Measurement of ATP production and respiratory chain enzyme activities in mitochondria isolated from small muscle biopsy samples

A set of methods suitable for assessment of respiratory chain function in mitochondria isolated from 25mg of muscle is described. This set of methods includes determination of the mitochondrial ATP production rate (MAPR) and the activities of the respiratory chain complexes I, I+III, II+III, and IV and citrate synthase. MAPR is determined with an optimized version of a luminometric method previously described. The optimized method measures 50-220% higher activities than the original method. The highest MAPRs are recorded using the substrate combinations glutamate+succinate and N,N,N(1),N(1)-tetramethyl-1,4-phenyldiamine+ascorbate. The respiratory chain complex activities are determined with standard spectrophotometric methods, adapted to an automated photometer. The sensitivity in the determination of complex I, I+III, and II+III activities was increased considerably by pretreating the samples with saponin. The set of methods was evaluated on double biopsy samples from five healthy volunteers and showed coefficients of variation between 7 and 14% when citrate synthase was used as reference base. All of the various measures of mitochondrial function showed high correlation coefficients to each other (r=0.84-0.98; p<0.01). It is concluded that the set of methods is suitable for diagnosis of mitochondrial disorders in adults and small children.     

疏勒河流域气候变化和人类活动对植被NPP的相对影响评价

气候变化和人类活动是对陆地生态系统碳循环产生重要影响的两个因素,定量评估气候变化与人类活动对植被净初级生产力(NPP)的相对影响,对深入理解其驱动机制和控制荒漠化发展具有重要意义。以疏勒河流域为研究区,利用遥感和气象数据计算潜在NPP(PNPP)及其与实际NPP(ANPP)之间的差值,分别衡量了气候变化和人类活动对流域NPP的相对影响。研究结果表明:(1)2001—2015年疏勒河流域年ANPP整体呈缓慢增加趋势,与全国和西北地区相比,普遍较低,流域植被整体生产力水平不高。流域年ANPP空间分布呈现上游祁连山区和中下游绿洲区ANPP较高,而中下游荒漠戈壁区ANPP较低的分布格局。(2)2001—2015年流域年PNPP的变化趋势表明,降水量的变化是导致疏勒河流域植被退化加剧或缓解的主要气候驱动因素,但气温的变化对植被的影响较为复杂。(3)2001—2015年流域大部分地区植被退化系人类活动造成的,但人类活动的负向影响力在减弱。(4)气候变化和人类活动对植被NPP的相对影响均表现出明显的空间异质性,其中人类活动是疏勒河流域植被变化的主要驱动因素。     

Developmental downregulation of GABAergic drive parallels formation of functional synapses in cultured mouse neocortical networks

Networks of cortical neurons in vitro spontaneously develop synchronous oscillatory electrical activity at around the second week in culture. However, the underlying mechanisms and in particular the role of GABAergic interneurons in initiation and synchronization of oscillatory activity in developing cortical networks remain elusive. Here, we examined the intrinsic properties and the development of GABAergic and glutamatergic input onto presumed projection neurons (PNs) and large interneurons (L-INs) in cortical cultures of GAD67-GFP mice. Cultures developed spontaneous synchronous activity already at 5-7 days in vitro (DIV), as revealed by imaging transient changes in Fluo-3 fluorescence. Concurrently, spontaneous glutamate-mediated and GABA(A)-mediated postsynaptic currents (sPSCs) occured at 5 DIV. For both types of neurons the frequency of glutamatergic and GABAergic sPSCs increased with DIV, whereas the charge transfer of glutamatergic sPSCs increased and the charge transfer of GABAergic sPSCs decreased with cultivation time. The ratio between GABAergic and the overall charge transfer was significantly reduced with DIV for L-INs and PNs, indicating an overall reduction in GABAergic synaptic drive with maturation of the network. In contrast, analysis of miniature PSCs (mPSCs) revealed no significant changes of charge transfer with DIV for both types of neurons, indicating that the reduction in GABAergic drive was not due to a decreased number of functional synapses. Our data suggest that the global reduction in GABAergic synaptic drive together with more synaptic input to PNs and L-INs during maturation may enhance rhythmogenesis of the network and increase the synchronization at the level of population bursts.     

