Effects of selenium and serum on selenoprotein W in cultured L8 muscle cells

When rat L8 muscle cells were cultured to examine the effects of serum and selenium concentration on selenoprotein W levels and glutathione peroxidase (GPX) activities, no significant differences (P > 0.05) were found in selenoprotein W levels and GPX activities during differentiation. With three different forms of selenium, selenoprotein W levels and GPX activities were shown to increase in L8 myotubes cultured in media with these selenocompounds. Selenite was utilized more efficiently than selenocysteine for both selenoprotein W and GPX activity, but selenium as selenomethionine was less available. Both the protein content and mRNA levels for selenoprotein W were affected by the selenium content of the media. Northern blot data indicated that the expression of selenoprotein W mRNA increased significantly when L8 myotubes were cultured with selenium (P > 0.05). L8 myotubes cultured in 10% calf serum (CS) versus 2% CS with or without addition of 10 m selenium indicated that the increase of selenoprotein W level in L8 myotubes cultured with higher serum concentration (10% CS) is due to the higher selenium concentration in media rather than serum itself.     

Selenoprotein expression and function—Selenoprotein W

Selenoprotein W (SeW) is a small selenoprotein (85 to 88 amino acids) first identified in sheep suffering from selenium deficiency. The levels are highest in muscle, heart (except rodents) spleen and brain. The deduced amino acid sequence has been obtained for mice, rats, monkeys, humans, sheep, pigs, fish and chickens. The sequences of SeW are identical in rats and mice as well as monkeys and humans. In all eight species of animals cysteine is present at residue number 9 and selenocysteine at residue number 13. Residue number 37 is cysteine in six species of animal with fish and chickens as the exceptions. Of those examined, the rodent SeW is the only one containing four cysteines whereas the others contain only two cysteines. Glutathionylaltion has been shown for SeW from rats and monkeys but has not been confirmed for this selenoprotein from the other six animals. The biological function of SeW has not been definitely identified. Evidence has been obtained that it can serve as an antioxidant, responds to stress, involved in cell immunity, specific target for methylmercury, and has thioredoxin-like function.     

Effects of long-term selenium yeast supplementation on selenium status studied in the rat

To investigate the selenium status during long-term dietary supply of selenium yeast, 30-day-old male rats were fed for 379 days a methionine-adequate low-selenium diet supplemented with 0.2 mg Se/kg (selenium-adequate diet) or 1.5 mg Se/kg (high-selenium diet) in the form of selenium yeast that contained 60% of the element as l-selenomethionine. Their selenium load was determined at several intervals by neutron activation analysis of the selenium concentrations in the main selenium body pools, skeletal muscle and liver. After 64 days the tissue selenium concentrations plateaued in both groups and then stayed at that level. Compared with the selenium-adequate group, elevated tissue selenium concentrations were found in the high-selenium group, but the increase by a factor of 3.5 in the muscle and by a factor of 2.3 in the liver was smaller than the 7.5-fold increase in the selenium intake. In the selenium-adequate group about 50% of the muscle selenium and 30% of the liver selenium and in the high-selenium group about 85% of the muscle selenium and 70% of the liver selenium were estimated to be present in non-selenoprotein forms. During selenium depletion the liver glutathione peroxidase activity in the high-selenium group remained unaffected for 4 weeks and then decreased more slowly than that in the selenium-adequate group. From these results it can be concluded that selenium incorporated from the selenium yeast diet into non-selenoprotein forms can serve as an endogenous selenium source to maintain selenoprotein levels in periods of insufficient selenium supply.     

Selenium and Zinc Concentrations in Kidney,Liver And Muscle of Cattle from Different Parts of Norway

Cattle slaughtered in four different parts of Norway have been examined with respect to selenium and zinc content in kidney, liver and muscle. Highest selenium concentrations were found in kidney and lowest in muscle. In spite of extensive use of standardized concentrates, geographic differences were detected with regard to selenium tissue levels, animals from the southeastern inland region having the lowest levels. According to other workers, this region has low-selenium humus soils, and selenium responsive diseases among young ruminants have been of considerable importance, especially when concentrates had not been given during winter feeding. The recorded tissue selenium levels are compared to other workers’ proposals for normal values. All animals examined in this study seem to be well within healthy limits. Kidney, liver and muscle from cattle are good sources of selenium with respect to human nutrition. As far as zinc concentrations are concerned, muscle has the highest and kidney the lowest levels. Geographic differences were found, and individuals from the midland and northern coastal regions have the highest zinc tissue levels. Cattle from the northern coastal region seems to have especially high zinc concentrations in the organs.     

