JAR human placental choriocarcinoma cells actively synthesize,take up and release polyamines

Uptake of polyamines has been investigated extensively in many cells, but not in placenta, where the polyamine– polyamine oxidase system is supposed to have an immunoregulatory function in pregnancy. Due to the importance of the transfer in this tissue, we have started this study. JAR human placental choriocarcinoma cells in monolayer at confluency were used as a model for measuring the key enzymes of polyamine synthesis and interconversion, rate of uptake and efflux, and the polyamine content. Polyamines were taken up by JAR cells and released by an independent mechanism. Ornithine decarboxylase and spermidine acetyltransferase activities and the rate of transport in and out of the cell were much higher than in other cells, such as L1210 cells. However the systems used for uptake and release appear in many respects to be similar to those observed in L1210 cells, but different from others. The uptake appears to be regulated by an inhibitory protein. Moreover, protein kinase C appears to be involved in the process. The efflux also is regulated as in L1210 cells, through control of H⁺ and Ca²⁺ concentration. In conclusion, this study shows that, in JAR cells, ornithine decarboxylase and spermidine acetyltransferase activities were much higher than in other cells, and so was the rate of transport in and out of the cells. As a result, a much higher polyamine content was observed.     

Response and tolerance of root border cells to aluminum toxicity in soybean seedlings

Root border cells (RBCs) and their secreted mucilage are suggested to participate in the resistance against toxic metal cations, including aluminum (Al), in the rhizosphere. However, the mechanisms by which the individual cell populations respond to Al and their role in Al resistance still remain unclear. In this research, the response and tolerance of RBCs to Al toxicity were investigated in the root tips of two soybean cultivars [Zhechun No. 2 (Al-tolerant cultivar) and Huachun No. 18 (Al-sensitive cultivar)]. Al inhibited root elongation and increased pectin methylesterase (PME) activity in the root tip. Removal of RBCs from the root tips resulted in a more severe inhibition of root elongation, especially in Huachun No. 18. Increasing Al levels and treatment time decreased the relative percent viability of RBCs in situ and in vitro in both soybean cultivars. Al application significantly increased mucilage layer thickness around the detached RBCs of both cultivars. Additionally, a significantly higher relative percent cell viability of attached and detached RBCs and thicker mucilage layers were observed in Zhechun No. 2. The higher viability of attached and detached RBCs, as well as the thickening of the mucilage layer in separated RBCs, suggest that RBCs play an important role in protecting root apices from Al toxicity.     

植物根边缘细胞的抗逆性研究进展

综述了近几年来国内外有关植物根边缘细胞抗逆性方面的研究,重点概述植物根边缘细胞对生物与非生物胁迫的响应及其相应的抗性机理。在生物胁迫下,边缘细胞能吸引和固定病原根结线虫,排斥或约束致病性细菌,可作为真菌感染的假目标,减少或避免各种病原菌对根尖的伤害。在非生物胁迫下,边缘细胞通过分泌粘液、诱导ROS产生刺激细胞死亡以抵抗铝毒,并通过其数量的改变来调节高温、高浓度CO2等多种生理反应。最后在当前植物根边缘细胞研究的基础上,提出了今后的研究方向。     

大豆（Glycine max L.）边缘细胞对铝毒的生理生态响应

以大豆浙春2号(铝耐性)和华春18号(铝敏感)为材料,研究了铝胁迫下附着于根尖边缘细胞(即原位边缘细胞)的存活率、根伸长抑制率和PME活性变化以及Al3 对离体后边缘细胞存活率、黏液层厚度的影响。结果表明:铝胁迫下附着于根尖的边缘细胞比离体边缘细胞有更高的活性,前者Al3 处理24h后,其成活率仍能达到74%以上,而后者Al3 处理12h,浙春2号和华春18号边缘细胞的活性在400μmol/L时分别只有44.58%和26.16%;前后两者细胞活性都有随着Al3 浓度升高和处理时间的延长,边缘细胞活性呈越来越低的变化趋势,而离体边缘细胞Al3 处理6h时,相对于敏感性品种而言,高浓度Al3 (≥200μmol/L)有利于铝耐性品种的边缘细胞存活。随Al3 浓度的提升,果胶甲基酯酶(PME)活性增加,根伸长受抑加剧,敏感品种PME活性及铝造成的根伸长抑制均高于耐性品种;同时,Al3 对黏液的产生有一定的影响,黏液层的厚度随Al3 浓度成正向变化趋势。不同Al3 浓度及处理时间下,耐性品种都比敏感品种边缘细胞有较高的活性、分泌较多的黏液。以上结果说明Al3 对边缘细胞具有一定的毒害效应,果胶甲基酯化程度、根伸长受抑及边缘细胞黏液的分泌是根冠对Al毒胁迫反应的结果。     

