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The objectives of this study are to examine hepatic gene expression changes caused by GH transgenesis and enhanced growth. This is the first use of cDNA microarrays to study the influence of GH transgenesis on liver gene expression in a non-mammalian vertebrate, and the first such study using sexually immature animals. Three groups of coho salmon were examined: GH transgenic on full ration (T), GH transgenic on restricted ration (R), and control non-transgenic (C). Specific growth rates for weight in T were approximately eightfold higher than in C, and fourfold higher than in R. Differential gene expression in T, R, and C samples was determined using approximately 3500 and 16,000 gene microarrays, and R and C samples were compared on a different approximately 4000 gene microarray. The use of multiple microarray platforms increased the overall proportion of the hepatic transcriptome considered in these studies. Cross-platform comparisons identified genes behaving similarly between studies. For example, genes encoding a precerebellin-like protein and complement component C3 were downregulated in R relative to C (R < C) in two microarray studies, and hemoglobins alpha and beta were R > C in all three studies. Comparisons of informative gene lists within and between studies inferred causes of altered gene expression. For example, ten genes, including 78 kDa glucose-regulated protein, glycerol-3-phosphate dehydrogenase, hemoglobins alpha and beta, and a C-type lectin, were likely induced by GH transgenesis due to their presence in both T > C and R > C gene lists. Eleven genes, including hepcidin, nuclear protein p8, precerebellin-like, transketolase, and fatty acid-binding protein, were present in both T < C and R < C gene lists and were, therefore, likely suppressed by GH transgenesis. A large number of salmonid genes identified in these studies are involved in iron homeostasis, mitochondrial function, carbohydrate metabolism, cellular proliferation, and innate immunity. Pentose phosphate pathway genes phosphogluconate dehydrogenase, transaldolase, and transketolase, were dysregulated in GH transgenic samples relative to control samples. Changes in the expression of genes involved in maintaining hemoglobin levels (heme oxygenase, hemoglobins alpha and beta, Kruppel-like globin gene activator, hepcidin) in R and T fish indicate a need for additional hemoglobin in the transgenic fish, perhaps due to higher metabolic rate required for enhanced growth.
J Mol Endocrinol 2006 Oct
PMID:Multiple microarray platforms utilized for hepatic gene expression profiling of GH transgenic coho salmon with and without ration restriction. 1703 44

This study examined whether adult bovine muscle satellite cells from 30-month-old Hanwoo cattle are multipotential. The satellite cells were found to have the potential to proliferate and differentiate into myoblasts with the formation of multinucleated cells. In addition, treatment with the peroxisome proliferator activating receptor-gamma (PPARgamma) agonist, rosiglitazone, promoted their trans-differentiation into adipocytes with significant increases in glycerol accumulation and glycerol-3-phosphate dehydrogenase activity. Western blot analysis revealed that increased levels of the adipocyte fatty acid-binding protein, PPARgamma and of CCAAT/enhancer-binding protein were closely related to rosiglitazone-induced differentiation of the cells. These findings demonstrate that satellite cells from adult Hanwoo cattle are multipotent, and that their trans-differentiation into adipocytes can be induced by rosiglitazone.
Mol Cells 2006 Oct 31
PMID:Satellite cells isolated from adult Hanwoo muscle can proliferate and differentiate into myoblasts and adipose-like cells. 1708 78

