黄瓜种子萌发过程中Ca2+分布的变化

采用细胞化学方法,研究了黄瓜种子中贮藏Ca2+的分布特点及其在萌发过程中的变化动态.干种子的子叶细胞中贮藏有大量的蛋白体、油脂体,Ca2+沉淀颗粒大量分布于胞质、胞间隙以及细胞质膜上.大多数蛋白体中有1至数个圆球形或椭圆体形含Ca2+的球状晶体.相比之下,胚芽和胚根细胞中Ca2+较少.种子萌发早期,子叶中的贮藏钙及晶体溶解释放出的Ca2+部分转运到生长发育中的胚芽和胚根中.随着萌发的继续,胚根和胚芽细胞中的Ca2+不会持续增多,反而下降.     

铝胁迫下小麦根尖分生细胞中Ca2+分布变化

运用透射电镜细胞化学方法对铝胁迫下小麦根尖分生细胞中Ca2+分布的变化进行了观察,在正常生长条件下,Ca2+广泛分布于细胞质、细胞核、细胞间隙中,特别是液泡中有大量的Ca2+沉淀颗粒;在Al3+胁迫条件下,细胞质、细胞核中Ca2+沉淀颗粒明显减少,分布发生改变,细胞质中液泡增多,但其中Ca2+沉淀颗粒明显减少.结果表明,Al3+不但抑制了根尖细胞对Ca2+的吸收,而且引起细胞中原有Ca2+分布的变化,这很可能引起细胞功能的紊乱,进而影响根系的生长.     

脱硫废弃物对碱胁迫下水稻叶片钙分布、Ca2+-ATPase活性及抗氧化特征的影响

为了探讨脱硫废弃物提高水稻抗盐碱的作用机制,采用盆栽法,研究脱硫废弃物对碱胁迫下水稻幼苗叶片总钙含量、Ca2+分布、细胞膜Ca2+-ATPase活性及活性氧含量等的变化.结果表明:对照处理的细胞中钙颗粒零星分布于细胞壁和叶绿体中,添加脱硫废弃物和CaSO4处理的细胞质膜、细胞间隙、细胞壁和液泡中有大量的钙颗粒分布;随着脱硫废弃物和CaSO4添加量的增加,叶片总钙含量增加,质膜和液泡膜Ca2+-ATPase活性呈上升趋势,质膜透性、MDA含量和活性氧O2-产生速率呈下降趋势,SOD、POD等保护酶活性升高.添加脱硫废弃物在一定程度上能够减缓碱胁迫对水稻造成的细胞伤害,起主要作用的物质可能是其主要成分CaSO4.     

温度逆境交叉适应对葡萄叶片膜脂过氧化和细胞钙分布的影响

 研究了高温锻炼对低温胁迫下和低温锻炼对高温胁迫下葡萄(Vitis vinifera)叶片中丙二醛（MDA）、谷胱甘肽(GSH)和抗坏血酸(AsA)含量变化以及细胞中Ca2+分布的影响。结果表明: 高（低）温胁迫使正常生长的叶片丙二醛含量升高, GSH和AsA含量下降,低（高）温锻炼预处理能减少MDA含量,提高GSH和AsA含量,抑制了由于温度胁迫引起MDA含量升高和GSH和AsA下降趋势。常温下葡萄叶肉细胞的Ca2+主要分布于液泡、细胞间隙中;高温胁迫和低温胁迫后,细胞质中聚集大量Ca2+沉淀颗粒,液泡中和细胞间隙Ca2+沉淀颗粒减少,叶绿体超微结构被破坏,Ca2+稳态平衡遭到破坏。高温锻炼后细胞质出现大量的Ca2+沉淀颗粒,主要来源于细胞间隙,低温锻炼后细胞质也出现大量的Ca2+沉淀颗粒,主要来源于液泡,两者的叶绿体超微结构都完整;高温锻炼的叶片经过低温胁迫和低温锻炼的叶片经过高温胁迫后,细胞间隙和液泡内Ca2+沉淀颗粒增加,细胞质中Ca2+沉淀颗粒很少,叶绿体较完整,Ca2+稳态平衡得以维持。推测高低温锻炼能够通过Ca2+启动抗逆基因表达和维持细胞中Ca2+稳态平衡来交叉适应低高温的胁迫。     

