过表达MyBPC1对牛骨骼肌卫星细胞增殖分化的影响

旨在探究肌球蛋白结合蛋白C1（myosin binding protein C1,MyBPC1）对牛骨骼肌卫星细胞增殖与成肌分化的影响,为进一步研究MyBPC1在细胞分化和肌肉发育过程中的调控作用提供依据。本研究利用西门塔尔胎牛原代牛骨骼肌卫星细胞体外诱导成肌分化模型模拟牛骨骼肌的生长发育过程。采用qRT-PCR和Western blot检测MyBPC1的细胞时序表达谱。试验分为两组。在RNA水平每组4个重复,每个重复20 μL;在蛋白水平每组3个重复,每个重复15 μg。采用qRT-PCR和Western blot检测牛骨骼肌卫星细胞转染MyBPC1的过表达效果,并进一步检测细胞增殖期标志因子Pax7、Ki67以及细胞分化期标志因子MyHC、MyOG的表达变化情况,观察牛骨骼肌卫星细胞肌管形成状态。结果,MyBPC1在牛骨骼肌卫星细胞分化前后表达水平存在极显著差异,牛骨骼肌卫星细胞诱导分化后MyBPC1的mRNA和蛋白表达量均极显著高于增殖期（P<0.01）。过表达MyBPC1后,细胞分化形成的肌管数量明显多于对照组,增殖标志因子Pax7的mRNA水平和蛋白表达水平无显著差异,分化标志因子MyHC的mRNA水平和蛋白表达水平极显著高于对照组（P<0.01）。过表达MyBPC1可以促进牛骨骼肌卫星细胞体外成肌分化,为进一步开展MyBPC1对牛骨骼肌卫星细胞的调控机制奠定基础。     

Suppression of MyoD induces spontaneous adipogenesis in skeletal muscle progenitor cell culture

The degree of intramuscular adipose tissue accumulation is one of the factors affecting meat quality. Accumulation of adipocytes is also observed under the pathological condition of skeletal muscle such as muscular dystrophy and sarcopenia. The origin of adipocytes seen in skeletal muscle is mesenchymal progenitor cells that can give rise to both adipocytes and fibroblasts. In the present study, we demonstrated that siRNA-mediated suppression of MyoD expression in rat skeletal muscle progenitor cell culture, which comprises both myogenic satellite cells and mesenchymal progenitor cells, resulted in diminished myotube formation and an unexpected spontaneous appearance of white adipocytes. Suppressing myomaker expression also resulted in complete absence of myotube formation without reducing MyoD expression, but no adipogenesis was seen in this scenario, indicating that decline in MyoD expression rather than decreased myotube formation is necessary to induce adipogenesis. In addition, spontaneous adipogenesis induced by suppressing MyoD expression in culture was inhibited by the conditioned medium from control culture, indicating that anti-adipogenic factor(s) are secreted from MyoD-positive myogenic cells. These results indicate the presence of regulatory mechanism on adipogenesis by myogenic cells.     

MSTN基因对牛肌动蛋白细胞骨架调节通路的影响

为探究肌肉生长抑制素（myostatin,MSTN）对牛骨骼肌生长发育的作用机制,本研究前期利用定量蛋白质组学与磷酸化蛋白质组学分析野生型蒙古牛（MG.WT）和MSTN^+/-蒙古牛（MG.MSTN^+/-）腿臀肌肌肉组织中蛋白质水平和磷酸化修饰水平的差异变化,使用已建立的牛骨骼肌卫星细胞体外诱导成肌分化模型,检测设计合成的MSTN siRNA （si-MSTN）干扰效果;采用实时荧光定量PCR和Western blotting方法检测转染si-MSTN的增殖期（GM）和分化第3天（DM3）牛骨骼肌卫星细胞中肌动蛋白细胞骨架调节通路相关基因的mRNA和蛋白水平的表达变化,研究敲低MSTN表达对肌动蛋白细胞骨架调节通路的影响。结果显示,在MSTN^+/-蒙古牛肌肉组织中共鉴定到16个肌动蛋白细胞骨架调节通路相关基因表达丰度上调;转染si-MSTN细胞中的MSTN表达水平极显著降低（P<0.01）;在转染si-MSTN的GM期牛骨骼肌卫星细胞中,肌动蛋白细胞骨架调节通路相关基因ENAH、ACTN4和Cdc42的mRNA水平均显著升高（P<0.05）,PFN1、RhoA和ACTN4的蛋白水平均显著或极显著升高（P<0.05;P<0.01）;在转染si-MSTN的DM3牛骨骼肌卫星细胞中,ENAH、CFL1、SCIN和Cdc42基因mRNA水平均显著升高（P<0.05）,RhoA基因mRNA水平极显著升高（P<0.01）,PFN1和ACTN4的蛋白水平均显著升高（P<0.05）。结果表明,干扰MSTN可以促进肌动蛋白细胞骨架调节通路相关基因的表达,探明了MSTN可能通过介导肌动蛋白细胞骨架调节通路影响牛骨骼肌卫星细胞增殖和成肌分化的分子机制,为进一步研究MSTN对牛成肌分化的调控机制提供参考。     

