Picomolar nitric oxide signals from central neurons recorded using ultrasensitive detector cells

Nitric oxide (NO) is a widespread signaling molecule with potentially multifarious actions of relevance to health and disease. A fundamental determinant of how it acts is its concentration, but there remains a lack of coherent information on the patterns of NO release from its sources, such as neurons or endothelial cells, in either normal or pathological conditions. We have used detector cells having the highest recorded NO sensitivity to monitor NO release from brain tissue quantitatively and in real time. Stimulation of NMDA receptors, which are coupled to activation of neuronal NO synthase, routinely generated NO signals from neurons in cerebellar slices. The average computed peak NO concentrations varied across the anatomical layers of the cerebellum, from 12 to 130 pm. The mean value found in the hippocampus was 200 pm. Much variation in the amplitudes recorded by individual detector cells was observed, this being attributable to their location at variable distances from the NO sources. From fits to the data, the NO concentrations at the source surfaces were 120 pm to 1.4 nm, and the underlying rates of NO generation were 36-350 nm/s, depending on area. Our measurements are 4-5 orders of magnitude lower than reported by some electrode recordings in cerebellum or hippocampus. In return, they establish coherence between the NO concentrations able to elicit physiological responses in target cells through guanylyl cyclase-linked NO receptors, the concentrations that neuronal NO synthase is predicted to generate locally, and the concentrations that neurons actually produce.     

黄土高原北部紫花苜蓿草地退化过程与植物多样性研究

以陕西省神木县六道沟小流域为研究区域,探讨了黄土高原北部紫花苜蓿人工草地退化过程中植物多样性的变化.结果表明,在黄土高原北部森林草原气候区,紫花苜蓿人工草地的退化演替过程可以划分为3个阶段:紫花苜蓿草地阶段(1～6年)、人工草地向天然草地演替的过渡阶段(6～10年)与长芒草次生天然草地阶段(10年以上).在1～30年的演替过程中,调查样方内共出现高等植物32种,分属于13科28属,其中90%以上出现在前6年.植被演替过程中累积出现的植物科属种数的动态变化可以用对数函数进行较好地描述.在群落演替过程中,物种丰富度指数、多样性指数和Pielou均匀度指数的变化趋势基本一致,在演替前期(第1个阶段)增加较快,在第2个阶段(过渡阶段)达到最高,然后有所降低并渐趋稳定.种植紫花苜蓿可显著地加快植被的自然演替进程,这与人工草地水分消耗强烈加速了土壤的旱化过程密切相关.发展人工草地是黄土高原北部加速天然植被恢复与发展畜牧业、增加农民收入的有机结合点,是西部生态环境建设中生态效益与经济效益兼顾的良好范式.     

Small molecule AKAP-protein kinase A (PKA) interaction disruptors that activate PKA interfere with compartmentalized cAMP signaling in cardiac myocytes

A-kinase anchoring proteins (AKAPs) tether protein kinase A (PKA) and other signaling proteins to defined intracellular sites, thereby establishing compartmentalized cAMP signaling. AKAP-PKA interactions play key roles in various cellular processes, including the regulation of cardiac myocyte contractility. We discovered small molecules, 3,3'-diamino-4,4'-dihydroxydiphenylmethane (FMP-API-1) and its derivatives, which inhibit AKAP-PKA interactions in vitro and in cultured cardiac myocytes. The molecules bind to an allosteric site of regulatory subunits of PKA identifying a hitherto unrecognized region that controls AKAP-PKA interactions. FMP-API-1 also activates PKA. The net effect of FMP-API-1 is a selective interference with compartmentalized cAMP signaling. In cardiac myocytes, FMP-API-1 reveals a novel mechanism involved in terminating β-adrenoreceptor-induced cAMP synthesis. In addition, FMP-API-1 leads to an increase in contractility of cultured rat cardiac myocytes and intact hearts. Thus, FMP-API-1 represents not only a novel means to study compartmentalized cAMP/PKA signaling but, due to its effects on cardiac myocytes and intact hearts, provides the basis for a new concept in the treatment of chronic heart failure.     

