细胞外基质在植物发育中的作用

植物细胞壁是由纤维素和果胶交联的多糖和蛋白质构成的既彼此独立,又相互作用的三维动力学网络。和动物的细胞外基质一样,植物细胞壁中的许多成分积极地参与植物细胞发育过程的调节,它们以某种方式将信息传递给细胞,调节细胞的行为,以便对各种外界环境作出相应的反应。因此细胞壁不再是一种环绕植物细胞的惰性结构,比起细胞壁,植物细胞外基质这一名词更能反映出这一动力学的特性。     

植物的细胞壁

绝大多数植物细胞的质膜外都有细胞壁,这是区别予动物细胞的显著特征之一。由于细胞壁的存在,使原生质体的膨胀受到限制,细胞成熟后,使其形态和大小变为固定。细胞壁有保护作用,厚而硬的细胞壁还有支持植物器官的机械作用,同时,细胞壁能影响植物组织的吸收、蒸腾、运输和分泌等功能。     

细胞壁与细胞的发育

细胞壁除了起着机械支持、参与物质运输和防御反应等功能外,与细胞的发育密切相关。它们可作为信号分子,促进植物细胞的分裂增殖,决定细胞的分化方向,参与细胞识别过程等。概述了近年来细胞壁调节细胞发育的新进展。     

水分亏缺下细胞延伸生长与细胞膨压和细胞壁特性的关系

在简要介绍植物细胞延伸生长的生物物理模型的基础上,综述了水分亏缺下植物细胞延伸生长与细胞膨压、细胞壁伸展性和细胞壁塑变阈值的关系,阐述了植物细胞壁调节在作物抗旱性中的作用。     

植物细胞壁结构及成像技术研究进展

植物细胞壁作为细胞外复杂交联网络,为植物细胞生长、发育以及适应环境变化提供机械支撑,具有调节植物形态、抵抗胁迫、运输水分等功能。除此之外,植物光合作用积累的生物质大部分贮藏在细胞壁中,因此,研究细胞壁的成分和纳微结构对更好的利用植物能源具有重要意义。植物细胞壁的结构研究是当今植物界研究的前沿热点之一。随着新型成像技术的发展,近年来关于细胞壁成分和纳微结构的研究取得了阶段性的进展。本文就植物细胞壁的成分、结构、成像技术和力学性质进行了总结与展望,以期为植物细胞壁的相关研究提供新思路。     

基质金属蛋白酶与心肌重塑

细胞外基质参与和促进了心肌重塑的过程,基质金属蛋白酶是调节细胞外基质重要的酶,基质金属蛋白酶在心肌重塑过程表达变化可分为三个时相,其活性受到信号传导途径、炎症因子和活性氧/活性氮的调节,基质金属蛋白酶可能作为心肌梗塞等疾病治疗的靶标     

微丝骨架和活性氧在调节气孔运动中的作用及机制

气孔运动调节植物的光合作用和蒸腾作用,对植物的生长发育和干旱等非生物胁迫的响应都起到重要的作用。保卫细胞能够通过感知胞内和胞外多种信号调节气孔开度,因此,保卫细胞已经成为植物细胞信号转导研究中广泛应用的细胞模型。该文对保卫细胞中微丝骨架和活性氧对气孔运动的调节作用、微丝骨架在调节细胞壁与质膜间联系中的作用进行了综述,最后分析了微丝骨架通过ROS(reactive oxygen species)调节保卫细胞壁–质膜联系参与气孔运动调控的可能机制。     

植物细胞壁研究进展

植物细胞壁是一种复杂的网状结构,其成分包含纤维素、半纤维素、果胶和少量的结构蛋白等。在植物细胞生长过程中,细胞能产生伸展素蛋白,打断纤维素和半纤维素之间的氢键,引起细胞膨压驱动的细胞壁扩张。成熟细胞壁扩张性的丧失是由于细胞壁硬化作用而对扩张性蛋白的作用不敏感造成的,细胞壁成熟过程中很多不同的连接会同时发生,当细胞壁基质多聚体分子之间的连接增加到一定的程度。细胞壁的伸长就会被完全抑制。     

白芷细胞外钙调素结合蛋白（ECBP21）免疫组化及金标定位分析

ECBP21是我们从白芷悬浮培养细胞外纯化及cDNA克隆的一种钙调素结合蛋白(CaMBP),亦是植物中首次报道的细胞外CaMBP。本实验以大肠杆菌表达的重组ECBP21蛋白为抗原,制备了高效价特异性抗体;随后,利用免疫组化及金标定位技术,研究了ECBP21蛋白组织特异性分布及亚细胞定位。免疫组化结果表明：ECBP21在白芷各组织中均有分布,但在叶、花、花序轴中较多,而在根中较少,并且ECBP21在细胞中多分布于细胞壁区域;免疫胶体金电镜定位结果显示：在白芷花序轴细胞中金颗粒主要分布在细胞壁,表明ECBP21蛋白主要定位于细胞壁区域,从而首次为细胞外CaMBP(ECBP21)的胞外存在提供了直观证据,并为进一步研究其在植物生长发育中的功能提供了初步信息。     