Transformed roots of Artemisia annua exhibit an unusual pattern of border cell release

Summary Border cells from Artemisia annua were examined from hairy roots grown in shake flasks, culture plates, a bubble column reactor, and a nutrient mist (aeroponic) reactor. When well-hydrated roots were subjected to shear, border cells were first released as an agglomerate and did not disperse for several hours. Staining with neutral red and fluorescein diacetate (FDA) showed that both agglomerates and dispersed cells were alive. It was determined that FDA is cleaved by pectin methylesterase (PME) and that PME may not be particularly active in the released agglomerates until the border cells disperse. Untransformed roots isolated from A. annua plants showed no border cell agglomerate formation and border cells readily dispersed. These results suggest that our hairy root clone is deficient in border cell release perhaps resulting from the transformation process.     

Effect of sodium fluoride on the root apex border cells in one-day-old wheat seedlings

The number of border (scaled off) cells (BCs) was determined in the root apex of 1-day-old wheat (Triticum aestivum L.) seedlings. Microscopic examination of cytological root preparations showed that in 24 h the number of BCs in the gel sheath of the root apex was 30–40 per root. When the gel sheath was preparatively removed, their number per root increased twice. It is assumed that the subpopulation of BCs directly associated with the root apex differs from the subpopulation of BCs freely accommodated in the gel sheath. The number of BCs was the same in the roots with low and high natural growth rates. NaF (1–20 mM) suppressed growth of wheat seedling roots; the viscosity of the gel sheath increased (by 3–5 times), and the number of BC rose with the most pronounced increment in the size of the BC subpopulation directly associated with the root apex. Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 4, pp. 530–538.     

Passage of sugars across the plasmalemma of carrot callus cells

In pieces of carrot callus, the regions exterior to the plasmalemma were washed out in running tap-water within 15 min. The effect of temperature on sugar efflux and influx, suggests that the passage of sugars across the plasmalemma occurs mainly, if not entirely, by passive diffusion. The results of other workers are considered, and we conclude that the reported apparent active uptake across the plasmalemma may in reality have been due to active uptake across the tonoplast. Estimates of the apparent free space were unaffected by the sugar used, or the temperature, and we suggest that the apparent free space includes most, if not all of the metabolic compartment.     

Uptake and release of tetracycline by cultured carrot cells

Summary The nature of tetracycline uptake by carrot cell suspension cultures is described. Tetracycline enters the cells by diffusion and the intracellular level of the antibiotic increases with the amount added. Exposure of carrot cells to high levels of tetracycline for a limited time (24 hr) followed by the removal of the drug and the resuspension of the cells in drug-free medium does not affect cell growth and has no inhibitory effect on protein synthesis (¹⁴C-leucine incorporation).     

Interrelationship between the growth rate of wheat roots,their excretory activity,and the number of border cells

Excretion of proteins and carbohydrates, the number of root border cells (BC), and the effect of different concentrations of NaF (1, 5, 10, and 20 mM) on the growth rate of the roots were investigated in the seedlings of wheat (Triticum aestivum L.) with low (LGR) and high growth rate (HGR). At the early stage of growth (1 day), the rate of protein excretion to the medium was much greater in the LGR roots; as to carbohydrates, the difference between the LGR and HGR roots was less pronounced. When added to the germination medium, NaF suppressed root growth and induced excretion of high-molecular-weight proteins into the medium; this phenomenon was more pronounced in the HGR-roots. The number of BC did not depend on the rate of protein excretion into the medium. The population of BC was the same in the HGR and LGR roots. When 1 mM NaF was added to the medium, the number of BC in the rhizosphere equally increased in both HGR and LGR roots. The elevation of NaF concentration did not affect the number of BC.     