荔枝雌蕊发育过程中钙分布变化与细胞程序性死亡

应用焦锑酸钾沉淀法研究了荔枝雌花和雄花雌蕊发育过程中钙的分布变化。在大孢子母细胞阶段,雌花近珠孔内珠被细胞和花柱细胞的钙沉淀颗粒主要分布在细胞壁和细胞间隙,少部分在液泡;雌花花柱维管细胞中含有很多的钙沉淀颗粒;在雄花的近珠孔内珠被细胞钙沉淀颗粒大多在液泡中;雄花花柱细胞和维管细胞中钙沉淀颗粒很少。大孢子母细胞减数分裂后,雌花雌蕊继续发育,雄花雌蕊败育。雌花维管中的钙沉淀颗粒数量减少,可能被转运到将要发生花粉萌发和受精的部位。雌花近珠孔内珠被细胞壁的钙沉淀颗粒分布增加,花柱细胞从上（近柱头）到下（近子房）钙沉淀颗粒量递增。雄花近珠孔内珠被细胞发生程序性死亡：液泡中的钙进入细胞核启动细胞程序性死亡,核周隙与质膜腔形成连续的通道,钙在核与细胞质之间的流动不受限制;在特定的时间段,钙沉淀颗粒出现在线粒体、过氧化物体和线型内质网的外膜上。钙在细胞中重新分布可能触发和调节细胞程序性死亡的进程。缺乏钙沉淀颗粒的雄花花柱细胞迅速解体。     

脱硫废弃物对碱胁迫下水稻叶片钙分布、Ca2+-ATPase活性及抗氧化特征的影响

为了探讨脱硫废弃物提高水稻抗盐碱的作用机制,采用盆栽法,研究脱硫废弃物对碱胁迫下水稻幼苗叶片总钙含量、Ca2+分布、细胞膜Ca2+-ATPase活性及活性氧含量等的变化.结果表明： 对照处理的细胞中钙颗粒零星分布于细胞壁和叶绿体中,添加脱硫废弃物和CaSO4处理的细胞质膜、细胞间隙、细胞壁和液泡中有大量的钙颗粒分布;随着脱硫废弃物和CaSO4添加量的增加,叶片总钙含量增加,质膜和液泡膜Ca2+-ATPase活性呈上升趋势,质膜透性、MDA含量和活性氧O2〖SX(B-*3〗-〖〗·〖SX)〗产生速率呈下降趋势,SOD、POD等保护酶活性升高.添加脱硫废弃物在一定程度上能够减缓碱胁迫对水稻造成的细胞伤害,起主要作用的物质可能是其主要成分CaSO4.     

灵武长枣果实发育过程中阿拉伯半乳糖蛋白组织化学分布

为揭示灵武长枣（Ziziphus jujuba ‘Lingwu Changzao’）果实发育过程中阿拉伯半乳糖蛋白（AGPs）的分布规律,以膨大前期、快速膨大期、着色期和完熟期灵武长枣果实为试验材料,通过组织化学和免疫荧光定位的方法,研究了不同发育时期果实AGPs的分布特征。结果显示：βGlcY-AGPs形成的棕红色沉淀和MAC204抗体识别的抗原在各时期果实的外果皮及相邻的内部数层排列紧密的中果皮小细胞的细胞壁和细胞内部均有分布。中果皮大型卵圆形薄壁细胞的细胞壁和细胞内在膨大前期均有βGlcY-AGPs形成的棕红色沉淀和MAC204抗体识别的抗原分布,而在快速膨大期、着色期和完熟期均主要分布于薄壁细胞的细胞壁上,大部分细胞内部无分布;随着果实发育成熟,中果皮薄壁细胞间隙拉大排列更加松散,出现细胞破裂,βGlcY-AGPs形成的棕红色沉淀和MAC204抗体识别的抗原分布逐渐减少。各时期果实维管束中维管束鞘、木质部、韧皮部、形成层的所有细胞的细胞壁和细胞内部都分布有βGlcY-AGPs形成的棕红色沉淀和MAC204抗体识别的抗原,维管束数量和大小随果实发育及体积的进一步增大逐渐减少,βG...     

