伍氏游仆虫(Euplotes woodruffi)大核原基中的多线染色体

用染色体铺展技术,光镜和透射电镜技术,和有色光分光光度计量术等观察了伍氏游仆虫在２３—２５℃培养液中接合后体大核原基多线染色体的发生、发展及碎解的全过程。在发育到４—５小时的接合后体中,球形的大核原基中开始出现染色体。此时的染色体大多为颗粒状,有些染色体已从颗粒状延长而成平均３μｍ的短杆。染色体数初步计算为２ｎ＝７０—９０。约经２０分钟,染色体延长并多线化。再过约５—６小时,多线染色体发育到最高峰。细长丝状的多线染色体上带和间带分化明显。有些染色体的末端有扇面状或小球形结构。各别染色体还显示了类似于着丝点的结构。这一时期的大核原基体积最大,ＤＮＡ含量达到最高值,接近营养大核ＤＮＡ含量的一半。然后多线染色体碎解,各多线染色体为纤维膜横割为大小不一的片段,ＤＮＡ含量也随之降至最高值的１／１０。     

蒜有丝分裂前期和前中期染色体的螺旋结构

本文研究了蒜(Allium sativum)根端细胞有丝分裂前期和前中期染色体的螺旋结构及其形成过程。在光镜和电镜下都看到前期核内存在螺旋化的染色线。从早前期到晚前期染色线的螺旋化是逐渐进行的。开始只有部分染色线螺旋化,螺幅直径约为1.2-1.5微米,以后螺幅增大,达1.8～2微米,并且螺旋结构变得更为紧密。前中期染色体中可见由直径600 nm左右的染色线形成的螺旋结构,螺旋比晚前期更加紧密。本文对中期染色体的高层次结构进行了讨论。     

多线染色体

介绍了多线染色体形态结构,生理功能及多线染色体在生物的细胞遗传,昆虫的系统分类,遗传防治、基因图的筑建,特别是在遗传工程研究应用中的进展。     

介绍一种摇蚊唾腺染色体永久封片的制作方法

多线染色体是体细胞分裂间期染色质丝多次复制但不分开形成的。虽然摇蚊多线染色体早在1881年就已被发现,但由于人们缺乏对其遗传学的了解,并未用它做深入的遗传学研究。随着真核生物基因表达研究的深入,多线染色体的斑带及疏松结构再次引起遗传学家的兴趣。摇蚊幼虫个体大、廉价易操作,其唾腺多线染色体明显,故成为理想的遗传学实验材料。对摇蚊唾腺染色体染色方法进行了一些探索。     

水稻染色体DNA的交变脉冲电泳分析

从水稻完整的细胞核中释放出来的染色体DNA在交变脉冲电泳申表现为一种“240kb单位”的形式。这种单位,经限制性内切酶消化,可产生连续分布的大至1500kb的染色体DNA片段。文章讨论了“240kb单位”作为水稻染色体DNA基本结构单位的可能性。     

兴义维蚋多线染色体研究（英文）

首次在国内对兴义维蚋Simulium (Wilhelmia) xingyiense的多线染色体进行研究, 并提供其多线染色体标准图。选取兴义维蚋的成熟幼虫, 用改良苯酚品红染色法进行唾腺多线染色体制备, 并进行测量、 描述及分析。结果表明： 兴义维蚋多线染色体数目为3对（2n=6）。Ⅰ号染色体具中央着丝粒, Ⅱ和Ⅲ号染色体均为亚中央着丝粒染色体。核仁组织者区位于Ⅰ号染色体短臂近着丝粒端。巴尔比尼氏环和双泡位于Ⅱ号染色体短臂近中央位置。3对染色体的着丝粒区可形成明显的染色中心。兴义维蚋多线染色体具有多态性的倒位, 倒位频率为0.64。兴义维蚋多线染色体的着丝粒、 核仁组织区、 巴氏环、 双泡等主要特征性结构的位置及形态恒定一致,可作为该种的重要鉴别特征。其多态性的倒位可为该蚋种在细胞水平上进行蚋类分类鉴别和系统发育等研究提供基础资料。     

