Morphological apoptotic characteristics of the post-meiotic micronuclei in Paramecium caudatum

In a previous study, the apoptotic degeneration of meiotic products outside the paroral region of Paramecium caudatum was indirectly demonstrated by means of “apofluor” staining. In this experiment, conjugating pairs and exconjugants of P. caudatum were stained with either “apofluor” or carbol fuchsin or both to find some direct evidence to demonstrate the apoptotic characteristics of this process. As a result, asynchronous meiotic nuclear degeneration was observed. Furthermore, a number of additional meiotic nuclei were found. Disintegrating/dividing meiotic nuclei outside the paroral region were observed, which might be the origin of these additional meiotic nuclei. Condensed chromatin and disintegrated chromatin attached to the nuclear membrane were also observed in degenerating nuclei, which are the typical morphological characteristics of apoptosis. Comparison of the cells stained by the above two methods indicated that “apofluor”-stained meiotic nuclei could not be detected by carbol fuchsin in some cells, which suggests a time lag between meiotic nuclear DNA degradation and their eventual disappearance. In this study, some direct evidence was found to show that the meiotic nuclear degeneration in P. caudatum is of apoptotic nature, which further confirmed our previous study (Yang et al. 2007) and indicated that morphological apoptotic characteristics discovered in multicellular organisms do exist in unicellular eukaryotic ciliate protozoa.     

Myostatin regulates glucose metabolism via the AMP-activated protein kinase pathway in skeletal muscle cells

Myostatin (Mstn) is a secreted growth factor predominately expressed in skeletal muscle that negatively regulates skeletal muscle mass. Recent studies have indicated that loss function of myostatin not only increases muscle mass but also improves insulin sensitivity in vivo. In the present report, we demonstrated that myostatin regulates glucose metabolism by promoting glucose consumption and glucose uptake, increasing glycolysis, and inhibiting glycogen synthesis in skeletal muscle cells. Microarray analysis revealed that myostatin upregulates several genes involved in regulating glucose metabolism such as Glut1, Glut4, Hk2, and IL-6. Further investigation of the molecular basis of these phenomena revealed that AMP-activated protein kinase (AMPK), a key component for maintaining energy homeostasis, was activated by myostatin for promotion of glycolysis. Taken together, these findings provide the first experimental evidence that myostatin regulates glucose metabolism through the AMPK signal pathway in muscle cells. Importantly, our findings highlight that continued investigation of the metabolic function of myostatin is necessary for a comprehensive understanding of its active role in the regulation of skeletal muscle energy metabolism.     

中国暖温带若干灌丛群落多样性问题的研究

 暖温带灌丛的生物多样性的研究表明：1．灌丛群落的均一度指数同优势度指数的相关性较丰富度指数同变化度指数的相关性为高;在众多的多样性指数中,D4(Shannon信息度指数)和D6(Gini指数)因最大限度地包含了其它各类多样性指数的信息而被采用。2．灌丛群落的物种多样性指数值随着演替的进行,没有表现出降低的趋势。3．生物多样性较高的荆条灌丛和辽东栎萌丛的物种多度分布是对数分布,而生物多样性较低的鬼见愁灌丛和毛榛灌丛的物种多度分布是几何分布,处于中间的三桠绣线菊灌丛的物种多度分布是Broken-Stick分布;没有正态对数分布类型。     

冬季清凉峰山区小麂和野猪的生境选择及差异

从2005 年11 月5 日至2006 年1 月21 日,为了评估小麂和野猪的生境选择及其差异,我们在浙江清凉峰国家级自然保护区核心区共设置了248 个样方群。研究发现,两者都选择平缓坡的灌丛植被、草本密度小、离住宅较近和离隐蔽物距离适中的生境,表现了它们的生境选择有一定的重叠性。但是它们也表现出生境选择的一些差异,小麂选择乔木密度适中、灌丛密度大和郁闭度适中的生境,而野猪选择乔木密度小的生境,随机地使用各类灌丛密度和郁闭度的生境。小麂还选择北坡、离水源距离较远的生境,野猪只是随机地使用不同的坡向和离水源距离的生境。分析两者共存的机制,我们发现小麂采取隐藏策略而野猪采取逃跑策略来避敌和利用资源,即它们之间存在与反捕食策略相关的生境选择差异。
     

坐骨神经结扎后大鼠背根神经节和脊髓CGRP表达的变化

目的研究大鼠坐骨神经结扎后降钙素基因相关肽(calcitoningene-relatedpeptide,CGRP)表达变化。方法SD大鼠随机分为假手术对照组和坐骨神经结扎组,实验组结扎后分别存活1、3、5、7、14、21和28d(n=8),免疫荧光(双标法)和免疫组织化学(SABC法)观察术后不同时间点CGRP和NGF在坐骨神经、背根神经节(dorsalrootganglion,DRG)和脊髓的表达变化,Westernblot结合图像分析技术对不同时间的变化进行定量测定。结果术后1d结扎远端坐骨神经内NGF大量堆积,持续到28d仍高于正常。结扎后7dDRG内CGRP阳性细胞百分率减少,持续到28d仍低于正常;结扎后14d脊髓后角CGRP下降,28d仍低于正常,各时间点脊髓前角CGRP表达未见明显变化。结论神经结扎可导致DRG和脊髓后角的CGRP表达下调,可能与靶源性的NGF来源减少有关。     

