同步辐射(软X射线)辐照玉米的细胞学效应研究

利用同步辐射(软X射线)辐照玉米自交系H65和H14D种子,研究其M1代的细胞学效应,并以60Co-γ射线作对照。结果表明,软X射线辐照处理后,不仅能够诱发玉米M1代根尖细胞内核畸变和染色体畸变,而且还能够诱发染色体多种类型的变异,其变异频率随辐照剂量的增加而增大,辐照剂量与细胞总畸变率呈正相关。软X射线对玉米根尖细胞的有丝分裂具有明显地抑制作用,辐照剂量与细胞分裂指数呈负相关。软X射线辐照的细胞学效应与γ射线基本相似,但在诱发的细胞畸变率和染色体变异类型上存在一定的差异。两个供试品系对辐射的敏感性为H14D>H65。     

~(60)钴γ-线离体照射人淋巴细胞诱发的染色体畸变:剂量-效应关系及其与 X-线的比较研究

为了把外周血淋巴细胞中所观察到的染色体畸变量用来估算活体照射的剂量,需要确定离体照射血样本的剂量-效应曲线。继X-线离体照射的刻度实验后,我们又研究了~(60)钴γ-线诱发染色体畸变的剂量-效应曲线,并且比较了这两种辐射的效应,所得的结论如下:1.根据人淋巴细胞的染色体畸变进行生物剂量测定,只有在标准条件下确定的离体剂量-效应曲线才能得出有意义的结果,也就是说,应尽可能地与事故照射时所出现的条件相同。我们的研究表明,细胞遗传学的剂量测定应在37℃照射未受PHA~**刺激的全血,培养的时间应少于54小时。当比较各实验室符合上述条件的剂量-效应曲线时,显然有差异。因此,我们提议在细胞遗传学技术“标准化”之前,各实验室应建立自己的刻度曲线。2.按照世界卫生组织的标准,在分析剂量-效应关系时,以最小二乘方对实验资料作泊森方差和加权回归分析,拟以恰当的模式。如X-线一样,~(60)钴γ-线诱发的双着丝粒体和着丝粒环资料最适于二次多项式,a=0(回归线通过原点),分别为Y_(γ-线)=(0.59±0.94)×10~(-4)D+(4.77±0.40)×10~(-6)D~2和Y_(x-线)=(0.51±0.21)×10~(-3)D+(5.02±0.77)×10~(-6)D~2。这一方程的物理含义是,一些互换畸变由一次击中所产生,而另一些则是由二个分开的击中相互作用所致。当把两种辐射的系数b 和c 作比较时,主要差别在于导致不对称互换畸变的b 项。根据该指数方程.,畸变量的直线成分(即由一击损伤所致)与剂量率无关,而λ值(b/c)即为一击和二击事件的提供量相等时的剂量,此值分别为λ_(γ-线)=12拉德,而λ_(X-线)=100拉德。由此可以设想,在从事~(60)钴γ-线超暴光例子的生物剂量测定时,与X-线相比较,更应考虑到剂量率的问题。双着丝粒体和着丝粒环资料,同样也可拟以幂函数,此时,Y_(γ-线)=1.64×10~(-5)D~(1.79±0.11),而Y_(X-线)=6.50×10~(-5)D~(1.61±0.05)。然而应当指出,资料拟以二次多项式比幂函数更好些,当从畸变量外推至低剂量时尤其是这样。3.鉴于各种辐射诱发的畸变量彼此不一,所以从辐射防护出发,必须分别地确定剂量-效应曲线。这里的工作表明,~(60)钴γ-线诱发畸变的效能比X-线要低。~(60)钴γ-线相对于180kVX-线诱发互换畸变的RBE 不是一个单一值,其RBE 值变动在0.12和0.89之间。从RBE-剂量关系可以看出,在所用的剂量范围(24～488拉德)内,RBE 值随~(60)钴γ-线剂量的增加而上升,随后即趋向于饱和。     