Characterization and expression of chicken selenoprotein W

As an essential trace element, selenium (Se) deficiency results in White Muscle Disease in livestock and Keshan disease in humans. The main objectives of this study were to clone and characterize the chicken selenoprotein W (SeW) gene and investigate SeW mRNA expression in chicken tissues. The deduced amino acid (AA) sequence of chicken SeW contains 85 AAs with UAG as the stop codon. Like all SeW genes identified in different species, chicken SeW contains one well-conserved selenocysteine (Sec) at the 13th position encoded by the UGA codon. The proposed glutathione (GSH)-binding site at the Cys₃₇ of SeW is not conserved in the chicken, but Cys₉ and Sec₁₃, with possible GSH binding, are conserved in SeWs identified from all species. There are 23–59% and 50–61% homology in cDNA and deduced AA sequences of SeW, respectively, between the chicken and other species. The predicted secondary structure of chicken SeW mRNA indicates that the selenocysteine insertion sequence element is type II with invariant adenosines within the apical bulge. The SeW mRNA expression is high in skeletal muscle followed by brain, but extremely low in other tissues from chickens fed a commercial maize-based diet. The SeW gene is ubiquitously expressed in heart, skeletal muscle, brain, testis, spleen, kidney, lung, liver, stomach and pancreas in chickens fed a commercial diet supplemented with sodium selenite. These results indicate that dietary selenium supplementation regulates SeW gene expression in the chicken and skeletal muscle is the most responsive tissue when dietary Se content is low.     

Developmental changes in the level of glucocorticoid receptors in chick-embryo tissues

The concentrations of glucocorticoid receptors were assayed in various chick embryo tissues by a cytosol charcoal-dextran method using [3H]dexamethasone as ligand. The highest levels of receptors were found in muscle on developmental day 15-16. The order of maximum binding of dexamethasone in various tissues was muscle greater than heart greater than skin greater than tendon greater than kidney greater than cartilage greater than liver. However, marked variation in the level of receptors was found even in the same tissue during development of the chick-embryo. The highest levels of receptors were generally found on days 15-17. For example, in tendon the difference between the highest and lowest level of receptors was about 6-7 fold. Thus the results show that specific glucocorticoid receptors may be found in various tissues of developing chick-embryos.     

Zinc,cadmium, mercury and selenium in polar bears (Ursus maritimus) from Central East Greenland

Muscle, liver, and kidney tissues from 38 polar bears (Ursus maritimus) caught in the Scoresby Sound area, Central East Greenland, were analysed for zinc, cadmium, mercury and selenium. In general, cadmium concentrations were low in muscle, liver and kidney tissue, with geometric means (g.m.) of 0.022 (range: <0.015–0.085), 0.841 (range: 0.092–3.29) and 13.1 (range: 1.04–115) g Cd/g wet weight (ww) respectively. This finding can be explained by low cadmium levels in the blubber of ringed seals. The concentration of mercury in muscle tissue was low (g.m. 0.071; range: 0.039–0.193 g Hg/g ww), whereas concentrations in liver and kidney tissue were relatively high (liver: g.m. 7.87; range: 1.35–24.8 g Hg/g ww, and kidney: g.m. 15.2; range: 1.59–66.6 g Hg/g ww). Mercury and cadmium were positively correlated with age in liver and kidney. Zinc was positively correlated with age in kidney, and selenium was correlated with age in liver. Contrary to other marine mammals, polar bears had higher mercury levels in the kidneys than in the liver. In all three tissues polar bears had significantly lower cadmium levels than ringed seals from the same area. Mercury levels were likewise significantly lower in the muscle tissue of polar bears than in ringed seals, whereas levels in the liver and kidney were significantly higher. The previous geographic trend for cadmium and mercury found in Canadian polar bears could be extended to cover East Greenland as well. Hence cadmium levels were higher in Greenland than in Canada, while the opposite was the case for mercury. Greenland polar bears had higher mercury and cadmium contents in livers and kidneys than polar bears from Svalbard. The mercury levels in muscle and liver tissue from polar bears from East Greenland were twice as high as found in bears from western Alaska, but half the levels found in northern Alaska. Cadmium and zinc were partially correlated in kidney tissue, and this was found for mercury and selenium as well. Cadmium and zinc showed molar ratios close to unity with the highest concentrations occurring in kidney tissue, while the levels of zinc exceeded cadmium in muscle and liver tissue by up to several decades. Mercury and selenium showed molar ratios close to unity in liver and kidneys.     