We assessed the daily patterns of parameters involved in energy metabolism in plasma and brain of rainbow trout. Where daily rhythms were found, we analyzed the potential influence of feeding. Immature rainbow trout were randomly distributed in 3 groups: fish fed for 7 days, fish fasted for 7 days, and fish fasted for 7 days and refed for 4 days. On sampling day, fish of fed and refed groups were fed at 11.00 h, and all fish were sampled from each treatment group using the following time schedule: 14.00, 18.00, 21.00, 00.00, 04.00, 07.00, 10.00 and 14.00 h. The results obtained from metabolic parameters assessed in plasma and brain can be grouped into three different categories, such as (i) those displaying no 24 h changes in fed fish such as plasma lactate, protein or acetoacetate levels, as well as brain amino acid and protein levels, and lowKm(glucose) hexokinase, and aspartate aminotransferase activities, (ii) those displaying 24 h changes that were apparently dependent on feeding since they disappeared in fasted fish such as the case of plasma cortisol, glucose and triglyceride levels, as well as brain glycogen, glucose, and lactate levels, and pyruvate kinase and hexokinase IV activities, and (iii) those parameters displaying 24 h changes apparently not dependent on feeding such as plasma amino acids, brain acetoacetate levels as well as several enzyme activities measured in brain such as glucose 6-phosphate dehydrogenase, alpha-glycerophosphate dehydrogenase, glutamate dehydrogenase, and lactate dehydrogenase-oxidase. In general, 24 h changes dependent on feeding indicate an increased use of glucose in brain several hours post-feeding whereas those changes not dependent on feeding were characterized by reduced levels/activity at the night period suggesting a metabolic depression in brain during darkness.
Comp Biochem Physiol A Mol Integr Physiol 2007 Feb
PMID:Daily changes in parameters of energy metabolism in brain of rainbow trout: dependence on feeding. 1712 77

We assessed the daily patterns of parameters involved in energy metabolism in liver, white muscle, and gills of rainbow trout. Where daily rhythms were found, we analyzed the potential influence of feeding. Immature rainbow trout were randomly distributed in 3 groups: fish fed for 7 days, fish fasted for 7 days, and fish fasted for 7 days and refed for 4 days. On sampling day, fish of fed and refed groups were fed at 11.00 h, and all fish were sampled from each treatment group using the following time schedule: 14.00, 18.00, 21.00, 00.00, 04.00, 07.00, 10.00 and 14.00 h. The results obtained from metabolic parameters can be grouped into four different categories, such as i) those displaying no daily changes in any group assessed in liver (acetoacetate and lactate levels), white muscle (protein levels, and low Km (glucose) hexokinase (HK) and HK-IV activities) and gills (protein levels), ii) those displaying no 24 h changes in fed fish but in refed or fasted fish in liver (glucose, glycogen, amino acid and protein levels, and HK-IV activity), white muscle (glycogen and amino acid levels) and gills (glucose levels), iii) those displaying 24 h changes that were apparently dependent on feeding since they disappear in fasted fish in liver (Low Km (glucose) HK, lactate dehydrogenase (LDH-O), glucose 6-phosphatase (G6Pase), fructose 1,6-bisphosphatase (FBPase) , alpha-glycerophosphate dehydrogenase (G3PDH), glutamate dehydrogenase (GDH) and aspartate aminotransferase (Asp-AT) activities), white muscle (glucose levels, and pyruvate kinase (PK), LDH-O, G3PDH and Asp-AT activities) and gills (glycogen and lactate levels, and Low Km (glucose) HK, HK-IV, LDH-O and Asp-AT activities), and iv) those parameters displaying 24 h changes apparently not dependent on feeding in liver (lactate levels and PK activity) and gills (amino acid levels, and PK and GDH activities). In general, most 24 h changes observed were dependent on feeding and can be also related to daily changes in activity.
Comp Biochem Physiol A Mol Integr Physiol 2007 Jun
PMID:Daily changes in parameters of energy metabolism in liver, white muscle, and gills of rainbow trout: dependence on feeding. 1731 50

Nitric oxide (NO) is an important chemical messenger controlling many physiological functions, involving cell proliferation, and differentiation. The purpose of this study was to investigate the effect of NO on adipocyte differentiation using a murine preadipocyte cell line, 3T3-L1. The treatment with a NO donor, 1-hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene (NOC18), reduced some markers of adipocyte differentiation such as glycerol-3-phosphate dehydrogenase activity, and intracellular lipid accumulation. To examine whether these effects of NOC18 on adipocyte differentiation markers are due to its cytotoxity, lactate dehydrogenase (LDH) release from the cells were measured. NOC18 did not affect LDH release into the culture medium. Thus, the suppressive actions of NO donor were unlikely to result from its cytotoxicity. Peroxisome proliferator-activated receptor (PPAR) gamma is a critical transcription factor for adipocyte differentiation and adipocyte fatty acid binding protein (aP2) gene is one of its targets. Protein expression of PPARgamma was not diminished by NOC18 treatment, although mRNA expression of aP2 was reduced. Electrophoretic mobility shift assay showed that NOC18 interfered with the DNA binding activity of PPARgamma. Therefore, the present experiment suggest that NO suppresses adipocyte differentiation through suppressing the transcriptional activity of PPARgamma, without suppressing its expression level.
Mol Cell Biochem 2007 Jun
PMID:Nitric oxide suppresses preadipocyte differentiation in 3T3-L1 culture. 1754 9