~(60)Co-γ射线辐照对豇豆M1代生长发育的影响

用~(60)Co-γ射线对3个豇豆品种(天畅9号、詹豇215、早生王)干种子进行200 Gy、400 Gy、~(60)0 Gy、800 Gy和1 000 Gy辐照处理,研究不同剂量对豇豆M1代生长发育的影响。结果表明:低剂量下效应不明显,高剂量对生育进程有延后和抑制作用,发芽势、发芽率、出苗率、生长速率及叶绿素含量下降,叶片变小,分枝数、有效花梗数、结荚数减少;豇豆不同品种对γ射线敏感性不同,天畅9号、詹豇215、早生王的半致死剂量分别为234 Gy、490 Gy、553 Gy。     

~(60)Coγ线引起上海真厉螨染色体畸变的研究

本文首次报道用~(6O)Coγ线照射一种革螨——上海真厉螨引起的染色体畸变的研究。用~(6O)Coγ线(剂量1—50Krad)照射雌性革螨,引起的染色体畸变类型有:染色体裂隙、断片、微小体、环形染色体、粉碎化和多倍体,染色体断片是最常见的畸变类型,并观察到微核的形成。染色体畸变率随照射剂量增加而增高,辐射剂量与畸变率之间存在密切相关(相关系数为0.85,P<0.025),配得曲线回归方程为Y=3.27+14.49lg(X+1)。     

60Co线引起伤害真厉螨染色体畸变的研究

本文首次报道用60Coγ线照射一种地革螨-上海真厉螨引起的染色体畸变的研究.用60Coγ线(剂量1-50Krad)照射雌性革螨,引起的染色体裁畸变类型有: 染色提裂隙、断片、微小体、环形染色体、粉碎化和多倍体,染色体断片是最常见的畸变类型,并观察到微核的形成。染色体畸变率随照射剂量增加而增高,辐射剂量与畸变率之间存在密切相关（相关系数为0.85, P<0.025), 配得曲线回归方程为Y=3.27+14.49lg(X+1).     

60Coγ射线辐照花魔芋球茎的早期诱变效应研究

本试验开展了花魔芋早期诱变效应研究。通过^60Coγ射线辐照后的植株苗期发育观察和根尖细胞学检测分析,结果表明魔芋辐射诱变效应显著：辐照诱发核畸变和染色体畸变,且变异频率与剂量呈二次曲线关系;低剂量时对细胞分裂有刺激作用;辐照抑制芽体发育、苗期生长,抑制效应随剂量增加而加大,直至产生致死作用。依据花魔芋的辐射敏感性,建立了魔芋辐照诱变体系,以催芽球茎为诱变材料,诱变剂量范围为0Gy～50Gy,剂量率为1Gy／min,中等适宜剂量为7Gy～10Gy,致死剂量50Gy。     

14MeV中子照射离体人血细胞诱发的染色体畸变:剂量-效应关系及其与X-、γ-线的比较研究

继X-、γ-线在离体条件下的“刻度”实验之后,我们又在“标准”条件下研究了14MeV中子诱发人淋巴细胞染色体畸变的剂量-效应关系,并着重比较了这三种辐射的效应。所得主要结果如下: 1.具有高LET的快中子诱发染色体畸变的类型,与具有低LET的X-线和γ-线大致是相似的。     

提高辐射诱变育种效果及辐射遗传效应的研究：Ⅲ作物辐射遗传效应 …

试验结果证明：１．γ辐射处理可导致水稻和蚕豆芽中可溶性蛋白质与同工酶谱的变化;２．γ射线诱发蚕豆根尖细胞畸变率、断片率、微核率、染色体畸变率与辐照剂量的关系均符合模式Ｙ＝ａ＋ｂｘ＋ｃｘ＾２,呈抛物线变化趋势;３．微核率、断片率与核畸变率、染色体畸变率呈正相关。从而作者认为微核率、断片率可以作为检测染色体辐射效应的可靠指标;４．γ射线照射可引起蚕豆根尖细胞染色体带型的变化。     

平阳霉素诱发人精子染色体畸变——评价化学物质致断性的一种离体测试系统

人精子经已知的致断剂平阳霉素处理后与去透明带地鼠卵受精,继而制备人精子染色体进行核型分析。平阳霉素20μg、40μg、60μg/ml各剂量组均诱发出了多种类型的染色体结构畸变;其畸变精子率依次为39％、44％、52.4％,其断裂均数依次为1.90、2.70、3.86,与对照组的畸变精子率4％和断裂均数0.07分别进行比较,差异非常显著(P<0.01)并存在明显的剂量依赖关系。本研究为检测化学物质对人精子中染色体的致断作用提供了一种新的离体测试系统。     

