Hormonal regulation of metabolism in hepatocytes of the ureogenic teleostopsanus beta

Short-term exposure of isolated toadfish hepatocytes to high concentrations (100 nM) of glucagon, glucagon-like peptide (GLP) or epinephrine significantly increases the rate of lactate gluconeogenesis (1.3-fold) and glycogenolysis (5- to 7-fold). Half-maximal responsiveness to GLP is reached at about 2 nM for gluconeogenesis and 6 nM for glycogenolysis, while the value for glycogenolysis activated by catfish glucagon is 28 nM. Cells do not to respond to 5 nM epinephrine. Norepinephrine, urotensin II and leucine-enkephalin, each applied at 100 nM, increase the rate of glycogenolysis by 1.3 to 1.5-fold. All other hormones tested (vasotocin, isotocin, VIP, methionine-enkephalin, ovine prolactin, -endorphin, APY, salmon insulin) failed to affect metabolic flux through glycogenolysis or gluconeogenesis. None of the hormones altered the rate of urea synthesis or the rate of lactate oxidation by hepatocytes. Although toadfish hepatocytes are responsive to hormonal stimuli, they do not appear to be a useful model to study evolutionary trends in short-term hormonal regulation of urea synthesis. However, the obvious differences in mechanisms of control of urea synthesis in this species compared with ureogenic amphibians and mammals open an intriguing avenue for research.     

草鱼3个6-磷酸葡萄糖酶催化亚基的基因表达分析及高糖饲料对其表达的影响

为了探讨6-磷酸葡萄糖酶催化亚基(glucose-6-phosphatase catalytic subunit,G6PC)在草鱼(Ctenopharyngodon idellus)糖代谢中的作用,采用同源序列比对的方式,在草鱼基因组中获取了3个g6pc基因的序列,通过序列比对和进化树分析,将其分别命名为g6pca、g6pcb1和g6pcb2,其编码的氨基酸序列与斑马鱼(Danio rerio)、虹鳟(Oncorhynchus mykiss)和人(Homo sapiens)具有较高的同源性,相似度分别为85%～94%、64%～83%和54%～66%。同线性分析表明g6pc在草鱼染色体上的分布与斑马鱼等高度相似,表明草鱼g6pc基因在进化中具有较高的保守性。利用RT-PCR检测3个基因在鳃、脂肪、脑、心脏、肝、肾、前肠、中肠、后肠和肌肉10个组织中的表达,结果显示, g6pca在脑和肝中表达量较高,脂肪组织次之; g6pcb1在肝中表达量最高,中肠次之; g6pcb2在心脏表达量最高,其次为脂肪组织。同时探讨了高糖饲料对草鱼不同g6pc亚型转录水平的影响,以及不同浓度葡萄糖和胰岛素刺激草鱼肝细胞(L8824)后对g6pca转录水平的影响。结果显示,饲喂高糖饲料(7周)后,与对照组相比,草鱼肝脏g6pca mRNA水平显著升高, g6pcb1和g6pcb2 mRNA水平无显著变化。离体情况,与5 mmol/L葡萄糖组相比,15 mmol/L葡萄糖显著增加了L8824的g6pca mRNA水平,且1 mol/L胰岛素可以抑制这种作用; 30 mmol/L葡萄糖对L8824 g6pca mRNA水平无显著性影响。本研究表明,草鱼g6pc发生加倍后存在功能分化,高糖可以诱导g6pca mRNA的表达,而对g6pcb1和g6pcb2的转录水平无影响,其具体功能还需进一步研究。     

Propionate absorbed from the colon acts as gluconeogenic substrate in a strict carnivore, the domestic cat (Felis catus)