果蝇唾腺多线染色体研究进展及其在遗传学教学中的应用

果蝇唾腺多线染色体是细胞遗传学的3大经典染色体之一,从1934年至今因其具有显著的特点已经作为一个优异的模型用在不同的遗传学研究中。果蝇唾腺多线染色体最大的贡献就是为研究间期染色体结构和基因的表达调控提供了一个非凡的视角;另外,果蝇唾腺多线染色体还可以用于解释一些特殊的遗传现象,例如剂量补偿效应和花斑位置效应。文章一方面就以上进展作一简要总结,另一方面尝试将这一典型案例系统地用于遗传学教学中,引导和激发学生学习遗传学的兴趣。     

摇蚊马氏管和直肠多线染色体的初步研究

从发育观点观察了摇蚊成蚊马氏管、直肠细胞中的多线染色体,特别是第Ⅳ染色体自幼虫至成蚊的动态变化过程。结果表明:染色体在个体发育中的变化是有规津的,并与细胞的分化和活动密切相关的;染色体泡和多线分化反映着细胞的结构与功能。本文特别强调了研究双翅目昆虫消化道后部器官在发生遗传学上的重要意义。     

甜菜夜蛾细胞分裂期染色体的观察

以精巢组织为材料,采用空气干燥法制备染色体标本,对甜菜夜蛾Spodopteraexigua有丝分裂和减数分裂染色体形态行为进行了研究。结果表明：甜菜夜蛾的染色体数目为n=31;染色体为弥散着丝粒染色体,2条染色体上存在次缢痕;晚偶线期出现染色体互锁现象;从早粗线期到晚粗线期联会复合体逐渐伸长;终变期同源染色体形成环状、端部交叉、尾尾相对的结构。     

红翅皱膝蝗减数分裂染色体的螺旋与轴结构

用低渗处理和苯酚品红染色,在经过卡诺液(甲醇3∶冰醋酸1)固定和未经固定的红翅皱膝蝗减数分裂染色体上都看到了螺旋结构。观察和测量结果表明,每条染色单体都是由430nm左右的染色线螺旋形成的。由染色线到染色体的压缩率为4∶1。低渗处理后固定的材料经过银染,则显示了染色体轴结构。同样,未经低渗处理直接固定的材料银染时也出现了轴结构。银染的轴结构位于每个染色单体的中央,并贯穿整个染色单体。在光镜下,这个轴并不是直径均一的棒状结构,而似乎是由许多大小相近的颗粒相连而成。本文对染色体结构的有关模型、骨架和轴结构的真实性以及轴和螺旋的关系等问题进行了讨论。     

G_2期后期染色质结构及染色体高级结构形成的研究

用适量BrdU处理中华大蟾蜍外周血淋巴细胞,能以较高频率得到形态多样的染色质(体)结构。本文以两栖类和人类细胞为材料,采用Feulgen染色、Ag-NOR染色、DAPI荧光染色及放射自显影等方法,证实了其染色质性质,初步讨论了其产生原因,并将其命名为:“G,期后期染色质”。在此基础上,进一步从形态学角度初步分析了从G_2期后期到M期染色质转变为染色体的动态过程,提出不同染色体形成其高级结构是非同步的,有可能存在染色体包裹顺序的设想。     

Nucleosomes in metaphase chromosomes.

Previous studies of the structure of metaphase chromosomes have relied heavily on electron micrography and have revealed the existence of a 10-nm unit fiber that is thought to generate the native 23-30-nm fiber by higher order folding. The structural relationship of these metaphase fibers to the interphase fiber remains obscure. Recent studies on the digestion of interphase chromatin have revealed the existence of a regularly repeating subunit of DNA and histone, the nucleosome that generates the appearance of 10-nm beads connected by a short fiber of DNA seen on electron micrographs. It was therefore of interest to probe the structure of the metaphase chromosome for the presence of nucleosomal subunits. To this end metaphase chromosomes were prepared from colchicine-arrested cultures of mouse L-cells and were subjected to digestion with stayphylococcal nuclease. Comparison of the early and limit digestion products of metaphase chromosomes with those obtained from interphase nuclei indicates that although significant morphologic changes occur within the chromatin fiber during mitosis, the basic subunit structure of the chromatin fiber is retained by the mitotic chromosome.     