Estimation of fine root production, mortality and turnover with Minirhizotron in Larix gmelinii and Fraxinus mandshurica plantations

Fine root turnover is a major pathway for car-bon and nutrient cycling in forest ecosystems. However, to estimate fine root turnover, it is important to first understand the fine root dynamic processes associated with soil resource availability and climate factors. The objectives of this study were: (1) to examine patterns of fine root production and mortality in different seasons and soil depths in the Larix gmelinii and Fraxinus man-dshurica plantations, (2) to analyze the correlation of fine root production and mortality with environmental factors such as air temperature, precipitation, soil temperature and available nitrogen, and (3) to estimate fine root turn-over. We installed 36 Minirhizotron tubes in six mono-specific plots of each species in September 2003 in the Mao'ershan Experimental Forest Station. Minirhizotron sampling was conducted every two weeks from April 2004 to April 2005. We calculated the average fine root length, annual fine root length production and mortality using image data of Minirhizotrons, and estimated fine root turnover using three approaches. Results show that the average growth rate and mortality rate in L. melinii were markedly smaller than in F. mandshurica, and were high-est in the surface soil and lowest at the bottom among all the four soil layers. The annual fine root production and mortality in F. mandshurica were significantly higher than in L. gmelinii. The fine root production in spring and summer accounted for 41.7% and 39.7% of the total annual production in F. mandshurica and 24.0% and 51.2% in L. gmelinii. The majority of fine root mortality occurred in spring and summer for F. mandshurica and in summer and autumn for L. gmelinii. The turnover rate was 3.1 a-1 for L. gmelinii and 2.7 a-1 for F. mandshurica. Multiple regression analysis indicates that climate and soil resource factors together could explain 80% of the varia-tions of the fine root seasonal growth and 95% of the seasonal mortality. In conclusion, fine root production and mortality in L. gmelinii and F. mandshurica have dif-ferent patterns in different seasons and at different soil depths. Air temperature, precipitation, soil temperature and soil available nitrogen integratively control the dynamics of fine root production, mortality and turnover in both species.     

Changes in photosynthetic capacity and antioxidant enzymatic systems in micropropagated <Emphasis Type="Italic">Zingiber officinale</Emphasis> plantlets during their acclimation

Ginger (Zingiber officinale Rosc.) plantlets were propagated in vitro and acclimated under different photosynthetic photon flux densities (60 and 250 μmol m⁻² s⁻¹ = LI and HI, respectively). Increases in chlorophyll (Chl) content and Chl a/b ratio were found under both irradiances. In vitro plantlets (day 0) exhibited a low photosynthesis, but chloroplasts from in vitro leaves contained well developed grana and osmiophillic globules. Photoinhibition in leaves formed in vitro was characterized by decrease of photochemical efficiency and quantum efficiency of photosystem 2 photochemistry in HI treatment during acclimation. The new leaves formed during acclimation in both treatments showed a higher photosynthetic capacity than the leaves formed in vitro. Also activities of antioxidant enzymes of micropropagated ginger plantlets changed during acclimation.     

