四膜虫大核基质中含有NuMA类似蛋白

NuMA (Nuclear mitotic apparatus protein)是细胞核基质中的一种高分子量蛋白质,具有一个特殊的细胞周期依赖性(cell cycle- dependent distribution)。在多细胞动物中有关NuMA的研究已经很深入,但是目前尚未见原生动物中NuMA蛋白的报道。四膜虫(Tetrahymena)是一种单细胞真核生物,属原生动物(Protozoa)纤毛虫纲(Ciliata),具有较特殊的分类地位和分子生物学研究背景。本实验结果表明,四膜虫大核基质中含有NuMA类似蛋白,不仅进一步验证了NuMA蛋白存在的普遍性,而且是对四膜虫中大核基质成分研究的补充。     

CHANGES IN THE BASE COMPOSITION OF NUCLEAR RIBONUCLEIC ACID OF NEURONS DURING A SHORT PERIOD OF ENHANCED PROTEIN PRODUCTION

Nuclei from isolated nerve cells were sampled by microdissection. The content and composition of the nuclear RNA was studied and compared with that of the cytoplasmic RNA of Deiters' nerve cells of rabbits. Analyses were made of control nerve cells and of cells in which an enhanced RNA and protein production had been induced by chemical means, tricyano-amino-propene, for 60 minutes. The nuclear RNA content of the control nerve cells was 56 µµg, i.e. 3 per cent of the total RNA content of the nerve cell. The base ratios were: adenine 21.3, guanine 26.6, cytosine 30.8, uracil 21.3. Purine-pyrimidine analyses showed that the nuclear RNA differed significantly from the cytoplasmic RNA in having higher adenine and uracil values. The guanine and cytosine values were high, however, and the ratio G/C was 0.86 as compared with 1.16 for the cytoplasmic RNA. The composition of the nuclear RNA was interpreted as reflecting the extraordinarily strong development of the nucleolus in these neurons. During the 60 minutes of enhanced neuronal RNA production (+25 per cent) the guanine value increased and the uracil value decreased significantly in the nuclear RNA. In the cytoplasmic RNA the guanine value also increased although not so much as the nuclear guanine. The cytoplasmic cytosine value decreased. The result indicated that the production of the characteristic cytoplasmic RNA had been influenced by the change in the nuclear RNA     

不同品种水稻种子贮藏蛋白质组分的差异

水稻是我国的主要粮食作物,它的蛋白质含量为6—14%。在水稻种子蛋白质中,谷蛋白占80%,球蛋白占10%,醇溶蛋白占5%,清蛋白占5%。据研究,水稻种子清蛋自主要由分子量16800的亚基组成,球蛋白由10种不同分子量的亚基通过疏水交互作用相结合,谷蛋白由分子量38000、25000和16000三种亚基通过双硫键相结合。但是,不同品种水稻种子贮藏蛋白质组分有何差异?研究报道的很少。本文简要报道不同品种水稻种子(籼稻和粳稻)贮藏蛋白质组分的差异,为     

蚕豆叶片细胞中IAA的胶体金免疫电镜定位

利用胶体金免疫电镜技术对蚕豆(Vicia faba L.)叶片细胞中的IAA定位进行了研究。幼嫩叶片的叶肉细胞中金颗粒主要分布在细胞核和叶绿体中,细胞质及细胞壁也有金颗粒标记。成熟叶片的叶肉细胞中金颗粒主要分布在叶绿体和细胞质,细胞壁也有少量金颗粒标记,液泡中没有发现金颗粒标记。成熟叶片小叶脉的韧皮细胞发现有大量的金颗粒标记,金颗粒主要标记在传递细胞的细胞壁中。小叶脉的维管束鞘细胞中也有很多的金颗粒标记,金颗粒主要分布在叶绿体、细胞质及细胞壁中。幼嫩叶片组织不进行IAA的固定或用正常兔IgG代替IAA抗体染色的对照,很难发现金颗粒标记。对IAA在组织及亚细胞中的定位及其生理意义进行了讨论。     

