herein demonstrated differences in the effects of 24 h of fasting on the expression of genes involved in skeletal muscle proteolysis between broiler and layer chicks. The mRNA levels of proteolysis-related genes were analyzed in the pectoralis major muscle of 14-day-old chicks after 0 or 24 h of fasting. The mRNA levels of ubiquitin ligases such as atrogin-1 and muscle RING finger-1 (MuRF-1) as well as transcription factor forkhead box class O (FOXO) 1 were significantly increased by fasting in broiler and layer chicks, suggesting that the FOXO1-induced ubiquitin-proteasome system, a major proteolytic system in skeletal muscles, was activated by fasting in both chicks. The mRNA levels of atrogin-1 were significantly lower in broiler chicks than in layer chicks after fasting. Furthermore, the mRNA levels of insulin-like growth factor-1 were significantly decreased by fasting in layer chicks, but not in broiler chicks. The mRNA levels of FOXO3 were significantly increased by fasting in layer chicks, but not in broiler chicks. Therefore, the ubiquitin-proteasome system did not appear to have been fully upregulated in broiler chicks. These results suggest that differences in the expression of genes related to the ubiquitin-proteasome system in skeletal muscle proteolysis between broiler and layer chicks during food deprivation are one of the causes of the high growth rate in broiler chickens., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Various lines of evidence suggest that appetite-related neuropeptides in the hypothalamus are regulated by adiposity signals such as leptin and insulin in mammals. In the present study, we examined age-dependent changes in the weight of abdominal fat and hypothalamic mRNA levels of neuropeptide Y (NPY, an orexigenic neuropeptide) and proopiomelanocortin (POMC, a precursor of anorexigenic neuropeptides) in growing chickens at 7, 14, 21 and 28 days of age. Hypothalamic NPY mRNA levels were significantly (P < 0.05) decreased after 14 days of age, whereas hypothalamic POMC mRNA levels were significantly (P < 0.05) increased at 28 days of age. The percentage of abdominal fat was significantly increased after 14 days of age in chickens. We next examined the correlation of hypothalamic NPY and POMC mRNA levels and several parameters at 28 days of age. There were no significant correlations between hypothalamic mRNA levels of NPY or POMC and the percentage of abdominal fat. These findings suggest that the gene expressions of NPY and POMC do not depend on adiposity in chickens, at least in 28-day-old layer chickens., (© 2014 Japanese Society of Animal Science.)

Glucagon-related peptides, such as glucagon-like peptide (GLP)-1, GLP-2 and oxyntomodulin (OXM), are processed from an identical precursor proglucagon. In mammals, all of these peptides are suggested to be involved in the central regulation of food intake. We previously showed that intracerebroventricular administration of chicken OXM and GLP-1 significantly suppressed food intake in chicks. Here, we show that central administration of chicken GLP-2 potently suppresses food intake in chicks. Male 8-day-old chicks (Gallus gallus domesticus) were used in all experiments. Intracerebroventricular administration of chicken GLP-2 significantly suppressed food intake in chicks. Plasma glucose concentration was significantly decreased by chicken GLP-2, whereas plasma nonesterified fatty acid concentration was significantly increased. Intracerebroventricular administration of chicken GLP-2 did not affect plasma corticosterone concentration. In addition, the anorexigenic effect of GLP-2 was not reversed by the corticotropin-releasing factor (CRF) receptor antagonist α-helical CRF, suggesting that CRF is not a downstream mediator of the anorexigenic pathway of GLP-2 in chicks. Intracerebroventricular administration of an equimolar amount of GLP-1 and GLP-2, but not OXM, significantly suppressed food intake in both broiler and layer chicks. All our findings suggest that GLP-2 functions as a potent anorexigenic peptide in the brain, as well as GLP-1, in chicks., (© 2014 Japanese Society of Animal Science.)