草酸处理对热胁迫下辣椒叶片膜透性和钙分布的影响

研究了外源草酸对热胁迫下辣椒叶片中细胞膜相对透性、谷胱甘肽(GSH)和抗坏血酸(AsA)含量变化以及Ca2+分布的影响.结果表明热胁迫使叶肉细胞膜相对透性升高,草酸处理则减轻升高幅度.热胁迫使叶片中GSH和AsA含量下降;草酸处理则使二者在热胁迫下含量下降幅度较小.常温下辣椒叶肉细胞的焦锑酸钙沉淀颗粒主要分布于液泡、胞间隙和叶绿体中,热胁迫下液泡、细胞间隙中减少,但在细胞核和细胞质中出现;经过草酸处理的叶肉细胞,焦锑酸钙沉淀颗粒在胞间隙中明显增多,液泡中减少.     

黄瓜种子萌发过程中Ca~(2 )分布的变化

采用细胞化学方法 ,研究了黄瓜种子中贮藏Ca2 的分布特点及其在萌发过程中的变化动态。干种子的子叶细胞中贮藏有大量的蛋白体、油脂体 ,Ca2 沉淀颗粒大量分布于胞质、胞间隙以及细胞质膜上。大多数蛋白体中有 1至数个圆球形或椭圆体形含Ca2 的球状晶体。相比之下 ,胚芽和胚根细胞中Ca2 较少。种子萌发早期 ,子叶中的贮藏钙及晶体溶解释放出的Ca2 部分转运到生长发育中的胚芽和胚根中。随着萌发的继续 ,胚根和胚芽细胞中的Ca2 不会持续增多 ,反而下降     

钙对亚适温弱光下黄瓜幼苗Ca2+分布及叶绿素荧光参数的影响

以'津优3号'黄瓜幼苗为试验材料,采用焦锑酸钙沉淀的电镜细胞化学方法,研究了CaCl2预处理对亚适温(昼/夜18℃/12℃)弱光(100 μmol·m-2·s-1)下黄瓜幼叶细胞中Ca2+分布、Ca2+-ATP酶活性及叶绿素荧光参数的影响.结果显示:正常温光条件(CK)下,黄瓜幼叶细胞Ca2+主要存在于液泡和液泡膜上,细胞质中含量较低;经亚适温和弱光处理7 d后,叶片细胞质和细胞膜中形成较大的钙沉淀颗粒,液泡中的Ca2+颗粒聚集成团,膜组织边缘模糊,Ca2+-ATPase活性降低;胁迫前用CaCl2预处理的细胞质中Ca2+颗粒略有增加,且分布较均匀,膜组织完整,Ca2+-ATPase活性与CK差异不显著;而经LaCl3、EGTA和CPZ预处理的Ca2+多呈大颗粒状聚积在细胞质、液泡膜或细胞壁上,Ca2+-ATPase活性大幅度下降.在7 d亚适温弱光处理后,各处理黄瓜叶片的Fv/Fm变化不大,而ΦPSⅡ、qP和ETR显著降低;与水预处理相比,叶片ΦPSⅡ、qP和ETR在CaCl2处理下显著增加,而在EGTA和CPZ处理下显著减小,LaCl3处理的无显著变化.研究表明,亚适温弱光处理能打破黄瓜幼苗细胞内的Ca2+平衡,使其膜组织受到一定程度破坏;CaCl2可维持胞内较高的Ca2+-ATP酶活性,保持Ca2+平衡,保护细胞膜组织结构完整,并参与了光合作用光能捕获和光合效率的调控,能有效减轻亚适温弱光对黄瓜幼苗光合作用的不良影响.     

