Metabolic activities of partially degenerated hypertrophic chondrocytes: gene expression of hyaluronan synthases

Ultrastructural aspects of hypertrophic chondrocytes in hamster and mouse epiphysial cartilage were examined in relation to their metabolic activities. With the hypertrophic change, cytoplasmic vacuolization proceeded leaving the partially intact endoplasmic reticulum (ER). In the hypertrophic cells, cytoplasmic hyaluronan was stained with the biotinylated hyaluronan-binding region (b-HABR) of aggrecan, and mRNAs of hyaluronan synthase (Has 1, Has 2 and Has 3) were detected by in situ hybridization. When the epiphysial cartilage was cultured in the presence of 35S, 3H-GlcNAc, 3H-proline or 14C-palmitic acid, vacuolated late hypertrophic chondrocytes were labeled with these radioactive precursors. The evidence indicates that late hypertrophic chondrocytes are metabolically active, which appears to be essential for the enlargement of chondrocytes.     

温度对梯棱羊肚菌抗氧化酶活及其基因表达的影响

大田栽培条件下,环境温度无法精确调控,温度胁迫是影响羊肚菌生长发育的重要因素.抗氧化酶和抗氧化活性物质是羊肚菌抵御逆境胁迫的重要因子.温度胁迫下,羊肚菌菌丝会通过增加相应酶活性来减少活性氧的积累,降低对细胞的损伤.作者研究了不同温度对梯棱羊肚菌菌丝生长、抗氧化酶(超氧化物歧化酶SOD、过氧化氢酶CAT、谷胱甘肽还原酶G...     

Dietary protein modulates digestive enzyme activities and gene expression in red tilapia juveniles

It is known that the level of dietary protein modulates the enzymatic activity of the digestive tract of fish; however, its effect at the molecular level on these enzymes and the hormones regulating appetite has not been well characterised. The objective of this study was to evaluate the effect of CP on the activity of proteases and the expression of genes related to the ingestion and protein digestion of juveniles of red tilapia (Oreochromis sp.), as well as the effects on performance, protein retention and body composition of tilapia. A total of 240 juveniles (29.32 ± 5.19 g) were used, distributed across 20 tanks of 100 l in a closed recirculation system. The fish were fed to apparent satiety for 42 days using four isoenergetic diets with different CP levels (24%, 30%, 36% and 42%). The results indicate that fish fed the 30% CP diet exhibited a higher growth performance compared to those on the 42% CP diet (P < 0.05). Feed intake in fish fed 24% and 30% CP diets was significantly higher than that in fish fed 36% and 42% CP diets (P < 0.05). A significant elevation of protein retention was observed in fish fed with 24% and 30% CP diets. Fish fed with 24% CP exhibited a significant increase in lipid deposition in the whole body. The diet with 42% CP was associated with the highest expression of pepsinogen and the lowest activity of acid protease (P < 0.05). The expression of hepatopancreatic trypsinogen increased as CP levels in the diet increased (P < 0.05) up to 36%, whereas trypsin activity showed a significant reduction with 42% CP (P < 0.05). The diet with 42% CP was associated with the lowest intestinal chymotrypsinogen expression and the lowest chymotrypsin activity (P < 0.05). α-amylase expression decreased with increasing (P < 0.05) CP levels up to 36%. No significant differences were observed in the expression of procarboxypeptidase, lipase or leptin among all the groups (P > 0.05). In addition, the diet with 42% CP resulted in a decrease (P < 0.05) in the expression of ghrelin and insulin and an increase (P < 0.05) in the expression of cholecystokinin and peptide yy. It is concluded that variation in dietary protein promoted changes in the metabolism of the red tilapia, which was reflected in proteolytic activity and expression of digestion and appetite-regulating genes.     

Metabolic load and heterologous gene expression

The expression of a foreign protein(s) in a recombinant host cell or organism often utilizes a significant amount of the host cell's resources, removing those resources away from host cell metabolism and placing a metabolic load (metabolic drain, metabolic burden) on the host. As a consequence of the imposed metabolic load, the biochemistry and physiology of the host may be dramatically altered. The numerous physiological changes that may occur often lowers the amount of the target foreign protein that is produced and eventually recovered from the recombinant organism. In this review the physiological changes to host cells, the causes of the phenomenon of metabolic load, and several strategies to avoid some of the problems associated with metabolic load are elaborated and discussed.     