International trading markets of meat require the animal's age information to prevent cross-contamination of ineligible meat products. Individual livestock age is either evaluated from physiological features or verified by breeding history. However, it remains impossible to perform age verification on meat when a suspicion of error occurred in the importing country. To investigate an age-related protein in skeletal muscle of livestock, we compared protein expression among chicken pectoralis major of different ages. Results indicated that the level of expression of chicken HSPB1, one of the small heat shock proteins, was increased in aged muscles. On the other hand, other heat shock proteins, heat shock factors, and myosin heavy chain isoform did not change the expression levels in aged chicken muscle. In addition, we identified that αB-crystallin interacted with HSPB1 in aged chicken muscle. These results suggest that HSPB1 protein forms complexes with αB-crystallin in aged chicken muscle and suppose to become the candidate of age-related bio-marker for verifying the age of chicken meat.

The utilization of copra meal as a feed ingredient is limited because it contains a high level of mannan. However, recent findings indicate that the effect of copra meal on growth performance in broiler chickens can be improved by the supplementation of mannanase in the diet. In the present study, we examined the effect of mannanase-hydrolyzed copra meal (MCM) on growth performance and muscle protein metabolism in growing broiler chickens (Gallus gallus domesticus). Forty 8-day-old male broiler chicks were assigned to two groups (four birds in each pen, five replicates) and fed either a commercial diet (as a control diet) or a diet containing MCM at 0.2% until 22 days of age. Dietary MCM significantly increased the weights of body, breast muscle, and thighs in chickens, whereas the weights of abdominal adipose tissue and liver were not affected. Cumulative feed intake was significantly increased by MCM. Dietary MCM significantly decreased plasma 3-methylhistidine level. The messenger RNA and protein levels of muscle protein metabolism-related factors were not altered by MCM. These findings suggest that the growth-promoting effect of MCM is related to the suppression of muscle proteolysis in growing broiler chickens., (© 2014 Japanese Society of Animal Science.)

Central administration of proglucagon-derived peptides, glucagon, glucagon-like peptide-1 (GLP-1), and oxyntomodulin (OXM), suppresses food intake in both mammals and birds. Recent findings suggest that GLP-1 receptor is involved in the anorexigenic action of OXM in both species. However, mammalian (bovine) OXM was used in chicken studies, even though the amino acid sequence and peptide length of chicken OXM differ from those of bovine OXM. In the present study, we examined the effect of chicken OXM on food intake and plasma components in chicks to investigate the mechanisms underlying the OXM effect. Male 8-day-old chicks (Gallus gallus domesticus) were used in all experiments. Intracerebroventricular administration of chicken OXM significantly suppressed food intake in chicks. Plasma concentrations of glucose and corticosterone were significantly increased by chicken OXM. These phenomena were also observed after bovine OXM injection in chicks. In contrast, central administration of chicken GLP-1 significantly decreased plasma glucose concentration and did not affect plasma corticosterone concentration. We previously showed that central administration of chicken glucagon significantly increased plasma concentrations of glucose and corticosterone in chicks. All our findings suggest that the mechanism underlying the anorexigenic action of OXM is similar to that of glucagon in chicks., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Glucagon-related peptides such as glucagon, glucagon-like peptide-1, and oxyntomodulin suppress food intake in mammals and birds. Recently, novel glucagon-like peptide (GCGL) was identified from chicken brain, and a comparatively high mRNA expression level of GCGL was detected in the hypothalamus. A number of studies suggest that the hypothalamus plays a critical role in the regulation of food intake in mammals and birds. In the present study, we investigated whether GCGL is involved in the central regulation of food intake in chicks. Male 8-day-old chicks (Gallus gallus) were used in all experiments. Intracerebroventricular administration of GCGL in chicks significantly suppressed food intake. Plasma glucose level was significantly decreased by GCGL, whereas plasma corticosterone level was not affected. Central administration of a corticotrophin-releasing factor (CRF) receptor antagonist, α-helical CRF, attenuated GCGL-suppressed food intake. It seems likely that CRF receptor is involved in the GCGL-induced anorexigenic pathway. All our findings suggest that GCGL functions as an anorexigenic peptide in the central nervous system of chicks., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Based on a recent study indicating that enzymatically synthesized glycogen (ESG) possesses a dietary, fiber-like action, we hypothesized that ESG can reduce the risk of obesity. In this study, the antiobesity effects of ESG were investigated in a model of diet-induced obesity. Male Sprague-Dawley rats were divided into 4 groups and fed a normal or high-fat diet, with or without 20% ESG, for 4 weeks. Body weight, food intake, lipid deposition in the white adipose tissues and liver, fecal lipid excretion, and plasma lipid profiles were measured. At week 3, the body fat mass was measured using an x-ray computed tomography system, which showed that ESG significantly suppressed the high-fat diet-induced lipid accumulation. Similar results were observed in the weight of the adipose tissue after the experiment. Moreover, ESG significantly suppressed the lipid accumulation in the liver but increased fecal lipid excretion. The plasma concentrations of triacylglycerol and nonesterified fatty acid were lowered after a high-fat diet, whereas the total bile acid concentration was increased by ESG. However, the hepatic messenger RNA (mRNA) levels of enzymes related to lipid metabolism were not affected by ESG. Conversely, the mRNA levels of long-chain acyl-CoA dehydrogenase and medium-chain acyl-CoA dehydrogenase were up-regulated by ESG in the muscle. These results suggest that the combined effects of increased fecal lipid excretion, increased mRNA levels of enzymes that oxidize fatty acids in the muscle, and increased total bile acid concentration in the plasma mediate the inhibitory effect of ESG on lipid accumulation., (© 2013.)