Proteome analysis of human metaphase chromosomes

DNA is packaged as chromatin in the interphase nucleus. During mitosis, chromatin fibers are highly condensed to form metaphase chromosomes, which ensure equal segregation of replicated chromosomal DNA into the daughter cells. Despite >1 century of research on metaphase chromosomes, information regarding the higher order structure of metaphase chromosomes is limited, and it is still not clear which proteins are involved in further folding of the chromatin fiber into metaphase chromosomes. To obtain a global view of the chromosomal proteins, we performed proteome analyses on three types of isolated human metaphase chromosomes. We first show the results from comparative proteome analyses of two types of isolated human metaphase chromosomes that have been frequently used in biochemical and morphological analyses. 209 proteins were quantitatively identified and classified into six groups on the basis of their known interphase localization. Furthermore, a list of 107 proteins was obtained from the proteome analyses of highly purified metaphase chromosomes, the majority of which are essential for chromosome structure and function. Based on the information obtained on these proteins and on their localizations during mitosis as assessed by immunostaining, we present a four-layer model of metaphase chromosomes. According to this model, the chromosomal proteins have been newly classified into each of four groups: chromosome coating proteins, chromosome peripheral proteins, chromosome structural proteins, and chromosome fibrous proteins. This analysis represents the first compositional view of human metaphase chromosomes and provides a protein framework for future research on this topic.     

Scanning electron microscopy of the centromeric region of L-cell chromosomes after treatment with Hoechst 33258 combined with 5-bromodeoxyuridine

When mouse L-cells were treated with a combination of 5-bromodeoxyuridine (BrdUrd) and Hoechst 33258, the metaphase chromosomes revealed undercondensation of the chromatin fibers in the sister centromeres. The application of the osmium-thiocarbohydrazide technique to the air-dried chromosome preparations made it possible to elucidate the ultrastructure of the undercondensed centromeric region at the level of the 30 nm chromatin fiber. Scanning electron microscopy revealed that the undercondensed region consisted of a coiled fiber with a diameter of about 400 nm, and a gyre diameter of approximately 600 nm. The coiled fiber was composed of the 30 nm chromatin fiber loops. These findings indicate that a continuous coiled structure, which is the final higher order structure of the condensed chromatin fiber, exists throughout the entire length of the mouse L-cell metaphase chromosome.     

Cell cycle-specific changes in the ultrastructural organization of prematurely condensed chromosomes

Prematurely condensed chromosomes (PCC) of HeLa cells synchronized in different phases of the cell cycle were analyzed by high-resolution scanning electron microscopy. The purpose of this study was to examine changes in the arrangement of the basic 30-nm chromatin fiber within interphase chromosomes associated with progression through the cell cycle. These studies revealed that highly condensed metaphase chromosomes and early G1-PCC consisted of tightly packed looping fibers. Early to mid G1-PCC were more extended and exhibited gyres suggestive of a despiralized chromonema. Further attenuation of PCC during progression through G1 was associated with a gradual transition from packed looping fibers to single extended longitudinal fibers. This process occurs prior to the initiation of DNA synthesis which appears to be localized within single longitudinal fibers. Following replication of a chromosome segment, extended longitudinal fibers were rapidly reorganized into packed looping fiber clusters concomitant with the formation of a multifibered chromosome axis. This results in the characteristic “pulverized” appearance of S-PCC when viewed by light microscopy. Subsequently, adjacent looping fiber domains coalesce, resulting in the uniformly packed, looping fiber arrangement observed in G2-PCC. Spiralization of the chromonema during the G2-mitotic transition results in the formation of highly compact metaphase chromosomes.     

草鱼染色体的包装（间期—中期）

以草鱼ＺＣ７９０１细胞株为材料,观察鱼类细胞从间期染色质到中期染色体的包装过程。主要通过（１）分裂期与间期细胞融合,诱导染色体早熟凝集;（２）染色体“伸长”处理;（３）培养细胞的低渗处理;（４）染色质辅展等方法,制作染色体标本,进行扫描和透射电镜观察。观察表明,鱼类染色质的基本结构与哺乳类细胞相同,也是直径约１０ｎｍ的核丝。染色体的色装有两种形式：一种是多级螺旋化形成直径约３００ｎｍ的染色单体,     

