Metabolic Enzyme Alterations and Astrocyte Dysfunction in a Murine Model of Alexander Disease With Severe Reactive Gliosis

Alexander disease (AxD) is a rare and fatal neurodegenerative disorder caused by mutations in the gene encoding glial fibrillary acidic protein (GFAP). In this report, a mouse model of AxD (GFAP^Tg;Gfap^+/R236H) was analyzed that contains a heterozygous R236H point mutation in murine Gfap as well as a transgene with a GFAP promoter to overexpress human GFAP. Using label-free quantitative proteomic comparisons of brain tissue from GFAP^Tg;Gfap^+/R236H versus wild-type mice confirmed upregulation of the glutathione metabolism pathway and indicated proteins were elevated in the peroxisome proliferator-activated receptor (PPAR) signaling pathway, which had not been reported previously in AxD. Relative protein-level differences were confirmed by a targeted proteomics assay, including proteins related to astrocytes and oligodendrocytes. Of particular interest was the decreased level of the oligodendrocyte protein, 2-hydroxyacylsphingosine 1-beta-galactosyltransferase (Ugt8), since Ugt8-deficient mice exhibit a phenotype similar to GFAP^Tg;Gfap^+/R236H mice (e.g., tremors, ataxia, hind-limb paralysis). In addition, decreased levels of myelin-associated proteins were found in the GFAP^Tg;Gfap^+/R236H mice, consistent with the role of Ugt8 in myelin synthesis. Fabp7 upregulation in GFAP^Tg;Gfap^+/R236H mice was also selected for further investigation due to its uncharacterized association to AxD, critical function in astrocyte proliferation, and functional ability to inhibit the anti-inflammatory PPAR signaling pathway in models of amyotrophic lateral sclerosis (ALS). Within Gfap⁺ astrocytes, Fabp7 was markedly increased in the hippocampus, a brain region subjected to extensive pathology and chronic reactive gliosis in GFAP^Tg;Gfap^+/R236H mice. Last, to determine whether the findings in GFAP^Tg;Gfap^+/R236H mice are present in the human condition, AxD patient and control samples were analyzed by Western blot, which indicated that Type I AxD patients have a significant fourfold upregulation of FABP7. However, immunohistochemistry analysis showed that UGT8 accumulates in AxD patient subpial brain regions where abundant amounts of Rosenthal fibers are located, which was not observed in the GFAP^Tg;Gfap^+/R236H mice.     

The Mu Transpososome Through a Topological Lens

ABSTRACT

Phage Mu is the most efficient transposable element known, its high efficiency being conferred by an enhancer DNA element. Transposition is the end result of a series of well choreographed steps that juxtapose the enhancer and the two Mu ends within a nucleoprotein complex called the ‘transpososome.’ The particular arrangement of DNA and protein components lends extraordinary stability to the transpososome and regulates the frequency, precision, directionality, and mechanism of transposition. The structure of the transpososome, therefore, holds the key to understanding all of these attributes, and ultimately to explaining the runaway genetic success of transposable elements throughout the biological world. This review focuses on the path of the DNA within the Mu transpososome, as uncovered by recent topological analyses. It discusses why Mu topology cannot be analyzed by standard methods, and how knowledge of the geometry of site alignment during Flp and Cre site-specific recombination was harnessed to design a new methodology called ‘difference topology.’ This methodology has also revealed the order and dynamics of association of the three interacting DNA sites, as well as the role of the enhancer in assembly of the Mu transpososome.     

毛乌素沙地优势克隆半灌木生物量配置对小尺度植被盖度变异的响应

克隆植物根据其构型可以分为游击型和密集型。游击型克隆植物的间隔子长,分株在水平空间的扩展范围大,可以利用更大空间范围内的资源,其通过克隆生理整合作用发生的非局部反应的能力强。由此可以得出的推论之一是,小生境斑块的环境发生变化,生长于其中的密集型克隆植物的反应可能会更灵敏。这种反应可能会体现在生物量以及配置格局的变化上。以毛乌素地区沙生半灌木群落中两种优势克隆植物羊柴(H edy sarum laeve)和油蒿(A rtem isia ord osica)为研究对象,前者是典型的游击型克隆植物,后者是密集型克隆植物。采取野外调查的方式,观测在不同植被盖度的小生境斑块内二者地上生物量分配格局的变化情况,并结合二者的克隆构型和生活史特征试图探讨产生这种格局的原因。结果表明:羊柴的地上各部分生物量对植被盖度变化的响应不如油蒿敏感。这或者是因为羊柴的游击型克隆构型决定其可以跨越小尺度斑块实现克隆生理整合,可以利用不同小生境斑块的资源导致的。油蒿只能利用小生境斑块内的资源,当小生境斑块的条件改变,其生物量以及配置方式也随之发生相应的变化。在繁殖方式上,羊柴的有性繁殖结构以及有性繁殖投资显著小于油蒿。在资源有限的条件下,对一种繁殖方式的投资常常会削弱另一种繁殖方式。羊柴主要依靠克隆繁殖,这或者符合并支持配置理论的观点。     

