短尾鮠精子细胞和精子的超微结构研究     

张汉峰  魏刚  黄林  代大临  陈永强  张运明 《西南大学学报(自然科学版)》2005,27(5):696-699

用透射电镜观察了短尾鮠精子细胞和精子的形态以及细胞核、拟染色体和线粒体等的结构变化规律。结果表明,随着精子细胞的发育,核质凝聚程度逐渐增强。精子细胞时期细胞器较丰富,到精子细胞后期,细胞器的形态和数量发生了较大变化。短尾鮠的精子具有椭圆的头部和复杂的中片;近侧中心粒和远侧中心粒靠近核的中央;鞭毛呈9+2轴丝结构,并具有由外膜折迭形成的波浪形的鳍状结构。  相似文献   

环境因子对鲍和牡蛎精子运动能力及受精率的影响   总被引：2，自引：0，他引：2  

李霞  刘淑范  康蕾  徐丹霞 《大连水产学院学报》2002,17(1):25-30

研究了环境因子如K^ 、Na^ 、Ca^2 、Mg^2 、Cl^-、SO4^2-以及蔗糖、pH、氨海水对皱纹盘鲍（Haliotis discus Hannai）和太平洋牡蛎（Creastrea gigas）精子运动及受精率的影响。精子在0.4-0.5mol/L的NaCl、KCl溶液中运动能力为1005，但受精能力较差或完全不受精。在替代海水中无Ca^2 的情况下，牡蛎不受精，但鲍有较低的受精能力。Mg^2 作用不明显。在0.5-0.6mol/L蔗糖溶液中鲍精子能运动，且有较低的受精能力，但牡蛎精子完全不能运动和受精。氨海水对精子运动有一定的刺激作用。在pH为6.0-8.0时精子运动能力较强，但受精的pH适宜范围为7.0-8.0，同时鲍和牡蛎在精子运动和受精方面有明显差异。  相似文献   

稀土元素铈（Ce）对雄性小鼠精子畸形率和因清酮分泌的影响   总被引：18，自引：0，他引：18  

刘玉  陈祖义  王元兴 《南京农业大学学报》2001,24(1):77-80

研究两种剂量和两种染毒方法对雄性小鼠性腺的影响，结果发现200和800mg.kg^-1.d^-1的随饲料摄入，雄性小鼠业子畸形率均显高于对照，且与摄入时间和剂量呈依赖性关系，后期（30和45d)尤为明显，小鼠睾丸重略有下降，但差异不显，以铈200mg.kg^-1腹腔注射急性染毒，对小鼠睾酮分泌有一定的影响，小鼠血清睾酮浓度较对照为低，而以200和800mg.kg^-1.d^-1剂量随饲料摄入（45D）的小鼠，血清中睾酮浓度变化不明显。  相似文献   

钙离子对猪精子蛋白酪氨酸磷酸化及活力影响   总被引：2，自引：2，他引：0  

赵娜  甄林青  姜红菊  严国祥  李新红 《上海交通大学学报(农业科学版)》2015,(2):66-72

研究主要探究钙离子(Ca2+)对猪精子蛋白酪氨酸磷酸化及活力的作用。利用计算机辅助精子活力分析仪(CASA)检测、蛋白免疫印迹测定不同Ca2+浓度对精子活力指标及蛋白酪氨酸磷酸化水平的影响,并对精子样品的酪氨酸磷酸化蛋白的分布情况进行观察。结果显示,当Ca2+浓度≤1.0mmol/L时,精子活力和蛋白酪氨酸磷酸化水平增高;然而,当Ca2+浓度≥3.0mmol/L时,则明显抑制精子活力和蛋白酪氨酸磷酸化水平,尤其分子量约为27和34kDa的高丰度蛋白磷酸化程度明显降低(P0.05)。高浓度Ca2+(4.0mmol/L)明显抑制鞭毛处蛋白酪氨酸磷酸化。因此,细胞外钙离子对猪精子活力和蛋白磷酸化起到重要调节作用,低浓度Ca2+促进精子活力及蛋白酪氨酸磷酸化,高浓度Ca2+(≥3.0mmol/L)抑制精子活力及蛋白酪氨酸磷酸化。本研究初步明确了Ca2+浓度对猪精子蛋白磷酸化及精子活力的影响,为进一步研究精子获能提供参考。  相似文献   