In six normal‐weight and six obese cats, the metabolic effect of propionate absorbed from the colon was assessed. Two colonic infusions were tested in a crossover design with intervals of 4 weeks. The test solution contained 4 mmol sodium propionate per kg ideal body weight in a 0.2% NaCl solution. Normal saline was given as control solution. Solutions were infused into the hindgut over 30 min. Blood samples were obtained prior to and at various time points after starting the infusion. As body condition did not affect evaluated parameters, all data were pooled. Plasma glucose concentrations showed differences neither over time nor during or after infusion with propionate or control. Plasma amino acid concentrations rose over time (p < 0.001), but were similar for both infusions. Plasma propionylcarnitine rose markedly towards the end of the propionate infusion and decreased afterwards (p < 0.001), whereas 3‐hydroxy‐3‐methylglutarylcarnitine was lower 30 (p = 0.005) and 60 min (p = 0.032) after ending propionate infusions and acetylcarnitine tended to fall at the same time points (p = 0.079; p = 0.080), suggesting inhibition of gluconeogenesis from pyruvate and amino acids, but initiation of propionate‐induced gluconeogenesis. In conclusion, propionate absorbed from the colon is hypothesized to act as gluconeogenic substrate, regardless of the cat’s body condition.     

Metabolic actions of glucagon-family hormones in liver

This review addresses direct and indirect metabolic actions of hormones co-encoded in the preproglucagon gene of fishes. Emphasis is placed on a critical analysis of the effects of glucagon and glucagon-like peptide (GLP) and the current knowledge of the respective modes of action is reviewed. In mammals GLPs exert no direct metabolic actions. In fish liver, GLP and glucagon act on similar targets of intermediary metabolism by enhancing flux through glycogenolysis, lipolysis and gluconeogenesis. Increases in substrate oxidation are not uniform. Hormonal activation of glycogen phosphorylase and triglyceride lipase and inhibition of pyruvate kinase are implicated in these actions. Hormone-dependent hyperglycemia, depletion of hepatic glycogen and increases in free fatty acids are noticeablein vivo. Glucagon also activates hepatic amino acid uptake and ammonia excretion. Glucagon actions are accompanied by large increases in hepatic cAMP and increased phosphorylation of pyruvate kinase. Metabolic effects measured after GLP administration are associated with minor, if any, increases in cAMP and effects on pyruvate kinase are variable. We hypothesize that different hepatic receptors with differing modes of intracellular message transduction are involved in glucagon and GLP actions while targetting identical metabolic routes. Responses of different species of fish cover a wide spectrum, and variation of response with the circannual cycle of experimental animals makes comparisons of results, even within one species, difficult.     

Effect of sorghum grain supplementation on glucose metabolism in cattle and sheep fed temperate pasture

The aim of this work was to evaluate the effect of sorghum grain supplementation on plasma glucose, insulin and glucagon concentrations, and hepatic mRNA concentrations of insulin receptor (INSR), pyruvate carboxylase (PC), and phosphoenolpyruvate carboxykinase (PCK1) mRNA and their association with nutrient intake, digestion and rumen volatile fatty acids (VFA) in cattle and sheep fed a fresh temperate pasture. Twelve Hereford × Aberdeen Angus heifers and 12 Corriedale × Milchschaf wethers in positive energy balance were assigned within each species to one of two treatments (n = 6 per treatment within specie): non‐supplemented or supplemented with sorghum grain at 15 g/kg of their body weight (BW). Supplemented cattle had greater plasma glucose concentrations, decreased plasma glucagon concentrations and tended to have greater plasma insulin and insulin‐to‐glucagon ratio than non‐supplemented ones. Hepatic expression of INSR and PC mRNA did not differ between treatments but PCK1 mRNA was less in supplemented than non‐supplemented cattle. Supplemented sheep tended to have greater plasma glucagon concentrations than non‐supplemented ones. Plasma glucose, insulin, insulin‐to‐glucagon ratio, and hepatic expression of INSR and PC mRNA did not differ between treatments, but PCK1 mRNA was less in supplemented than non‐supplemented sheep. The inclusion of sorghum grain in the diet decreased PCK1 mRNA but did not affect PC mRNA in both species; these effects were associated with changes in glucose and endocrine profiles in cattle but not in sheep. Results would suggest that sorghum grain supplementation of animals in positive energy balance (cattle and sheep) fed a fresh temperate pasture would modify hepatic metabolism to prioritize the use of propionate as a gluconeogenic precursor.     

