盘基网柄菌(Dictyostelium discoideum)细胞的分化及其调控

本文综述了盘基网柄菌(Dictyostelium dis-coideum)发育过程中细胞类型的诱导和分化,细胞外cAMP及其四种位于细胞表面的受体及PKA(蛋白激酶A)、GSK-3(糖原合成酶激酶)和STATa等在网柄菌发育过程中的作用。     

盘基网柄菌细胞分化及细胞比例的调控

本文综述了盘基网柄菌(Dictyosteliumdiscoideum)发育过程中调控细胞分化及细胞比例的一些信号分子,包括分化诱导因子(DIF-1、SDF-2)、糖原合成酶激酶(GSK-3)、环状亮氨酸拉链蛋白(rZIP)等,介绍了这些信号分子的功能及其作用机制。     

细胞色素c在盘基网柄菌细胞凋亡中的作用

细胞色素c在细胞凋亡中发挥着重要的作用,其作用机理在高等真核生物及低等真核生物酵母中已经比较清楚,但在盘基网柄菌(Dictyostelium discoideum)中的作用却没有相关报道.所以我们用western blot和实时荧光定量PCR的方法分别测定了盘基网柄菌前柄细胞和前孢子细胞中细胞色素c的含量及表达量的变化...     

gp150蛋白在盘基网柄菌发育中的作用及粘附分子间关系的分析

饥饿的盘基网柄菌进入多细胞发育期 ,在发育早期 ,AK12 7细胞 (gp150突变细胞 )能表达DdCAD 1和 gp80两种粘附分子 ,但它们不足以促进细胞继续发育 ,发育停留在细胞疏松结合阶段。粘附分子 gp150调节的细胞与细胞间的粘着影响了细胞丘“突出”的形成 ,由此影响了盘基网柄菌多细胞发育的形态发生。TL93细胞 (DdCAD 1和gp80突变细胞 )能完成发育。主要原因是在细胞流发育阶段就表达了gp150分子 ,在细胞粘着的功能上有替代DdCAD 1和 gp80的作用。因此 gp150蛋白对盘基网柄菌多细胞发育有着不可或缺的作用     

盘基网柄菌发育中的细胞粘附分子及其信号转导

在盘基网柄菌发育早期,DdCAD-1和csA调节了变形虫细胞间的粘着,调控该过程的机制类似于胚胎发育中上皮细胞层的闭合。完成网柄菌发育的一个必需分子是gpl50异嗜性粘附分子。盘基网柄菌β-连环蛋白同源物Aardvark(Aar)的缺乏使细胞间失去粘着连接,Aar也有信号转导功能,调控了前孢子细胞基因的表达。因此,细胞间的粘着是盘基网柄菌发育的一个重要组成部分,并与调控形态发生过程的信号转导有密切相互作用关系。     

新型重组糖蛋白表达载体--盘基网柄菌

近年来,盘基网柄菌作为异源重组糖蛋白表达载体的研究受到了学术界的重视,已经有多种具有生物活性的复杂糖蛋白成功地得到了表达。通过对表达产物的研究发现,盘基网柄菌具有各种翻译后加工机制,例如磷酸化、酰基化及形成GPI(糖基磷脂酰基醇)锚点等,具有类似于高等动物的糖基化修饰能力。与哺乳动物细胞表达载体相比较,盘基网柄菌具有培养成本低廉、细胞生长迅速及易于大规模培养的优势。盘基网柄菌有可能发展成为一种有重要应用前景的糖蛋白表达载体系统。     

盘基网柄菌——研究致病机制的宿主模型

盘基网柄菌作为致病菌宿主模型的研究主要有:筛选致病菌株及相应突变菌株毒性;鉴别对致病菌易感性和抗性的突变细胞宿主;宿主细胞的有效标记、已完成的基因组计划以及宿主细胞与致病菌间信号转导通路的相互作用;这些都表明盘基网柄菌是致病机制研究的理想宿主模型。     

PKA在盘基网柄菌(Dictyostelium discoideum)多细胞发育中的作用

在盘基网柄菌(Dictyosteliumdiscoideum)多细胞发育中,蛋白激酶A(proteinkinaseA,PKA)发挥多重作用.细胞聚集阶段,PKA调节腺苷酰环化酶的活性,中转cAMP,诱导dut、pdi等一些发育早期的基因表达;参与启动聚集后的细胞分化和形态构成,增强GBF活性,激活前孢子细胞特有基因的表达;它还精密调控前柄细胞特有基因ecmB的表达,准确启动拔顶发育,诱导孢柄和孢子的成熟.子实体形成后,PKA又是维持孢子休眠和保证孢子有效萌发的必需因子.在PKA调控下,盘基网柄菌有条不紊地完成整个发育过程.     