The aim of this study was to analyze the expression patterns of key genes involved in lipid metabolism in response to feeding in chicks. A total of 18 thirteen day-old male chicks were fasted for 12h. The mRNA levels of the genes in the liver and white adipose tissue were analyzed after 0, 2, and 4h of refeeding. The mRNA levels of sterol regulatory element-binding protein (SREBP) 1, liver X receptor α, peroxisome proliferator-activated receptor (PPAR) γ, acetyl-CoA carboxylase α and fatty acid synthase were significantly increased after 2h of refeeding. In contrast, the mRNA levels of PPARα and carnitine palmitoyltransferase 1a were significantly decreased after 2h of refeeding. The mRNA level of acyl-CoA oxidase was significantly decreased after 4h of refeeding. The mRNA levels of cholesterol metabolism-related genes such as SREBP2 and 3-hydroxy-3-methylglutaryl-CoA reductase were significantly increased after 2h of refeeding. In the white adipose tissue, the mRNA level of PPARγ was significantly increased after 2h of refeeding, whereas the mRNA level of adipose triglyceride lipase was significantly decreased after 4h of refeeding. These results demonstrated that expression of lipid metabolism-related genes is regulated by short-term refeeding in chicks., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The aim of this study was to analyze the expression patterns of key genes involved in lipid metabolism in response to short term fasting in chicks (Gallus gallus). The mRNA level of the genes was analyzed after 0, 2, and 4 h of fasting in the liver and white adipose tissue. In the liver, the mRNA level of peroxisome proliferator-activated receptor α was significantly increased after 2 h of fasting. The mRNA levels of carnitine palmitoyltransferase 1a and acyl-CoA oxidase were significantly increased after 4 h of fasting. In contrast, the mRNA levels of sterol regulatory element-binding protein 1, acetyl-CoA carboxylase α, and fatty acid synthase were significantly decreased after 4 h of fasting. The mRNA levels of cholesterol metabolism-related genes such as 3-hydroxy-3-methylglutaryl-CoA reductase and cholesterol 7α-hydroxylase were significantly decreased after 4 h of fasting. In the white adipose tissue, the mRNA level of adipose triglyceride lipase was significantly increased after 4 h of fasting. In contrast, the mRNA levels of peroxisome proliferator-activated receptor γ and lipoprotein lipase were significantly decreased after 4 h of fasting. These results demonstrated that the gene expression of lipid metabolism-related genes is regulated by short term fasting in both the liver and WAT in chicks., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Clothianidin (CTD) is a neonicotinoid developed in the 1990s as an insecticide having selective toxicity, but it was later found to cause reproductive abnormalities in rats through oxidative stress. There is an attempt to preserve endangered animals, including the Japanese crested ibis, in Japan. However, there is a concern that neonicotinoid affects the reproduction of this bird, since it is used in its habitat. CTD toxicity in the birds is poorly understood, so we investigated whether or not the daily oral administration of CTD has any deleterious effects on the reproductive functions of mature male quails as experimental animals. The animals were randomly divided into four groups of 6 or 7 quails each, treated orally with 0, 0.02, 1 or 50 mg CTD/kg body weight (Control, CTD0.02, CTD1 and CTD50). After that the males bred with untreated females to estimate the egg weights, and rates of fertilization and normal development, the testes, liver and spleen were examined histologically. Vacuolization and the number of germ cells having fragmented DNA in seminiferous tubules, and the number and size of vacuoles in hepatocytes increased dose-dependently. There were no significant differences in egg weights and fertilization rates between the groups, but some eggs of the CTD1 and CTD50 groups failed to develop, and embryonic length decreased dose-dependently. Thus, it was found that CTD affected the reproduction of the male quail through the fragmentation of germ cells and the inhibition or delay of embryonic development.

