Effect of fertilizers on Cd accumulation and subcellular distribution of two cosmos species (Cosmos sulphureus and Cosmos bipinnata)

Addition of fertilizer amendments is regarded as an ideal approach to enhancing phytoextraction. However, there is little information available on the influence of common fertilizers on Cd accumulation of two newly reported Cd accumulators, Cosmos sulphureus and Cosmos bipinnata (C. sulphureus and C. bipinnata). The effects of N (CO(NH₂)₂), NP (CO(NH₂)₂ + Ca(H₂PO₄)₂), and NPK (CO(NH₂)₂ + Ca(H₂PO₄)₂ + KCl) fertilizers on Cd accumulation and subcellular distribution of C. sulphureus and C. bipinnata were studied in a 70-d pot experiment. The results showed that Cd uptake of C. sulphureus and C. bipinnata with NPK fertilizer was significantly higher than control, N, and NP fertilizers, especially 3.8- and 4.7-fold higher than control (p < 0.05). Compared with C. bipinnata, C. sulphureus achieved higher biomass and Cd uptake in aboveground parts under fertilizer treatments, especially NPK fertilizer. The Cd subcellular distribution revealed that segregation of Cd to Cd-rich granules (MRG) might play an important role in Cd detoxification in both species. C. sulphureus is more likely than C. bipinnata to separate the Cd in MRG and reduce the partition in the heat-denatured protein fraction, especially with NPK fertilizer. Therefore, C. sulphureus combined with NPK fertilizers could be an effective method to remediate Cd-polluted farmland soils in China.     

Phosphorylation states greatly regulate the activity and gating properties of Cav3.1 T-type Ca2+ channels

Ca_v3.1 T-type Ca²⁺ channels play pivotal roles in neuronal low-threshold spikes, visceral pain, and pacemaker activity. Phosphorylation has been reported to potently regulate the activity and gating properties of Ca_v3.1 channels. However, systematic identification of phosphorylation sites (phosphosites) in Ca_v3.1 channel has been poorly investigated. In this work, we analyzed rat Ca_v3.1 protein expressed in HEK-293 cells by mass spectrometry, identified 30 phosphosites located at the cytoplasmic regions, and illustrated them as a Ca_v3.1 phosphorylation map which includes the reported mouse Ca_v3.1 phosphosites. Site-directed mutagenesis of the phosphosites to Ala residues and functional analysis of the phospho-silent Ca_v3.1 mutants expressed in Xenopus oocytes showed that the phospho-silent mutation of the N-terminal Ser18 reduced its current amplitude with accelerated current kinetics and negatively shifted channel availability. Remarkably, the phospho-silent mutations of the C-terminal Ser residues (Ser1924, Ser2001, Ser2163, Ser2166, or Ser2189) greatly reduced their current amplitude without altering the voltage-dependent gating properties. In contrast, the phosphomimetic Asp mutations of Ca_v3.1 on the N- and C-terminal Ser residues reversed the effects of the phospho-silent mutations. Collectively, these findings demonstrate that the multiple phosphosites of Ca_v3.1 at the N- and C-terminal regions play crucial roles in the regulation of the channel activity and voltage-dependent gating properties.     

Modeling interactions between voltage-gated Ca2+ channels and KCa1.1 channels

High voltage-activated (HVA) Cav channels form complexes with KCa1.1 channels, allowing reliable activation of KCa1.1 current through a nanodomain interaction. We recently found that low voltage-activated Cav3 calcium channels also create KCa1.1-Cav3 complexes. While coimmunoprecipitation studies again supported a nanodomain interaction, the sensitivity to calcium chelating agents was instead consistent with a microdomain interaction. A computational model of the KCa1.1-Cav3 complex suggested that multiple Cav3 channels were necessary to activate KCa1.1 channels, potentially causing the KCa1.1-Cav3 complex to be more susceptible to calcium chelators. Here, we expanded the model and compared it to a KCa1.1-Cav2.2 model to examine the role of Cav channel conductance and kinetics on KCa1.1 activation. As found for direct recordings, the voltage-dependent and kinetic properties of Cav3 channels were reflected in the activation of KCa1.1 current, including transient activation from lower voltages than other KCa1.1-Cav complexes. Substantial activation of KCa1.1 channels required the concerted activity of several Cav3.2 channels. Combined with the effect of EGTA, these results suggest that the Ca²⁺ domains of several KCa1.1-Cav3 complexes need to cooperate to generate sufficient [Ca²⁺]_i, despite the physical association between KCa1.1 and Cav3 channels. By comparison, Cav2.2 channels were twice as effective at activating KCa1.1 channels and a single KCa1.1-Cav2.2 complex would be self-sufficient. However, even though Cav3 channels generate small, transient currents, the regulation of KCa1.1 activity by Cav3 channels is possible if multiple complexes cooperate through microdomain interactions.     

