Location of RAD51-like protein during meiotic prophase in Eimeria tenella

This study focuses on reporting events in Eimeria tenella oocysts from early to late prophase I in terms of RAD51 protein in association with the synaptonemal complex formed between homologous chromosomes. The aim of the study was the sequential localization of RAD51 protein, which is involved in the repair of double-strand breaks (DSBs) on the eimerian chromosomes as they synapse and desynapse. Structural Maintenance of Chromosome protein SMC3, which plays a role in synaptonemal complex formation, was labeled to identify initiation and progress of chromosome synapsis and desynapsis in parallel with the appearance and disappearance of RAD51 foci. Antibodies directed against RAD51 and cohesin subunit SMC3 proteins were labeled with either fluorescence or colloidal gold to visualize RAD51 protein foci and synaptonemal complexes. RAD51 protein localization during prophase I was studied on meiotic chromosomes spreads obtained from oocysts at different points in time after the start of sporulation. The present findings showed that foci detected with the antibody directed against RAD51 protein first appeared at the pre-leptotene stage before homologous chromosomes began pairing. Subsequently, the foci were detected in association with the lateral elements at the precise sites where synapsis were in progress. These findings lead us to suggest that in E. tenella, homologous chromosome pairing was a DSB-dependent mechanism and reinforced the participation of RAD51 protein in meiotic homology search, alignment and pairing of chromosomes.     

哺乳动物卵母细胞非整倍体产生机制的研究进展

雌性生殖细胞进行减数分裂时易发生染色体分离错误而产生非整倍体卵母细胞,其受精后会产生非整倍体胚胎,导致出生缺陷或胚胎致死,是影响哺乳动物繁殖的重要因素。卵母细胞在第一次减数分裂前期发生同源染色体联会,此时DNA双链断裂引发重组。重组时缺乏交叉、重组事件数量的减少及交叉靠近端粒或着丝粒导致染色体发生同向分离或不分离,从而产生非整倍体卵母细胞。减数分裂期间,当染色体的端粒共向于同一极或没有完全附着在纺锤体微管上时,纺锤体组装检查点（spindle assembly checkpoint,SAC）被激活,E3泛素连接酶APC/Cyclome （APC/C）沉默,保护分离酶抑制蛋白（securin）和细胞周期蛋白B （cyclin B）不被降解,从而抑制分离酶和染色体的分离。直到所有染色体与纺锤体实现稳定的双极定向并正确排列到赤道板上,SAC关闭,染色体正确分离。卵母细胞中SAC蛋白缺失,导致SAC不能有效地监测端粒在纺锤体上的正确附着,发生染色体分离错误,从而产生非整倍体卵母细胞。因此,通过现代分子技术手段解析非整倍体卵母细胞所涉及的机制是保护哺乳动物生育的重要目标。作者主要介绍了卵母细胞减数分裂的特点,详细阐述了卵母细胞非整倍体发生的染色体分离错误的分子机制,以期为开发卵母细胞非整倍体的治疗手段提供参考。     

Roles of Cohesin and Condensin in Chromosome Dynamics During Mammalian Meiosis

Meiosis is a key step for sexual reproduction in which chromosome number is halved by two successive meiotic divisions after a single round of DNA replication. In the first meiotic division (meiosis I), homologous chromosomes pair, synapse, and recombine with their partners in prophase I. As a result, homologous chromosomes are physically connected until metaphase I and then segregated from each other at the onset of anaphase I. In the subsequent second meiotic division (meiosis II), sister chromatids are segregated. Chromosomal abnormality arising during meiosis is one of the major causes of birth defects and congenital disorders in mammals including human and domestic animals. Hence understanding of the mechanism underlying these unique chromosome behavior in meiosis is of great importance. This review focuses on the roles of cohesin and condensin, and their regulation in chromosome dynamics during mammalian meiosis.     

Fertile male tortoiseshell cat with true chimerism 38,XY/38,XY

The tortoiseshell coat colour is characteristic to female cats, and its occurrence in tomcats is very rare and associated with chromosome abnormalities (additional copy of X chromosome). The aim of this study was identification of the genetic basis of a case of tortoiseshell colour in a fertile Maine coon tomcat. Cytogenetic and molecular genetic studies were carried out with painting molecular probes (WCPP) specific to the X and Y sex chromosomes as well as a DNA microsatellite panel for the parentage verification of cats. Cytogenetic analysis revealed only a single set of sex chromosomes typical for male – 38,XY. The results of the microsatellite polymorphism obtained from DNA showed three alleles in locus FCA201 and four alleles in loci FCA149 and FCA441 in different tissues (blood, hair roots and testicles). Based on these results, the case was diagnosed as a true chimerism 38,XY/38,XY. To the best of our knowledge, this is the first case of a 38,XY/38,XY chimera diagnosed in cats, confirmed by genetic analysis.     