外源Ca~(2 )预处理对高温胁迫下辣椒叶片细胞膜透性和GSH、AsA含量及Ca~(2 )分布的影响

以辣椒 (Capsicum annuum)幼苗的叶片为材料 ,研究了外源 Ca2 预处理对热胁迫下细胞质膜透性和谷胱甘肽 (GSH)、抗坏血酸 (As A)含量变化及 Ca2 分布的影响。结果表明 :外源 Ca2 预处理能减轻热胁迫引起的细胞膜破坏 ,能够减少叶片中 GSH和 As A的破坏。热胁迫后 ,Ca2 具有从胞外转运到胞质内和叶绿体中的趋势 ;外施Ca2 预处理能够明显增加细胞间隙、液泡和叶绿体中的 Ca2 颗粒密度 ,能够稳定热胁迫下叶肉细胞膜和叶绿体的超微结构。结果表明 ,外施 Ca2 预处理可能通过改变细胞内外的 Ca2 分布 ,减轻热胁迫对叶肉细胞的伤害     

Changes of the Level of Ca2+ in Cells of Rice Seedlings Under Low Temperature Stress

The changes of Ca2+ localization in ceils of rice (Oryza sativa L. ) seedlings under chilling stress were investigated with calcium antimonate precipitate-electromicroscopic-cyto- chemical methods. When rice seedlings grew at the optimum temperature, it was shown that the deposits of calcium antimonate, being the indicator for Ca2+ localization, mainly concen-trated within the vacuoles and intercellular spaces, and that there was also some Caz+ deposits in plastid, mitochondria, cytoplasm and nucleus. This indicates that under the normal condition, the vacuoles are the main pool of Ca2+ in plant cells, and that there is quite an amount of Ca2+ in the intercellular spaces. On the contrary, the free Ca2+ in cytoplasm and nucleus is very low under the normal condition. When the rice seedlings were treated at the temperature of 1 ℃ for 24 h, there nearly appeared a ring of well arranged Ca2+ precipitates in the inner side of plasmalemma. Meanwhile, the level of Ca2+ in cytoplasm and nucleus increased considerably. When the chilling stress of 1 ℃ continued for 48 h, a great amount of Ca2+ distributed within the cytoplasm and nucleus, and there was also a large quantity of Ca2+ deposits on vacuolar membranes and envelope of plastid. However, the ultrastructures of the cells remained normal. Based on the above observations, the authors proposed that the increase of Ca2+ in cytoplasm and nucleus under chilling stress might be related to the ulterior changes of physiological-biochemical processes.     

Ultrastructural study of the Ca2+-dependent ATPase activity in rat adenohypophyseal cells

Ca-dependent ATPase activity in the rat anterior pituitary was demonstrated in 50-microns tissue slices of aldehyde-fixed tissue with the medium of Takano et al. (Cell Tissue Res. 243:91. 1986).--The outer surface of the plasma membrane of the parenchymal as well as the folliculo-stellate cells was lined with lead precipitate. The reaction deposit was particularly well localized in intercellular spaces both between two parenchymal cells, and between a parenchymal and a folliculo-stellate cell. A fine reaction deposit was also seen in the endoplasmic reticulum and Golgi apparatus of some parenchymal cells. Elimination of Ca2+ from the tissue and the substrate medium drastically reduced the amount of reaction product. If ATP was omitted or replaced by sodium beta-glycerophosphate, no reaction product was seen. Changing the Ca2+ concentration or addition of Mg2+ to the standard medium caused a decrease in reaction intensity. Substitution of Mg2+ for Ca2+ resulted, again in well-localized lead deposition which we attribute to the activity of another enzyme. We suggest that the activity we described in the membrane of glandular cells may correspond to the enzyme involved in the long-term regulation of intracellular Ca2+ level.     

