Involvement of Ca2+-ATPase in suppressing the appearance of bovine helically motile spermatozoa with intense force prior to cryopreservation

In cattle, cryopreserved spermatozoa are generally used for artificial insemination (AI). Many of these specimens exhibit helical movement, although the molecular mechanisms underlying this phenomenon remain unclear. This study aimed to characterize helically motile spermatozoa, investigate the involvement of Ca²⁺-ATPase in suppressing the appearance of these spermatozoa prior to cryopreservation, and examine the potential of helical movement as an index of sperm quality. In the cryopreserved semen, approximately 50% of spermatozoa were helically motile, whereas approximately 25% were planarly motile. The helically motile samples swam significantly faster than those with planar movement, in both non-viscous medium and viscous medium containing polyvinylpyrrolidone. In contrast, in non-cryopreserved semen, planarly motile spermatozoa outnumbered those that were helically motile. Fluorescence microscopy with Fluo-3/AM and propidium iodide showed that flagellar [Ca²⁺]_i was significantly higher in cryopreserved live spermatozoa than in non-cryopreserved live ones. The percentage of non-cryopreserved helically motile spermatozoa was approximately 25% after washing, and this increased significantly to approximately 50% after treatment with an inhibitor of sarcoplasmic reticulum Ca²⁺-ATPases (SERCAs), “thapsigargin.” Immunostaining showed the presence of SERCAs in sperm necks. Additionally, the percentages of cryopreserved helically motile spermatozoa showed large inter-bull differences and a significantly positive correlation with post-AI conception rates, indicating that helical movement has the potential to serve as a predictor of the fertilizing ability of these spermatozoa. These results suggest that SERCAs in the neck suppress the cytoplasmic Ca²⁺-dependent appearance of helically motile spermatozoa with intense force in semen prior to cryopreservation.     

Effects of acrosomal conditions of frozen-thawed spermatozoa on the results of artificial insemination in Japanese Black cattle

The purposes of this study were to examine the relationship between male artificial insemination (AI) fertility and sperm acrosomal conditions assessed by new and conventional staining techniques and to identify possible reproductive dysfunctions causing low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions in Japanese Black bulls. We investigated individual differences among bulls in the results concerning (1) acrosomal conditions of frozen-thawed spermatozoa as assessed by not merely peanut agglutinin-lectin staining (a conventional staining technique) but also immunostaining of acrosomal tyrosine-phosphorylated proteins (a new staining technique), (2) routine AI using frozen-thawed spermatozoa as assessed by pregnancy diagnosis, (3) in vivo fertilization of frozen-thawed spermatozoa and early development of fertilized eggs as assessed by superovulation/AI-embryo collection tests and (4) in vitro fertilization of frozen-thawed spermatozoa with oocytes. The percentages of frozen-thawed spermatozoa with normal acrosomal conditions assessed by the abovementioned staining techniques were significantly correlated with the conception rates of routine AI, rates of transferable embryos in superovulation/AI-embryo collection tests and in vitro fertilization rates. These results are consistent with new suggestions that the distribution of acrosomal tyrosine-phosphorylated proteins as well as the acrosomal morphology of frozen-thawed spermatozoa are AI fertility-associated markers that are valid for the prediction of AI results and that low conception rates in AI using frozen-thawed spermatozoa with poor acrosomal conditions result from reproductive dysfunctions in the processes between sperm insemination into females and early embryo development, probably failed fertilization of frozen-thawed spermatozoa with oocytes.     

人为刺激对海菜花生长及形态特征的影响

海菜花(Ottelia acuminate)的花葶具有较高食用和药用价值,为了探究人为刺激对其生长及形态特征的影响,同时为了保护和恢复高原湖泊特有雌雄异株的珍稀沉水植物,通过实际观察和测量,采用海菜花营养生长及生殖生长的不同部位形态特征作为对比,选取植株的整体叶面积、叶柄、叶片数、花葶总数、长度、佛焰苞内花朵数目、佛焰苞基座大小指标,在种植密度和摘叶留葶处理的人为干扰作用下,比较这些指标在雌雄植株间的差异情况,通过单因子方差分析检测其差异的显著性,为人工高效种植、生产优质海菜花提供理论依据。结果表明,从营养生长方面来说,在2种人为因素影响下,雄株无论整体还是单片叶面积水平均与雌株有显著相关关系,而叶片数量、叶柄与植株性别间不存在相关关系;从生殖生长的角度上来说,佛焰苞内花朵数目与性别有显著相关关系,雌株佛焰苞内花朵数小于等于7朵,雄株则大于等于10朵;雄株能够在花期内保持最大花葶数以保证同花期内的雌株受粉,雌株的花葶虽然数量较少,但质量更好,经人为摘叶后质量更佳,佛焰苞更大,随性别分化明显,而花葶长度可以在种植密度不同时,成为对比鉴定其性别特征的辅助手段。从经济效益角度出发,首选种植雌株且一定程度上合理的密植,可以达到高产高效的目的。     

