Effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine on differentiating mouse N2a neuroblastoma cells

The effect of the neurotoxin 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine (MPTP) was investigated in mouse N2a neuroblastoma cells, induced to differentiate by serum withdrawal and addition of dibutyryl cyclic AMP, over a 24-h period. Addition of MPTP (10 microM) during differentiation caused a change in cell morphology characterised by an inhibition of axon outgrowth, in the absence of cell death. Biochemical characterisation by western blotting revealed that MPTP had no significant effects on the levels of actin, alpha-tubulin, or total heavy-chain neurofilament (NF-H). However, NF-H phosphorylation appeared to increase following MPTP treatment when blots were probed with the phosphorylation state-specific antibodies RMd09 and Ta51. In addition, indirect immunofluorescence analysis revealed an accumulation of phosphorylated NF-H in the cell perikaryon, suggesting that altered NF-H distribution was associated with the observed effects of MPTP on cell morphology. These changes may represent a useful in vitro marker of MPTP neurotoxicity within a simple differentiating neuronal cell model system.     

Moderate superoxide production is an early promoter of mitochondrial biogenesis in differentiating N2a neuroblastoma cells

Reactive oxygen species (ROS) have been widely considered as harmful for cell development and as promoters of cell aging by increasing oxidative stress. However, ROS have an important role in cell signaling and they have been demonstrated to be beneficial by triggering hormetic signals, which could protect the organism from later insults. In the present study, N2a murine neuroblastoma cells were used as a paradigm of cell-specific (neural) differentiation partly mediated by ROS. Differentiation was triggered by the established treatments of serum starvation, forskolin or dibutyryl cyclic AMP. A marked differentiation, expressed as the development of neurites, was detected by fixation and staining with coomassie brilliant blue after 48 h treatment. This was accompanied by an increase in mitochondrial mass detected by mitotracker green staining, an increased expression of the peroxisome proliferator-activated receptor gamma (PPARγ) coactivator 1-alpha (PGC-1α) and succinate dehydrogenase activity as detected by MTT. In line with these results, an increase in free radicals, specifically superoxide anion, was detected in differentiating cells by flow cytometry. Superoxide scavenging by MnTBAP and MAPK inhibition by PD98059 partially reversed differentiation and mitochondrial biogenesis. In this way, we demonstrated that mitochondrial biogenesis and differentiation are mediated by superoxide and MAPK cues. Our data suggest that differentiation and mitochondrial biogenesis in N2a cells are part of a hormetic response which is triggered by a modest increase of superoxide anion concentration within the mitochondria.     

Mutagenic activation of 2-amino-3-methylimidazo[4,5-f]-quinoline (IQ) and 2-amino-3,4-dimethylimidazo[4,5-f]-quinoline (MeIQ) by subcellular fractions and cells isolated from small intestine,kidney and liver of the rat

The mutagenic activity of the pyrolysis products 2-amino-3-methylimidazo[4,5-f]-quinoline and 2-amino-3,4-dimethylimidazo[4,5-f]-quinoline in Salmonella typhimurium TA98 using rat intestinal and renal subcellular fractions as activation systems was approximately 1 and 5 revertants per nmol, respectively. This was 1,000 times less than the activity with a subcellular fraction from rat liver. The mutagenic activity of both compounds was considerably increased using intestinal, renal and hepatic preparations isolated from PCB (Aroclor 1254)-pretreated rats, compared to preparations from control animals. In addition, both compounds displayed a moderate direct-acting mutagenic activity at concentrations above 10^-5 M. Isolated cells from small intestine, kidney and liver incubated in nucleopore chambers were able to convert both compounds into products which mutated bacteria outside the chambers. The concentrations of chemicals required to yield responses of a similar magnitude were approximately 3 orders of magnitude higher in the intestinal and renal systems compared to the hepatic system. The formation of metabolites mutagenic for Salmonella typhimurium by hepatic subcellular and cellular systems was shown to be superior to the respective intestinal and renal systems.Abbreviations AHH arylhydrocarbon hydroxylase - IQ 2-amino-3-methylimidazo[4,5-f]-quinoline - MC 3-methylcholanthrene - MeIQ 2-amino-3,4-dimethylimidazo[4,5-f]-quinoline - PCB polychlorinated biphenyls (Aroclor 1254) - S9 the 9,000 g supernatant tissue fraction - TCDD 2,3,7,8-tetrachlorodibenzo-p-dioxin     