重金属铬对体外培养猪精子活力及蛋白磷酸化水平的影响   总被引：1，自引：1，他引：0  

甄林青  赵娜  王立蕊  李新红 《上海交通大学学报(农业科学版)》2015,(3):1-8

为探究铬元素在猪精子体外获能培养过程中,对猪精子活力、蛋白磷酸化信号通路的影响,研究采取八头杜洛克猪新鲜精液,分别在体外获能培养液中添加不同浓度的6价铬离子(0、0.1、0.5、1、2、10、100μmol/mL)以及双丁酰环腺苷酸(dbcAMP)、异丁基黄嘌呤(IBMX)、PKA抑制剂(H-89)等cAMP-PKAs信号通路调节因子进行调控培养2h。利用精子活力计算机检测(CASA)、蛋白质免疫印迹(WB)技术,分析测定猪精子体外铬暴露获能培养过程中精子活力、蛋白磷酸化水平,并用相应试剂盒检测细胞中甘油醛-3-磷酸脱氢酶(GAPDH)活性、腺苷-3′,5′-环化一磷酸(cAMP)及ATP水平。结果表明:铬离子对猪精子活力、蛋白磷酸化水平有抑制作用,且随铬离子浓度增加抑制作用增强。当铬浓度达到2μmol/mL时精子活力MOT(58.4%)显著低于获能培养组(73.6%)(P0.05);10μmol/mL铬处理明显抑制蛋白磷酸化水平,且细胞内GAPDH活性、cAMP及ATP水平显著降低(P0.05)。而10μmol/mL铬处理中分别添加葡萄糖(5.0μmol/mL)及dbcAMP(1.0 mmol/L)和IBMX(0.1 mmol/L),能缓解铬对蛋白酪氨酸磷酸化(PTP)、蛋白丝氨酸/苏氨酸磷酸化(P-PKAs)及GAPDH活性的抑制作用。结果提示铬可能是通过干扰糖代谢及cAMP/PKA信号通路影响猪精子活力及蛋白磷酸化水平。本研究首次探究了体外培养过程中铬离子对家畜精子活力的影响,并初步探究了其作用机理,为进一步研究铬离子引起繁殖毒性的分子机制及家畜繁育提供一定的理论基础。  相似文献   

Embryonic and larval development of a hybrid between kelp grouper Epinephelus moara ♀ × giant grouper E. lanceolatus ♂ using cryopreserved sperm          下载免费PDF全文

Zhang‐Fan Chen  Yong‐Sheng Tian  Peng‐Fei Wang  Jiang Tang  Jiang‐Chun Liu  Wen‐Hui Ma  Wen‐Sheng Li  Xiao‐Mei Wang  Jie‐Ming Zhai 《Aquaculture Research》2018,49(4):1407-1413

Hybridization was used to take advantage of desirable traits in offspring. In the present study, we applied the cryopreserved Epinephelus lanceolatus sperm into interspecific hybridization with E. moara. Successful hybridization between these two species was achieved and cultured in 23–24°C seawater (34‰). There was no difference in survival rate between hybrid (E. moara ♀ × E. lanceolatus ♂) and non‐hybrid (E. moara ♀ × E. moara ♂) at 24 hrs post hatch (HPH), but less hybrid (14.35 ± 8.02%) hatched than non‐hybrid (93.60 ± 1.65%), which might be due to irregularity cell cleavage and skeletal deformities from the formation of embryonic body to the later embryonic development. Similar phenomenon was found in hybrid embryos from fertilization with fresh sperm, indicating that species variations between parents, rather than cryopreserved sperm, resulted in deformities in embryos. Mean hatch time of the hybrid was 1 hr faster than that of E. moara. The hybrid was 1.95 ± 0.06 mm in total length when newly hatched and reached 38.00 mm at 58 days post hatch (DPH), which showed faster growth than E. moara (recorded in the previous study). Considering its faster growth, E. moara × E. lanceolatus hybrid was a potential breeding production in aquaculture. Over 212,000,000 larvae have been produced and launched in the market since 2015. The results of this study also shed some lights on further comparative studies in grouper hybrid performance.  相似文献   