芦苇叶水提物的化感活性分析及潜在化感成分的筛选

采用不同溶剂对芦苇﹝Phragmites australis ( Cav.) Trin. ex Steud.﹞叶片水提物进行萃取,并以小麦( Triticum aestivum Linn.)和萝卜( Raphanus sativus Linn.)种子为实验材料对不同萃取物的化感效应进行检测;采用薄层层析和柱层析对抑制作用最强的正丁醇萃取物进行进一步分离,并采用GC-MS法对生物活性较高的组分进行组成成分分析;在此基础上,选择相对含量高并具有代表性的潜在化感成分进行生物活性检测,以期筛选出芦苇叶中的潜在化感成分。结果显示：随质量浓度(20、100和500 mg·L-1)提高,芦苇叶水提物的石油醚、二氯甲烷、乙酸乙酯、正丁醇和水萃取物对小麦和萝卜种子萌发的抑制作用均逐渐增强,其中正丁醇萃取物的抑制作用最强。在正丁醇萃取物的11个组分中,Fr.5、Fr.6、Fr.7、Fr.9和Fr.10组分均能显著抑制萝卜或小麦幼苗的生长,经质量浓度500 mg·L-1各组分处理液处理后萝卜或小麦幼苗的株高、根长及单株鲜质量均显著低于对照(P<0.05)。采用GC-MS法从Fr.5、Fr.6、Fr.7、Fr.9和Fr.10组分中分别鉴定出11、15、15、12和22种成分,分别占各组分总相对含量的83.02%、91.31%、87.36%、97.92%和94.34%,主要成分包括糖类、醇类、有机酸类、酮类、酰胺类和酯类。对14种潜在化感成分生物活性的检测结果显示这些成分对小麦幼苗生长有明显的抑制作用,其中,经质量浓度20 mg·L-1油酸酰胺、棕榈酸甲酯、亚油酸、2-苯乙胺、2-甲基烯丙醇和4-羟基-3-甲氧基苦杏仁酸处理后,小麦幼苗的株高、根长及单株鲜质量显著低于对照。综合分析结果显示：芦苇叶水提物具有较强的化感活性,其潜在的化感成分为油酸酰胺、棕榈酸甲酯、亚油酸、2-苯乙胺、2-甲基烯丙醇和4-羟基-3-甲氧基苦杏仁酸。     

Substrate availability regulates the suppressive effects of Canada goldenrod invasion on soil respiration

基质有效性调节加拿大一枝黄花入侵对土壤呼吸的抑制作用 外来植物入侵不仅会降低河边近岸湿地生态系统植被多样性,而且会改变湿地生态系统的地下碳过程。外来入侵植物加拿大一枝黄花(Solidago canadensis L.)已广泛入侵我国东南部地区,但加拿大一枝黄花入侵对入侵地生态系统地下土壤碳循环过程的影响却知之甚少。本研究通过野外原位观测实验和温室模拟入侵实验,探究外来植物加拿大一枝黄花入侵对入侵地土壤呼吸的影响规律及其驱动因素。野 外原位观测实验开展于2018年7月21日至12月15日,期间每周测定样地土壤呼吸。温室模拟入侵实验开展于2019年7月15日至12月15日,期间每月1日与15日上午测定土壤呼吸、自养呼吸和异养呼吸。土壤呼吸、自养呼吸和异养呼吸通过静态箱结合深埋根系隔离法测定。野外原位观测实验和温室模拟入侵实验结果均显示,加拿大一枝黄花的入侵降低了土壤二氧化碳的排放通量。加拿大一枝黄花入侵对土壤呼吸的抑制作用可能归因于其入侵引起的土壤可利用底物质量与数量的变化,表明外来入侵植物加拿大一枝黄花可通过改变植物释放基质以及与本地植物和/或土壤微生物争夺土壤有效基质而影响土壤碳循环。这些研究结果对于评估外来入侵植物对入侵地地下碳动态的影响以及对全球变暖的贡献具有重要意义。     

Impaired long-term memory and long-term potentiation in N-type Ca2+ channel-deficient mice