Effects of changes in turbidity and phosphate influx on the ecosystem of the Ems estuary as obtained by a computer simulation model

In the Ems estuary the gradients in the concentration of nutrients and in turbidity (the factors that mainly determine the amount of carbon assimilated by phytoplankton) are steep. The effects of changing the turbidity in the estuary and the amount of phosphate discharged by the rivers Ems and Westerwoldsche Aa were analysed, using the simulation model developed by BOEDE (Biological Research Ems-Dollard estuary). The results of several sensitivity runs were compared with the standard run.A 50% reduction of turbidity led to a strong increase in phytoplanktonbiomass, especially in the inner parts of the estuary where turbidity is high. On average, the effects are two to three times larger for the inner part than for the outer part of the estuary. When the turbidity doubles the opposite occurs resulting in a significant decrease of phytoplankton biomass in the upper reaches. In the lower reaches of the estuary a 50% reduction in the river discharge of phosphate is largely compensated for by changes in phosphate transport from the North Sea. This results in a nearly unchanged primary production in the lower reaches as compared with the standard run.In the upper reaches a 50% reduction of phosphate loads results in a strongly reduced primary production.In general, the zooplankton groups (copepods and microzooplankton) are influenced less than the phytoplankton. Benthic fauna is hardly influenced, except for filter feeders; which are strongly affected by the total density of the particles, a parameter which also is directly related to turbidity.     

Effects of toxin II from the scorpion Androctonus australis Hector on sodium current in neuroblastoma cells and their modulation by oleic acid

The effects of toxin II (AaH II) isolated from the scorpion Androtonus australis Hector on sodium current in neuroblastoma X glioma NG 108-15 hybrid cells were analysed under patch clamp conditions in the whole cell configuration. AaH II (70 nM)_induced a maintained sodium current, as well as increasing both fast and slow inactivation time constants and the amplitude of the peak current. This latter effect occurred via a shift of the activation-voltage curve towards negative voltage values by about 9 mV. Oleic acid (5 M), which had no effect on I_Na under control conditions, decreased the AaH II-induced maintained current. It also reversed, or prevented the increase of the peak current induced by AaH II. However, it neither prevented nor modified the AaH II-induced increase in inactivation time constants. The binding of the toxin to its specific site and the number of binding sites for AaH II were not significantly modified by oleic acid. The oleic acid-induced effects could not be related to the activation of protein kinase C since PMA, a potent activator of this enzyme, did not produce oleic acid-like effects. From these results, it is concluded that AaH II has several independent effects on sodium channels, some of which could be modulated by the lipid environment of sodium channels in the membrane.     

Effects of ascorbic acid and ascorbic acid 2-phosphate, a long-acting vitamin C derivative, on the proliferation and differentiation of human osteoblast-like cells

In order to investigate the effect of ascorbic acid (AsA) and ascorbic acid 2-phosphate (Asc 2-P), a long-acting vitamin C derivative, on the growth and differentiation of human osteoblast-like cells, we supplemented the culture medium of MG-63 cells with various concentrations (0.25 to 1 mM) of these factors. Asc 2-P significantly stimulated nascent cell growth at all concentrations in the presence of fetal bovine serum (FBS). On the other hand, AsA showed a growth repressive effect depending on its concentration, and that of FBS. Asc 2-P also increased expression of osteoblast differentiation markers, such as collagen synthesis and alkaline phosphatase (ALP) activity. These stimulative activities of Asc 2-P were attenuated by inhibitors of collagen synthesis, indicating that these effects were dependent on collagen synthesis. Electron micrographs of the cells showed the formation of a three-dimensional tissue-like structure endowed with a mature extracellular matrix in the presence of Asc 2-P.     