Evaluation of different extenders for the cold storage of meagre (Argyrosomus regius) semen          下载免费PDF全文

Mariana Santos  Florbela Soares  Márcio Moreira  José Beirão 《Aquaculture Research》2018,49(8):2723-2731

Meagre (Argyrosomus regius) is considered a potential candidate for aquaculture diversification in southern Europe. The main objective of this experiment was to develop a cold storage protocol for meagre semen to facilitate artificial reproduction techniques. Three extenders (non‐activating medium, 0.9% NaCl, and 0.9% NaCl with glycine and glucose) in three different sperm:extender dilutions (1:4, 1:9 and 1:19) were tested in a full factorial design. The quality parameters assessed along the storage time were the sperm motility, viable sperm percentage, adenosine triphosphate (ATP) content, and bacterial growth. The 0.9% NaCl and 0.9% NaCl with glycine and glucose extenders and the 1:4 and 1:9 dilutions maintained a higher sperm motility and a higher sperm linearity for a longer period time. Sperm viability was maintained at a higher value over a longer period with the 0.9% NaCl and 0.9% NaCl with glycine and glucose extenders. Sperm motility and viability appeared to be the main parameters showing the loss of semen quality during cold storage. Meagre semen demonstrated an ability to be stored for up to 10 days at 4°C when using 0.9% NaCl in a 1:4 dilution. These results contribute to a better understanding of the causes of fish semen quality deterioration during cold storage.  相似文献   

A Strategy for Sperm Cryopreservation of Atlantic Salmon,Salmo salar,for Remote Commercial‐scale High‐throughput Processing          下载免费PDF全文

Huiping Yang  E Hu  John T. Buchanan  Terrence R. Tiersch 《Journal of the World Aquaculture Society》2018,49(1):96-112

Sperm cryopreservation is an essential tool for long‐term storage of genetic resources for aquaculture fishes. The goal of this study was to develop an efficient and streamlined protocol for high‐throughput processing for sperm cryopreservation in Atlantic salmon, Salmo salar. The objectives were to evaluate: (1) osmolality of blood serum for determining extender osmolality, (2) effects of extenders for fresh sperm dilution and refrigerated storage, (3) effects of methanol and dimethyl sulfoxide (DMSO) on fresh sperm motility, and (4) motility and fertility after thawing. In this study, sperm samples were collected at a hatchery site in Canada and shipped to a freezing site located 2200 miles (3550 km) away in the USA. Evaluation of three extenders indicated that Mounib solution was suitable for diluting dry sperm for sample processing. Ten percent of methanol or DMSO was less toxic to sperm cells than was 15% within 30 min. Further testing with methanol at 5, 10, and 15%, and sperm solution : extender dilutions (v:v) of 1:1, 1:3, and 1:19 (at concentrations of 5 × 10⁷, 3 × 10⁸, and 1 × 10⁹ cells/mL) indicated that methanol at 5 and 10% showed less toxicity to fresh sperm within 1 h at sperm:extender dilutions of 1:1 and 1:3. Post‐thaw motility of sperm cryopreserved with 10% methanol was significantly higher than that with 10% DMSO, and fertility reflected those results (0–1% in DMSO vs. 38–55% in methanol). Further evaluation of sperm cryopreservation with 10 and 15% methanol at sperm dilution ratios of 1:1, 1:3, and 1:19 indicated that post‐thaw motility in 10% methanol was significantly higher than that in 15% methanol, and post‐thaw fertility in 10% methanol at 1:1 and 1:3 dilution ratios had fertilization rates similar to that of fresh sperm controls. Sperm samples from 12 males cryopreserved with 10% methanol showed male‐to‐male variation in post‐thaw motility (0–36%). Overall, a simplified standard protocol was established for cryopreservation of shipped sperm of Atlantic salmon using extender without egg yolk and yielded satisfactory post‐thaw motility and fertilization rates. This procedure can be readily adopted by aquaculture facilities to take advantage of high‐throughput cryopreservation capabilities at remote service centers. Most importantly, this approach lays the groundwork for an alternative commercial model for commercial‐scale production, quality control, and development of industrial standards. Control of male variability and sperm quality remain important considerations for future work.  相似文献   