提高辐射诱变育种效果及辐射遗传效应的研究Ⅲ作物辐射遗传效应的研究

试验结果证明：１．γ辐射处理可导致水稻和蚕豆芽中可溶性蛋白质与同工酶谱的变化;２．γ射线诱发蚕豆根尖细胞畸变率、断片率、微核率、染色体畸变率与辐照剂量的关系均符合模式Ｙ＝ａ＋ｂｘ＋ｃｘ￣２,呈抛物线变化趋势;３．微核率、断片率与核畸变率、染色体畸变率呈正相关。从而作者认为微核率、断片率可以作为检测染色体辐射效应的可靠指标;４．γ射线照射可引起蚕豆根尖细胞染色体带型的变化。     

Control of daughter centriole formation by the pericentriolar material

Controlling the number of its centrioles is vital for the cell, as supernumerary centrioles cause multipolar mitosis and genomic instability. Normally, one daughter centriole forms on each mature (mother) centriole; however, a mother centriole can produce multiple daughters within a single cell cycle. The mechanisms that prevent centriole 'overduplication' are poorly understood. Here we use laser microsurgery to test the hypothesis that attachment of the daughter centriole to the wall of the mother inhibits formation of additional daughters. We show that physical removal of the daughter induces reduplication of the mother in S-phase-arrested cells. Under conditions when multiple daughters form simultaneously on a single mother, all of these daughters must be removed to induce reduplication. The number of daughter centrioles that form during reduplication does not always match the number of ablated daughter centrioles. We also find that exaggeration of the pericentriolar material (PCM) by overexpression of the PCM protein pericentrin in S-phase-arrested CHO cells induces formation of numerous daughter centrioles. We propose that that the size of the PCM cloud associated with the mother centriole restricts the number of daughters that can form simultaneously.     

Effects of 4-hydroxyandrostenedione and hyperstimulation with pregnant mare serum gonadotrophin on early embryonic development in rats

A possible role of high oestradiol levels in mediating the adverse effects of hyperstimulation with pregnant mare serum gonadotrophin (PMSG) on early embryonic development in the rat was investigated using an aromatase inhibitor, 4-hydroxyandrostenedione (4-OHA), to inhibit endogenous oestradiol production. Three experiments were conducted in this study. In the first, varying doses of 4-OHA were administered either concurrently with human chorionic gonadotropin (hCG) to pro-oestrus female rats hyperstimulated at early di-oestrus stage with 20 IU PMSG or alone into nonhyperstimulated pro-oestrus females. At high doses of 1000, 2000, or 5000 microg/rat, 4-OHA substantially improved the survival of embryos in hyperstimulated females, while low doses of 100 and 500 microg/rat were ineffective. The protective effect of 4-OHA on embryo count was optimum at 2000 microg. When administered alone, only the highest dose of 5000 microg/rat 4-OHA increased embryo count. In the second experiment, higher doses of PMSG were studied (30 or 40 IU), with or without 5000 microg/rat 4-OHA given at the time of hCG injection. PMSG proved to be more detrimental with increasing dose, and 5000 microg/rat 4-OHA was able to rescue embryos from death in the 30, but not 40, PMSG group. In the third experiment, the influence of the timing of 4-OHA treatment on its ability to improve the embryo count in hyperstimulated females was examined by introducing 4-OHA 24 h earlier, rather than at the time of hCG treatment. The results showed the importance of timing of 4-OHA administration, as 5000 microg/rat 4-OHA was able to restore embryo survival in the 40 PMSG hyperstimulated group only when it was administered 24 h before hCG injection. Together, these results highlighted that 4-OHA, when administered at the appropriate time and dose, could reverse the negative effects of hyperstimulation from PMSG on early embryonic development. This may be due to its potent aromatase inhibiting properties that lead to the suppression of oestrogen production, thereby alleviating the supraphysiological level of oestradiol, which is typically present in PMSG-treated females. Interestingly, 4-OHA treatment on its own was able to positively influence embryo count when given at a high dose of 5000 microg/rat, and this may be associated with its weak androgenic properties. In conclusion, this study supports the hypothesis that excessive oestradiol is responsible for the negative effects of hyperstimulation with PMSG on early embryonic development.     