荞麦、高粱根系分泌物对玉米根边缘细胞和根生长的影响

植物的根系分泌物是植物根系与周围环境之间的化学媒介,通过传递特定的信息,调节根际微环境,影响周围植物的生长。玉米(Zea mays L.)和荞麦(Fagopyrum esculentum Moench)是农作物间套作体系中典型的不能搭配的组合,其障碍因素尚不清楚。以玉米为受体植物,采用根悬空培养的方法,研究了荞麦、高粱(Sorghum bicolor(L.) Moench)根系分泌物对玉米根边缘细胞和根生长的影响。结果发现,玉米根边缘细胞离体培养条件下,用荞麦根系分泌物中的小分子物质处理4、8 h显著诱导边缘细胞凋亡、死亡,细胞活率分别比对照降低了71.6%和72.3%;荞麦根系分泌物中的小分子物质对玉米根产生氧化胁迫,诱导根SOD、POD和CAT活性分别比对照高22.6%、33.9%和107.2%,根中超氧阴离子(O₂^-·)和脯氨酸含量分别比对照高33.9%和49.8%;荞麦根系分泌物中小分子物质的胁迫使根细胞膜透性增大,与对照相比升高80.0%,丙二醛(MDA)含量比对照升高31.5%;荞麦根系分泌物中小分子物质诱导根内源激素(IAA)含...     

Temperature dependence of solute transport and enzyme activities in hog renal brush border membrane vesicles

The temperature dependence of sodium-dependent and sodium-independent d-glucose and phosphate uptake by renal brush border membrane vesicles has been studied under tracer exchange conditions. For sodium-dependent d-glucose and phosphate uptake, discontinuities in the Arrhenius plot were observed. The apparent activation energy for both processes increased at least 4-fold with decreasing temperature. The most striking change in the slope of the Arrhenius plot occurred between 12 and 15°C. The sodium-independent uptake of d-glucose and phosphate showed a linear Arrhenius plot over the temperature range tested (35–5°C). The behavior of the transport processes was compared to the temperature dependence of typical brush border membrane enzymes. Alkaline phosphatase as intrinsic membrane protein showed a nonlinear Arrhenius plot with a transition temperature at 12.4°C. Aminopeptidase M, an extrinsic membrane protein exhibited a linear Arrhenius plot. These data indicate that the sodium-glucose and sodium-phosphate cotransport systems are intrinsic brush border membrane proteins, and that a change in membrane organization alters the activity of a variety of intrinsic membrane proteins simultaneously.     

Aluminum-induced cell death of barley-root border cells is correlated with peroxidase- and oxalate oxidase-mediated hydrogen peroxide production

The function of root border cells (RBC) during aluminum (Al) stress and the involvement of oxalate oxidase, peroxidase and H₂O₂ generation in Al toxicity were studied in barley roots. Our results suggest that RBC effectively protect the barley root tip from Al relative to the situation in roots cultivated in hydroponics where RBC are not sustained in the area surrounding the root tip. The removal of RBC from Al-treated roots increased root growth inhibition, Al and Evans blue uptake, inhibition of RBC production, the level of dead RBC, peroxidase and oxalate oxidase activity and the production of H₂O₂. Our results suggest that even though RBC actively produce active oxygen species during Al stress, their role in the protection of root tips against Al toxicity is to chelate Al in their dead cell body.     

Root organogenesis from single cells released from the root cap of Medicago sp.

Root border cells were isolated from alfalfa seedlings, and incubated in culture medium with growth regulators. Alfalfa seedlings yielded 1500±100 cells per root, and initial viability of the cells was 95±5%. Multiple cell divisions occurred in the border cells within two weeks. Cell clusters transferred to solidified medium containing growth regulators developed into rapidly growing, friable callus. When transferred to growth regulator-free medium, some of the calluses generated normal roots.Abbreviations BRD cells border cells - SHDN Schenk & Hildebrandt salts medium with growth regulators - SHO Schenk & Hildebrandt salts medium without growth regulators - NAA I-naphthaleneacetic acid     

Modulation of insulin release by adenosine A1 receptor agonists and antagonists in INS-1 cells: the possible contribution of 86Rb+ efflux and 45Ca2+ uptake