Shape-dependent regulation of differentiation lineages of bone marrow-derived cells under cyclic stretch

Multipotent stem cells are considered as a key material in regenerative medicine, and the understanding of the heterogeneity in the differentiation potentials of bone marrow-derived cells is important in the successful regenerative tissue repair. Therefore, the present study has been performed to investigate how the differentiation of post-harvest, native bone marrow-derived cells is regulated by cyclic stretch in vitro. Bone marrow-derived cells were obtained from mouse femur of both hind limbs and categorized into the following five categories: amebocytes, round cells, spindle cells, stellate cells and others. The cells were seeded on a silicone-made stretch chamber, and subjected to cyclic stretch with an amplitude of 10% at a frequency of 1 Hz for 7 days for cell shape analysis and for 3 days for the analysis of the expression of marker proteins of osteogenic (osteocalcin), vascular smooth muscle (α-smooth muscle actin and smooth muscle myosin heavy chain) and neurogenic (neurofilament) differentiation. When disregarding the differences in the cell shapes, there was an overall trend that the application of 10% cyclic stretch inhibited osteogenic and neurogenic differentiation, but enhanced smooth muscle differentiation. Close examinations revealed that round cells were influenced the most by cyclic stretch (significant up- or down-regulation in all the four marker protein expressions) while amebocytes and spindle cells were only influenced by cyclic stretch for vascular smooth muscle and/or neurogenic differentiation. As far as the authors know, this is the first study reporting the shape-related differences in the fate decision criteria for mechanical strain in bone marrow-derived cells.     

土壤易氧化有机碳对西双版纳热带森林群落演替的响应

土壤易氧化有机碳（Readily oxidizable carbon,ROC）作为土壤中易被氧化且活性较高的有机碳,能够敏感反映群落植被环境与土壤环境的早期变化。为探明土壤ROC时空变化对热带森林次生演替的响应,以西双版纳热带森林不同次生演替阶段（白背桐群落、野芭蕉群落与崖豆藤群落）为研究对象,采用高锰酸钾氧化法测定并分析土壤ROC时空动态特征,探究这些变化与土壤微生物量碳及理化性质之间的相互关系。结果表明：（1）不同次生演替阶段热带森林土壤ROC含量存在显著差异,其大小顺序为：野芭蕉群落（11.38 mg/g） > 崖豆藤群落（10.5 mg/g） > 白背桐群落（9.72 mg/g）;（2）不同次生演替阶段热带森林土壤ROC含量的月份变化趋势基本一致,均表现为6月显著高于12月,且各月份间差异显著;（3）不同次生演替阶段热带森林土壤ROC含量随着土层深度增加而递减,且不同土层间差异显著;（4）土壤有机碳、微生物量碳、全氮、水解氮和铵态氮显著影响土壤ROC含量的时空变化,而pH值与土壤ROC显著负相关。因此,土壤ROC对西双版纳热带森林群落演替具有敏感的响应,土壤总有机碳、微生物量碳、全氮、水解氮、铵态氮及pH是土壤ROC时空变化的主控因素。     

Regulation of Calmodulin- and Dopamine-Stimulated Adenylate Cyclase Activities by Light in Bovine Retina

Abstract: Neural retina from most species contains 3,4-dihydroxyphenylethylamine (dopamine) receptors coupled to stimulation of adenylate cyclase activity. It has been demonstrated that release of dopamine from its neurons and subsequent occupation of dopamine receptors is increased by light. In this study, we have shown that adenylate cyclase activity in bovine retina is highly responsive to the endogenous Ca²⁺-binding protein, cal-modulin, and that calmodulin can increase dopamine-sen-sitive adenylate cyclase activity in bovine retina. We further demonstrate that both dopamine- and calmodulin-stimulated adenylate cyclase activities can be regulated by alterations in light. Bovine retinas were dissected from the eye under a low-intensity red safety light, defined as dark conditions, and incubated for 20 min in an oxygenated Krebs Henseleit buffer under either dark or light conditions. The retinas were then homogenized and adenylate cyclase activity measured in a paniculate fraction washed to deplete it of endogenous Ca²⁺ and calmodulin. Activation of adenylate cyclase activity by calmodulin, dopamine, and the nonhydrolyzable GTP analog, gua-nosine-5′-(β,γ-imido)triphosphate (GppNHp), was significantly (60%) greater in paniculate fractions from retinas that had been incubated under dark conditions as compared to those incubated under light conditions. Basal, Mn²⁺-, and GTP-stimulated adenylate cyclase activities were not altered by changes in lighting conditions. Calmodulin could increase the maximum stimulation of adenylate cyclase by dopamine in retinas incubated under either dark or light conditions, but the degree of its effect was greater in retinas incubated under light conditions. Activation of adenylate cyclase by calmodulin, dopamine, and GppNHp in paniculate fractions from retinas incubated under light conditions was indistinguishable from the activation obtained when retinas were incubated in the dark in the presence of exogenous dopamine. These results suggest that an increased release of dopamine occurs in light. The decreased response of adenylate cyclase to exogenous dopamine can then be explained by a subsequent down-regulation of dopamine receptor activity. The down-regulation of dopamine receptor activity can also regulate activation of adenylate cyclase by GppNHp and calmodulin. The results suggest that dopamine, calmodulin, and GppNHp are modulators of a common component of adenylate cyclase activity, and this component is regulated by light.     