逆境条件下小麦种子活力与种子萌发相关酶活性及其基因表达的关系

分析不同基因型小麦品种逆境萌发过程中种子萌发相关酶活性及基因表达差异,明确在逆境条件下,种子活力与种子萌发相关酶活性及基因表达量的关系.通过标准发芽试验和逆境(冷浸、人工老化、干旱胁迫)发芽试验,测定4个小麦品种种子活力、萌发过程中可溶性总糖和可溶性蛋白含量、α-淀粉酶活性、半胱氨酸蛋白酶活性及相关基因表达量.结果表明:干旱、人工老化和冷浸胁迫3种逆境对种子活力都有一定影响.不同萌发条件下,可溶性总糖含量呈现先小幅度升高后小幅度降低再迅速升高的趋势;而可溶性蛋白含量随着萌发时间的延长呈现逐渐下降的趋势.α-淀粉酶活性整体呈现逐渐升高的趋势,但在冷浸胁迫处理后,豫农949和轮选061的α-淀粉酶活性在萌发60 h后出现下降.半胱氨酸蛋白酶活性整体呈先降低后升高的趋势,但在干旱胁迫条件下,豫农949、豫麦49-198和轮选061的半胱氨酸蛋白酶活性呈现先升高后降低再升高的趋势.不同逆境萌发条件下,α-AMY(α-淀粉酶基因)表达量整体呈先上升后下降的趋势.冷浸胁迫处理后,轮选061的α-AMY表达量高于对照,在其他逆境萌发条件下,4个品种的α-AMY表达量均低于对照;人工老化处理后,长4738的CP(半胱氨酸蛋白酶基因)表达量与对照差异不显著,在其他逆境萌发条件下,4个品种的CP表达量均高于对照.种子萌发期间,不同萌发条件下α-淀粉酶和半胱氨酸蛋白酶活性与其基因表达并没有直接关系,α-淀粉酶活性与可溶性总糖含量达到显著正相关,半胱氨酸蛋白酶活性与可溶性蛋白含量的相关性不显著.在标准发芽条件下,α-淀粉酶活性与活力指数呈显著正相关,而在逆境萌发过程中,其相关性不显著.冷浸胁迫处理后,半胱氨酸蛋白酶活性与活力指数呈显著正相关,但在标准发芽、干旱胁迫、人工老化处理后,其相关性不显著.     

Metabolic effects of hemoglobin gene expression in plants

Hemoglobin (Hb) genes are ubiquitous in plants. Several classes have been identified and are expressed during infection by nitrogen-fixing symbionts, as a result of tissue hypoxia, during seed germination, and in developing (e.g. meristematic) tissues. The induction of the Hb gene by hypoxia is linked to a decrease in ATP levels and is mediated by Ca(2+). Numerous investigations have led to the conclusion that the main function of hypoxically-induced Hb is to metabolize nitric oxide (NO) formed as a by-product of nitrate/nitrite reduction. In this function, Hb serves as a part of an NO dioxygenase system, using traces of oxygen to convert NO to nitrate. It operates in conjunction with a methemoglobin reductase protein, which reduces the oxidized form of Hb (methemoglobin) formed in the course of the NO dioxygenase reaction. The complete reaction serves to maintain the cellular energy and redox state. Plant hemoglobins may also function to modulate effects of plant hormones that employ NO as a downstream signal transduction component.     

Spatial and temporal gene expression patterns occur during corm development.

We investigated gene expression patterns that occur during taro corm development. Two-dimensional gel electrophoresis identified several different prevalent proteins that accumulate during corm development. Microsequencing studies indicated that some of these proteins are related to taste-modifying proteins, such as curculin and miraculin, and proteins found in other storage organs, such as sporamin and the Kunitz trypsin inhibitor. A curculin-encoding cDNA clone, designated as TC1, was identified that corresponds to a highly prevalent 1-kb corm mRNA. The TC1 mRNA accumulates during corm development, is more prevalent in corm apical than basal regions, and is either absent, or present at low concentrations, in other vegetative organs such as the leaf and root. In situ hybridization experiments showed that the TC1 mRNA is highly concentrated in corm storage parenchyma cells and is absent, or present in reduced concentrations, in other corm cells and tissues. Our results show that corm development is associated with the differentiation of specialized cells and tissues, and that these differentiation events are coupled with the temporal and spatial expression of corm-specific genes.     