Ultracytochemical localization of Ca2+ during the phloem ganglion development in Phyllostachys edulis

Ultracytochemical localization of Ca2+ was investigated using the potassium pyroantimonate precipitation method during the development of phloem ganglion.The result showed that Ca2+ was mainly localized in the cell wall and intercellular spaces in the initiating phase.With the development of the phloem ganglion,the distribution of Ca2+ transferred to the vacuole,and the Ca2+ deposits in the cell wall and intercellular space decreased.At the later stage of the developmental phase.Ca2+ was distributed in the tonoplast and vacuole phagocytosis,and the vacuole became the main calcium storage in this phase.At the early stage of maturation of the phloem ganglion,most of the phloem ganglion cells'vacuoles cracked,and the cytoplastic Ca2+ content increased in large number.In the mature phloem ganglion,not only were there a few Ca2+ localized in the cytoplast of mature cells,but also in the differentiating cells in the vacuoles.Ca2+ was distributed in the tonoplast and vacuole contents;initiating cells almost had no Ca2+.In general,Ca2+ concentration in mature phloem ganglion cells was at a low level.The results indicated that the changes in Ca2+ distribution evoked the phloem ganglion generation,and Ca2+ regulated the physiological function of the phloem ganglion.     

低温胁迫下董棕（Garyota urens L.）幼苗叶肉细胞内Ca2+水平及细胞超微结构的变化

用焦锑酸钙沉淀的电镜细胞化学方法,研究了低温胁迫下董棕（Garyota urensL.） 幼苗叶肉细胞内Ca2+水平的变化。研究结果表明,未经低温处理的董棕幼苗叶肉细胞,焦锑 酸钙沉淀颗粒大量出现在液泡和细胞间隙中,细胞壁中也可见少量沉淀,而细胞基质中则看 不到焦锑酸钙沉淀;经2 ℃ 48 h低温处理后,细胞基质和细胞膜上焦锑酸钙沉淀增加,而液泡和细胞间隙中的焦锑酸钙沉淀则显著减少,并且超微结构已初步显示出寒害的特征,叶绿体外膜部分破损,类囊体片层稀疏且排列不规则,光合速率明显下降等;经2℃ 120 h低温处理后,细胞间隙内的焦锑酸钙沉淀极少,有的也紧贴在细胞外壁上,而细胞基质和细胞膜上则分布有非常多的焦锑酸钙沉淀,在核基质和液泡中也可见到少量的焦锑酸钙沉淀,并且超微结构遭到了显著破坏,叶绿体结构完全被破坏,核膜与液泡膜严重破损,内部结构模糊,细胞只表现为呼吸作用,不进行光合作用。表明Ca2+的区域性分布的变化与植物抗寒性存在一定关系。     

Coordinated intercellular calcium waves induced by noradrenaline in rat hepatocytes: dual control by gap junction permeability and agonist.

Calcium-mobilizing agonists induce intracellular Ca2+ concentration ([Ca2+]i) changes thought to trigger cellular responses. In connected cells, rises in [Ca2+]i can propagate from cell to cell as intercellular Ca2+ waves, the mechanisms of which are not elucidated. Using fura2-loaded rat hepatocytes, we studied the mechanisms controlling coordination and intercellular propagation of noradrenaline-induced Ca2+ signals. Gap junction blockade with 18 alpha-glycyrrhetinic acid resulted in a loss of coordination between connected cells. We found that second messengers and [Ca2+]i rises in one hepatocyte cannot trigger Ca2+ responses in connected cells, suggesting that diffusion across gap junctions, while required for coordination, is not sufficient by itself for the propagation of intercellular Ca2+ waves. In addition, our experiments revealed functional differences between noradrenaline-induced Ca2+ signals in connected hepatocytes. These results demonstrate that intercellular Ca2+ signals in multicellular systems of rat hepatocytes are propagated and highly organized through complex mechanisms involving at least three factors. First, gap junction coupling ensures coordination of [Ca2+]i oscillations between the different cells; second, the presence of hormone at each hepatocyte is required for cell-cell Ca2+ signal propagation; and third, functional differences between adjacent connected hepatocytes could allow a 'pacemaker-like' intercellular spread of Ca2+ waves.     