浅议居住区人工湿地设计的几个问题

通过对国内居住区人工湿地进行分析,指出国内居住区人工湿地设计存在植物配植不够合理、冬季保温措施不当及后期管理养护不力等问题,并针对问题提出了相应的改进措施。     

驼背鲈室内池塘育苗技术研究

[目的]改进驼背鲈的育苗方法。[方法]选择3龄以上的驼背鲈亲鱼进行海水网箱养殖,用绒毛膜促性腺激素（HCG）和释放激素（LRH—A3）混合悬浊液进行催产。通过向育苗池中定时定量加入高级虾片和EM茵的方法进行驼背鲈的育苗。[结果]驼背鲈仔鱼在水泥池水温22～26℃、盐度28％-32％、pH7．9～8．7的条件下,经30d培育,平均鱼体全长达3．3cm,仔鱼变态发育成幼鱼。[结论]驼背鲈可在室内池塘通过亲鱼人工催产获得受精卵,EM菌＋高级虾片方法是经济有效的驼背鲈育苗方法。     

盐渍地人工苜蓿草地生产力与土壤改良研究

[目的]为改进和利用河西走廊盐渍化土地提供依据。[方法]在临泽县盐渍地种植苜蓿,测定1～5年生人工苜蓿草地的生产力,并研究苜蓿对盐渍化土地的改良效果。[结果]在盐渍地种植苜蓿后,苜蓿的主根长、主根直径和侧根数随着生长年限的增加而增加。从种植到第5年,盐渍地苜蓿的鲜草产量呈逐年增加的趋势。盐渍地种植苜蓿后,土壤有机质含量不断增加,而土壤盐离子含量不断下降。有机质含量的增加速度和盐离子含量的下降速度均在苜蓿种植后的生长前期较慢而生长后期较快。[结论]苜蓿的生产力与灌水量、灌水次数有关。一个生育周期应至少灌水4次,种植苜蓿的适宜灌水量为4275m3/hm2。     

半夏小块茎的形态发生及人工种子制作

离体培养中，经过小块茎发生途径从半夏叶柄诱导了完整植株。用实体解剖、连续切片和三维结构重组等方法观察分析得出：小块茎顶端的叶原基由于不对称生长逐渐将顶端分生组织复盖，随后叶原基的尖端和两侧生长延缓，生长中心转移到背轴面中央，结果导致叶原基长帽状结构。不同发育程度的雏叶和叶原基总是相互叠在一起顶端分生组织复盖于下面。小块茎枯端形成芽的同时，在顶芽附近分化根。     

地中海水牛人工授精效果研究

[目的]为了解意大利地中海水牛冻精的授精效果及其在我国生长适应情况。[方法]利用国内首次引进意大利地中海水牛冻精与我国现有的摩拉、尼里——拉菲和本地水牛进行人工授精,试验选取自然发情的摩拉水牛20头,尼里水牛16头,本地水牛32头,年龄在2.5~9岁之间,采用直肠把握子宫角深部输精法,配种后40 d应用B超进行早期妊娠诊断怀孕情况。[结果]：摩拉、尼里——拉菲和本地水牛受胎率分别为50.00%、75.00%、56.25%,平均人工授精配种受胎率为58.82%,品种间人工授精受胎率差异不显著（P〉0.1）。[结论]表明引进地中海水牛冻精开展杂交组合,改良我国现有的水牛品种,提高其生产性能是切实可行的。     

药用植物人工诱导多倍体育种的研究进展

多倍体是一种有效的育种途径,多倍体药用植物具有器官巨大,产量高,具有重要的经济价值和药用价值。回顾了近年来人工诱导多倍体的原理和方法,并总结了药用植物人工诱导多倍体的研究进展以及应该注意的几个关键问题。     

The Use of Sex-Sorted Stallion Semen in Embryo Transfer Programs

Sorting stallion semen into two separate populations of enriched X- or Y-bearing sperm can be done successfully. For this, stallion semen can be shipped to a sorting facility, but the mare must be in close to the sorting laboratory. Fertility rates when using 20-40 million sperm are an acceptable 60% per insemination. The procedure can be implemented in embryo transfer programs, with no deleterious effect on the pregnancy rate or embryonic death.