铜绿假单胞菌色素代谢相关基因的研究

首次应用Mu转座重组技术研究铜绿假单胞菌色素合成与调控的机制。通过一系列的表型筛选,得到8株色素合成能力改变的突变子。经基因克隆、核苷酸测序研究,证明转座子分别插入到hmgA、ptsP、sucC、phzS、phzF1五个基因中。hmgA基因转座失活导致酪氨酸分解代谢中间产物尿黑酸的积累,后四种情况转座突变显著地影响了铜绿假单胞菌最重要的色素绿脓素（pyocyanin）的合成,其中PhzS和phzF1是绿脓素合成过程中的结构基因,ptsP基因是1个磷酸转移酶系统的重要组分,sucC基因的产物是三羧酸循环中的琥珀酰辅酶A合成酶,对后两个基因在色素合成的调控方面可能起到重要作用的报道尚属首例。     

毛乌素沙地南缘沙丘生物结皮对凝结水形成和蒸发的影响

在水分极度匮乏的荒漠生态系统,凝结水是除降雨之外最重要的水分来源之一,它对荒漠生态系统结构、功能和过程的维持产生重要的影响。为探明半干旱沙区生物结皮表面的凝结水形成和蒸发特征,采用自制的微型蒸渗计(直径7 cm、高5 cm的PVC管)实验观测了不同类型地表(裸沙、浅灰色藻类结皮、黑褐色藻类结皮和苔藓结皮)对凝结水形成和蒸发的影响。结果表明:(1)观测期间共有20次凝结水形成记录,除降雨天气外,几乎每天都能观测到水分凝结现象;(2)不同类型地表凝结水总量依次为(1.998±0.075),(2.326±0.083),(2.790±0.058)和(3.416±0.068) mm,生物结皮表面的凝结水总量显著大于裸沙(P < 0.05);随生物结皮的发育,不同类型生物结皮表面的凝结水总量呈增加的趋势,凝结水总量之间差异显著(P < 0.05);观测期间不同类型地表日平均凝结水量依次为(0.100±0.003),(0.116±0.004),(0.140±0.002)和(0.171± 0.003) mm,不同类型地表日平均凝结水量之间差异极显著(P < 0.01);(3)凝结水形成过程的观测结果显示,凝结水19:00开始形成,23:00-凌晨1:00形成不明显,1:00-7:00继续形成,除浅灰色藻类结皮外,太阳升出后在黑褐色藻类结皮和苔藓结皮表面继续形成少量的凝结水;凝结水7:30开始蒸发,10:30到11:00之间结束蒸发,凝结水在裸沙和浅灰色藻类结皮中的保持时间显著大于黑褐色藻类结皮和苔藓结皮中的保持时间(P < 0.05);(4)凝结水的形成受大气温度、地表温度、空气相对湿度和大气地表温度差等气象因素的影响,但其形成过程不与某一个气象因素呈简单的线性关系。     

Opioid receptor selectivity profile change via isosterism for 14-O-substituted naltrexone derivatives

Isosterism is commonly used in drug discovery and development to address stability, selectivity, toxicity, pharmacokinetics, and efficacy issues. A series of 14-O-substituted naltrexone derivatives were identified as potent mu opioid receptor (MOR) antagonists with improved selectivity over the kappa opioid receptor (KOR) and the delta opioid receptor (DOR), compared to naltrexone. Since esters are not metabolically very stable under typical physiological conditions, their corresponding amide analogs were thus synthesized and biologically evaluated. Unlike their isosteres, most of these novel ligands seem to be dually selective for the MOR and the KOR over the DOR. The restricted flexibility of the amide bond linkage might be responsible for their altered selectivity profile. However, the majority of the 14-N-substituted naltrexone derivatives produced marginal or no MOR stimulation in the ³⁵S-GTP[γS] assay, which resembled their ester analogs. The current study thus indicated that the 14-substituted naltrexone isosteres are not bioisosteres since they have distinctive pharmacological profile with the regard to their opioid receptor binding affinity and selectivity.     

毛乌素沙地半固定沙丘油蒿种群对土壤湿度空间异质性的响应

将统计学和地统计学方法相结合,探讨了毛乌素沙地半固定沙丘土壤湿度空间异质性与油蒿种群空间异质性之间的关系. 结果表明,在块尺度（80 m×80 m）上,油蒿密度和生物量的空间异质性受制于土壤湿度的空间异质性,尤其油蒿生物量的空间异质性与土壤湿度的空间异质性呈显著正相关关系.表明在半固定沙丘阶段,土壤湿度的空间异质性决定油蒿种群的空间异质性. 由于沙丘地形对降水量的再分配在土壤湿度分布中的作用,土壤湿度和油蒿种群空间异质性相互作用的结果为地形的缀块性→土壤湿度分布的缀块性→油蒿种群分布的缀块性→油蒿生物量和密度的缀块性.     