The organ distribution of selenium in German adults

The selenium concentrations were determined in liver, kidney, skeletal muscle, heart, brain, prostate, testis, bile, lung, and spleen of German traffic accident victims. In addition the nitrogen and phosphorus contents were determined in the same organs and tissues. On a per-weight unit basis, the highest selenium concentration was found in kidney. However, this corresponds to only 4% of the total body selenium. Most of the whole body selenium (50%) is present in skeletal muscle, which thus appears to act as a selenium storage organ. However, there is also evidence that selenium is required for muscle function. In plasma and interstitial fluid, .450 mg of Se, or 7.5% of the total body selenium is present. A comparison of the organ Se concentrations of the German traffic accident victims with the selenium concentrations of the same human organs as reported in different countries indicates that the organ concentrations of West Germans are comparable to that of the population of New Zealand, a low-Se country, and significantly lower than that observed in the organs of American, Canadian, and especially Japanese subjects. The international comparison of the organ selenium concentrations also revealed that the selenium uptake of kidney is higher at low- and adequate dietary Se intakes and lower if the dietary Se supply is high, as is the case for Japanese subjects. Estimates of the daily excretion of selenium with the bile indicate that the amounts are three times higher than the daily urinary losses and in the same order of magnitude as the daily dietary selenium intakes. Enterohepatic reabsorption of selenium from the bile appears to be a significant mechanism of conserving dietary selenium and to maintain Se balance at comparatively low dietary Se intakes.     

Characterisation of cDNA clones for rat muscle carbonic anhydrase III

cDNA clones for rat muscle carbonic anhydrase III have been isolated from a gt-11 library and sequenced. Comparison with human CAIII cDNA showed about 90% homology to rat. The rat clones were used to estimate mRNA from liver and muscle on Northern blots and showed that the sexual dimorphism of CAIII in rat liver relates to a difference in mRNA levels.     

Tissue-specific expression atlas of murine mitochondrial tRNAs

Mammalian mitochondrial tRNA (mt-tRNA) plays a central role in the synthesis of the 13 subunits of the oxidative phosphorylation complex system (OXPHOS). However, many aspects of the context-dependent expression of mt-tRNAs in mammals remain unknown. To investigate the tissue-specific effects of mt-tRNAs, we performed a comprehensive analysis of mitochondrial tRNA expression across five mice tissues (brain, heart, liver, skeletal muscle, and kidney) using Northern blot analysis. Striking differences in the tissue-specific expression of 22 mt-tRNAs were observed, in some cases differing by as much as tenfold from lowest to highest expression levels among these five tissues. Overall, the heart exhibited the highest levels of mt-tRNAs, while the liver displayed markedly lower levels. Variations in the levels of mt-tRNAs showed significant correlations with total mitochondrial DNA (mtDNA) contents in these tissues. However, there were no significant differences observed in the 2-thiouridylation levels of tRNA^Lys, tRNA^Glu, and tRNA^Gln among these tissues. A wide range of aminoacylation levels for 15 mt-tRNAs occurred among these five tissues, with skeletal muscle and kidneys most notably displaying the highest and lowest tRNA aminoacylation levels, respectively. Among these tissues, there was a negative correlation between variations in mt-tRNA aminoacylation levels and corresponding variations in mitochondrial tRNA synthetases (mt-aaRS) expression levels. Furthermore, the variable levels of OXPHOS subunits, as encoded by mtDNA or nuclear genes, may reflect differences in relative functional emphasis for mitochondria in each tissue. Our findings provide new insight into the mechanism of mt-tRNA tissue-specific effects on oxidative phosphorylation.     

Effects of acclimation salinity on the expression of selenoproteins in the tilapia,Oreochromis mossambicus

Selenoproteins are ubiquitously expressed, act on a variety of physiological redox-related processes, and are mostly regulated by selenium levels in animals. To date, the expression of most selenoproteins has not been verified in euryhaline fish models. The Mozambique tilapia, Oreochromis mossambicus, a euryhaline cichlid fish, has a high tolerance for changes in salinity and survives in fresh water (FW) and seawater (SW) environments which differ greatly in selenium availability. In the present study, we searched EST databases for cichlid selenoprotein mRNAs and screened for their differential expression in FW and SW-acclimated tilapia. The expression of mRNAs encoding iodothyronine deiodinases 1, 2 and 3 (Dio1, Dio2, Dio3), Fep15, glutathione peroxidase 2, selenoproteins J, K, L, M, P, S, and W, was measured in the brain, eye, gill, kidney, liver, pituitary, muscle, and intraperitoneal white adipose tissue. Gene expression of selenophosphate synthetase 1, Secp43, and selenocysteine lyase, factors involved in selenoprotein synthesis or in selenium metabolism, were also measured. The highest variation in selenoprotein and synthesis factor mRNA expression between FW- and SW-acclimated fish was found in gill and kidney. While the branchial expression of Dio3 was increased upon transferring tilapia from SW to FW, the inverse effect was observed when fish were transferred from FW to SW. Protein content of Dio3 was higher in fish acclimated to FW than in those acclimated to SW. Together, these results outline tissue distribution of selenoproteins in FW and SW-acclimated tilapia, and indicate that at least Dio3 expression is regulated by environmental salinity.     