PI/Rh123双染和流式细胞术作用下不同盐度对葡萄牙牡蛎精子质量及受精的影响   总被引：1，自引：0，他引：1  

杜俊鹏  王昭萍  崔玉婷  李阳春  杨洋  李鹏飞  柳逸群 《水产学报》2018,42(11):1737-1746

以葡萄牙牡蛎精子为对象,采用碘化丙啶(PI)/罗丹明123(Rh123)双染与流式细胞术(FCM),研究了不同盐度胁迫(10、15、20、25、30、35)对精子质量的影响,并对受胁迫精子进行授精实验。结果显示,随着盐度升高,精子的存活率、运动时间和活力先升高后降低;葡萄牙牡蛎精子的适宜盐度为20～35,活力较高的盐度范围为25～35;通过双染和FCM检测了精子质膜完整性和线粒体活性,发现盐度胁迫先对精子质膜造成损伤,后对线粒体活性造成伤害;低盐胁迫(10、15)对精子损伤严重,胁迫15 min时质膜受损比例高达67.26%±2.35%。人工授精结果显示,低盐胁迫下精子受精率和卵裂率显著降低,且卵裂阶段出现畸形及分裂停止等现象,表明精子质量不仅严重影响了受精过程,还对卵裂造成一定影响。本实验不仅探究了PI/Rh123双染和FCM检测牡蛎精子质膜完整性和线粒体活性的可行性,还结合显微观察全面评价了精子质量,为牡蛎精子质量的研究提供了理论基础。  相似文献   

Development of an in vitro oviduct epithelial explants model for studying sperm–oviduct binding in the buffalo          下载免费PDF全文

KK Saraf  A Kumaresan  S Nayak  S Chhillar  L Sreela  S Kumar  UK Tripathi  TK Datta  TK Mohanty 《Reproduction in domestic animals》2017,52(4):687-691

In this study, we developed an in vitro model for studying sperm–oviduct binding in the buffalo. Oviduct explants were prepared by overnight culture of epithelial cells in TCM‐199 medium under 5% CO₂ at 38.5 °C. Cryopreserved spermatozoa from buffalo bulls (n = 4) were incubated with the oviduct explants, and the sperm–oviduct explants complex was stained with JC‐1. The effect of sperm concentration (2, 3 and 4 million), size of the oviduct explants (<0.2, 0.2–0.3, 0.3–0.4 and >0.4 mm²) and time of incubation (1 hr and 4 hr) on binding index (BI—number of sperm bound to unit area of explants) was studied. No significant difference was observed in the BI among <0.2, 0.2–0.3 and 0.3–0.4 mm² size of explants; however, the BI decreased significantly (p < .05) when the size of explants exceeded 0.4 mm². The BI decreased significantly (p < .05) when the sperm concentration was increased to 4 million, while the duration of incubation did not have any significant effect on the BI. The interaction of bulls with explants size, sperm concentration and incubation time was not significant. The developed assay has the potential to be used as an in vitro model for studying sperm–oviduct binding in the buffalo.  相似文献