马鹿染色体核型分析

采用常规细胞遗传学分析手段，对青海省西宁动物园饲养的马鹿染色体核型进行分析，结果：马鹿染色体数目为２ｎ＝６８，染色体臂数ＮＦ＝７０（♀）、７１（）；常染色体类型为２条中（Ｍ）着丝粒，６４条近端或端（Ａ）着丝粒；性染色体类型Ｘ为近端（Ａ），Ｙ为近中（ｓＭ）着丝粒；公母鹿核型式为６８，ＸＹ和６８ＸＸ。     

rob(13/17)纯合子猪的联会复合体分析

采用表面铺展法和硝酸银染色技术对rob(13/17)纯合子猪减数分裂的配对行为进行了联会复合体（synaptonemal complex,SC）的电镜观察和分析。结果表明，rob(13/17）纯合子猪的SC配对行为正常，未见三价体和侧生组分形成。SC的形成开始于偶线期，成熟于粗线期，解体于双线期。在减数分裂前期，性染色体轴呈强嗜银性，配对明显落后于常染色体。根据减数分裂前期性染色体的形态和行为，把性染色体的SC分为4个类型。     

Wapl localization on the synaptonemal complex, a meiosis-specific proteinaceous structure that binds homologous chromosomes, in the female mouse.

The wings apart-like (Wapl) protein is required to hold sister chromatids together in mitotic heterochromatin in Drosophila melanogaster. It is localized on the synaptonemal complex (SC), a meiosis-specific structure connecting one pair of sister chromatids to the homologous pair in mouse pachytene spermatocytes. The human Wapl is a cohesin-binding protein that facilitates cohesin's timely release from chromosome arms during prophase. The objective of the present study was to determine the subcellular localization of the mouse Wapl on female meiotic chromosomes at pachynema. The pachytene oocytes were isolated from foetal ovaries at 18.5 dpc and double immunostained with anti-synaptonemal complex protein 2 (SYCP2) and anti-Wapl. In the pachytene oocytes examined, mouse Wapl was colocalized with SYCP2 on the SC. Our results further implicated that Wapl might play a crucial role in meiotic chromosome remodelling at early meiosis.     

高羊茅雄性不育株花粉母细胞减数分裂染色体行为观察

采用卡宝品红染色制片法,对高羊茅(Festuca arundinacea)雄性不育植株花粉母细胞减数分裂过程及异常行为进行观察,结果发现,高羊茅减数分裂进程与小花大小、花药长度、色泽有较为密切的关系。减数分裂周期具有不同步性,同一制片中可观察到2～4个不同时期的分裂相,这种现象是高羊茅在进化过程中环境适应的一种表现,有利于增强种群繁殖稳定性。本研究还发现,减数分裂过程中存在大量单价体、染色体桥、落后染色体、不均等分离、微核、三分体等异常现象,初步分析确定这些小孢子异常分裂是导致高羊茅花粉败育的重要原因之一。     

布尔山羊的染色体核型分析

采用外周血淋巴细胞培养法，对布尔山羊的染色体核型进行了分析。结果表明，布尔山羊的二倍体染色体数目为2n=60，公羊核型为60，XY；母羊核型为60，XX。共有29对常染色体和1对性染色体。所有常染色体均为端部着丝点染色体；X染色体为第二大的端部着丝点染色体，Y染色体为最小的天是唯一的中部着丝点染色体。研究发现，布尔山羊存在1.7％的三倍体和5.9％的四倍体。     

荒漠猫染色体的首次研究

采用外周血淋巴细胞培养和常规染色体制备分析技术，对青海省珍稀兽类荒漠猫染色体进行首次研究，结果显示：荒漠猫的染色体数目为２ｎ＝３８，常染色体形态类型为１０Ｍ＋１０ｓＭ＋１２ｓＴ＋４Ｔ（或Ａ），性染色体形态类型Ｘ为ｓＭ，Ｙ为ｓＴ，染色体总臂数ＮＦ＝７２，公母猫核型式为３８，ＸＹ和３８，ＸＸ。     

Distribution of the sex chromosome during mouse spermatogenesis in testis tissue sections