Regulation and function of avian selenogenome

Background

Selenium (Se) is an essential micronutrient required by avian species. Dietary Se/vitamin E deficiency induces three classical diseases in chicks: exudative diathesis, nutritional pancreatic atrophy, and nutritional muscular dystrophy.

Zinc,cadmium, mercury and selenium in minke whales,belugas and narwhals from West Greenland

Summary Samples of muscle, liver and kidney from 24 minke whales (Balaenoptera acutorostrata), 43 belugas (Delphinapterus leucas), and 98 narwhals (Monodon monoceros) were analyzed for zinc, cadmium, mercury, and selenium. Highly significant age accumulation of mercury was found. A lower level of significance of age accumulation of cadmium in belugas and narwhals is probably due to the fact that some of the highest cadmium concentrations are in subadults and young adults. The maximum concentrations of cadmium and mercury are very high: 1.68, 73.7, and 125 g cadmium, and 9.88, 42.8, and 4.61 g mercury per g wet weight of narwhal muscle, liver and kidney, respectively. The cadmium concentrations are correlated in the three organs, as are mercury and to a lesser extent selenium concentrations. The concentrations of mercury and selenium in liver are highly correlated.     

Molecular cloning and chromosomal localization of a human skeletal muscle PP-1γ1 cDNA

Type-1-protein phosphatase (PP-1) activity is reduced in skeletal muscle from human subjects with insulin resistance (Kida et al. 1990). This reduced phosphatase activity probably leads to the abnormal insulin action for glucose storage observed in insulin-resistant subjects. In the present study, a human homolog of rat liver PP-11 cDNA was isolated from human skeletal muscle. The nucleotide sequence contains a 957-nucleotide open reading frame encoding an amino acid sequence identical to that encoded by rat liver PP-11 cDNA. Northern blot analysis shows PP-11-specific mRNA is expressed in human heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas. PP-11 was localized to human Chromosome 12.     

L-type glycogen synthase. Tissue distribution and electrophoretic mobility

We previously reported (Kaslow, H.R., and Lesikar, D.D.FEBS Lett. (1984) 172, 294-298) the generation of antisera against rat skeletal muscle glycogen synthase. Using immunoblot analysis, the antisera recognized the enzyme in crude extracts from rat skeletal muscle, heart, fat, kidney, and brain, but not liver. These results suggested that there are at least two isozymes of glycogen synthase, and that most tissues contain a form similar or identical to the skeletal muscle type, referred to as "M-type" glycogen synthase. We have now used an antiserum specific for the enzyme from liver, termed "L-type" glycogen synthase, to study its distribution and electrophoretic mobility. Immunoblot analysis using this antiserum indicates that L-type glycogen synthase is found in liver, but not skeletal muscle, heart, fat, kidney, or brain. In sodium dodecyl sulfate-polyacrylamide gels of crude liver extracts prepared with protease inhibitors, rat L-type synthase was detected with electrophoretic mobility Mapp = 85,000. In contrast, the M-type enzyme in crude skeletal muscle extracts with protease inhibitors was detected with Mapp = 86,000 and 89,000. During purification of L-type synthase, apparent proteolysis can generate forms with increased electrophoretic mobility (Mapp = 75,000), still recognized by the antiserum. These M-type and L-type antisera did not recognize a protein with Mapp greater than phosphorylase. The anti-rat L-type antisera recognized glycogen synthase in blots of crude extracts of rabbit liver, but with Mapp = 88,000, a value 3,000 greater than that found for the rat liver enzyme. The anti-rat M-type antisera failed to recognize the enzyme in blots of crude extracts of rabbit muscle. Thus, in both muscle and liver, the corresponding rat and rabbit enzymes are structurally different. Because the differences described above persist after resolving these proteins by denaturing sodium dodecyl sulfate electrophoresis, these differences reside in the structure of the proteins themselves, not in some factor bound to the protein in crude extracts.     

Selenium-regulated hierarchy of human selenoproteome in cancerous and immortalized cells lines

Background

Selenoproteins (25 genes in human) co-translationally incorporate selenocysteine using a UGA codon, normally used as a stop signal. The human selenoproteome is primarily regulated by selenium bioavailability with a tissue-specific hierarchy.

Translation regulation of mammalian selenoproteins

Background

Interest in selenium research has considerably grown over the last decades owing to the association of selenium deficiencies with an increased risk of several human diseases, including cancers, cardiovascular disorders and infectious diseases. The discovery of a genetically encoded 21^st amino acid, selenocysteine, is a fascinating breakthrough in molecular biology as it is the first addition to the genetic code deciphered in the 1960s. Selenocysteine is a structural and functional analog of cysteine, where selenium replaces sulfur, and its presence is critical for the catalytic activity of selenoproteins.