植物细胞核基质(Nuclear Matrix)的形态结构及其NuMA蛋白成分的研究

应用细胞选择性抽提并结合DGD包埋去包埋剂电镜技术对植物细胞核基质的形态结构进行了观察。结果显示胡萝卜悬浮培养细胞、银杏花粉细胞和精子细胞的细胞核内存在一个非染色质性的纤维蛋白网络体系。免疫荧光染色结果说明植物细胞核基质中含有与动物NuMA多抗交叉反应的多肽。免疫印迹反应显示胡萝卜悬浮培养细胞核基质蛋白与NuMA蛋白多抗的阳性反应条带为74KD和76 KD;银杏花粉细胞只有78 KD一条阳性带。以动物核基质NuMA蛋白保守杆状区的cDNA片段作为探针,与白菜子叶总DNA进行Southern杂交的结果表明植物细胞基因组中含有动物NuMA蛋白cDNA的同源序列。     

THE SYNTHESIS OF PHOSPHOLIPIDS IN THE NUCLEUS AND NUCLEAR MEMBRANE OF SYNCHRONIZED HeLa CELLS

HeLa cells were synchronized by a double thymidine block and pulse labeled at different stages of the cell cycle with ³H-choline. The specific activity of phospholipids extracted from the cell, the nucleus and the nuclear membrane showed a progressive increase from S to G₁; the incorporation of choline into phospholipids of asynchronous cells showed a specific activity intermediate between the values of S and G₁ cells. Similar results were obtained when ³²phosphorus was used as a precursor instead of choline. Thin layer chromatographic analysis of phospholipids extracted from cells in S and from cells in G₁ failed to show any difference in the distribution of radioactivity among the various phospholipid classes. Choline uptake by HeLa cells in different phases of the cell cycle did not show significant variations. However, during the synchronization process, shortly after the addition of excess thymidine, an increased uptake of choline by cells and an increased incorporation of choline into phospholipids were found. The results indicate that some of the changes occurring in phospholipids synthesis may not be cell cycle dependent, but may be the effect of the synchronizing process.     

THE INFLUENCE OF DIETARY PROTEIN DEFICIENCY ON HAEMOPOIETIC CELLS IN THE MOUSE

These experiments examined the effect of a diet limited only in protein (4% by weight) on haemopoietic stem cells in mice. This diet places severe restrictions on growth and cell proliferation and this was reflected in lower numbers of colony forming units (CFUs) and in vitro colony forming cells (CFCs). Differences were apparent in the response of different organs to this stress; for instance, the incidence of spleen CFUs fell sharply from around 40/mg spleen tissue to 1 -4/mg spleen tissue after 3 weeks on a low protein diet. This selective loss did not occur in bone marrow where total CFUs remained proportional to cellular content. Yet a third pattern was shown by thymus CFUs–although the numbers were low these increased from 16/thymus in normal mice to 132/thymus in deprived mice. This was the only organ examined which showed an increase. The effects of a return to a high protein (18 %) diet showed that the spleen was the most responsive organ. By day 5 after the return to 18% protein the spleen contained as many CFUs per million cells as the bone marrow. During this time the content of CFU in the spleen had increased some 50-fold whereas bone marrow CFUs only doubled. The spleen assumes the major reconstitutive role during the refeeding process.     

CHANGES IN THE PROTEIN COMPOSITION OF MOUSE BRAIN MYELIN DURING DEVELOPMENT

Abstract— Myelin was isolated from the brains of mice at various ages by a procedure involving a final purification on a continuous CsCl gradient. Myelin protein accumulated throughout development, increasing from 0.25 mg of protein/brain at 8 days of postnatal age to 3.5 mg of protein/brain at 300 days, although the rate of accumulation was greatest at about 21 days of age. Quantitative studies of the protein composition of these samples were carried out, utilizing discontinuous polyacrylamide gel electrophoresis in buffers containing sodium lauryl sulphate. Mouse brain myelin, contained (in order of increasing molecular weight) two basic proteins, an uncharacterized doublet, proteolipid protein, and a group of high molecular weight proteins. There were marked changes in the quantitative distribution of these proteins with increasing postnatal age. The basic protein fraction of total myelin protein increased from about 18 per cent at 8 days to 30 per cent at 300 days of age. Proteolipid protein increased even more dramatically, from 7 to 27 per cent in the same time interval. These chemical studies were correlated with ultrastructural investigations, both of the developing myelin sheath in situ and the isolated myelin obtained from mice of various ages. A hypothesis, relating the observed changes in protein composition of myelin during development to its mode of formation, is developed. Another subcellular fraction, separated from myelin, by virtue of its greater density in a CsCl gradient, was also studied. It was a vesicular, membranous fraction present at a level of 0.35 mg of protein/brain at all ages and was related to myelin in terms of protein composition.     