Voltage-dependent N-type Ca(2+) channels, along with the P/Q-type, have a crucial role in controlling the release of neurotransmitters or neuromodulators at presynaptic terminals. However, their role in hippocampus-dependent learning and memory has never been examined. Here, we investigated hippocampus-dependent learning and memory and synaptic plasticity at hippocampal CA3-CA1 synapses in mice deficient for the alpha(1B) subunit of N-type Ca(2+) channels. The mutant mice exhibited impaired learning and memory in the Morris water maze and the social transmission of food preference tasks. In particular, long-term memory was impaired in the mutant mice. Interestingly, among activity-dependent long-lasting synaptic changes, theta burst- or 200-Hz-stimulation-induced long-term potentiation (LTP) was decreased in the mutant, compared with the wild-type mice. This type of LTP is known to require brain-derived neurotrophic factor (BDNF). It was found that both BDNF-induced potentiation of field excitatory postsynaptic potentials and facilitation of the frequency of miniature excitatory postsynaptic currents (mEPSCs) were reduced in the mutant. Taken together, these results demonstrate that N-type Ca(2+) channels are required for hippocampus-dependent learning and memory, and certain forms of LTP.     

Chronic social defeat stress-induced enhancement of T-type calcium channels increases burst-firing neurons in the ventral subiculum

The ventral subiculum (vSub), a representative output structure of the hippocampus, serves as a main limbic region in mediating the brain's response to stress. There are three subtypes of subicular pyramidal neurons based on their firing patterns: regular-spiking (RS), weak-bursting (WB) and strong-bursting (SB) neurons, located differently along proximal–distal axis. Here, we found that chronic social defeat stress (CSDS) in mice increased the population of SB neurons but decreased RS neurons in the proximal vSub. Specific blockers of T-type calcium channels inhibited the burst firings with a concomitant reduction of afterdepolarization, suggesting that T-type calcium channels underlie the burst-spiking activity. Consistently, CSDS increased both T-type calcium currents and expression of Cav3.1 proteins, a subtype of T-type calcium channels, in the proximal vSub. Therefore, we conclude that CSDS-induced enhancement of Cav3.1 expression increased bursting neuronal population in the vSub, which may contribute to stress-related behaviors.     

The expression of Cav3.1 on T-type calcium channels of rats with subarachnoid hemorrhage

To study delayed cerebral vasospasm (DCVS) induced by subarachnoid hemorrhage (SAH), 60 healthy Sprague Dawley (SD) rats were randomly divided into 5 groups (12 rats in each group), namely sham operation group, blood injection model group, nimodipine group, flunarizine hydrochloride group, and normal group. Then, the physiological parameters were detected, and after the rats were killed under anesthesia, the degree of nerve injury, vasospasm as well as the therapeutic effect of drugs were evaluated by Western Blot (WB). Neurological impairment (NI), endothelial contraction and spasm were obvious in rats following blood injection. The expression of Cav3.1 on T-type calcium channels was significantly higher in the blood injection model group than in the sham operation group along with the normal group. Moreover, Cav3.1 mRNA was expressed in all groups. The Cav3.1 expression in blood injection model group and two drug groups were significantly higher than that in sham operation group and lower than that in blood injection model group. Vasospasm was improved in two drug groups, which indicated that calcium channel antagonists nimodipine and flunarizine hydrochloride had a certain therapeutic effect on DCVS in rats. The decrease in body weight and food intake of the two groups of rats treated with drugs decreased, and the delayed vasospasm was improved, but the expression of Cav3.1 was not changed significantly, indicating nimodipine and flunarizine hydrochloride had a therapeutic effect on delayed vasospasm in rats, but Cav3.1 expression on calcium channels was not affected.     

蛇床子素对L-和N-型钙通道的影响

目的：比较蛇床子素对不同钙通道亚型的作用差异。方法：首先在tsA201细胞上瞬时转染Cavl．2,Cav1.3,Cav2．2e[37a],和Cav2．2e[37b]通道,然后采用全细胞膜片钳技术,记录tsA201细胞上的钙电流,并观察蛇床子素对各种钙通道亚型的影响。结果：蛇床子素可以浓度依赖性抑制Cav1．2和Cav1.3电流,抑制的半有效浓度分别为162．1μmol·L-1和56．2μmol·L-1．此外,蛇床子素对Cav2．2通道也有一定的抑制作用,在300μmol·L-1。的浓度下,抑制38％的Cav2．2e[37a]电流和61％的Cav2．2e[37b]电流,蛇床子素对钙电流的抑制是快速可逆的。蛇床子素在各个测试电位水平均能抑制上述四种钙通道电流,但不改变电流的激活阂值和最大峰值电流的激活电压。结论：蛇床子素以浓度依赖的方式抑制多种钙通道亚型并表现出不同的亲和力。