Functional characterization of kurtz, a Drosophila non-visual arrestin, reveals conservation of GPCR desensitization mechanisms

The arrestins are a family of molecules that terminate signaling from many different G protein-coupled receptors, by inhibiting the association between receptor and downstream effectors. We recently employed a human betaarrestin2-GFP fusion protein to explore the dynamics of different neuropeptide receptors in Drosophila and have previously used a betaarrestin translocation assay to identify ligands at orphan receptors. Here, we report that the Drosophila arrestin encoded by kurtz functions in a similar fashion and can be employed to investigate GPCR-arrestin associations. Specifically, a GFP-krz fusion protein, upon co-expression with various Drosophila peptide receptors, an amine receptor, and a mammalian peptide receptor translocates to the plasma membrane in specific response to ligand application. This molecular phenotype is exhibited in a mammalian cell line as well as in a Drosophila cell line. Notably, the details of receptor-arrestin associations in terms of endocytotic patterns are functionally conserved between the mammalian arrestins and kurtz. Furthermore, we report that kurtz mutants exhibit hypersensitivity to osmotic stress, implicating GPCR desensitization as an important feature of the endocrine events that shape this stress response.     

Differential binding to dorsal and ventral cell surfaces of fibroblasts: effect on collagen phagocytosis

Matrix remodeling by phagocytic fibroblasts is essential for growth and development but the regulatory processes are undefined. We evaluated the impact of spreading on the binding step of collagen phagocytosis with a novel culture system that more closely replicates phagocytosis in vivo than previous models. 3T3 cells were plated on collagen-coated beads, thereby loading only ventral surfaces (adhesion with spreading), or were allowed to spread on collagen films and then loaded with beads on their dorsal surfaces (adhesion without spreading). Ventral surfaces bound three-fold more beads than dorsal surfaces which was accompanied by accelerated phagosomal maturation. Arp3 and cortactin, markers of the actin-associated spreading machinery, strongly accumulated around ventrally but not dorsally loaded beads, suggesting that spreading contributes to enhanced binding of ventral surfaces. Further, ventral surfaces exhibited two-fold more free alpha2beta1 integrins, the major collagen receptors. Notably, compared to cells spread on collagen substrates, spreading cells exhibited a three-fold higher alpha2beta1 mobile fraction which was correlated with limited engagement of ventral receptors by actin filaments. Thus integrin ligation by actin filaments regulates the mobility of collagen receptors which in turn mediates the enhanced binding of collagen beads on spreading surfaces.     

The pro-apoptotic protein Bim is a convergence point for cAMP/protein kinase A- and glucocorticoid-promoted apoptosis of lymphoid cells

The mechanisms by which cAMP mediates apoptosis are not well understood. In the current studies, we used wild-type (WT) S49 T-lymphoma cells and the kin(-) variant (which lacks protein kinase A (PKA)) to examine cAMP/PKA-mediated apoptosis. The cAMP analog, 8-CPT-cAMP, increased phosphorylation of the cAMP response element-binding protein (CREB), activated caspase-3, and induced apoptosis in WT but not in kin(-) S49 cells. Using an array of 96 apoptosis-related genes, we found that treatment of WT cells with 8-CPT-cAMP for 24 h induced expression of mRNA for the pro-apoptotic gene, Bim. Real-time PCR analysis indicated that 8-CPT-cAMP increased Bim RNA in WT cells in <2 h and maintained this increase for >24 h. Bim protein expression increased in WT but not kin(-) cells treated with 8-CPT-cAMP or with the beta-adrenergic receptor agonist isoproterenol. Both apoptosis and Bim expression were reversible with removal of 8-CPT-cAMP after <6 h. The glucocorticoid dexamethasone also promoted apoptosis and Bim expression in S49 cells. In contrast, both UV light and anti-mouse Fas monoclonal antibody promoted apoptosis in S49 cells but did not induce Bim expression. 8-CPT-cAMP also induced Bim expression and enhanced dexamethasone-promoted apoptosis in human T-cell leukemia CEM-C7-14 (glucocorticoid-sensitive) and CEM-C1-15 (glucocorticoid-resistant) cells; increased Bim expression in 8-CPT-cAMP-treated CEM-C1-15 cells correlated with conversion of the cells from resistance to sensitivity to glucocorticoid-promoted apoptosis. Induction of Bim appears to be a key event in cAMP-promoted apoptosis in both murine and human T-cell lymphoma and leukemia cells and thus appears to be a convergence point for the killing of such cells by glucocorticoids and agents that elevate cAMP.