THE PROTEIN COMPOSITION OF ISOLATED MYELIN1

—Three fractions, each containing markedly different proteins, was obtained from myelin: (1) The first fraction was obtained as an insoluble residue when myelin was extracted with neutral chloroform-methanol (CM, 2:1, v/v). It was digestible with trypsin and had an amino acid composition similar to that of the acidic proteolipid protein of Wolfgkam (1966). (2) The second fraction was obtained as a precipitate by the addition of various electrolytes (KCl, NaCl, CaCl₂, MgCl₂ or HCl) to the CM (2:1 v/v) extract. This fraction consisted mainly of a basic protein which exhibited an electrophoretic mobility and amino acid composition indistinguishable from those of the basic protein obtained from white matter (Martensson and LeBaron, 1966). This procedure provided for a simple and rapid isolation of the basic protein from myelin. Depending on the conditions of precipitation, this fraction was either free of lipid or contained tri- and diphosphoinositide. The effects of different ions at differing concentrations and the yield and nature of the precipitate have been studied. (3) A third fraction remained in solution in CM (2:1, v/v) after the addition of the electrolyte. It comprised the bulk of the myelin lipids and a protein fraction which was resistant to digestion with trypsin and had an amino acid composition similar to the classical proteolipid protein of Folch-Pi and Lees (1951). The possibility of a salt-type bonding between the basic protein and the polyphosphoinositides is discussed, and values for tri- and diphosphoinositide in bovine myelin are given.     

逆境中沙冬青叶片细胞叶绿体的结构(简报)

沙冬青(Ammopiptanthus mongolicus)是亚洲中部一种超旱生植物,生长在荒漠、半荒漠地带,主要分布于我国内蒙古、宁夏、甘肃等地,是北方地区极少有的常绿阔叶植物,也是国家二级保护植物。沙冬青体内含有黄花木素、拟黄花木素等强生物碱,是重要的药用植物,也是极好的固沙、绿化荒丘植物。它耐热、旱、盐碱,更耐寒,能抵抗-30℃至-40℃的低温,所以也是植物抗逆性研究的理想材料。     

FINE STRUCTURE OF NUCLEAR CRYSTALS IN LEAVES OF TWO DIANTHUS SPECIES

Examination of leaf cells of Dianthus barbatus with the election microscope revealed the presence of crystals in the nuclei, but not in the cytoplasm or in other organelles. They were at first thought to be viral crystalline inclusions, but no evidence could be adduced to support this possibility. The crystals were found in the nuclei of cells from all parts of the leaf, including parenchyma cells of the vascular elements. They were usually 0.5–2 μ long, and occasionally up to 5 μ. In longitudinal section they appeared as electron-dense striations, about 10–15 mμ wide, alternating with electron-transparent bands, 6–8 mμ wide. In cross section they had a honeycomb appearance, resulting from closely packed hexagonal elements, with electron-transparent centers. Similar crystals were found in nuclei of D. chinensis leaf cells, but not in other members of the Caryophyllaceae, including two varieties of D. caryophyllus, D. deltoides, D. arenarius, Silene acaulis, Lychnis chalcedonica, Saponaria calibrica, and Stellaria media. This appears be the first fine structural report of a lattice inclusion in nuclei of cells of healthy higher plants.     