GIANT CENTRIOLE FORMATION IN SCIARA

Although somatic tissues of Sciara contain 9-membered centrioles, germ line tissues develop giant centrioles with 60–90 singlet tubules disposed in an oval array. Some 9-membered centrioles still may be seen in second instar spermatogonia. Each of these centrioles is associated with a larger "daughter" or secondary centriole at right angles to it. Most centrioles of second instar spermatogonia consist of 20–50 singlet tubules arranged in an oval, sometimes associated with an even larger secondary centriole. The more recently formed centriole of a pair is distinguishable from its partner by a concentric band of electron-opaque material inside its tubules. If a pair of centrioles at right angles to each other is pictured as a "T" formed by two cylinders, the secondary centriole is always the stem of the T; the primary centriole is the top. The two centrioles are oriented at the pole of the mitotic spindle so that the tubules of the primary centriole are parallel to the spindle axis. Each daughter cell receives a pair of centrioles and, during interphase, each of these centrioles gives rise to a new daughter centriole. A Golgi area of characteristic morphology is found in association with centrioles shortly after two new ones have formed. We conclude that in Sciara a centriole may give rise to a daughter morphologically different from itself. Whether the daughter is a 9-membered or giant centriole depends on the tissue type and stage of development.     

Spatial relationships between the anterior centriole and the mitotic center during interphase in the amoebae of the myxomycete Physarum polycephalum

Amoebae of the Myxomycete Physarum polycephalum in the interphase state typically contain only one proflagellar apparatus in which the anterior kinetosome (anterior centriole) is attached to the microtubule organizing center 1 (mtoc 1). We built strains possessing more than one mtoc 1 and a variable number of anterior centrioles to allow the appearance of new structures. In 8% of the amoebae of these strains, the 1:1 attachment between the anterior centriole and the mtoc 1 is not always respected. In nine cases studied using tridimensional reconstructions from ultrastructural thin sections, the pattern of attachment was more complex. A mtoc 1 could be linked to several anterior centrioles, and/or reciprocally an anterior centriole could be linked to several mtoc 1. In one case, an anterior centriole was not linked to a mtoc 1 and in three cases, a single centriole exhibited anterior and posterior characteristics. These observations suggest that (1) each pair of centrioles constitutes a morphological and physiological entity that is distinct from the mitotic center (mtoc 1); (2) the attachment of the anterior centriole to the mtoc 1 occurs at the end of each mitosis; (3) there is an inductory process during the morphogenesis of the link between the anterior centriole and the mtoc 1; (4) the anterior characteristics of a centriole can be present in the absence of the link with the mtoc 1; (5) the anterior and posterior characteristics of a centriole are not exclusive of each other, ruling out the existence of a lineage corresponding to the anterior centriole and a lineage corresponding to the posterior centriole; and (6) the differences between anterior and posterior centrioles result from a maturation process.     

The Chernobyl fallout in Salzburg/Austria and its effect on blood chromosomes

The radioactive fallout of the Chernobyl accident caused an increase in radiation dose of 20 to 110 per cent over the normal environmental burden to the inhabitans of Salzburg City in Austria (in a distance of about 1300 km from the accident). The structural chromosome aberration in the lymphocytes of the peripheral blood of 15 test-persons have been investigated one year after the accident. From two of these we know also the aberration frequencies before the accident which were significantly lower. The results from all test persons were pooled according to their Cs137 and Cs134 content, measured by whole body counter. Their mean additional blood doses from the incorporated caesium plus the external fallout radiation were 0.23, 0.36 and 0.55 mGy/yr. The aberration frequencies increased with dose. The slope of the best straight-line through the points was 2.0 +/- 0.7 total chromosome type aberrations in 100 metaphases per mGy/yr. This result fits in well with former investigations of persons with individually calculated radiation burden from the environment. The sharp increase with dose at this low level is not compatible with values extrapolated from high doses. The usual dose assessment based on chromosome aberrations extrapolated from high to low doses is therefore not possible in the range considered in this investigation.     

