Spectral karyotyping (SKY) of mouse meiotic chromosomes.

The spectral karyotyping procedure of in situ hybridization with chromosome-specific probes assigns a unique colour code to each of the 21 mouse mitotic chromosomes. We have adapted this procedure to meiotic prophase chromosomes, and the results show that each of the pachytene or metaphase I bivalents can be identified. This technique has the potential to recognize synaptic anomalies and chromosome-specific structural and behavioural characteristics. We confirm these potentials by the recognition of the heterologous synapsis of the X and Y chromosomes and by the variances of synaptonemal complex lengths for each of the colour-coded bivalents in eight prophase nuclei.     

Spectral Karyotyping to Study Chromosome Abnormalities in Humans and Mice with Polycystic Kidney Disease

Conventional method to identify and classify individual chromosomes depends on the unique banding pattern of each chromosome in a specific species being analyzed ^{1, 2}. This classical banding technique, however, is not reliable in identifying complex chromosomal aberrations such as those associated with cancer. To overcome the limitations of the banding technique, Spectral Karyotyping (SKY) is introduced to provide much reliable information on chromosome abnormalities.SKY is a multicolor fluorescence in-situ hybridization (FISH) technique to detect metaphase chromosomes with spectral microscope ^{3, 4}. SKY has been proven to be a valuable tool for the cytogenetic analysis of a broad range of chromosome abnormalities associated with a large number of genetic diseases and malignancies ^{5, 6}. SKY involves the use of multicolor fluorescently-labelled DNA probes prepared from the degenerate oligonucleotide primers by PCR. Thus, every chromosome has a unique spectral color after in-situ hybridization with probes, which are differentially labelled with a mixture of fluorescent dyes (Rhodamine, Texas Red, Cy5, FITC and Cy5.5). The probes used for SKY consist of up to 55 chromosome specific probes ^7-10.The procedure for SKY involves several steps (Figure 1). SKY requires the availability of cells with high mitotic index from normal or diseased tissue or blood. The chromosomes of a single cell from either a freshly isolated primary cell or a cell line are spread on a glass slide. This chromosome spread is labeled with a different combination of fluorescent dyes specific for each chromosome. For probe detection and image acquisition,the spectral imaging system consists of sagnac interferometer and a CCD camera. This allows measurement of the visible light spectrum emitted from the sample and to acquire a spectral image from individual chromosomes. HiSKY, the software used to analyze the results of the captured images, provides an easy identification of chromosome anomalies. The end result is a metaphase and a karyotype classification image, in which each pair of chromosomes has a distinct color (Figure 2). This allows easy identification of chromosome identities and translocations. For more details, please visit Applied Spectral Imaging website (http://www.spectral-imaging.com/).SKY was recently used for an identification of chromosome segregation defects and chromosome abnormalities in humans and mice with Autosomal Dominant Polycystic Kidney Disease (ADPKD), a genetic disease characterized by dysfunction in primary cilia ^11-13. Using this technique, we demonstrated the presence of abnormal chromosome segregation and chromosomal defects in ADPKD patients and mouse models ¹⁴. Further analyses using SKY not only allowed us to identify chromosomal number and identity, but also to accurately detect very complex chromosomal aberrations such as chromosome deletions and translocations (Figure 2).     

黄扯旗鱼核型分析方法及核型特征

本研究的目的在于建立黄扯旗鱼(Pristella maxillaris)的核型分析方法,以了解该物种的染色体数目、形态及分类等遗传学特征。比较头肾-PHA法和胚胎制片法制备黄扯旗鱼染色体样品的异同。结果发现,头肾-PHA法难度较大,操作复杂且制备的分裂相细胞数目较少;而胚胎法操作简单,用时短,制备的分裂相细胞稳定且数目多。随后基于胚胎制片法利用Photoshop CS5和image J软件进行了黄扯旗鱼的核型分析。统计结果表明,黄扯旗鱼核型为2n=52=6m+12sm+34t,NF=70,且未发现多倍体、异型性染色体及随体染色体的现象。     