Due to the lack of specific agonists and antagonists the role of adenosine receptor subtypes with respect to their effect on the insulin secretory system is not well investigated. The A1 receptor may be linked to different 2nd messenger systems, i.e. cAMP, K+- and 45Ca2+ channel activity. Partial A1 receptor agonists are going to be developed in order to improve diabetes (increase in insulin sensitivity, lowering of FFA and triglycerides). In this study newly synthesized selective A1 receptor agonists and antagonists were investigated thereby integrating three parameters, insulin release (RIA), 45Ca2+ uptake and 86Rb+ efflux (surrogate for K+ efflux) of INS-1 cells, an insulin secretory cell line. The presence of A1-receptors was demonstrated by Western blotting. The receptor nonselective adenosine analogue NECA (5-N-ethylcarboxyamidoadenosine) at high concentration (10 microM) had no effect on insulin release and 45Ca2+ uptake which could be interpreted as the sum of effects mediated by mutual antagonistic adenosine receptor subtypes. However, an inhibitory effect mediated by A1 receptor agonism was detected at 10 nM NECA and could be confirmed by adding the A1 receptor antagonist PSB-36 (1-butyl-8-(3-noradamantyl)-3-(3-hydroxy-propyl)xanthine). NECA inhibited 86Rb+ efflux which, however, did not fit with the simultaneous inhibition of insulin secretion. The selective A1 receptor agonist CHA (N6-cyclohexyladenosine) inhibited insulin release; the simultaneously increased Ca2+ uptake (nifedipine dependent) and inhibition of 86Rb+ efflux did not fit the insulin release data. The CHA effect (even the maximum effect at 50 microM) can be increased by 10 microM NECA indicating that CHA and NECA have nonspecific and physiologically non-relevant effects on 86Rb+ efflux in addition to their A1-receptor interaction. Since PSB-36 did not influence the NECA-induced inhibition of 86Rb+ efflux, the NECA effect is not mediated by potassium channel-linked A1 receptors. The nonselective adenosine receptor antagonist caffeine increased insulin release which was reversed by CHA as expected when hypothesizing that both act via A1 receptors in this case. In conclusion, stimulation of A1 receptors by receptor selective and nonselective compounds reduced insulin release which is not coupled to opening of potassium channels (86Rb+ efflux experiments) or inhibition of calcium channels (45Ca2+ uptake experiments). It may be expected that of all pleiotropic 2nd messengers, the cAMP system (not tested here) is predominant for A1 receptor effects and the channel systems (K+ and Ca2+) are of minor importance and do not contribute to insulin release though being coupled to the receptor in other tissues.     

不同发育期反枝苋对黄瓜根缘细胞的化感作用

以黄瓜为实验材料,采用悬空气培养法研究了不同发育期反枝苋水浸提液对根缘细胞的化感效应.结果表明:在反枝苋水浸提液作用下,黄瓜根果胶甲基酯酶(PME)活性升高,但随着水浸提液浓度的升高,这种促进效应逐渐降低;而黄瓜根缘细胞存活率随着水浸提液浓度的升高而下降.相同发育期反枝苋不同部位的化感作用差异不显著,不同发育时期反枝苋化感作用差异较为明显,其化感作用以幼苗期最强,现蕾期次之,成熟期最弱.推测化感作用是反枝苋发育初期取得竞争优势,迅速占领生态位的原因之一.     

Hesperetin impairs glucose uptake and inhibits proliferation of breast cancer cells

The flavanone hesperetin is known to decrease basal glucose uptake, although the inhibitory mechanism is largely unknown. Here, we used MDA‐MB‐231 breast cancer cells to investigate the molecular pathways affected by hesperetin. The results indicate that the suppression of glucose uptake is caused by the down‐regulation of glucose transporter 1 (GLUT1). Hesperetin was also found to inhibit insulin‐induced glucose uptake through impaired cell membrane translocation of glucose transporter 4 (GLUT4). In addition, the phosphorylation of the insulin receptor‐beta subunit (IR‐beta) and Akt was suppressed. Hesperetin also decreased cellular proliferation, which is likely due to the inhibition of glucose uptake. Cancer cells are highly dependent on glucose and hesperetin may, therefore, have potential application as an anticancer agent. Copyright © 2012 John Wiley & Sons, Ltd.     

Localization of brush border cytoskeletal proteins in gastric oxynticopeptic cells from the bullfrog Rana catesbeiana

The contribution of brush border cytoskeletal proteins (actin, villin, fimbrin, and brush border myosin-1) to organization of the cytoskeletal network underlying apical plications of oxynticopeptic cells was examined by immunohistochemical techniques in frozen sections of gastric mucosa from the bullfrog, Rana catesbeiana. Apical localization of F-actin with phalloidin in oxynticopeptic cells inhibited with cimetidine revealed small, punctate domains within the apical cytoplasm that were consistent with the presence of short microvilli revealed by electron microscopy. Localization of F-actin in cells stimulated with forskolin was limited to a wide continuous band of cytoplasm corresponding to the location of numerous long surface folds. Inhibition of protein synthesis with cycloheximide did not prevent acid secretion or formation of actin filaments within surface folds in stimulated oxynticopeptic cells, suggesting that the formation of filaments does not require actin synthesis. Staining of gastric mucosae with fluorescent DNase-1 demonstrated that oxynticopeptic cells possess an unusually large pool of non-filamentous actin. Taken together, these results suggest that actin-filament formation in stimulated cells occurs by polymerization of an existing pool of non-filamentous actin. Localization of antibodies specific for villin and fimbrin revealed that these proteins were present within intestinal absorptive cells and gastric surface and neck cells but were not present within inhibited or stimulated oxynticopeptic cells. Brush border myosin-1, present in intestinal absorptive cells, was not present in gastric epithelium. Thus, we propose that actin-containing projections in oxynticopeptic cells are not organized like intestinal microvilli and that filament formation occurs after stimulation by modulating intracellular pools of filamentous and non-filamentous actin.     