Ion activities in the lateral intercellular spaces of gallbladder epithelium transporting at low external osmolarities

The ion activities in the lateral spaces of the unilateral preparation of the gallbladder of Rana catesbiana were measured by double-barrelled ion-selective microelectrodes. The bladders were bathed in a saline solution with a low osmolarity (62 mOsm) containing, in mM: 27 Na+, 27 Cl-, 2 K+, 1 Ca++, 4 HCO3-. Working at reduced osmolarities had the advantage of an increased volume transport and of widened intercellular spaces. The reference barrel recorded an electrical potential of +2.7 mV in the spaces; they contained a solution similar to the external solution. The electrodes recorded a Na+ concentration of 27 mM, a K+ concentration of 1.7 mM, a Ca++ concentration of 0.69 mM and a Cl- concentration of 28.5 mM. In the spaces there was a lower resistance between the tip of the electrode and the serosal bath than that recorded with the tip in the lumen, and injection of fluorescent dye (11 A diameter) via the electrodes did not stain the cells. The concentrations in the secretion were similar to those in the spaces. The intracellular compartment had an apparent K+ concentration of 95 mM, and the concentrations of Na+ and Cl- were both about 5 mM. These data indicate that when the gallbladder is bathed with hypotonic solutions and is transporting fluid at approximately three or four times the normal rate, there are no significant osmotic gradients between the lumen and the lateral spaces. It is suggested that transcellular transport of water is implemented by a combination of high osmotic permeabilities across both mucosal and serosal cell membranes and low reflection coefficients (for K+ salts) at the serosal cell membranes.     

Potassium pyroantimonate and osmium-zinc iodide reactivity in the tubular epithelium of the opisthonephric kidney of the sea lamprey,Petromyzon marinus L

Pyroantimonate precipitate indicates that the epithelium of the proximal tubule is the only segment of the tubular nephron of the fresh water lamprey where large accumlations of cations are distributed. Unusually large amounts of reaction product are located within the lateral intercellular spaces and within vesicles closely associated with the plasma membrane at the lateral and basal surfaces. This technique suggests the continuity of these vesicles with the plasma membrane and alludes to the possibility of an endomembranous system of vesicles and the intercellular spaces as vehicles for ion transport. Lateral intercellular spaces of proximal tubules of lower vertebrates may play a different role in kidney function that their counterparts in higher vertebrates. Osmium-zinc iodide has a specificity for certain cells within the proximal, intermediate, and distal segments, but no structural differences are noted when these cells are compared to unstained cells. Smooth endoplasmic reticulum remains unstained in the distal segment but the stain has a strong affinity for elements of the Golgi apparatus, lysosomes, and the nuclear envelope of all cell types. This technique does not suggest a structural or functional similarity between cells of the distal segment and the chloride cells of the gills of teleosts.     

The mechanism mediating regenerative intercellular Ca2+ waves in the blowfly salivary gland.

Intercellular Ca2+ signaling in intact salivary glands of the blowfly Calliphora erythrocephala was studied by fluorimetric digital imaging combined with microinjection of putative messenger molecules. Iontophoretic injection of D-myo-inositol 1,4, 5-trisphosphate (InsP3) into salivary gland cells evoked regenerative intercellular Ca2+ waves that spread through the impaled cell and several rows of surrounding cells. Ca2+ increases induced by microinjection of Ca2+ ions were confined to the injected cells and their nearest neighbors. Depletion of intracellular Ca2+ stores by thapsigargin pre-treatment did not alter the time course of the Ca2+ increase caused by Ca2+ injection. However, activation of Ca2+ release became clearly evident when Ca2+ was injected in the presence of serotonin (5-HT). Under these conditions, injection of Ca2+ triggered intercellular Ca2+ waves that consecutively passed through >10 cells. The phospholipase C inhibitor U73122 blocked 5-HT-induced Ca2+ increases but did not affect InsP3-dependent Ca2+ spiking and intercellular Ca2+ wave propagation. The results demonstrate that propagation of agonist-evoked Ca2+ waves in the blowfly salivary gland requires supra-basal [InsP3] but does not depend on feedback activation of phospholipase C. We conclude that the intra- and intercellular transmission of these Ca2+ waves is mediated by diffusion of Ca2+ and Ca2+-induced Ca2+ release via the InsP3 receptor channel.