Na+和Cl- 在小麦叶肉细胞超微结构中的分布

小麦经200mmol NaCl溶液培养3天后,采用改进的焦锑酸钾方法对叶肉细胞中Na~+及Cl~-进行超微结构定位。电镜观察及电子探针X-射线显微分析表明,Cl~-主要分布在细胞间隙、细胞壁及细胞质膜中。用电子探针X~-射线能谱仪在这些部位中未探测出Na~+,提示Cl~-比Na~+更多地进入小麦的叶肉细胞。此外,在叶肉细胞的细胞核、线粒体及叶绿体中也可见到离子沉淀颗粒。经氯化钠溶液培养的小麦幼苗,其叶肉细胞的叶绿体、线粒体的超微结构受损,植株生长受到抑制。     

百合花粉母细胞核骨架的超微结构观察

参照动物细胞核骨架的研究方法,用整装电镜技术和DGD包埋-去包埋技术研究了选择性抽提的和完整的百合(Lilium davidii var. willmottiae (Wilson) Roffill)花粉母细胞。结果表明,住减数分裂前期Ⅰ,百合花粉母细胞核内存在一个精细的非染色质纤维——核骨架。该网络由5～15nm的纤维交织而成,广泛地分布于细胞核内。这些核骨架有的分布于染色体间,有的分布于染色体周围,并与染色体和核仁相连。随着减数分裂时间的推移,染色体(质)间核骨架纤维逐渐减少,染色体(质)周围的核骨架纤维逐渐增多,并与染色体内部的纤维结构相连,表明核骨架一方面为染色体拓扑变化提供一个空间支架,另一方面也可能参与了染色体骨架的构建。     

NUCLEAR MEMBRANES OF CULTURED MAMMALIAN CELLS IN THE PERIOD PRECEDING DNA SYNTHESIS

When kidney cells are cultured directly from the rabbit, the nuclear membranes undergo a change that can be measured as an increase in electrophoretic mobility. The change appears to begin immediately upon culture and is maximal 2 hours later, after which the mobility remains constant at the elevated level. Actinomycin D and p-fluorophenylalanine, but not EDTA or ionizing radiation, suppress the increase in nuclear electrophoretic mobility. With synchronously growing L cells, no change was detected in nuclei from cells taken during various parts of the division cycle.     

改变食物组成对大鼠胃肠道嗜铬细胞的影响

为研究食物组成对肠嗜铬（ＥＣ）细胞的影响，用免疫组织化学方法（ＰＡＰ法）对４组食物组成不同大鼠的胃肠道５－羟色胺免疫反应阳性（５－ＨＴＩＲ）细胞的数量和分布作了观察。发现食物改变会引起大鼠摄食量和胃肠形态的变化，但未见其生长发育异常。食物中米糠含量加倍后，大鼠５－ＨＴＩＲ细胞数量无显著增加，但分布型有明显的变化。食物中米糠含量或米糠和麦麸含量都减少到１０％后，大鼠５－ＨＴＩＲ细胞数量和分布都有明显     

ON THE FINE STRUCTURE AND COMPOSITION OF THE NUCLEAR ENVELOPE

Nuclei from nearly ripe eggs of Rana pipiens were isolated and cleaned in 0.1 M KCl. The whole nucleus was then digested to various degrees with ribonuclease or trypsin, followed by washing and fixation in either osmium tetroxide or potassium permanganate. The nuclear envelope was dissected off, placed on a grid, air dried, and compared with undigested controls in the electron microscope. Some envelopes were dehydrated, embedded in methacrylate, and sectioned. Annuli around "pores" are composed of a substance or substances, at least partially fibrillar, which is preserved by osmium but lost during permanganate fixation. Material within the "pores" is also preserved by osmium but partially lost after permanganate. No evidence of granules or tubules in the annuli was found in air dried mounts although a granular appearance could be seen in tangentially oriented thin sections. Thin sections of isolated envelopes give evidence of diffuse material within the "pores" as well as a more condensed diaphragm across their waists. In whole mounts of the envelope the total density within "pores" is relatively constant from "pore" to "pore." All material within "pores," including the condensed diaphragm, is removable by trypsin digestion. Wispy material from the "pore" structure projects into the nucleus and annular material extends into the cytoplasm. Both annular and diaphragm materials remain with the envelope when it is isolated and are thus considered a part of its structure, not merely evidences of material passing through. There is no evidence of ribonuclease-removable material in any part of the "pore" complex.