TIME SEQUENCE OF NUCLEAR PORE FORMATION IN PHYTOHEMAGGLUTININ-STIMULATED LYMPHOCYTES AND IN HELA CELLS DURING THE CELL CYCLE

The time sequence of nuclear pore frequency changes was determined for phytohemagglutinin (PHA)-stimulated human lymphocytes and for HeLa S-3 cells during the cell cycle. The number of nuclear pores/nucleus was calculated from the experimentally determined values of nuclear pores/µ² and the nuclear surface. In the lymphocyte system the number of pores/nucleus approximately doubles during the 48 hr after PHA stimulation. The increase in pore frequency is biphasic and the first increase seems to be related to an increase in the rate of protein synthesis. The second increase in pores/nucleus appears to be correlated with the onset of DNA synthesis. In the HeLa cell system, we could also observe a biphasic change in pore formation. Nuclear pores are formed at the highest rate during the first hour after mitosis. A second increase in the rate of pore formation corresponds in time with an increase in the rate of nuclear acidic protein synthesis shortly before S phase. The total number of nuclear pores in HeLa cells doubles from ~2000 in G₁ to ~4000 at the end of the cell cycle. The doubling of the nuclear volume and the number of nuclear pores might be correlated to the doubling of DNA content. Another correspondence with the nuclear pore number in S phase is found in the number of simultaneously replicating replication sites. This number may be fortuitous but leads to the rather speculative possibility that the nuclear pore might be the site of initiation and/or replication of DNA as well as the site of nucleocytoplasmic exchange. That is, the nuclear pore complex may have multiple functions.     

OBSERVATIONS ON SYNAPTONEMAL COMPLEXES IN PREMEIOTIC INTERPHASE OF WHEAT

Synaptonemal complexes (SCs) were found in stage 3, premeiotic (S phase) pollen mother cell (PMC) nuclei of wheat which were labeled with ³H-thymidine. Three nucleoli are present in PMC nuclei at the beginning of stage 3, premeiotic interphase (S3). During S3, nucleoli move toward the nuclear envelope and fuse to form one nucleolus near the end of the stage. PMC nuclei labeled with ³H-thymidine were serially sectioned to show that more than one nucleolus was present and that SCs were also present in these DNA synthetic nuclei. Entire S3 PMC nuclei were serially sectioned to show the presence of SCs and all three nucleoli. Entire leptotene nuclei were also serially sectioned and segments of SCs were found. It is concluded that the association of homologous chromosomes in S3 of wheat is an early step in SC formation which proceeds through leptotene and is completed in zygotene and pachytene. Thus there is evidence that the continuum of chromosome pairing in wheat starts much earlier than was once thought.     

植物细胞核的凹入和核液泡的形成

核质互作在细胞核中形成膜囊结构首先是在动物细胞中揭示的［１］。对植物细胞超微结构的研究亦发现有类似现象的存在［２—４］。在植物细胞核中形成的膜泡认为有两种形式：一是核被膜向核基质深度凹入,形成细胞质深入细胞核的状态,称之为“假包被（ｐｓｅｕｄｏ－ｉｎｃｌｕｓｉｏｎ）”［２,５］;另一种形式是细胞核内膜或内外双层核膜向核基质深度凹入,并最终脱离核被膜,在核基质中形成囊泡结构,称之为“核液泡（ｎｕｃｌｅａｒｖａｃｕｏｌｅ）”［３,４,６,７］。对细胞核质间通过核被膜在结构上的特殊作用形式缺少像对通过…     

PROPHASING OF INTERPHASE NUCLEI AND INDUCTION OF NUCLEAR ENVELOPES AROUND METAPHASE CHROMOSOMES IN HELA AND CHINESE HAMSTER HOMO- AND HETEROKARYONS