TIMP-3表达下调促进前脂肪细胞分化

细胞与细胞外基质的相互作用以及细胞外基质的重构在脂肪组织的形成过程中发挥了重要作用。细胞外基质的这一系列变化是由细胞分泌的蛋白酶及其抑制物调控的,其中基质金属蛋白酶(MMPs)是一类调控细胞外基质分解的蛋白酶家族。MMPs的活性受其四种组织抑制物调节,即TIMP1-4。以前对TIMP在脂     

Cell separation with horizontal cell electrophoresis under near-isopycnic conditions on a “density cushion”

This report describes an improvement made to the horizontal cell electrophoresis methodology. It involves using two liquid layers differing in density to produce an interface described as a "density cushion". The electrophoretic system that employed an anti-convective porous matrix to separate red blood cells (RBC) and charged dyes effectively was found to be unsuitable for some other mammalian cells. The "density cushion" method was found to be more versatile and applicable to studies on the separation of a variety of cell types. The experiments described show the differences between the electrophoretic mobilities of a human eosinophilic leukaemia cell line (Eol-1) and RBC, both with and without the modification of the cell surface properties.     

Polarization and Migration of Hematopoietic Stem and Progenitor Cells Rely on the RhoA/ROCK I Pathway and an Active Reorganization of the Microtubule Network

Understanding the physiological migration of hematopoietic progenitors is important, not only for basic stem cell research, but also in view of their therapeutic relevance. Here, we investigated the role of the Rho kinase pathway in the morphology and migration of hematopoietic progenitors using an ex vivo co-culture consisting of human primary CD34⁺ progenitors and mesenchymal stromal cells. The addition of the Rho kinase inhibitor Y-27632 led to the abolishment of the uropod and microvillar-like structures of hematopoietic progenitors, concomitant with a redistribution of proteins found therein (prominin-1 and ezrin). Y-27632-treated cells displayed a deficiency in migration. Time-lapse video microscopy revealed impairment of the rear pole retraction. Interestingly, the knockdown of ROCK I, but not ROCK II, using RNA interference (RNAi) was sufficient to cause the referred morphological and migrational changes. Unexpectedly, the addition of nocodazole to either Y-27632- or ROCK I RNAi-treated cells could restore their polarized morphology and migration suggesting an active role for the microtubule network in tail retraction. Finally, we could demonstrate using RNAi that RhoA, the upstream regulator of ROCK, is involved in these processes. Collectively, our data provide new insights regarding the role of RhoA/ROCK I and the microtubules in the migration of stem cells.     

Cell electrophoresis — a method for cell separation and research into cell surface properties

In this paper, we discuss the application of various methods of cell electrophoresis in research into cell surface properties (analytical methods), and the separation of uniform cell subpopulations from cell mixtures (preparative methods). The emphasis is on the prospects of the development of simplified and versatile methodologies, i.e. microcapillary cell electrophoresis and horizontal cell electrophoresis under near-isopycnic conditions. New perspectives are considered on the use of analytical and preparative cell electrophoresis in research on cell differentiation, neoplastic transformation, cell-cell interactions and the biology of stem cells. Paper authored by participants of the international conference: XXXIV Winter School of the Faculty of Biochemistry, Biophysics and Biotechnology of Jagiellonian University, Zakopane, March 7–11, 2007, “The Cell and Its Environment”. Publication cost was covered by the organisers of this meeting.     

Expanisns

Biochemical dissection of the “acid-growth” process of plant cell walls led to the isolation of a new class of wall loosening proteins, called expansins. These proteins affect the rheology of growing walls by permitting the microfibril-matrix network to slide, thereby enabling the wall to expand. Molecular sequence analysis suggests that expansins might have a cryptic glycosyl transferase activity, but biochemical results suggest that expansins disrupt noncovalent bonding between microfibrils and the matrix. Recent discoveries of a new expansin family and gene expression in fruit, meristerms and cotton fibers have enlarged our view of the developmental functions of this group of wall loosening proteins.     

酸性磷酸酶法检测体外培养细胞数

利用小鼠成纤维细胞系(NIH3T3)、小鼠骨髓瘤细胞系(SP2/0)、人大肠癌细胞系(LO-VO)和人白血病细胞系(K562),评价酸性磷酸酶(APA)法用于检测体外各类型细胞的增殖和杀伤作用。用直线回归分析光吸收度与每孔活细胞数的关系。结果表明,APA法能准确地反映检测的活细胞数(相关系数均>0.99)。本方法不仅能很好地检测表皮生长因子对细胞的增殖作用,也能够检测顺铂对体外细胞的杀伤作用。结果表明APA法简单、灵敏,可以用于上皮和间质等贴壁和悬浮生长的细胞计数。     

An overview of the KIN1/PAR-1/MARK kinase family

Members of the KIN1/PAR-1/MARK kinase family are conserved from yeast to humans and share a similar primary structural organization. Several kinases of this family appear to be at the crossroads of various biological functions including cell polarity, cell cycle control, intracellular signalisation, microtubules stability and protein stability. Here we present an overview of known roles of KIN1/PAR-1/MARK kinases including pEg3 a newly identified member which is regulated during the cell cycle and is a potential regulator of the cell cycle progression. Some common modes of action can be deciphered for this protein kinase family.     