小叶锦鸡儿灌丛下土壤水分对降雨的响应

以科尔沁沙地主要固沙灌木小叶锦鸡儿为研究对象,在其生长季次降雨21.5 mm后180 h内,利用TDR和微渗仪测量小叶锦鸡儿灌丛下不同部位的土壤含水量和土壤蒸发,并计算该灌丛下不同部位储水量和水量平衡.结果表明:降雨结束后初期,灌丛枝干的茎流作用使其根部的土壤含水量明显高于其他部位;灌丛根部水分的入渗速率大于灌丛中部和灌丛外缘.因冠幅的庇荫作用,灌丛下蒸发量小于灌丛外裸露沙地.水量平衡表明:小叶锦鸡儿灌丛下降雨后前期蒸散量明显高于灌丛外裸露沙地,与灌丛下根系的分布有直接关系.     

枣园生态系统中害虫与天敌群落演替规律的研究

通过对太谷地区枣园害虫天敌群落的系统调查,并应用最优分割法.主分量分析法和1维和2维排序法,探讨了引起枣园害虫和天敌群落演替的主要类群及其演变规律.结果表明,群落具有明显的主导因子和时序演替格局,枣园害虫和天敌群落在前3个主分量上的变动较大,且跟随效应明显;枣园害虫和天敌群落的演替在时间过程中明显分为4个阶段:枣树发芽前、枣树展叶开花期、枣树结果期、枣果着色成熟期;利用主分量分析法,明确了主要害虫及其天敌的种类.     

褪黑素对谷氨酸致痫大鼠海马cAMP水平的影响

目的探讨褪黑素(melatonin, MT)对谷氨酸(glutamate,Glu)致痫大鼠海马cAMP水平的影响.方法　随机将健康SD雄性大鼠分为A、B、C、D 4组,每组10只,分别为对照组、Glu组、MT Glu组和Luzidole MT Glu组,观察并记录动物行为学及EEG改变,应用放射免疫方法检测各组动物脑内cAMP水平.结果行为学观察和EEG显示,B组和D组大鼠均出现痫性发作,并出现频发性痫性放电,C组大鼠痫性发作不明显,无频发性痫性放电出现;放射免疫分析结果显示,B组和D组海马cAMP含量均较对照组显著的升高(P<0.05);C组较B组和D组cAMP水平明显降低(P<0.05),与A组无明显差异性(P>0.05).结论MT对Glu致痫大鼠有抑痫作用,此作用是通过其受体调节海马内cAMP水平来实现的.     

Effects of Restoration on Plant Species Richness and Composition in Scandinavian Semi-Natural Grasslands

Plant species richness in rural landscapes of northern Europe has been positively influenced by traditional management for millennia. Owing to abandonment of these practices, the number of species‐rich semi‐natural grasslands has decreased, and remaining habitats suffer from deterioration, fragmentation, and plant species decline. To prevent further extinctions, restoration efforts have increased during the last decades, by reintroducing grazing in former semi‐natural grasslands. To assess the ecological factors that might influence the outcome of such restorations, we made a survey of semi‐natural grasslands in Sweden that have been restored during the last decade. We investigated how plant species richness, species density, species composition, and abundance of 10 species that are indicators of grazing are affected by (1) the size of the restored site, (2) the time between abandonment of grazing and restoration, (3) the time elapsed since restoration, and (4) the abundance of trees and shrubs at the restored site. Only two factors, abundance of trees and shrubs and time since restoration, were positively associated with total species richness and species density per meter square at restored sites. Variation in species composition among restored sites was not related to any of the investigated factors. Species composition was relatively similar among sites, except in mesic/wet grasslands. The investigated factors had small effects on the abundance of the grazing‐indicator species. Only Campanula rotundifolia responded to restoration with increasing abundance and may thus be a suitable indicator of improved habitat quality. In conclusion, positive effects on species richness may appear relatively soon after restoration, but rare, short‐lived species are still absent. Therefore, remnant populations in surrounding areas may be important in fully recreating former species richness and composition.