Preliminary experiments on mechanical stretch-induced activation of skeletal muscle satellite cells in vivo

We have shown in vitro that mechanical stretch triggers activation of quiescent satellite cells of skeletal muscle to enter the cell cycle through an intracellular cascade of events including nitric oxide (NO) synthesis that results in the release of hepatocyte growth factor (HGF) from its extracellular association and its subsequent presentation to signaling receptors. In order to explore the activation mechanism in vivo, stretch experiments were conducted in the living animal using our suspension model developed. This system used the weight of the hind portion of rats to stretch the inside muscles of the left hind limb suspended for a period of 0.5–2.0 h. At the end of the stretch period, the rats received an intraperitoneal injection of bromodeoxyuridine followed by immunocytochemistry for its incorporation as an index of satellite cell activation in vivo. Depending on the period of stretch, bromodeoxyuridine labeling was increased significantly over the contralateral unstretched leg or control muscle from untreated rats. A stretched muscle extract prepared from the 2 h stretched tissue by incubating it in PBS, showed the active form of HGF as revealed by immunoblotting and it could stimulate the activation of unstretched satellite cells. Also, administering NO synthase inhibitor L‐NAME prior to muscle stretch abolished the stretch activation of satellite cells. Therefore, the results from these experiments demonstrate that stretching muscle triggers NO synthesis and HGF release, which could activate satellite cells in vivo.     

A method for mapping the distribution of willow at a catchment scale using bi-seasonal SPOT5 imagery

This study showed that seasonal imagery acquired at specific stages of phenology can be used to improve the mapping accuracy of invasive willow at a catchment scale. SPOT5 XI (10 m) satellite imagery was acquired for early autumn and winter to represent the phenological stages of leaf cover and leaf fall respectively. Four classification regimes were evaluated using single‐ and bi‐seasonal composite imagery to determine the most accurate method. Significant spectral noise was found in willow populations, especially in the winter image, due to the effects of undergrowth exposure, shadowing, topography and boundary‐mixed pixels. Two noise reduction techniques were applied to the bi‐seasonal composite image to improve the classification results. The noise‐reduced bi‐seasonal composite image was classified using the spectral angle mapper (SAM) algorithm before importation into a geographical information system. Aerial photography was used to reduce the errors of commission associated with misclassification of pastures. The class accuracy achieved for willow using the method described in this study was 77.5% (Kappa =0.87). The high cost of eradicating willow means that managers must establish priorities for control; this technique can provide a powerful tool for prioritizing control programmes and for monitoring results at a catchment scale.     

Bigeye tuna (Thunnus obesus) vertical movements in the Azores Islands determined with pop-up satellite archival tags

Movement patterns of 17 bigeye tuna (Thunnus obesus) near the Azores Islands were analyzed between April and May 2001 and 2002 using pop‐up satellite archival tags. Despite short attachment durations (1 to 21 days, 8.2 days on average), their vertical movements revealed much shallower distribution of bigeye tuna in comparison with previous studies in the tropical Pacific and tropical Atlantic. Depth and temperature histograms were unimodal, although overall depth distribution during the day was deeper than during the night due to daily incursions in deeper waters. Although generalized additive models showed significant non‐linear relationships with weight of the fish and sea level anomaly (as a proxy for variability of thermocline depth), the effect of these variables on bigeye depth appeared minor, suggesting that vertical movements of bigeye in the Azores during the spring migration may be influenced by food availability in upper water layers.     

SPOT5卫星遥感信息在森林资源调查中的应用现状与解决思路

介绍利用SPOT5卫星遥感在云南省森林资源调查中的应用情况,总结所取得的成果经验。分析存在的包括对遥感的内涵认识不足、特殊地类不易分辨、树种组成难判读、蓄积量不能直接判读、龄组和郁闭度判读因子粗放等问题,提出了解决问题的思路和建议。     

森林资源调查中Spot5卫星影像的处理方法

利用Spot5卫星进行森林资源调查时,首先要对遥感图像进行处理。文章介绍了处理环境对硬件和软件的要求。其处理过程包括影像配准、分辨率融合、影像拼接、色彩增强、分幅输出等。对其中的一些问题提出了个人见解。     

利用91卫图助手软件实现林业矢量数据坐标系的转换

以永平县林业矢量数据为例,利用91卫图助手软件和ArcGIS软件,通过北京54坐标系矢量数据在各坐标系下叠加、坐标系转换参数的计算、矢量数据坐标系的转换等步骤,阐述实现矢量数据坐标系WGS84、北京54、西安80和国家2000坐标系相互转换的方法,以实现各类自然资源矢量数据的共享。     

卫星遥感图像应用处理新模式的研究

卫星遥感图像增强与处理的研究旨在通过卫星遥感图像的处理,增强图像的显示质量,提高森林资源判读效果。通过应用ERDAS、ENYI遥感软件对TM、SPOT卫星数据的研究,实现资源调查卫片自主生产,降低了成本,提高了工作效率。