Dietary licorice fravonoid oil (LFO) significantly decreased hepatic cholesterol and plasma lipoprotein cholesterol levels in high-fat diet rats. It significantly suppressed hydroxymethylglutaryl-CoA synthase activity and increased cholesterol 7α-hydroxylase activity. The low density lipoprotein receptor mRNA level was significantly increased by LFO. These results suggest that dietary LFO improves cholesterol metabolism in obese animals.

The effects of dietary coenzyme Q10 (CoQ10) on cholesterol metabolism in laying hens were investigated. Dietary CoQ10 significantly reduced egg yolk cholesterol content and suppressed hepatic hydroxymethylglutaryl-CoA reductase (HMGR) activity. It is therefore likely that CoQ10 acts as an HMGR inhibitor in the livers of laying hens, which in turn results in a reduction in egg-yolk cholesterol.

We investigated the mechanism underlying central glucagon-induced hyperglycemia and anorexia in chicks. Male 8-day-old chicks (Gallus gallus) were used in all experiments. Intracerebroventricular administration of glucagon in chicks induced hyperglycemia and anorexia from 30 min after administration. However, the plasma insulin level did not increase until 90 min after glucagon administration, suggesting that glucose-stimulated insulin secretion from pancreatic beta cells may be suppressed by central glucagon. The plasma corticosterone concentration significantly increased from 30 min to 120 min after administration, suggesting that central glucagon activates the hypothalamic pituitary adrenal (HPA) axis in chicks. However, central administration of corticotropin-releasing factor (CRF), which activates the HPA axis in chicken hypothalamus, significantly reduced not only food intake but also plasma glucose concentration, suggesting that CRF and the activation of the HPA axis are related to the glucagon-induced anorexia but not hyperglycemia in chicks. Phentolamine, an α-adrenergic receptor antagonist, significantly attenuated the glucagon-induced hyperglycemia, suggesting that glucagon induced hyperglycemia at least partly via α-adrenergic neural pathway. Co-administration of phentolamine and α-helical CRF, a CRF receptor antagonist, significantly attenuated glucagon-induced hyperglycemia and anorexia. It is therefore likely that central administration of glucagon suppresses food intake at least partly via CRF-induced anorexigenic pathway in chicks., (Copyright © 2012 Elsevier Inc. All rights reserved.)