家蚕联会复合体组型分析

作者以表面铺张——硝酸银染色技术制备标本,从亚显微水平对雌、雄家蚕联会复合体(Synptonemal Complex,SC)的行为及组型进行观察和分析。在减数分裂前期,雌、雄家蚕SC的形态和行为均无明显差异。SC的形成起始于偶线期,成熟于粗线期,消失开始于双线期。在粗线期可见28条清晰的SC,在各SC均未见有相当于着丝粒区域的分化结构。无论在精母细胞或卵母细胞中,均未发现异形双价体。从早粗线期到晚粗线期,SC的平均总长由205.5μm伸长至348.9μm。作者根据10个细胞的测量及分析结果,绘制了家蚕SC组型模式图,并就家蚕的性决定进行了讨论。     

内江猪染色体G带及其定量分析

采用外周血淋巴细胞培养技术和胰酶法对内江猪染色体进行了G显带,并运用扫描显微分光光度法对染色体G带进行了系统的定量分析,获得了染色体核型及G带带纹的精确数据。包括每条单倍染色体的着丝粒指数、长度、面积、积分光密度;每条G带深染带纹的宽度、面积、积分光密度、带峰值等,并绘出了带吸收光密度柱形图和消光三维图。     

宜昌百合根尖染色体C-带分析

利用Giemsa C-带方法对宜昌百合（Lilium leucanthum(Baker) Baker）进行研究。结果表明：宜昌百合（L. leucanthum）的染色体数目为2n＝2x=24,单套染色体的条带总数目为21条。其带型公式为：2n＝24＝6C+2CI+2I+2CI⁺+2CI⁺+4I⁺+2I⁺+2T⁺+2I+S。宜昌百合（L. leucanthum）每条染色体上都显示出显著的特征带,且带纹的深浅差异明显。宜昌百合（L. leucanthum）的强带主要集中在着丝点及附近区域。通过Giemsa C-带方法可以将宜昌百合（L. leucanthum）的每条染色体区分开。     

Chromosome counts inPosidonia (Posidoniaceae)

The five species ofPosidonia occuring in Western Australia all have a diploid chromosome number of 20.     

Chromosome counts inCalea (Asteraceae-Heliantheae)

Chromosome numbers are reported for six taxa ofCalea, among them tetraploidC. septuplinervia, the only known polyploid inCalea s. str. The base chromosome number ofCalea is interpreted as x = 19.     

Rapid localization of transgenes in mouse chromosomes with a combined Spectral Karyotyping/FISH technique

We explored the feasibility of combined Spectral Karyotyping (SKY) and Fluorescence In Situ Hybridization (FISH) as means to rapidly map a chromosomally integrated renin/green fluorescent protein (GFP) fusion gene construct (Ren-GFP) in the transgenic mouse, Tg(Ren-GFP)1Kwg. A sequential hybridization with SKY probes followed by FISH gave consistently satisfactory results, demonstrating that multiple copies of the Ren-GFP transgene in this transgenic mouse line are integrated into a single chromosomal site of Chromosome (Chr) 4, most probably in the juxta-centromeric euchromatic region consisting of the A2-A3 domain. Chr 4 as a sole carrier of the transgene also was confirmed by co-hybridization to p1 BAC clone DNA containing telomeric sequences specific for mouse Chr 4 and the Ren-GFP construct in pGEM4Z vector. The hemizygosity of the Ren-GFP transgene is maintained not only in bone marrow cells, but also in lung cells proliferating in vitro, indicating that stable integration of the Ren-GFP transgene into chromosomal DNA was established at a very early embryonic stage. We conclude that the SKY/FISH technique is a reliable and facile method for establishing the integration site of a transgene. As such, this protocol has obvious advantages over traditional backcross methods in terms of time, cost and labor for determining the chromosomal location of transgenes.     

Chromosome counts inCephalotus (Cephalotaceae)

Cephalotus follicularis is uniformly n = 10 in all populations surveyed.     

Karyotyping and cytogenetic mapping of Atlantic cod (Gadus morhua Linnaeus, 1758)

The Atlantic cod (Gadus morhua) is an important natural resource for northern societies and is now also considered to be a promising candidate for aquaculture. In recent years, much effort has been directed towards the development of genomic tools, and genome initiatives for Atlantic cod have been established. Despite the growing attention devoted to the Atlantic cod genomics, basic aspects of its genome structure and organization remain unknown. Thus, the present work aims to study cytogenetic features of the Atlantic cod as a contribution to the knowledge of this species' genome. The Atlantic cod displays a diploid number of 46 chromosomes, with a karyotypic formula 16 m/sm + 30 st/t. Conventional karyotyping was improved by chromosomal mapping of two classes of repetitive sequences. 18S rDNA clusters were assigned to pairs 2 and 4; small amounts of 18S rDNA clusters were occasionally detected on pair 5. These findings could not be related to the geographical origin of the specimens, but were consistent with the variability of these repeated genes in fish in general. 5S ribosomal gene clusters, apparently corresponding to a single 5S rDNA class, were detected on twelve chromosomes (pairs 11, 12, 14, 17, 20 and 21). The present update of the existing but meagre information on the karyotype of Atlantic cod, plus the first physical mapping of repetitive genes in this species herein, opens the way for an integrated approach that combines genetic and physical mapping with the assembly of the genome of this commercially important species.     