In a previous study, we found interaction of gymnemic acid (GA) with glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a key enzyme in glycolysis. We now examined interaction of GA with glycolytic and related enzymes. We found that (1) GA induced a band smearing of glycerol-3-phosphate dehydrogenase (G3PDH) as well as that of GAPDH in SDS-PAGE, (2) GA diminished the G3PDH band detected by an antibody to phosphoserine, and (3) GA inhibited the G3PDH activity. The GA-induced smearing of the G3PDH band was diminished by prior incubation of GA with gamma-cyclodextrin. GA gave no effects on the electrophoretic and phosphoserine bands of other glycolytic enzymes. NAD and NADH diminished the GA-induced smearing of the G3PDH and GAPDH bands in different concentration-dependent manner. Pretreatment of G3PDH with heated SDS-containing buffer or pretreatment with hydroxylamine diminished the GA-induced smearing of G3PDH. Deacylation of GA by alkaline hydrolysis diminished the smearing of G3PDH band, thereby indicating that the acyl moieties of GA were necessary for the GA-induced smearing of G3PDH. These results indicated the interaction of GA with G3PDH, an enzyme involved in glycerol metabolism. These studies suggest that GA may have some pharmacological activities including antidiabetic activity and lipid lowering effects via interaction with GAPDH and G3PDH.
Mol Cell Biochem 2008 Mar
PMID:Gymnemic acid interacts with mammalian glycerol-3-phosphate dehydrogenase. 1808 92

The Forkhead transcription factor O 1 (FoxO1) gene plays an important role in the integration of hormone-activated signaling pathways with the complex transcriptional cascade that promotes clonal cell line differentiation. However, tissue expression of porcine FoxO1 and its function during porcine preadipocyte differentiation remained poorly understood. In the present study, we investigated tissue expressions of FoxO1 in pig by real time quantitative RT-PCR (qRT-PCR) and western blotting, and explored its role in porcine preadipocytes differentiation by RNA interference technique and qRT-PCR. FoxO1 gene expressions were highly in subcutaneous adipose and visceral adipose tissues, and higher in piglets than those in adults (P < 0.05). We showed that expression of endogenous FoxO1 in preadipocytes transfected with pBS/U6-siFoxO1-1748 expression vector was inhibited efficiently. After reducing expression of FoxO1, glycerol-3-phosphate dehydrogenase (GPDH) activity and triglyceride (TG) content increased from day 1 to 9, and the time-course expressions of several key adipogenic genes mRNA, including peroxisome proliferator-activated receptor gamma on day 3, 5, and 7, adipocyte fatty acid binding protein on day 1, 3, and 5, and sirtuin1 on day 1, 3, and 5, were increased significantly (P < 0.05). Lipoprotein lipase was unrelevant to FoxO1. By using insulin-like growth factor-I treating, expression of FoxO1 reduced at day 3 and 5 (P < 0.05), and significant differentiation of porcine preadipocyte with increasing number of filled-lipid cell and size of lipid droplets, GPDH activity and TG content were promoted. These results suggested that porcine FoxO1 gene took part in the regulation of adipose and was a negative transcription regulation factor in preadipocyte differentiation.
Mol Biol Rep 2009 Jan
PMID:Tissue expression of porcine FoxO1 and its negative regulation during primary preadipocyte differentiation. 1829 98