四种常见杂草根系及根边缘细胞对铝胁迫的响应

以2种禾本科杂草(升马唐、稗草)和2种菊科杂草(旱莲草、野茼蒿)为实验材料,通过砂培法研究不同科属杂草根部对铝胁迫的响应.结果表明:4种杂草根边缘细胞活性均随着铝胁迫浓度和时间呈显著下降的趋势,但禾本科杂草根系边缘细胞的活性高于菊科杂草,且活性的降低幅度较小;4种杂草根相对伸长率均随铝浓度和处理时间的增加呈递减趋势,但铝对旱莲草和野茼蒿根生长的抑制程度要明显高于升马唐和稗草;根系的铝含量、游离脯氨酸含量、MDA 含量和质膜透性均随铝处理浓度和处理时间的增加而增大,且在高铝浓度(1000 mg · L~(-1))时达到最大值,但升马唐和稗草根系的铝含量、游离脯氨酸含量、MDA含量和质膜透性均显著低于旱莲草和野茼蒿,且随着铝浓度的增加,禾本科杂草根系的游离脯氨酸含量及MDA含量的变化没有达到显著水平(P＞0.05).由此说明,铝毒对杂草造成的伤害随着浓度增加和时间延长而加重;升马唐和稗草的根系通过较高的根边缘细胞活性和根相对伸长率及较低的铝含量、游离脯氨酸含量、MDA含量和质膜透性来增加其对铝的耐性;2种禾本科杂草(升马唐、稗草)的耐铝性高于2种菊科杂草(旱莲草、野茼蒿).     

High doses of amphetamine augment, rather than disrupt, exocytotic dopamine release in the dorsal and ventral striatum of the anesthetized rat

High doses of amphetamine (AMPH) are thought to disrupt normal patterns of action potential-dependent dopaminergic neurotransmission by depleting vesicular stores of dopamine (DA) and inducing robust non-exocytotic DA release or efflux via dopamine transporter (DAT) reversal. However, these cardinal AMPH actions have been difficult to establish definitively in vivo. Here, we use fast-scan cyclic voltammetry (FSCV) in the urethane-anesthetized rat to evaluate the effects of 10 and 20 mg/kg AMPH on vesicular DA release and DAT function in dorsal and ventral striata. An equivalent high dose of cocaine (40 mg/kg) was also examined for comparison to psychostimulants acting preferentially by DAT inhibition. Parameters describing exocytotic DA release and neuronal DA uptake were determined from dynamic DA signals evoked by mild electrical stimulation previously established to be reinforcing. High-sensitivity FSCV with nanomolar detection was used to monitor changes in the background voltammetric signal as an index of DA efflux. Both doses of AMPH and cocaine markedly elevated evoked DA levels over the entire 2-h time course in the dorsal and ventral striatum. These increases were mediated by augmented vesicular DA release and diminished DA uptake typically acting concurrently. AMPH, but not cocaine, induced a slow, DA-like rise in some baseline recordings. However, this effect was highly variable in amplitude and duration, modest, and generally not present at all. These data thus describe a mechanistically similar activation of action potential-dependent dopaminergic neurotransmission by AMPH and cocaine in vivo. Moreover, DA efflux appears to be a unique, but secondary, AMPH action.     

高等植物根边缘细胞的发育调控及其生物学功能

植物根边缘细胞是从根冠表皮游离出来并聚集在根尖周围的一群特殊细胞,以前曾称为根冠脱落细胞.最近的证据表明,绝大多数物种边缘细胞具有生物学活性,其发育是受内外信号调控.边缘细胞一旦从根表皮游离后,其代谢活性大大上升、基因表达明显不同于根冠细胞.最近,与边缘细胞发育早期和晚期相关的两个基因PsUGT1和RCPME1分别被克隆和鉴定.边缘细胞能特异性地合成、分泌一系列的化学物质,包括花色素苷、抗生素、特异性酶类及其他化学物质能抑制或促进根际周围的细菌、真菌、病毒、线虫等的生长以及中和根际周围一些有毒化学物质如铝毒.因此,边缘细胞在植物生长发育过程中起着多种生物学功能.