用脉冲电场凝胶电泳分析棉病囊霉及其突变体的染色体DNA

采用交变脉冲电场凝胶电泳和碱变性交变脉冲电场凝胶电泳方法,分析了棉病囊霉酵母菌及其2个不同的突变菌株的核型,得知此菌株含有5条染色体 DNA,而2株突变体的染色体 DNA 都没有大片段的缺失或双链断裂,但其稳定性不如野生型菌株的 DNA,而且存在单链断裂等碱不稳定性位点.     

应用染色体涂染技术分析两例染色体结构异常 Analysis of the Structure of Abnormal Karyotypes by Using Chromosome Painting

为了确定两例细胞遗传学提示染色体结构异常的核型,应用通过显微切割技 术构建的人类18号和7号染色体探针池,分别对这两例病例的中期分裂相进行染色体涂染,结合显带染色体,确定两者核型分别为46,XY,t(3;18) (q12;q21)和46,XX,dir ins(1;7)(p3104;q34q36)。染色体涂染技术是染色体显带技术的重要补充和发展,为染色体结构异常提供了一种直观、准确的检测手段,在遗传咨询和产前诊断方面有重要作用。 Abstract:In this study,chromosome painting technique was performed to analyse the abnormal karyotypes of two carriers.Chromosome 18 and 7 specific libraries,which were generated by chromosome microdissection technique,were used as probe pools to hybridize the carriers metaphase chromosomes respectively.Unlabled human genomic DNA was used to inhibit the hybridization of sequences in the library that bind to mutiple chromosomes.Structure abnormality was detected clearly in metaphase.Combined with the banding chromosomes,we concluded that their karytypes were 46,XY,t(3;18)(q12;q21)and 46,XX,dir ins(1;7)(p3104;q34q36).Chromosome painting,as a direct and concise method in analysing chromosome structure abnormality,is an important complement and development of chromosome banding technique,and has important application in genetic counselling and prenatal diagnosis.     

Chromosome analysis ofHaemanthus andScadoxus (Amaryllidaceae)

Chromosome analysis of nine species ofHaemanthus (2n = 16) and four species ofScadoxus (2n = 18), using conventional stains, Quinacrine fluorescence and C-banding, has shown that the two genera do not possess significant amounts of constitutive heterochromatin. The two genera are closely related and differ in respect of a translocation which has resulted in the dysploid reduction in chromosome number from 2n = 18 inScadoxus to 2n = 16 inHaemanthus.     

Chromosome number ofSaruma oliver (Aristolochiaceae)

Chromosome number ofSaruma henryi, the only species of the genus, was counted for the first time. The species has 2n=52 chromosomes, and its chromosomal complement is characteristically composed of very small chromosomes. Based on chromosome data available for the family, comparisons indicate an isolated position ofSaruma in Aristolochiaceae.     

水晶兰属植物的染色体

对国产水晶兰属植物水晶兰（Monotropa uniflroa L．）和毛花松下兰（M．hypopitys var．hirsutea Roth）的花粉母细胞减数分裂进行了观察,它们的减数分裂中期Ⅰ的染色体数目均为n=24。结合前人所做的研究,确定该属的染色体基数为x=8,并对该属的染色体基数和倍性变异与地理分布区的关系进行了初步探讨。通过对鹿蹄草亚科和水晶兰亚科的染色体基数比较,结合两个亚科的生长习性和花药开裂方式的不同,作者赞同哈钦松系统将水晶兰亚科作为科的处理。     

Chromosome number inHalophytum ameghinoi (Halophytaceae)

The chromsome number and karyotype ofHalophytum ameghinoi (Speg.) Speg. was found to be 2n = 24 and 12m+10sm+2t, respectively. Meiosis was regular and 12 bivalents were observed. The relationships of this monotypic family are briefly discussed.