Identification of the fnx1+ and fnx2+ genes for vacuolar amino acid transporters in Schizosaccharomyces pombe

We have identified the Schizosaccharomyces pombe SPBC3E7.06c gene (fnx2(+)) from a homology search with the fnx1(+) gene involving in G(0) arrest upon nitrogen starvation. Green fluorescent protein-fused Fnx1p and Fnx2p localized exclusively to the vacuolar membrane. Uptake of histidine or isoleucine by S. pombe cells was inhibited by concanamycin A, a specific inhibitor of the vacuolar H(+)-ATPase. Amino acid uptake was also defective in the vacuolar ATPase mutant, suggesting that vacuolar compartmentalization is critical for amino acid uptake by whole cells. In both Deltafnx1 and Deltafnx2 mutant cells, uptake of lysine, isoleucine or asparagine was impaired. These results suggest that fnx1(+) and fnx2(+) are involved in vacuolar amino acid uptake in S. pombe.     

烟草花粉母细胞细胞融合过程中细胞骨架的超微结构观察

应用普通电镜和DGD去包埋技术 ,研究了烟草花粉母细胞中的细胞融合现象及细胞融合过程中细胞骨架的变化。观察发现 ,处于凝线期的花粉母细胞 ,其内含物 ,包括细胞器和染色质 ,主要通过胞质通道向相邻细胞发生转移。DGD去包埋观察发现 ,花粉母细胞中核骨架与细胞质内及胞间连丝和胞质通道内胞质骨架连接成一个整体。在整个细胞融合过程中 ,均有核骨架纤维与染色质相连。本文讨论了细胞骨架在细胞融合过程中的作用     

用非离子去垢剂抽提获得的小游仆虫皮层细胞骨架的构形

由扫描电镜术显示,应用非离子去垢剂抽提获得的小游仆虫(Euplotes grocilis)皮层细胞骨架是由非纤毛区皮层骨架、纤毛器骨架及其附属纤维等构成的三维结构网架。各类细胞骨架以纤维物质为基本成分组成纤维网、纤维层、纤维束和纤维薄片等不同形态单元。其中：非纤毛区皮层骨架以表面纤维网和表膜下纤维层为形态单元位于细胞的外周层;纤毛器骨架中的口围带骨架、口侧膜骨架、额腹横棘毛骨架按各自的分布图式在皮层内定位,成为主要的皮层骨架结构。尽管这些纤毛器骨架显示不同的形态,但却具有相同的建构特征,即都是由纤毛器的毛基体、纤毛器托架和骨架附属纤维相互联系镶嵌在一起形成的相对独立的结构单元。分析推测,游仆虫皮层表面纤维网使细胞表面形成区域化结构,它也可能与细胞表面各部分的联系及其细胞与环境的相互作用有关;纤毛器骨架中各个纤毛器的毛基体复合结构可能对纤毛器托架和骨架附属纤维等起到微管组织中心的作用。     

扁绒泡菌Physarum compressum原质团细胞核核骨架制备技术的研究

以扁绒泡菌显型原质团为材料,进行细胞核的提纯、核骨架的制备及电镜观察。结果表明：用2mol/L NaCl+TritonX-100/NP40可得到具有复合纤维的核骨架,而用Lis + TritonX-100/NP40得到的核骨架具有核纤层,不使用RNase得到了具有网状的细胞核骨架,RNA在核骨架的结构形态中起重要作用。阐述了原质团中游离细胞核在细胞生物学研究中的意义。     

Methyltransferase-inhibition interferes with neuronal differentiation of P19 embryonal carcinoma cells

We have analyzed the importance of substrate methylation by S-adenosylmethionine-dependent methyltransferases for neuronal differentiation of P19 embryonal carcinoma cells. We show that treatment of cells with methyltransferase inhibitor adenosine dialdehyde (AdOx) interferes with neuronal differentiation. Retinoic acid (RA) and AdOx co-treated cells had a decreased number of neurites and a flattened morphology compared with cells differentiated by RA. Also, the amount of neuronal class III tubulin (Tuj1) decreased from 76% to 9.6% with AdOx-treatment. Gene expression levels of wnt-1, brn-2, neuroD, and mash-1 were also down-regulated by AdOx-treatment. But AdOx-treatment did not up-regulate BMP-4 and GFAP genes. Treatment of RA decreased E-cadherin expression during neuronal differentiation. However, in AdOx/RA co-treated cells, E-cadherin expression was restored to the control level. Also, mRNA expression of N-cadherin decreased with AdOx-treatment. Taken together, these data show that methylation reactions might influence the cell-fate decision and neuronal differentiation of P19 cells.     

Opposing actions of Notch1 and VEGF in post-natal cardiac valve endothelial cells

The endothelium of the cardiac valves is unique compared the rest of the vasculature in its ability to undergo an endothelial-to-mesenchymal transformation (EMT) in vitro in response to transforming growth factor-β (TGF-β). EMT is a critical event during embryonic valve development, and both VEGF-A and Notch1 have been shown to function in this process. Here we investigate the effects of VEGF-A and Notch1 on EMT in clonal endothelial cell (EC) populations isolated from adult aortic valve leaflets. VEGF-A inhibited TGF-β-induced EMT. Endothelial growth, however, was not affected by VEGF-A or TGF-β. A positive role for Notch1 was revealed in three experiments: (1) TGF-β induced Notch1 mRNA in valve ECs, (2) a γ-secretase inhibitor of Notch1 signaling blocked EMT, and (3) overexpression of a ligand-independent form of Notch1 induced EMT. These results demonstrate, for the first time, that VEGF-A and Notch1 play opposing roles in regulating EMT in post-natal valve endothelium.     

Thalidomide inhibits epidermal growth factor-induced cell growth in mouse and human monocytic leukemia cells via Ras inactivation

The effect of thalidomide on epidermal growth factor (EGF)-induced cell growth was examined. Thalidomide inhibited EGF-induced cell growth in mouse and human monocytic leukemia cells, RAW 264.7, U937 and THP-1. Thalidomide inhibited EGF-induced phosphorylation of extracellular signal regulated kinase (ERK) 1/2, but not p38 and stress-activated protein kinase (SAPK)/JNK. The phosphorylation of MEK1/2 and Raf at Ser 338 as the upstream molecules of ERK 1/2 was also prevented by thalidomide. Further, it inhibited EGF-induced Ras activation through preventing the transition to GTP-bound active Ras. Thalidomide inhibited the Ras activation induced by lipopolysaccharide (LPS) and vascular endothelial growth factor (VEGF) as well as EGF. There was no significant difference in the expression and function of EGF receptor between thalidomide-treated and non-treated cells. Therefore, thalidomide was suggested to inhibit EGF-induced cell growth via inactivation of Ras.     

伊犁沙虎的骨骼系统研究

本文研究了伊犁沙虎（Teratoscincus scincus）的骨骼系统,对其各部分骨骼的形成、形态和位置作了详细的描述,为沙虎属的分类及演化研究提供骨骼方面的资料。