The present study was carried out to clarify the direct effect of fatty acids (FAs) on chick (Gallus gallus) adipocyte differentiation in the absence of dexmethasone (DEX), a commonly used as strong inducer for adipocyte differentiation. Adipocyte differentiation was initiated by maintaining confluent cell in serum-free medium supplemented with FAs. Upon exposure to FAs, glycerol-3-phosphate dehydrogenase activity (GPDH) as adipocyte differentiation marker rapidly increased, and was significantly higher in chick adipocyte than in control cell. The morphology of the FAs-treated cell changed from fibroblast-like to polygon, and the cells accumulated many cytoplasmic lipid droplets as estimated by Oil red O staining. Neither insulin nor bovine serum albumin, as substitutes for serum, had an effect on chick adipocyte differentiation. The FAs-treated cell had a higher protein and mRNA expression levels for peroxisome proliferator-activated receptor-gamma (PPARgamma), a master regulator of differentiation, compared with untreated cell. In FAs-treated cell, the mRNA expression levels of adipocyte-specific genes, such as CCAAT/enhancer binding protein-alpha (C/EBP alpha) and adipocyte fatty acid-binding protein (aP2) were higher than in control cell. These results indicated that FAs, but not DEX, are essential inducers for chick adipocyte differentiation by elevating PPARgamma expression.
Comp Biochem Physiol A Mol Integr Physiol 2008 Dec
PMID:Fatty acids but not dexamethasone are essential inducers for chick adipocyte differentiation in vitro. 1865 28

Exposure to environmental pollutants such as polychlorinated biphenyls (PCBs) is now taken into account to partly explain the worldwide decline of amphibians. PCBs induce deleterious effects on developing amphibians including deformities and delays in metamorphosis. However, the molecular mechanisms by which they express their toxicity during the development of tadpoles are still largely unknown. A proteomics analysis was performed on developing Xenopus laevis tadpoles exposed from 2 to 5 days postfertilization to either 0.1 or 1 ppm Aroclor 1254, a PCB mixture. Two-dimensional DIGE with a minimal labeling method coupled to nanoflow liquid chromatography-tandem mass spectrometry was used to detect and identify proteins differentially expressed under PCBs conditions. Results showed that 59 spots from the 0.1 ppm Aroclor 1254 condition and 57 spots from the 1 ppm Aroclor 1254 condition displayed a significant increase or decrease of abundance compared with the control. In total, 28 proteins were identified. The results suggest that PCBs induce mechanisms against oxidative stress (peroxiredoxins 1 and 2), adaptative changes in the energetic metabolism (enolase 1, glycerol-3-phosphate dehydrogenase, and creatine kinase muscle and brain types), and the implication of the unfolded protein response system (glucose-regulated protein, 58 kDa). They also affect, at least at the highest concentration tested, the synthesis of proteins involved in normal cytogenesis (alpha-tropomyosin, myosin heavy chain, and alpha-actin). For the first time, proteins such as aldehyde dehydrogenase 7A1, CArG binding factor-A, prolyl 4-hydroxylase beta, and nuclear matrix protein 200 were also shown to be up-regulated by PCBs in developing amphibians. These data argue that protein expression reorganization should be taken into account while estimating the toxicological hazard of wild amphibian populations exposed to PCBs.
Mol Cell Proteomics 2009 Apr
PMID:Protein expression profiling in the African clawed frog Xenopus laevis tadpoles exposed to the polychlorinated biphenyl mixture aroclor 1254. 1901 Dec 58

Oxidative damage to cellular macromolecules is believed to underlie the development of many pathological states and aging. The agents responsible for this damage are generally thought to be reactive oxygen species, such as superoxide, hydrogen peroxide, and hydroxyl radical. The main source of reactive species production within most cells is the mitochondria. Within the mitochondria the primary reactive oxygen species produced is superoxide, most of which is converted to hydrogen peroxide by the action of superoxide dismutase. The production of superoxide by mitochondria has been localized to several enzymes of the electron transport chain, including Complexes I and III and glycerol-3-phosphate dehydrogenase. In this chapter the current consensus view of sites, rates, mechanisms, and topology of superoxide production by mitochondria is described. A brief overview of the methods for measuring reactive oxygen species production in isolated mitochondria and cells is also presented.
Methods Mol Biol 2009
PMID:Reactive oxygen species production by mitochondria. 1951 74


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