miR-145靶向调控DAB2对前列腺癌PC3细胞迁移和侵袭能力的影响

已有研究表明, miR-145在多种肿瘤中低表达, 并与细胞增殖和转移相关。文章通过生物信息学分析并结合体外实验鉴定, 发现DAB2(Disabled homolog 2)为miR-145在肿瘤转移过程中累及的新靶点。DAB2一直被认为是一个重要的抑癌基因, 在多种肿瘤标本中表达低下。然而, 研究发现, 在具高侵袭能力的前列腺癌细胞株PC3中DAB2基因却呈较高水平表达。另外, 外源表达miR-145能显著下调 DAB2表达水平, 并抑制PC3细胞的迁移和侵袭能力, 且这种miR-145诱导的PC3细胞功能缺陷能被DAB2过表达修复。上述结果表明, miR-145能通过靶向调控DAB2而影响高侵袭前列腺癌细胞的迁移和侵袭能力。     

Effects of the protein kinase inhibitors, staurosporine and K-252a, on PGI2 production by rat liver cells (the C-9 cell line)

Staurosporine and K-252a, known inhibitors of several protein kinases, stimulated PGI2 production (measured as 6-keto-PGF1 alpha) in rat liver cells (the C-9 cell line). Preincubation of the rat liver cells with staurosporine or K-252a enhanced the PGI2 production stimulated by 12-O-tetradecanoylphorbol-13-acetate (TPA), platelet activating factor (PAF) and the Ca2(+)-ionophore A-23187, but not the PGI2 synthesis stimulated by exogenous arachidonic acid. These results suggest that phosphorylation of some proteins or certain amino acids on a protein can regulate arachidonic acid metabolism probably in the pathway leading to deesterification of phospholipids.     

潮汐作用下干湿交替对黄河三角洲盐沼湿地净生态系统CO2交换的影响

潮汐作用作为盐沼湿地独特的水文特征能在短时间内强烈影响盐沼湿地的碳平衡.利用涡度相关和微气象监测技术,对黄河三角洲盐沼湿地净生态系统CO₂交换(NEE)和环境因子进行监测,并同步监测潮汐变化,探究潮汐过程及潮汐作用下干湿交替对NEE的影响.结果表明: 潮汐过程促进了白天生态系统CO₂的吸收但未对夜晚CO₂的释放产生显著影响,潮汐淹水成为影响白天NEE的主要因子.干旱阶段和湿润阶段NEE的日平均动态均呈“U”型曲线,但干旱阶段NEE的变幅较小.干湿交替增强了白天生态系统CO₂的吸收,干旱阶段最大光合速率(A_max)、表观量子产量(α)和生态系统呼吸(R_eco)的均值均高于湿润阶段.此外,干湿交替减少了盐沼湿地夜晚NEE释放的同时增强了其温度敏感性.     

潮汐作用下干湿交替对黄河三角洲盐沼湿地净生态系统CO2交换的影响

潮汐作用作为盐沼湿地独特的水文特征能在短时间内强烈影响盐沼湿地的碳平衡.利用涡度相关和微气象监测技术,对黄河三角洲盐沼湿地净生态系统CO₂交换(NEE)和环境因子进行监测,并同步监测潮汐变化,探究潮汐过程及潮汐作用下干湿交替对NEE的影响.结果表明: 潮汐过程促进了白天生态系统CO₂的吸收但未对夜晚CO₂的释放产生显著影响,潮汐淹水成为影响白天NEE的主要因子.干旱阶段和湿润阶段NEE的日平均动态均呈“U”型曲线,但干旱阶段NEE的变幅较小.干湿交替增强了白天生态系统CO₂的吸收,干旱阶段最大光合速率(A_max)、表观量子产量(α)和生态系统呼吸(R_eco)的均值均高于湿润阶段.此外,干湿交替减少了盐沼湿地夜晚NEE释放的同时增强了其温度敏感性.