从感染黄瓜花叶病毒的烟草叶筛选抗病毒细胞突变体

用绿岛法从感染CMV的普通烟草(G140)叶筛选到抗病毒细胞突变体R~CMV2、6,其当代无性系群体病情指数较对照降低41.5-45.2%。Abstract:The 2 CMV-resistant-cell mutants,RCMV-2、6,were selected by using the method of culturing dark green island from CMV infected tobacco(N.tabacum cv G140)leaves.The disease in dex of RCMV-2、6′s was decreased by 41.5~45.2% in comparison with that of the control(G140).     

丝裂霉素C诱发人精子染色体畸变的研究

人精子经最终浓度分别为７．５,１５．３０μｇ／ｍｌ丝裂霉素Ｃ（ＭＭＣ）处理１ｈ后与去透明带地鼠卵受精,制备第１次卵裂中期相进行核型分析,以探讨ＭＭＣ对人精子的诱变效应。结果显示明显的量效关系。染色单体型畸变是诱发畸变的主要类型,但仍观察到一定数量染色体型畸变,表明ＭＭＣ作用于人精子与通常拟紫外线致断剂作用于体细胞诱变的结果并不完全一样。３６．８９％的诱发畸变为重接型,表明金黄地鼠卵母细胞内的修复系统能够修复ＭＭＣ所引起的人精子ＤＮＡ损伤     

Entry of immotile spermatozoa into zona-free hamster ova

We studied six men whose spermatozoa were immotile and possessed a variety of sperm tail structural abnormalities by electron microscopy. The semen of all six subjects had a normal percentage of oval forms and sperm undergoing capacitation and acrosome reaction. Despite the absence of motility, when incubated sperm from these subjects was added to a microdrop of medium containing zona pellucida-free hamster ova, sperm penetration or entry into the cytoplasm of from 1–9% of the eggs was evident with phase contrast microscopy. This latter finding suggests that, at least in this system, oocytes actively facilitate sperm incorporation. Penetration was absent when sperm of fertile men were rendered immotile, though still viable, by heat treatment.     

Fertilization by sperm injection in the rabbit

Whole rabbit spermatozoa and isolated sperm nuclei were microinjected directly into the ooplasm of hamster and rabbit ova. These injected sperm decondensed and formed male pronuclei during subsequent in-vitro culture. Injection of whole spermatozoa and sperm nuclei prepared by a protocol known to allow in-vitro capacitation of ejaculated spermatozoa yielded a significantly higher (P < 0.01) number of activated rabbit ova containing male pronuclei than did injection of uncapacitated epididymal sperm nuclei or ejaculated sperm nuclei. Rabbit ova fertilized by sperm injection were capable of undergoing normal-appearing cleavage division during 22 h of culture.     

喜树碱对人类巨细胞病毒诱发染色体畸变的频率的调节效应

本文评价了所选用的DNA修复抑制剂对人类巨细胞病毒(HCMV)诱发外周血淋巴细胞(PBLs)染色体畸变频率的影响。以拓扑酶Ⅰ的一种抑制剂——喜树碱(0.05—0.3μg/m1)处理HCMV感染的人类PBLs30小时,结果导致HCMV诱发的染色体损伤频率显著的协同性增加(P<0.O1)。另一方面以ADP核糖聚合酶的一种抑制剂——3—氨基苯酰胺(3—AB)(3—30μg/ml),或者拓扑酶Ⅱ的一种抑制剂——新霉素(3—30μg/ml)处理HCMV感染的PBLs30时小,染色体损伤频率未见明显增加。在喜树碱处理的HCMV感染细胞中,染色单体型断裂包括染色体交换是染色体畸变的主要类型,这提示HCMV感染与单链NDA断裂有关,这些发现还提示,HCMV感染不会造成通过3-AB或新霉素敏感途径修复的直接DNA损伤。     

Visualization and characterization of the acrosome reaction of human spermatozoa by immunolocalization with monoclonal antibody