植物中液泡膜H~＋-PPase的研究进展

液泡膜H＋-PPase是一种广泛存在于很多生物体内的H＋转运酶,主要介绍近年来该酶在植物抗逆性以及植物生长的调节作用上的研究进展。     

Regional mRNA expression of key gluconeogenic enzymes in the liver of dairy cows

Liver tissue was collected from eight random dairy cows at a slaughterhouse to test if gene expression of pyruvate carboxylase (PC), mitochondrial phosphoenolpyruvate carboxykinase (PEPCKm) and cytosolic phosphoenolpyruvate carboxykinase (PEPCKc) is different at different locations in the liver. Obtained liver samples were analysed for mRNA expression levels of PC, PEPCKc and PEPCKm and subjected to the MIXED procedure of SAS to test for the sampled locations with cow liver as repeated subject. Additionally, the general linear model procedure (GLM) for analysis of variance was applied to test for significant differences for mRNA abundance of PEPCKm, PEPCKc and bPC between the livers. In conclusion, this study demonstrated that mRNA abundance of PC, PEPCKc and PEPCKm is not different between locations in the liver but may differ between individual cows.     

Effect of dietary carbohydrate sources on growth performance and utilization for gibel carp (Carassius auratus gibelio) and Chinese longsnout catfish (Leiocassis longirostris Günther)

The nutritional function of monosaccharides, disaccharides and polysaccharides for omnivorous gibel carp and carnivorous Chinese longsnout catfish were investigated and the ability of these two species to utilize carbohydrates was compared. For each species, triplicate groups of fish were assigned to each of five groups of isoenergetic and isonitrogenous experimental diets with different carbohydrate sources: glucose, sucrose, dextrin, soluble starch (acid‐modified starch) and α‐cellulose. The carbohydrates were included at 60 g kg^?1 in Chinese longsnout catfish diets and at 200 g kg^?1 in gibel carp diets. A growth trial was carried out in a recirculation system at 27.8 ± 1.9 °C for 8 weeks. The results showed that fish with different food habits showed difference in the utilization of carbohydrate sources. For gibel carp, better specific growth rate (SGR) and feed efficiency (FE) were observed in fish fed diets containing soluble starch and cellulose, but for Chinese longsnout catfish, better SGR and FE were observed in fish fed diets containing dextrin and sucrose. Apparent digestibility coefficient of dry matter (ADC_d) and apparent digestibility coefficient of energy (ADC_e) were significantly affected by dietary carbohydrate sources in gibel carp. ADC_d and ADC_e significantly decreased as dietary carbohydrate complexity increased in Chinese longsnout catfish except that glucose diet had medium ADC_d and ADC_e. In both species, no significant difference of apparent digestibility coefficient of protein was observed between different carbohydrate sources. Dietary carbohydrate sources significantly affected body composition, and liver phosphoenolpyruvate carboxykinase (PEPCK), pyruvate kinase (PK), glucose 6‐phosphate dehydrogenase (G6PD) and malic enzyme (ME) activities also varied according to dietary carbohydrate complexity. Fish with different food habits showed different abilities to synthesize liver glycogen, and the liver glycogen content in gibel carp was significantly higher than in Chinese longsnout catfish. The influence of carbohydrate source on gluconeogenesis and lipogenesis was also different in the two fish species.     