During mammalian spermatogenesis, spermatogenic cells undergo mitotic division and are subsequently divided into haploid spermatids by meiotic division, but the dynamics of sex chromosomes during spermatogenesis are unclear in vivo. To gain insight into the distribution of sex chromosomes in the testis, we examined the localization of sex chromosomes before and after meiosis in mouse testis sections. Here, we developed a method of fluorescence in situ hybridization (FISH) using specific probes for the X and Y chromosomes to obtain their positional information in histological testis sections. FISH analysis revealed the sex chromosomal position during spermatogenesis in each stage of seminiferous epithelia and in each spermatogenic cell. In the spermatogonia and leptotene spermatocytes, sex chromosomes were distantly positioned in the cell. In the zygotene and pachytene spermatocytes at prophase I, X and Y chromosomes had a random distribution. After meiosis, the X and Y spermatids were random in every seminiferous epithelium. We also detected aneuploidy of sex chromosomes in spermatogenic cells using our developed FISH analysis. Our results provide further insight into the distribution of sex chromosomes during spermatogenesis, which could help to elucidate a specific difference between X and Y spermatids and sex chromosome-specific behavior.     

披碱草和野大麦杂种F1与BC1代细胞遗传学研究

对亲本披碱草(Elymus dahuricus)和野大麦(Hordeum brevisubulatum)及其杂种F₁和BC₁代体细胞染色体数和花粉母细胞(PMC)减数分裂染色体行为进行观察和分析。结果表明:亲本披碱草的染色体数为2n=42,野大麦为2n=28;正、反交杂种F₁染色体数均为35;BC₁代染色体发生了数目上的差异,变异范围为28~35,并出现很多非整倍性植株;披碱草的PMCMI二价体配对频率很高,野大麦PMCMI出版少量的四价体、三价体和单价体,但以二价体占优势;正反交杂种F₁减数分裂不规则,单价体的普遍存在是造成杂交不育的重要原因;BC₁代的PMCMI单价体频率较F₁代有所下降,二价体频率有所增高,有些株系的环状二价体占绝对优势,这为杂种育性恢复提供了细胞学依据。     

培育中的青海白猪染色体核型分析

采用外周血淋巴细胞培养方法和常规染色体制备分析技术．分析了青海白猪的染色体核型。结果表明：青海白猪染色体数目为2n=38条，染色体臂数NF为32，公猪染色体核型式为38，XY；母猪染色体棱型式为38，XX。核型模式：2n=♂/♀38/38（12M 8SM 4ST 12T）XY（M，M）/XX（M，M）     

哺乳动物卵母细胞纺锤体的形成及纺锤体检验点的作用机制

染色体精确分离是在纺锤体的正确组装和纺锤体检查点(spindle assembly checkpoint,SAC)的监控下完成的,对于哺乳动物卵母细胞来说,纺锤体的形成和SAC都是保证染色体精确分离的重要因素,如果染色体分离错误将直接导致自发性流产或其他出生缺陷。卵母细胞中心体缺失后,细胞依然能够依靠独立于中心体而围绕染色体成核的微管反向平行排列能形成双极纺锤体,即自我组装纺锤体。由微观组织中心(microtubue organizing center,MTOC)召集微管聚集,成熟促进因子(maturation promoting factor,MPF)维持两次减数分裂过程中纺锤体的形成过程,细胞静止因子(cytostatic factor,CSF)维持分裂中期结构,使纺锤体在染色体没有全部集合到赤道板时保持稳定。大体积的卵母细胞容易产生非整倍体,且卵母细胞中不含有中心体这一特殊性导致卵母细胞中是否存在SAC在很长一段时间内存在争议,但现在SAC是确保卵母细胞染色体精确分离的机制之一已被初步证明。在减数分裂中期染色体之间存在一种黏连,细胞会产生"等待-后期"信号抑制SAC活性,从而保持这种黏连稳定,直至所有染色体完成与纺锤体的连接,"等待-后期"信号失活,SAC启动,使染色体间的黏连失活,进而在纺锤体的作用下染色体分离。作者综述了减数分裂过程中纺锤体的特异性组装过程和纺锤体检查点的组成及作用机制,丰富了减数分裂的相关知识,并为减数分裂过程中非整倍体的形成机制提供依据。     

Determination of the correlation between stallion's age and number of sex chromosome aberrations in spermatozoa