A monoclonal antibody generated against hamster epididymal spermatozoa and recognizing an antigen within the acrosome was used in conjunction with FITC-antimouse immunoglobulin as a marker of the human acrosome during sperm development, capacitation, and the acrosome reaction. The specificity of binding of the monoclonal antibody was assessed using immunolocalization by epi-fluorescence and electron microscopy. Immunofluorescence revealed that antibody bound over the entire anterior acrosome in hamster and human spermatozoa. Ultrastructural localization indicated that antigen was predominantly present on the inner face of the outer acrosomal membrane and within the acrosomal content. Qualitative specificity was studied using a highly purified preparation of hamster acrosomes in an enzyme-linked immunosorbent assay. Since the antibody rapidly visualized human acrosomes, it was used to detect abnormal acrosome morphology of mature spermatozoa and to mark spermatids present in the ejaculate. During incubation in capacitating medium, changes in the immunofluorescence of live or methanol fixed spermatozoa were correlated with incubation interval and the ability of spermatozoa to fuse with zona-free hamster oocytes. Spermatozoa bound to zona-free hamster oocytes displayed no fluorescence, confirming that acrosome loss occurred before spermatozoa attached to the vitellus.     

Fertilization of cumulus-free,zona-intact mouse ova in vitro at high and low sperm concentrations

The effect of varying the sperm concentration between 2 × 10⁵ sperm/ml and 8 × 10⁶ sperm/ml on fertilization of cumulus-free, zona-intact F1 (CBA × C57BL) mouse ova by QS and F1 (CBA × C57BL) mouse spermatozoa was studied. The spermatozoa from both strains of mice exhibited optimal fertilization rates at 2 × 106 sperm/ml. However, at sperm concentrations greater than 4 × 106 sperm/ml and less than 1 × 106 sperm/ml, fertilization rates were significantly reduced. F1 spermatozoa were more susceptible to dilution than QS spermatozoa. A significant interaction between strain and sperm concentration indicated that the two strains produced different fertilization rates at different sperm densities. Extracts of epididymal fluid, medium from capacitated spermatozoa, or ampulla fluid did not improve the fertilization rate at 2 × 105 sperm/ml, but retaining the cumulus oophorus did. The decrease in fertilization rate at 8 × 106 sperm/ml can in part be attributed to a nondialysable inhibitor from the neat sperm preparation that appeared to be of epididymal origin.     

Evaluation of assay conditions for the zona-free hamster ova bioassay of boar sperm fertility

The ability to penetrate zona-free hamster ova may be a very useful test of fresh and frozen boar sperm fertility. These studies were designed to optimize assay conditions prior to evaluation of the accuracy of the bioassay in predicting boar sperm fertility. The ability to penetrate zona-free hamster ova was greater in sperm washed on a Percoll gradient than in sperm washed by dilution and centrifugation. Penetrating ability was greater in sperm from the sperm-rich fraction than from the whole ejaculate but did not differ among different aliquots of the sperm-rich fraction and did not decrease when the prewashing interval was increased from 15 to 85 min. Frequency of collection of ejaculates (1, 3, or 5 times per week) did not affect the penetrating ability of the sperm. Penetration rate was greater when sperm were coincubated with zona-free hamster ova at 39°C compared to 37°C. Sperm from an infertile boar had reduced penetrating ability compared to sperm from fertile boars (11% vs 93%, P < .001). These studies suggest that the zona-free hamster ova bioassay may be a useful assessment of fresh boar sperm fertility.     

Influence of physiological conditions on DNA repair and mutagenesis in higher plants

In barley ( Hordeum vulgare L.) and grass pea ( Lathyrus sativus L.), caffeine, an inhibitor of DNA repair activity, and Na₂ethylenediaminetetraacetate, an inhibitor of DNA-endonucleases, sharply decreased the excision repair of pyrimidine dimers induced in DNA by ultraviolet irradiation. An inhibitor of RNases, diethylpyro-carbonate, did not inhibit the process of excision, and in one experiment it even enhanced excision. Caffeine markedly increased the frequency of mutations and inhibited the growth of seedlings after UV-radiation. Such enhancement was greater with the higher UV fluence. Results of chemical inhibition were further confirmed by the suppression of repair by low temperatures: the frequency of chromatid aberrations induced with propyl methanesulfonate was increased more than 3 times and chromatid aberrations 1.5 times. Evidence for participation of repair enzymes in the modification of mutation processes was also obtained in the experiments which combined γ-irradiation and treatment with propyl methanesulfonate. Conditions favouring repair activity caused a drastic reduction in the frequency of aberrations, whereas with conditions preventing enzyme function the mutation frequency increased. In one of the experiments of this series we were able to demonstrate, with identical mutagenic treatment, that by changing post-mutagen conditions (wetting and drying of seeds, storage after mutagenic treatment) it was possible to alter the mutation frequency and to obtain below-additive, additive and synergistic mutational response.