聚氨酯泡沫半固定化培养盘基网柄菌

研究了聚氨酯泡沫应用于固定化盘基网柄菌的可行性,发现以简单处理过的聚氨酯泡沫为载体,能够高效实现盘基网柄菌的固定化培养。考察了载体粒径大小、载体量和摇床转速等对固定化培养的影响,在优化的培养条件和固定化条件下,盘基网柄菌的最大细胞密度是悬浮培养的2~4倍。     

盘基网柄菌发育机制研究进展

盘基网柄菌细胞与哺乳类细胞在行为、结构和信号通路方面,有很多相同或相似之处。用其作为研究发育机制的模型,为解决哺乳类发育中的一些难题可以发挥重要作用。该文详细讨论了分化诱导因子DIF-1、环磷酸腺苷c AMP、转录因子Srf A和Stk A、粘附分子gp150在盘基网柄菌发育过程中的作用及机制。关于DIF-1、c AMP和gp150信号通路间相互关系还缺乏系统研究,值得进一步深入研究。对这类问题进行深入研究可能有助于阐明多细胞动物起源及演化机制。     

Studies on the accumulation of the differentiation-inducing factor (DIF) in high-cell-density monolayers of Dictyostelium discoideum

A number of factors that have been shown to influence cell type determination in Dictyostelium discoideum were assessed for their effects on the accumulation of the stalk cell differentiation-inducing factor (DIF) in high-cell-density monolayers of strain V12-M2. DIF accumulation is markedly enhanced by low pH, butyrate, and the proton pump inhibitor diethylstilbestrol (DES), conditions that induce stalk cell formation in low-cell-density monolayers in the absence of added DIF. These results are discussed in relation to a model for cell type determination recently proposed by (J.D. Gross, M.J. Peacey, and R. Pogge Von Strandmann (1988, Differentiation, 38: 91-98). DIF accumulates in high-cell-density monolayers after the cells have become independent of cyclic AMP for stalk cell formation. This accumulation is greatly enhanced by the addition of cyclic AMP. This result may explain why cyclic AMP stimulates stalk cell formation in low-density monolayers in the presence of suboptimal levels of DIF, following preincubation in the presence of saturating levels of cyclic AMP (L. Kwong, A. Sobolewski, and G. Weeks, 1988, Differentiation 37, 1-6). Adenosine has no effect on DIF accumulation in high-cell-density monolayers.     

Purification and characterization of a T lymphocyte-derived differentiation-inducing factor for human promyelocytic cell line (HL-60) and its relationship to lymphotoxin

The human T cell leukemia virus type I (HTLV-I)-transformed T lymphocyte cell line MT-2 constitutively produces differentiation-inducing factor (DIF) for the human promyelocytic leukemia cell line HL-60. Purification and characterization of DIF derived from MT-2 were performed here to identify T cell-derived DIF. DIF was purified from conditioned medium of the MT-2 cell culture with serum-free medium by utilizing the sequential chromatographies of anion-exchange (mono-Q) column, gel filtration (superose-12) column, and hydrophobic (phenyl 5PW) column and finally the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In the SDS-PAGE, one major band with a molecular weight of 56,000 and one minor band with 15,000 were stained with Coomassie Brilliant Blue and both showed DIF activity after extraction from gels. DIF had an isoelectric point of 5.8. In all purification steps, DIF activity for HL-60 and cytotoxic activity to the sublines of mouse L-929 were not able to be separated. Further, DIF was neutralized by antibody against lymphotoxin (LT) but not by antibody against tumor necrosis factor. These results indicate that the major DIF activity derived from MT-2 is LT.     

A slow sustained increase in cytosolic Ca2+ levels mediates stalk gene induction by differentiation inducing factor in Dictyostelium.

During Dictyostelium stalk cell differentiation, cells vacuolate, synthesize a cellulose cell wall and die. This process of programmed cell death is accompanied by expression of the prestalk gene ecmB and induced by the differentiation inducing factor DIF. Using cell lines expressing the recombinant Ca2+-sensitive photoprotein apoaequorin, we found that 100 nM DIF increases cytosolic Ca2+ ([Ca2+]i) levels from approximately 50 to 150 nM over a period of 8 h. The Ca2+-ATPase inhibitor 2,5-di(tert-butyl)-1,4-hydroquinone (BHQ) induced a similar increase in [Ca2+]i levels and induced expression of the prestalk gene ecmB to the same level as DIF. The [Ca2+]i increases induced by DIF and BHQ showed similar kinetics and preceded ecmB gene expression by approximately 1-2 h. The Ca2+ chelator 1,2-bis(o-aminophenoxy)-ethane-N,N,N'N'-tetra-acetic acid (BAPTA) efficiently inhibited the BHQ-induced [Ca2+]i increase and blocked DIF-induced expression of the ecmB gene. These data indicate that the effects of DIF on stalk gene expression are mediated by a sustained increase in [Ca2-]i. Sustained [Ca2+]i elevation mediates many forms of programmed cell death in vertebrates. The Dictyostelium system may be the earliest example of how this mechanism developed during early eukaryote evolution.     