Fusing human HeLa metaphase cells with HeLa interphase cells resulted within 30 min in either of two phenomena in the resultant binucleate cell: either prophasing of the interphase nucleus or formation of a normal-appearing nuclear envelope around the metaphase chromosomes. The frequency of either occurrence was strongly dependent on environmental pH. At pH's of 6.6–8.0, prophasing predominated; at pH 8.5 nuclear envelope formation predominated. Additionally, the frequencies of the two events in multinucleate cells depended on the metaphase/interphase ratio. When the ratio was 0.33 nuclear envelope formation predominated; when it was 2.0 prophasing predominated. In their general features, the results with fused HeLa cells resembled those reported earlier with fused Chinese hamster Don cells. However, the results provided an indication that between pH 6.6 and 8.0 the HeLa metaphase cells possessed a much greater capacity than the Don metaphase cells to induce prophasing. Fusion of Don metaphase cells with HeLa interphase cells or of Don interphase cells with HeLa metaphase cells at pH 8.0 resulted in nuclear envelope formation or prophasing in each kind of heterokaryon. As in the homokaryons, the frequencies of the two events in the heterokaryons depended on the metaphase/interphase ratio. The statistics of prophasing and nuclear envelope formation in the homo- and heterokaryon populations were consistent with the notion that disruption or formation of the nuclear envelope depends on the balance attained between disruptive and formative processes.     

洋葱细胞核中类胀泡结构的数目,体积和分布

在电镜下观察了洋葱（Ａｌｌｉｕｍ ｃｅｐａ Ｌ．）根端分生组织细胞核的连续切片并进行了三维重组。发现：１． 类胀泡结构在细胞核中的数目与二倍染色体数有较大差异。在所观察的Ｓ期细胞核中,该结构数为４—１０ 个;在所观察的Ｇ２ 期细胞核中,该结构数为４—９ 个。２． 类胀泡结构的体积变化比较大,大多数的体积在０．１—０．４ μｍ ３ 之间。不同的Ｓ期细胞核中,类胀泡结构总体积较大者是总体积较小者的１．８ 倍;不同的Ｇ２ 期细胞核中,类胀泡结构总体积较大者是总体积较小者的２．１ 倍。３． 类胀泡结构有集中分布在细胞核内一定区域的倾向。讨论了类胀泡结构是否为着丝粒等问题     

缩球囊霉孢子萌发及细胞核在孢子和芽管中分布的研究*

本文研究了从本校分离的丛枝菌根真菌缩球囊霉Glomus constrictum的孢子萌发和萌发孢子的细胞核DAPI(46-diamidino-2-phenylindol)染色。结果显示,G. constrictum孢子直径为179.5-198.7μm ,顶生于产孢菌丝上。经表面消毒处理后,孢子在水琼脂平板上7天开始萌发。DAPI染色后,经稀释荧光计数,单个孢子细胞核数目约为5300,在孢子中无序分布, 细胞核直径约为9.9-11.2μm 。孢子萌发过程中,细胞核总数无明显变化,只是部分细胞核从孢子流向了萌发伸长的菌丝。     

间期和有丝分裂期HeLa细胞中核仁蛋白B23的分布和含量的差异

B23蛋白是真核细胞核仁的两种主要蛋白成份之一。已往的工作表明,细胞内B23蛋白的分布与rRNA合成速率和细胞的生长状况密切相关。本工作利用抗B23蛋白单克隆抗体,研究了被两种作用于微管的药物秋水仙酰胺(colcemid)和紫杉酚(taxol)阻断的有丝分裂期和间期HeLa细胞内B23蛋白的分布和含量的差异。结果发现有丝分裂期细胞内B23蛋白含量明显高于间期细胞,而且B23蛋白在两类细胞中的分布也有明显的不同。     

BEHAVIOR OF INTERPHASE EMBRYONIC NUCLEI TRANSPLANTED IN NUCLEAR MULTIPLICATION STAGE EMBRYOS OF DROSOPHILA MELANOGASTER

Interphase nuclei were transplanted from syncytial blastoderm into early cleavage embryos of Drosophila melanogaster. The transplanted nuclei, when exposed to host cytoplasm, were initiated to mitosis. During the period from 10 to 50 min after transplantation, the implanted nuclei and host nuclei were found not synchronous in their mitotic cycles. Synchrony was restored usually by the blastoderm stage.
About 5% of eggs with transplanted nuclei developed significantly faster than control eggs, resulting in premature blastoderm formation. This finding is discussed in relation to chimera formation and to embryonic development of grandchildless mutants.     