A molecular marker for centriole maturation in the mammalian cell cycle

The centriole pair in animals shows duplication and structural maturation at specific cell cycle points. In G1, a cell has two centrioles. One of the centrioles is mature and was generated at least two cell cycles ago. The other centriole was produced in the previous cell cycle and is immature. Both centrioles then nucleate one procentriole each which subsequently elongate to full-length centrioles, usually in S or G2 phase. However, the point in the cell cycle at which maturation of the immature centriole occurs is open to question. Furthermore, the molecular events underlying this process are entirely unknown. Here, using monoclonal and polyclonal antibody approaches, we describe for the first time a molecular marker which localizes exclusively to one centriole of the centriolar pair and provides biochemical evidence that the two centrioles are different. Moreover, this 96-kD protein, which we name Cenexin (derived from the Latin, senex for "old man," and Cenexin for centriole) defines very precisely the mature centriole of a pair and is acquired by the immature centriole at the G2/M transition in prophase. Thus the acquisition of Cenexin marks the functional maturation of the centriole and may indicate a change in centriolar potential such as its ability to act as a basal body for axoneme development or as a congregating site for microtubule-organizing material.     

Centrioles: some self-assembly required

Centrioles play an important role in organizing microtubules and are precisely duplicated once per cell cycle. New (daughter) centrioles typically arise in association with existing (mother) centrioles (canonical assembly), suggesting that mother centrioles direct the formation of daughter centrioles. However, under certain circumstances, centrioles can also selfassemble free of an existing centriole (de novo assembly). Recent work indicates that the canonical and de novo pathways utilize a common mechanism and that a mother centriole spatially constrains the self-assembly process to occur within its immediate vicinity. Other recently identified mechanisms further regulate canonical assembly so that during each cell cycle, one and only one daughter centriole is assembled per mother centriole.     

The de novo centriole assembly pathway in HeLa cells: cell cycle progression and centriole assembly/maturation

It has been reported that nontransformed mammalian cells become arrested during G1 in the absence of centrioles (Hinchcliffe, E., F. Miller, M. Cham, A. Khodjakov, and G. Sluder. 2001. Science. 291:1547-1550). Here, we show that removal of resident centrioles (by laser ablation or needle microsurgery) does not impede cell cycle progression in HeLa cells. HeLa cells born without centrosomes, later, assemble a variable number of centrioles de novo. Centriole assembly begins with the formation of small centrin aggregates that appear during the S phase. These, initially amorphous "precentrioles" become morphologically recognizable centrioles before mitosis. De novo-assembled centrioles mature (i.e., gain abilities to organize microtubules and replicate) in the next cell cycle. This maturation is not simply a time-dependent phenomenon, because de novo-formed centrioles do not mature if they are assembled in S phase-arrested cells. By selectively ablating only one centriole at a time, we find that the presence of a single centriole inhibits the assembly of additional centrioles, indicating that centrioles have an activity that suppresses the de novo pathway.     

Cep57 and Cep57L1 maintain centriole engagement in interphase to ensure centriole duplication cycle

Centrioles duplicate in interphase only once per cell cycle. Newly formed centrioles remain associated with their mother centrioles. The two centrioles disengage at the end of mitosis, which licenses centriole duplication in the next cell cycle. Therefore, timely centriole disengagement is critical for the proper centriole duplication cycle. However, the mechanisms underlying centriole engagement during interphase are poorly understood. Here, we show that Cep57 and Cep57L1 cooperatively maintain centriole engagement during interphase. Codepletion of Cep57 and Cep57L1 induces precocious centriole disengagement in interphase without compromising cell cycle progression. The disengaged daughter centrioles convert into centrosomes during interphase in a Plk1-dependent manner. Furthermore, the centrioles reduplicate and the centriole number increases, which results in chromosome segregation errors. Overall, these findings demonstrate that the maintenance of centriole engagement by Cep57 and Cep57L1 during interphase is crucial for the tight control of centriole copy number and thus for proper chromosome segregation.