几种动物染色体超微结构的研究

应用表面舒展技术、原位培养表面舒展技术和临界点干燥以及空气干燥等方法制备染色体标本,用FESEM和SEM观察了CHO、IB-RS-2哺乳动物细胞以及黄鳝肾细胞和鲫鱼血淋巴细胞的染色体。看到了染色体处于不同舒展状态的染色质纤维。在染色质纤维未完全展开排列紧密时,染色体臂的染色质纤维,缠绕排列有序,垂直于染色体纵轴,螺旋盘绕形成疏密程度不同的横纹。在纤维较为松散和完全松敌的状态下,可以看见直径约为300(?)的染色质纤维从有序到不完全有序到无序,弯扭、螺旋、缠绕,有些似“辐射环”状结构。在着丝点处可清楚地看到有二条纤维平行分别通连二染色单体臂,未见有染色体膜。初步比较了鱼类和哺乳类的染色质纤维,二者基本一致,但鱼类染色质纤维排列较哺乳动物的松散,类似“辐射环”状的结构较为明显。     

The basis of chromatin fiber assembly within chromosomes studied by histone-DNA crosslinking followed by trypsin digestion

To determine the structural basis of chromatin assembly that leads to chromosome formation in mitosis, crosslinks were introduced by formaldehyde between contiguous components within chromosomes. Crosslinked stable products were then observed by electronmicroscopy after non-cross-linked portions were briefly digested by trypsin to unfold chromosomes. — When the DNA-histone crosslink was the primary product, trypsin readily unfolded the whole chromosome structure while preserving the 250 Å unit chromatin fiber intact; only a single unit fiber was tracked within the centromere region connecting the arms of each chromatid. When a histone polymer was formed by a prolonged formaldehyde crosslinking, trypsin digestion gave rise to chromatin fibers interacting with others at certain distances, and the typical chromosome structure remained unchanged. Regardless of the degree of crosslinking, there were neither thick supercoiled unit fibers nor proteinaceous cores. — These results suggest that the fiber connection may represent, to some extent, the interacting sites of folded chromatin fibers in situ within chromosomes, and also that the 250 Å unit fibers are the sole, highest structural basis in chromosomes. Since virtually no appreciable histone digestion took place in the crosslinked chromosomes, the observation that even after DNA-histone crosslinking the fiber interacting sites were accessible to trypsin preferentially over other portions, may be consistent with our recent results that the exposed, lysine-rich tails of histones represent such interacting sites.     

Visualization and Characterization of High-Order Chromatin Fibers under Light Microscope during Interphase and Mitotic Stages in Plants

Using genomic in situ hybridization with genomic DNA, high-order chromatin fibers were successfully exhibited under a light microscope through the cell cycle in barley, rice, maize and field bean. From the interphase to prophase and metaphase of mitosis, the fibers were basically similar. Each was estimated to be around 200 nm in diameter, but the strength of signals was not the same along the fiber length. Through the cell cycle a series of dynamic distribution changes occurred in the fibers. In the interphase, they were unraveled. At the early prophase they were arranged with parallel and mirror symmetry. During late-prophase and metaphase, the fibers were bundled and became different visible chromosomes. The parallel coiling and mirror symmetry structures were visible clearly until the metaphase. In anaphase they disappeared. During telophase, in peripheral regions of congregated chromosome group, borderlines of the chromosomes disappeared and the fibers were unraveled. This demonstrated that mitotic chromosomes are assembled and organized by parallel and adjacent coiling of the fibers and the fibers should be the highest order structure for DNA coiling.     

Low angle x-ray diffraction studies of HeLa metaphase chromosomes: effects of histone phosphorylation and chromosome isolation procedure

To test whether gross changes in chromatin structure occur during the cell cycle, we compared HeLa mitotic metaphase chromosomes and interphase nuclei by low angle x-ray diffraction. Interphase nuclei and metaphase chromosomes differ only in the 30-40-nm packing reflection, but not in the higher angle part of the x-ray diffraction pattern. Our interpretation of these results is that the transition to metaphase affects only the packing of chromatin fibers and not, to the resolution of our method, the internal structure of nucleosomes or the pattern of nucleosome packing within chromatin fibers. In particular, phosphorylation of histones H1 and H3 at mitosis does not affect chromatin fiber structure, since the same x-ray results are obtained whether or not histone dephosphorylation is prevented by isolating metaphase chromosomes in the presence of 5,5'-dithiobis(2- nitrobenzoate) or low concentrations of p-chloromercuriphenylsulfonate (ClHgPhSO3). We also compared metaphase chromosomes isolated by several different published procedures, and found that the isolation procedure can significantly affect the x-ray diffraction pattern. High concentrations of ClHgPhSO3 can also profoundly affect the pattern.