Effects of simulated feeding by snow geese on Scirpus americanus rhizomes

Summary We simulated the feeding of Greater Snow Geese (Chen caerulescens atlantica) on the rhizomes of three-square bulrush (Scirpus americanus) in a tidal marsh along the St. Lawrence River estuary in Québec. During the spring staging period, aboveground biomass is unavailable and geese feed solely on rhizomes and overwintering buds. An experiment was designed to test the effect of three factors on subsequent growth of Scirpus: the intensity of removal (3 to 77% removal of belowground biomass), the number of bites (1, 2 or 3 sections removed) and the number of adventitious buds removed (1, 2 or 3). Rhizomes were dug out in May, treated and transplanted into 85-1 basins sunk in the marsh and filled with marsh soil freed of all plant material. Growth was observed weekly until the end of the growing season in August. Shoots and rhizomes were then collected, dried and weighed to obtain biomass estimates. The net above- and belowground production of Scirpus was inversely related to the initial rhizome biomass removed. At a high level of removal (>35%), the cumulative number of shoots was significantly reduced as early as two weeks after transplantation. The relative reduction in production of the treated rhizomes compared to the control plants was also related to the intensity of removal. An increased number of bites reduced production and the removal of an increased number of adventitious buds further amplified the effect of removal on rhizome production. These experimental results show that even low intensity of feeding by Snow Geese can reduce the production of Scirpus marshes.     

马尾松不同改良水平子代群体遗传多样性研究

该研究以马尾松三个不同改良水平的良种生产群体子代为材料,用天然群体作为对照,采用16对SSR引物对试验群体进行遗传多样性分析,并探究遗传改良对马尾松林分遗传多样性的影响。结果表明:马尾松天然群体、母树林子代、1代种子园子代及1.5代种子园子代的Shannon多样性指数(I)分别为0.53、0.53、0.53、0.46;观测杂合度(Ho)分别为0.36、0.36、0.39、0.35;期望杂合度(He)分别为0.32、0.32、0.33、0.27。在这三项主要遗传多样性指标上,马尾松母树林、1代种子园及1.5代种子园的子代之间无显著差异。由此说明,在广西马尾松的遗传改良进程中,遗传多样性并未因改良选择而受到明显的影响。良种人工林与天然林相比较,马尾松良种人工林在三项主要指标上无明显下降,说明广西三类主要的良种群体都具有较好的群体缓冲能力和个体缓冲能力。该研究结果对于科学制定马尾松育种策略具有重要意义,为马尾松高世代育种研究提供了重要的理论依据。     

PDGF-AA activates AKT and ERK signaling for testicular interstitial Leydig cell growth via primary cilia

Testes control the development of male reproductive system. The testicular interstitial Leydig cells (Leydig cells) synthesize testosterone for promoting spermatogenesis and secondary sexual characteristics. Type A platelet-derived growth factor (PDGF-AA) is one of the most important growth factors in regulating Leydig cell growth and function. Knockout of PDGF-AA or its congenital receptor PDGFR-α leads to poor testicular development caused by reducing Leydig cell numbers, supporting PDGF-AA/PDGFR-α signaling regulates Leydig cell development. Primary cilium is a cellular antenna that functions as an integrative platform to transduce extracellular signaling for proper development and differentiation. Several receptors including PDGFR-α are observed on primary cilia for initiating signaling cascades in distinct cell types. Here we showed that PDGF-AA/PDGFR-α signaling promoted Leydig cells growth, migration, and invasion via primary cilia. Upon PDGF-AA treatment, AKT and ERK signaling were activated to regulate these cellular events. Interestingly, active AKT and ERK were detected around the base of primary cilia. Depletion of ciliary genes (IFT88 and CEP164) alleviated PDGF-AA-activated AKT and ERK, thus reducing Leydig cell growth, migration, and invasion. Thus, our study not only reveals the function of PDGF-AA/PDGFR-α signaling in maintaining testicular physiology but also uncovers the role of primary cilium and downstream signaling in regulating Leydig cell development.     

降水变化与氮添加对晋北盐碱化草地土壤净氮矿化的影响

于2018年在晋北典型农牧交错带盐碱化草地设置增减降水和氮添加处理试验,2019年生长季(5-9月)采用原位顶盖PVC埋管法测定不同水、氮处理下土壤净氮矿化速率,研究盐碱化草地土壤净氮矿化速率对降水格局变化和氮沉降的响应.结果表明:土壤净氮矿化速率表现出明显的季节动态.单独增减降水(±50％)和氮添加(10g·m-2·...