Changes in antioxidant enzyme activities and gene expression in two muskmelon genotypes under progressive water stress

Responses of two muskmelon (Cucumis melo L.) genotypes (drought tolerant SC-15 and drought susceptible EC-564755) were analyzed at 0, 7, 14, and 21 d of progressive water stress. Although water deficit caused a significant decline in relative water content, the magnitude of reduction was lower in SC-15. Electrolyte leakage, hydrogen peroxide, and malonydialdehyde generation were higher in EC-564755, whereas accumulation of proline was higher in SC-15. Higher activities of antioxidant enzymes, such as catalase, superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, and glutathione reductase, and higher expression of the respective genes were recorded in SC-15 than in EC-564755. Expressions of DREB2C and DREB3 in SC-15 revealed a fluctuating pattern with down-regulation on days 7 and 21 of water stress, whereas up-regulation was observed on day 14. Concurrently, both genes in EC-564755 showed continuous down-regulation on days 7, 14, and 21 of water stress. Expressions of RD22 and dehydrin recorded on days 7, 14, and 21 were lower in SC-15. The cluster analysis showed that, these two genotypes had a clear distinction in physiological and biochemical properties and gene expressions under water stress and the genotype SC-15 had more efficient osmoprotectant mechanism than genotype EC-564755 under water deficit conditions.     

Spatial and temporal expression of a maize lipid transfer protein gene.

We studied the temporal and spatial pattern of lipid transfer protein (LTP) gene expression, as well as the localization of this protein, in maize. Using an LTP gene, we observed an accumulation of LTP mRNA in embryos and endosperms during seed maturation. LTP gene expression was also investigated in young seedlings. After germination, the level of LTP mRNA in the coleoptile increased, with a maximum at 7 days, whereas LTP mRNA levels were low in the scutellum and negligible in roots. The high levels of LTP mRNA found in coleoptiles and embryos were confirmed by in situ hybridization. Moreover, LTP gene expression appeared to be localized in the external cellular layers and around the leaf veins. Using immunogold methods, we also observed that LTP was distributed heterogeneously in the different cells of coleoptiles and leaves. The highest concentrations of LTP were found in the outer epidermis of the coleoptiles as well as the leaf veins. Together, our observations indicate that LTP gene expression is not only organ specific and time specific but also cell specific.     

Metabolic regulation and gene expression of root phosphoenolpyruvate carboxylase by different nitrogen sources

Alfalfa (Medicago sativa L.) N-sufficient plants were fed 1·5 mM N in the form of NO₃⁻, NH₄⁺ or NO₃⁻ in conjunction with NH₄⁺, or were N-deprived for 2 weeks. The specific activity of phosphoenolpyruvate carboxylase (PEPC) from the non-nodulated roots of N-sufficient plants was increased in comparison with that of N-deprived plants. The PEPC value was highest with NO₃⁻ nutrition, lowest with NH₄⁺ and intermediate in plants that were fed mixed salts. The protein was more abundant in NO₃⁻-fed plants than in either NH₄⁺- or N mixed-fed plants. Nitrogen starvation decreased the level of PEPC mRNA, and nitrate was the N form that most stimulated PEPC gene expression. The malate content was significantly lower in NO₃⁻-deprived than in NO₃⁻-sufficient plants. Root malate accumulation was high in NO₃⁻-fed plants, but decreased significantly in plants that were fed with NH₄⁺. The effect of malate on the desalted enzyme was also investigated. Root PEPC was not very sensitive to malate and PEPC activity was inhibited only by very high concentrations of malate. Asparagine and glutamine enhanced PEPC activity markedly in NO₃⁻-fed plants, but failed to affect plants that were either treated with other N types or N starved. Glutamate and citrate inhibited PEPC activity only at optimal pH. N-nutrition also influenced root nitrate and ammonium accumulation. Nitrate accumulated in the roots of NO₃⁻- and (NO₃⁻ + NH₄⁺)-fed plants, but was undetectable in those administered NH₄⁺. Both the nitrate and the ammonium contents were significantly reduced in NO₃⁻- and (NO₃⁻ + NH₄⁺)-starved plants. Root accumulation of free amino acids was strongly influenced by the type of N administered. It was highest in NH₄⁺-fed plants and the most abundant amides were asparagine and glutamine. It was concluded that root PEPC from alfalfa plants is N regulated and that nitrate exerts a strong influence on the PEPC enzyme by enhancing both PEPC gene expression and activity.