犬皮肤成纤维细胞的分离、培养及鉴定

目的探索和建立适用于犬皮肤成纤维细胞的体外分离、培养及鉴定的技术方法。方法采用组织贴块培养法和胰蛋白酶、胶原酶Ⅰ联合消化法对犬皮肤成纤维细胞进行体外培养、传代。并对所培养的细胞进行倒置显微镜观察和苏木素-伊红染色,观察成纤维细胞形态,并对培养细胞行波形蛋白免疫荧光染色。结果倒置相差显微镜下可见长梭形细胞生长,苏木素-伊红染色可见细胞呈漩涡状、平行排列,第5代细胞免疫荧光检测波形蛋白(vimentin)表达阳性。结论建立了高效快速分离和稳定培养成纤维细胞的方法,为诱导犬心房纤维化提供了充足的种子细胞。     

The plant cell body: a cytoskeletal tool for cellular development and morphogenesis

Summary Certain aspects of cellular behaviour in relation to growth and development of plants can be understood in terms of the cell body concept proposed by Daniel Mazia in 1993. During the interphase of the mitotic cell cycle, the plant cell body is held to consist of a nucleus and a perinuclear microtubule-organizing centre from which microtubules radiate into the cytoplasm. During mitosis and cytokinesis in meristematic cells, and also during the period of growth in post-mitotic cells immediately beyond the meristem, the plant cell body undergoes various characteristic morphological transformations, many of which are proposed as being related to changing structural connections with the actin-based component of the cytoskeleton and with specialized, plasma-membrane-associated sites at the cell periphery. In post-mitotic cells, these transformations of the plant cell body coincide with, and probably provide conditions for, the various pathways of development which such cells follow. They are also responsible, for the acquisition of new cellular polarities. Events in which the plant cell body participates include the formation of a mitotic spindle, phragmoplast, and new cell division wall, the rearrangement of a diffuse type of cell wall growth into tip growth (as occurs, e.g., during the initiation and subsequent development of root hairs), and the growth and division that occurs in reactivated vacuolate cells. If more evidence can be marshalled in support of the existence and properties of the plant cell body, then this concept could prove useful in interpreting the cytological bases of a range of developmental events in plants.Abbreviations CMT cortical microtubule - EMT endoplasmic microtubule - ER endoplasmic reticulum - MF microfilament - MT microtubule - MTOC microtubule-organizing centre - PPB preprophase band (of microtubules) - QC quiescent centre - VSC vesicle supply centre     

A Scalable Approach to Prevent Teratoma Formation of Human Embryonic Stem Cells

As the renewable source of all cell types in the body, human embryonic stem cells (hESCs) hold great promise for human cell therapy. However, one major bottleneck that hinders the clinic application of hESCs is that hESCs remaining with their differentiated derivatives pose cancer risk by forming teratomas after transplantation. NANOG is a critical pluripotency factor specifically expressed in hESCs but rarely in their differentiated derivatives. By introducing a hyperactive variant of herpes simplex virus thymidine kinase gene into the 3′-untranslated region of the endogenous NANOG gene of hESCs through homologous recombination, we developed a safe and highly scalable approach to efficiently eliminate the teratoma risk associated with hESCs without apparent negative impact on their differentiated cell types. As thymidine kinase is widely used in human gene therapy trials and is the therapeutic target of U. S. Food and Drug Administration-approved drugs, our strategy could be effectively applied to the clinic development of hESC-based human cell therapy.     

Molecular basis for T cell response induced by altered peptide ligand of type II collagen

Mounting evidence from animal models has demonstrated that alterations in peptide-MHC interactions with the T cell receptor (TCR) can lead to dramatically different T cell outcomes. We have developed an altered peptide ligand of type II collagen, referred to as A9, which differentially regulates TCR signaling in murine T cells leading to suppression of arthritis in the experimental model of collagen-induced arthritis. This study delineates the T cell signaling pathway used by T cells stimulated by the A9·I-A(q) complex. We have found that T cells activated by A9 bypass the requirement for Zap-70 and CD3-ζ and signal via FcRγ and Syk. Using collagen-specific T cell hybridomas engineered to overexpress either Syk, Zap-70, TCR-FcRγ, or CD3-ζ, we demonstrate that A9·I-A(q) preferentially activates FcRγ/Syk but not CD3-ζ/Zap-70. Moreover, a genetic absence of Syk or FcRγ significantly reduces the altered peptide ligand induction of the nuclear factor GATA3. By dissecting the molecular mechanism of A9-induced T cell signaling we have defined a new alternate pathway that is dependent upon FcRγ and Syk to secrete immunoregulatory cytokines. Given the interest in using Syk inhibitors to treat patients with rheumatoid arthritis, understanding this pathway may be critical for the proper application of this therapy.