用非放射性原位杂交和免疫金染色在豌豆间期染色质内显示的rDNA的分布

以异羟基洋地黄毒甙标记的ｒＲＮＡ为探针,通过与植物根尖分生组织区振动切片中的ｒＤＮＡ原位杂交,结合免疫金染色及银加强金处理研究了豌豆间期染色质中ｒＤＮＡ的分布。研究结果清楚地显示供试植物间期细胞内有１－５个显著的ｒＲＮＡ探针结合位点,其中显示４个结合位点的细胞占６０．６％,少于和多于４个结合位点的细胞分别上３７．９％和１．１４％。在显示４个结合位点的细胞核内,它们分别按照２：１、３：１和２：１：１     

RABL DISTRIBUTION OF INTERPHASE AND PROPHASE TELOMERES IN ALLIUM CEPA NOT ALTERED BY COLCHICINE AND/OR ULTRACENTRIFUGATION

Neither colchicine nor ultracentrifugation, singly or in sequence, significantly alters the normal Rabl distribution of interphase or prophase telomeres in root tip cells of Allium cepa L. The position of telomeres was determined by C-banding, which stains A. cepa chromosomes only at the telomeres. Centrifugation displaces mitotic figures toward one side of the cell, but otherwise their mitotic configurations are little changed. These light microscope results are interpreted to show that a) interphase and prophase telomeres are attached strongly to some component of the nuclear envelope; b) a colchicine-sensitive component apparently does not attach interphase and prophase telomeres to the nuclear envelope; and c) chromosomes at all stages of the cell cycle are attached to some structure, nuclear envelope, and/or spindle fibers.     

EFFECT OF METABOLIC INHIBITORS ON NUCLEAR PORE FORMATION DURING THE HELA S3 CELL CYCLE

The effect of various antimetabolites on nuclear pore formation was studied in synchronized HeLa S₃ cells. The nuclear size was determined by light microscopy and the pore number per unit area of nuclear surface by the freeze-etching technique and electron microscopy. It was found that the inhibition of DNA replication or ribosomal RNA synthesis has no effect on nuclear size increase or pore formation. However, the inhibition of ATP synthesis effectively stops nuclear pore formation. Cycloheximide blocks nuclear pore formation at the same time during G₁ phase of the cell cycle when nuclear size increase is blocked by high concentrations of actinomycin D. This suggests that certain proteins or other factors leading to pore formation and nuclear size increase are transcribed and synthesized at about 3–4 h after mitosis, i.e., about 1–2 h before S phase begins.     

芍药科分布格局及其形成的分析

芍药科Paeoniaceae为单属科,芍药属Paeonia L．共30种,隶属三个组,牡丹组Sect．Moutan DC．为该属的木本类型,有6种,分布在中国的一个局部地区,从云南的中部(景东24．4°N)和西藏东部(札囊,90°E)向东北至安徽的巢湖(117．7°E)和陕西延安(36．5°N)。6个种中滇牡丹P．delavayi分布最广,其余5种分布区都比较局限。北美芍药组Sect．Onaepis Lindley 仅2种,是该属最狭域的组,仅分布于北美西部,从太平洋沿岸的俄勒岗州至蒙大拿州,南起加利福尼亚州的圣迭戈(32．7°N),向北至华盛顿州与其它两组成间断分布。芍药组Sect．Paeonia是该属种类最多,分布范围最广的组,包括22种,从亚洲温带东部的色丹岛(146．6°E),至欧洲最西部的葡萄牙和非洲西北部的摩洛哥Demnat省,向南至我国云南的宁南,北缘进入北极圈。本文按照一般学者所能按受的演化趋势,并结合细胞学、生态学和分子系统学研究的初步结果,推论了该属性状演化趋势。结合在世界各区中种数的比较、各区特有种的比较和在中国的分布,得出下列初步结论：(1)牡丹组Sect．Moutan是芍药属中最原始的组;北美芍药组Sect． Onaepia既有一些原始性状,又有特化性状,它是直接从木本类型分化发生的类群,芍药组Sect．Paeonia 是该属最大,分化强烈、有若干复合体,又有不少特化的类型,因而它是芍药属中相对年轻、特化的类群。 (2)东亚区(尤其是中国中部)是芍药属的多样性中心,该区的中国东亚部分,特别是从云南至山西,陕西一线是现存芍药属原始类群分化发展的中心,地中海区是芍药属的种的分化中心,是次生分化中心。(3)随着Sect．Paeonia的成员从中国东亚部分向西、向北迁移,具四倍性和根纺锤状加粗等特化性状的种数增加。     

细胞器向核凹入与核液泡的形成和降解

利用电镜观察紫竹梅雄蕊行超薄切片,发现细胞器,如线粒体有向核凹入现象,其线粒体仍处在与细胞质相勾通的核沟槽里,但核沟槽能扩大核被膜表面积,以利用于胞核与胞质的物质交换和信息传递,当核沟槽颈部变窄时,核膜融合,核沟槽脱离核被膜,形成核液泡;随后,核液泡的内外两层膜相继降解,核液泡被消化溶入核基质中。     