中分辨率遥感像元尺度生物土壤结皮覆盖与植被及土壤间的交互关系

生物土壤结皮（Biological soil crust,BSC）作为干旱、半干旱地区健康生态系统的重要组成部分,具有固碳、固氮、降低土壤水分蒸发、减少土壤径流等多种生态功能,这些功能与BSC的发展阶段及覆盖度密切相关,使其在维系荒漠自然生态系统的稳定性中扮演重要角色。因此,结合遥感数据尺度定量研究影响BSC分布的环境因素是评估沙区生态系统稳定性并以此为依据进行沙化土地治理的重要基础性研究。影响BSC覆盖度的环境因子较为复杂,现有研究方法存在两方面局限,其一是多为小尺度定性分析,其二是多侧重于孤立地分析BSC与环境因子间的单向关系。利用结构方程模型（Structural Equation Modeling,SEM）中的非递归路径分析（Nonrecursive Path Analysis,NPA）对遥感30 m分辨率像元尺度BSC覆盖度、植被覆盖度、土壤pH值、盐度、有机质与粒度进行路径分析,旨在使用综合性方法从整体上阐明大尺度BSC分布与植被、土壤间的交互关系。结果表明：（1）在毛乌素沙地,BSC覆盖度受各环境因子综合影响,无法用单一变量说明。BSC覆盖度与植被覆盖度、土壤有机质、平均粒径和细颗粒占比呈极显著正相关（P<0.01）,与粗颗粒占比呈极显著负相关（P<0.01）。（2） BSC覆盖度与植被覆盖度通过有机质相互影响,BSC覆盖度对植被覆盖度有较大的正向直接影响,路径系数（Path Coefficient,PC）=0.43（P>0.05）,植被覆盖度对BSC覆盖度有交大的负向影响（PC=－0.22;P>0.05）。（3）土壤平均粒径和细颗粒占比均正向影响BSC的覆盖度,其中,平均粒径对BSC覆盖度的总体影响较大（PC=0.67;总效果值=0.590）,细颗粒占比对BSC覆盖度的间接影响较大（间接效果值=0.052）。（4）盐度对BSC覆盖度呈显著负向直接影响（PC=－0.41;P<0.05;总效果值=－0.398）,pH值对BSC覆盖度有极小的正向影响（总效果值=0.072）。上述研究结果可为遥感探测BSC、制定有效的荒漠生态系统保护与修复政策措施提供科学依据。     

Fine-scale spatial distribution of soil organic carbon and its fractions after afforestation with Pinus sylvestris and Salix psammophila in a semiarid desert of China

半干旱沙漠中樟子松和沙柳造林后土壤有机碳及其组分的小尺度空间分布 半干旱沙漠造林有助于改善土壤功能以及增加土壤有机碳(SOC)固定,但人们对造林后SOC及其不稳定(LOC)组分的小尺度空间分布了解甚少。本研究以毛乌素沙地东南缘樟子松(Pinus sylvestris)和沙柳 (Salix psammophila)为研究对象,量化了距离树体20、80、150和240 cm处SOC、LOC组分及其相关变量的小尺度空间分布。研究结果表明,沙柳和樟子松造林显著提高了SOC、总氮(TN)、可溶性有机碳 (DOC)、微生物碳(MBC)和易氧化有机碳(ROOC)含量;在距离树体20 cm处,0–100 cm土层樟子松SOC 储量比沙柳高27.21%;在距离树体80和150 cm处,沙柳SOC储量分别比樟子松高5.50%和5.66%;与流 沙地相比,在距离树体20、80、150 和240 cm处,沙柳和樟子松SOC储量显著增加了94.90%、39.50%、 27.10%和18.50%;沙柳和樟子松ROOC分别占SOC的14.09%和18.93%。总之,造林促进了半干旱流沙地SOC的积累,樟子松比沙柳分配更多的有机质到距离树体<80 cm范围内的土体中。     

Crystal structure of the C-terminal domain of Mu phage central spike and functions of bound calcium ion

Bacteriophage Mu, which has a contractile tail, is one of the most famous genus of Myoviridae. It has a wide host range and is thought to contribute to horizontal gene transfer. The Myoviridae infection process is initiated by adhesion to the host surface. The phage then penetrates the host cell membrane using its tail to inject its genetic material into the host. In this penetration process, Myoviridae phages are proposed to puncture the membrane of the host cell using a central spike located beneath its baseplate. The central spike of the Mu phage is thought to be composed of gene 45 product (gp45), which has a significant sequence homology with the central spike of P2 phage (gpV). We determined the crystal structure of shortened Mu gp45Δ1-91 (Arg92–Gln197) at 1.5 Å resolution and showed that Mu gp45 is a needlelike structure that punctures the membrane. The apex of Mu gp45 and that of P2 gpV contained iron, chloride, and calcium ions. Although the C-terminal domain of Mu gp45 was sufficient for binding to the E. coli membrane, a mutant D188A, in which the Asp amino acid residue that coordinates the calcium ion was replaced by Ala, did not exhibit a propensity to bind to the membrane. Therefore, we concluded that calcium ion played an important role in interaction with the host cell membrane.