The aim of this study was a cytogenetic analysis of stallions semen to find sex chromosome aberrations and to determine if there was an association between stallion's age and aberration frequency for the sex chromosomes. Sperm samples were collected from 22 stallions of various age from 3 to 23 years. Multicolour FISH was performed on each sample, using probes for the sex chromosomes and EGFR gene, localized on 4p12 in domestic horse. A total of 26199 sperm cells were analysed (from 1 070 to 1 532 per animal). Among the analysed cells, there were 50.318% with X chromosome, 48.543% with Y chromosome and 1.139% with aberrant chromosomes. The frequency of aberrations was: sex chromosomes nullisomy (0.466%), XY aneuploidy (0.454%), XX disomy (0.146%), YY disomy (0.041%), diploidy (0.024%) and trisomy XXY (0.008%). Additionally there was a correlation between the age of an animal and the frequency of sex chromosome aberration and a significant positive correlation between age and disomy of XY, XX, YY, trisomy of XXY, autosomal disomy was seen. A Correlation between the age of a stallion and the level of nullisomy was negative. The present study demonstrated that FISH technique is a powerful method to identify sex chromosome aberrations in equine spermatozoa and might be very helpful for a breeder during a selection for the best stallion.     

Male tortoiseshell cats in the United Kingdom

A questionnaire concerning the coat colour and sex of cats being vaccinated or neutered was sent to 2585 veterinary practices; 393 (15.2 per cent) were returned and information was obtained about 9816 cats. Of 4598 males, 20 were recorded as tortoiseshell (0.43 per cent). The frequency of the orange gene was 19.7 per cent assuming that male tortoiseshell cats had two X chromosomes. The chromosome complement and/or gonadal histology of 14 male tortoiseshell cats is described. Cytogenetic analysis of 11 animals revealed six with a 38,XX/38,XY complement, two with 39,XXY, two with 38,XX, and one with a 38,XY complement.     

Repeated Early Embryonic Loss in a Thoroughbred Mare with a Chromosomal Translocation [64,XX,t(2;13)]

Peripheral blood samples from a 13-year-old Thoroughbred mare were submitted for chromosome analysis. The mare had a poor reproduction record only producing four live foals during her 10 years as a broodmare. She had remained barren or experienced early embryonic loss in the other 6 years. Chromosomal analysis revealed the mare carried a rare nonreciprocal translocation involving chromosomes 2 and 13 [64,XX,t(2;13)]. Fluorescence in situ hybridization with probes specific for horse chromosomes 2 and 13 were used to confirm the nonreciprocal translocation. Both conventional and molecular cytogenetic techniques are important for identifying and characterizing chromosomal abnormalities in horses with poor reproductive performance, particularly in mares experiencing repeated early embryonic loss.     

过冷却处理家蚕卵对四倍体诱发率及后代性比的影响

探讨了家蚕盛产卵后不同时间的卵过冷却处理对四倍体诱发率和后代三倍体性比发生的影响。不同杂交组合材料盛产卵后不同时间卵所处的分裂期不同 ,影响到四倍体雌蚕的诱发率 ,处于第一分裂期的卵数越多 ,诱发率越高 ;诱发的四倍体与正常二倍体交配产生的三倍体蚕雌雄比例由 3∶1变为 5∶1是由于四倍体减数分裂中性染色体的行为 ,性比 3∶1时 ,仅是Z与Z、W与W之间的联会 ,其后随着这种联会增加 ,Z与W之间的联会也逐渐开始 ,从而使雌雄比例接近 5∶1。     

家蚕核仁和核仁染色体的初步研究

在家蚕的染色体制片中.以精巢为材料,采用涂片.Giemsa染色法,对一代杂种苏5×苏6的核仁和核仁染色体进行了观察和分析.结果表明,在间期细胞核内.大多数有一个核仁,少数有两个或三个核仁.在减数分裂粗线期至终变期的细胞核内,有一个核仁和三条贴附在核仁上的核仁染色体,这三条核仁染色体分别是第2号、第5号和第22号.有丝分裂中期的细胞核内未见核仁,但至少观察到四条染色体有明显的缢痕(Chromosome constriction),它们是两对与核仁形成的有关的核仁染色体.其他染色体均呈短杆状,未见明显缢痕.     

CYTOGENETICAL STUDIES OF INFERTILE PIGS

Cytogenetical studies were made on 6 infertile pigs. Post-mortem examination of the reproductive organs of 5 of these pigs showed them to be intersexes. Regardless of the degree of gonadal deviation from the normal, chromosome karyotype of cultured peripheral blood lymphocytes, bone marrow, liver and kidney, revealed that 4 of the intersexes had a 38, XX constitution. One intersex exhibited sex chromosome mosaicism (38XX/38XY) in lymphocytes from the peripheral blood and bone marrow and a normal male karyotype (38XY) in cultured liver and kidney cells. The sixth pig, a phenotypically normal boar, also had a 38XX/38XY chromosome constitution.