Metabolic enzyme activities in larvae of the African catfish,Clarias gariepinus: changes in relation to age and nutrition

The influence of ontogeny and nutrition on metabolic enzyme activities in larvae of the African catfish, Clarias gariepinus, was studied. After start of exogenous feeding, the larvae were reared for 10 days under three different nutritional conditions: Artemia nauplii, a dry starter diet, and starvation. The live feed gave the best growth (96 mg within 10 days) whereas the dry diet resulted in low growth (33 mg). This growth difference was reflected in larval RNA and DNA concentrations, but not in the levels of soluble protein. Enzymes representing the following aspects of metabolism have been analysed: NADPH generation (G6PDH, ME), glycolysis (PFK, PK), gluconeogenesis (FDPase), amino acid catabolism (GOT, GPT) and oxidative catabolism (CS). All enzymes were present from the start of exogenous feeding onwards, but their maximum specific activities displayed different developmental patterns. In catfish larvae fed on Artemia, G6PDH and ME activities steadily increased with age and weight of the larvae. CS levels remained, after an immediate enhancement upon onset of exogenous feeding, on a rather stable plateau. The amino acid-degrading enzymes GOT and GPT showed maximum levels at days 3–5 of feeding or at a body weight of 10–20 mg, but decreased thereafter. Activities of PFK, PK and FDPase showed low initial levels, and increased significantly with age and size. Based on the ontogenetic patterns of metabolic enzymes, in C. gariepinus larvae an early and a late developmental phase can be distinguished. During the early phase, the glycolytic and gluconeogenetic capacities are low, whereas they are enforced during the later phase. The oxidative capacity is high both during the early and the late phase. The metabolic changes in catfish development coincide with other major ontogenetic events, e.g., alterations of muscle organization, gill morphology, respiration and stomach structure and function. Rearing catfish larvae on a dry diet instead of Artemia partly altered the developmental pattern described: The ontogenetic elevation of CS, PFK and FDPase was delayed and the early peak in GOT and GPT activities was not realized. Particularly during the early developmental phase, the enzyme behaviour of the larvae fed on dry food was similar to that of starved larvae.Abbreviations CS citrate synthase - FDPase fructose-1,6-diphosphatase - GOT glutamate oxaloacetate transaminase - GPT glutamate pyruvate transaminase - G6PDH glucose-6-phosphate dehydrogenase - ME malic enzyme - PFK phosphofructokinase - PK pyruvate kinase     

Effects of cortisol on hepatic carbohydrate metabolism and responsiveness to hormones in the sea raven,Hemitripterus americanus

The sea raven, Hemitripterus americanus, is a sit-and-wait, low metabolic rate, marine teleost. The objective of this study was to determine i) whether cortisol implantation (50 mg. kg^-1) for 7 days altered hepatocyte metabolism, and hepatocyte responsiveness to epinephrine, glucagon and insulin, and ii) whether 8 weeks of food-deprivation modified the above response. Cortisol implantation significantly increased hepatocyte total glucose production and oxidation from alanine compared to the sham group. There was no cortisol effect on glycogen breakdown, suggesting that the activation of other pathways, including gluconeogenesis, are required to account for the increased glucose production. Epinephrine-mediated (10^-5M) glycogen breakdown and insulin-mediated (10^-8M) total glucose production were enhanced in hepatocytes of cortisol implanted sea ravens, but there were no change in any glucagon (10^-7M) effects. The enhanced glycogen breakdown in the absence of similar increases in total glucose production with epinephrine indicates mobilization of carbohydrate reserves for endogenous use by the liver. Food-deprivation for 8 weeks significantly decreased condition factor, plasma cortisol concentration and liver glycogen content in the sea raven, but had no effect on plasma glucose concentration. Hepatocyte total glucose production and flux rates from alanine increased significantly with food-deprivation. Moreover, food-deprivation increased responsiveness of total hepatocyte glucose production to the actions of glucagon and insulin, but not to epinephrine; none of these effects were modified by cortisol implantation. Our results indicate that cortisol in the sea raven exerts both a direct and an indirect or permissive effect on hepatocyte metabolism by modifying hepatocyte responsiveness to epinephrine and insulin stimulation. Cortisol implantation did not modify the effects of glucagon or food-deprivation in this species.