Artificial compounds differentially control Dictyostelium chemotaxis and cell differentiation

Differentiation-inducing factor-1 and -2 (DIF-1 and DIF-2) are small lipophilic signal molecules that control both cell differentiation and chemotaxis in the cellular slime mold Dictyostelium discoideum. In this study, we examined the effects of four amide derivatives of DIF-1 on stalk cell differentiation and chemotaxis. The DIF derivatives differentially affected cell differentiation and chemotaxis, suggesting the possible existence of at least three receptors for DIFs: one receptor responsible for stalk cell induction, and two receptors responsible for chemotaxis modulation. Furthermore, our results indicate that DIF derivatives can be utilized to analyze the DIF-signaling pathways.     

Multiple effects of differentiation-inducing factor on prespore differentiation and cyclic-AMP signal transduction in Dictyostelium

Abstract. The effects of the differentiation inducing factor (DIF) on several cAMP-induced responses in Dictyostelium were investigated. It was found that DIF reduces the apparent affinity of cell-surface cAMP receptors. DIF does not affect the cAMP-induced cGMP response, but it is a potent inhibitor of the cAMP-relay response. DIF also inhibits the induction of prespore differentiation by cAMP in aggregation-competent cells. We also compared the effects of DIF on cAMP-induced responses with those of the relay inhibitor, caffeine, and the morphogen, adenosine.     

Multiple effects of differentiation-inducing factor on prespore differentiation and cyclic-AMP signal transduction in Dictyostelium

Abstract. The effects of the differentiation inducing factor (DIF) on several CAMP-induced responses in Dictyostelium were investigated. It was found that DIF reduces the apparent affinity of cell-surface cAMP receptors. DIF does not affect the CAMP-induced cGMP response, but it is a potent inhibitor of the CAMP-relay response. DIF also inhibits the induction of prespore differentiation by cAMP in aggregation-competent cells. We also compared the effects of DIF on CAMP-induced responses with those of the relay inhibitor, caffeine, and the morphogen, adenosine.     

昼夜温差对番茄生长发育、产量及果实品质的影响

在3间人工气候室精密受控环境中,保持日平均温度为22 ℃,设置昼夜温差分别为6 ℃(25 ℃/19 ℃)、8 ℃(26 ℃/18 ℃)、10 ℃(27 ℃/17 ℃),研究昼夜温差对番茄生长的影响.结果表明: 番茄不同品种、不同生长时期适宜的昼夜温差条件不同.番茄开花前,与6 ℃温差相比,8 ℃温差可显著加快野生种醋栗番茄LA1781生长发育,使幼苗株高增加23.1%,出叶加快1～2片,开花提前7 d;10 ℃温差对LA1781苗期的促进作用与8 ℃温差相似.对栽培种普通番茄LA2397和LA0490来说,6 ℃温差使幼苗生长良好,8 ℃温差对幼苗无显著促进作用;与6 ℃温差相比,10 ℃温差对苗期生长及开花有抑制作用,使株高降低12.0%～18.3%,出叶慢2～3片,开花推迟2～4 d.10 ℃温差使3个品种番茄地上部分干质量增加25.2%～44.2%.番茄开花后,与6 ℃温差相比,10 ℃温差可显著提高LA1781的产量和果实品质,使果实数增加34.7%,单株产量增加92.1%,平均单果质量增加40.0%,果实可溶性糖含量增加16.3%,番茄红素含量增加95.6%.与6 ℃温差相比,LA2397和LA0490在8 ℃温差下产量和果实品质提高,番茄红素含量增加超过2倍;在10 ℃温差下产量略有降低(5.0%),果实含糖量降低,但果实尺寸和番茄红素含量增加.表明番茄苗期生长温差不宜过大,花果期适当增大昼夜温差可提高产量和果实品质,但温差过大易造成生长不良和减产.     

The effect of ammonia on stalk cell formation in submerged monolayers of Dictyostelium discoideum

It has been shown that ammonia inhibits stalk cell formation in monolayers of V12M2, and it was suggested that this inhibition was due to an antagonism of the differentiation-inducing factor (DIF) (Gross, J.D. et al., Nature, 303, 244-246, 1983). However, the results presented here indicate that ammonia inhibition is independent of DIF concentration, and that it occurs well in advance of the period of DIF requirement. Ammonia completely inhibits DIF accumulation and inhibits stalk cell differentiation, but there is no inhibition of prespore cell formation. These results imply the existence of an early ammonia-sensitive event that influences terminal cell type differentiation.