镉离子对菠菜叶绿体色素蛋白质复合物及激发能分配的影响

Cd~(2+)使叶绿体低温(77K)荧光发射光谱中 F686/F736及 F696/F736和激发光谱中F480/F436比值降低,说明 Cd~(2+)不利于激发能向 PSⅡ传递。SDS-PAGE 分析表明,Cd(2+)处理后叶绿体类囊体膜中光系统Ⅱ捕光叶绿素蛋白质复合物 LHCⅡ的部分寡聚体解聚成单体,且 LHCⅡ的总量也减少了。分析表明 Cd~(2+)使属于 LHCⅡ的多肽减少。已知LHCⅡ在光能吸收、传递以及激发能在两个光系统间的分配和调节方面起着重要作用,Cd~(2+)引起部分 LHCⅡ解聚和总量减少,必然导致由 LHCⅡ捕获和向光系统Ⅱ中心传递的能量减少。     

OXYGEN TENSION AND THE RATES OF MITOSIS AND INTERPHASE IN ROOTS

The object of this work was to determine the influence of a wide range of oxygen tensions upon the relative rates of respiration, mitosis, and interphase in pea root tips, compared with the normal rates of these processes in air. From the rates of disappearance of mitotic figures in excised tips kept in various oxygen tensions, the relative rates of mitosis were found to decrease gradually from 122 per cent in 100 per cent oxygen to 24 per cent in 0.0007 per cent oxygen. From the mitotic indices of intact seedlings, the relative rates of interphase were found to decrease sharply from 82 per cent in 10 per cent oxygen to 6 per cent in 5 per cent oxygen. The data on relative rates of respiration, mitosis, and interphase in root tips were compared, and it was shown that the three processes are perfectly distinct in their quantitative relationships to low oxygen tensions.     

CONTRAST BETWEEN THE ENVIRONMENTAL pH DEPENDENCIES OF PROPHASING AND NUCLEAR MEMBRANE FORMATION IN INTERPHASE-METAPHASE CELLS

In Chinese hamster Don cells, fusion of an interphase cell with a metaphase cell resulted either in prophasing of the interphase nucleus, including loss of the nuclear envelope (NE), or in the formation of a double membrane around the metaphase chromosomes. Only one of these phenomena occurred in a given interphase-metaphase (I–M) binucleate cell. At pH 7.4, there was about an equal probability that either event could occur amongst the population of I–M cells. The effect of pH changes in the medium containing the fused cells was examined. At pH 6.6, prophasing was the predominant event; at pH 8.0, membrane formation predominated. It was found that the rate of progression of a mononucleate cell from G₂ to metaphase was appreciably faster at pH 6.6 than at pH 8.0. Conversely, the progression from metaphase to G₁ was faster at pH 8.0 than at pH 6.6. These results with the mononucleate cells strengthen the hypothesis that structural changes in I–M cells are reflections of normal mitotic phenomena. Additional evidence for this hypothesis was produced by electron microscope examination after direct fixation in chrom-osmium. The double membrane around the chromosomes of the I–M cell was indistinguishable from the normal NE. The results obtained by varying the pH of the medium containing the fused cells provide an indication that disruption or formation of the NE of Don cells depends on the balance reached between disruptive and formative processes.     

SYNTHESIS OF RNA IN MAMMALIAN CELLS DURING MITOSIS AND INTERPHASE

Chinese hamster cells in the mitotic and G₁ phases of the growth cycle were incubated for 30 or 60 min in suspension tissue culture and pulse-labeled with tritiated uridine. After appropriate chases, washes, and extractions, it was found that all incorporation into the nucleic acid may be accounted for by those cells in interphase. An average of 410 counts was found for incorporation into the cell population (approximately 2.0 x 10⁵ cells) of which over 80% of the cells was initially in mitosis. The increasing number of cells leaving mitosis and entering interphase during the 30 min incubation was theoretically able to account for 470 counts. In addition, short-pulse labeling experiments have shown a consistent linear relationship between the percentage of cells in division and the incorporation of the isotope, which strongly suggests that, if 100% of the cells were in mitosis, the counts would be essentially zero. Thus, the entire label may be attributed to those cells in interphase where portions of the chromosomal material are known to be already extended.