Cu2+对泥鳅精子运动的影响

以泥鳅精子为受试材料,应用计算机辅助精子分析(CASA),研究不同浓度Cu2+暴露对运动型精子和静止型精子的毒性.结果 显示,运动型精子和静止型精子暴露在10 μM的Cu2+中,两者运动能力均受到明显抑制;在30 μM及以下浓度中运动型精子受影响程度较静止型精子大,而在更高浓度中出现相反结果.表明泥鳅精子对Cu2+毒性较敏感,不同能量代谢状态时其敏感性不同;Cu2+对泥鳅精子运动能力有较大影响,对淡水鱼具有潜在生殖毒性.     

青岛文昌鱼精子运动特征及计算机辅助分析

青岛文昌鱼的精子在精浆中不运动,在与精浆等渗的生理溶液中也不运动,把精液稀释到过滤海水或人工海水后,精子运动被激活。文昌鱼新鲜精液中精子浓度为(9.4±1.6)×1010/ml;在人工海水中,运动精子占总精子数的85.8%±6.9%,精子运动持续的平均时间是22.7±2.6min。文昌鱼精子经人工海水激活0.5min后,可观察到四种运动方式:(1)向前直线运动,其平均速度93.6±23.7μm/s,这类精子所占的百分比为27.8%±3.1%;(2)弧形曲线运动,平均速度55.6±18.9μm/s,所占的百分比为44.3%±2.5%;(3)左右摆动,平均前进速度27.4±13.4μm/s,所占百分比为14.7%±1.8%;(4)运动速度小于5μm/s的精子,视为不运动的精子,所占百分比为13.2%±1.8%。随着激活时间的延长,文昌鱼精子的运动状态发生改变,向前直线运动和弧形曲线运动的精子逐渐减少,而左右摆动和不运动的精子逐渐增多。     

腺嘌呤核苷脱氨酶及核苷运转系统对精子运动的调节作用

 腺嘌呤核苷(ADO)和它的类似物2-Cl-ADO对牛附睾尾部精子的运动有刺激作用,为了探讨ADO及其类似物对精子运动调节作用的机理,我们从ADO受体,核苷运转系统(NTS)及腺苷脱氨酶(ADA)三个方面对牛附睾尾部的精子进行了研究。我们发现腺苷受体不存在于牛精子膜上,但ADA和NTS以膜蛋白的形式结合在精子膜上,并对精子体内的ADO浓度起调节作用。我们的结论是ADO及其类似物对牛精子运动的调节作用是首先通过精子膜上的ADA和NTS影响精子体内的ADO浓度,进而ADO又通过调节钙离子浓度刺激牛精子运动。     

HgCl2对泥鳅精子运动的影响

为了检测不同浓度的HgCl2对泥鳅Misgurnus anguillicaudatus精子运动的影响,以泥鳅精子为实验材料,用含终浓度分别为0(对照)、1、5、10、15和20 μmoL/L的HgCl2待测液分别孵育0、2、4和6 h后激活,激活后立即在显微镜(Olympus IX81)下观察精子运动参数.为进一步探讨HgCl2对泥鳅精子运动影响的机制,用终浓度为20μmol/L的HgCl2保存液孵育泥鳅精子10 min,以含终浓度分别为0、0.1、1、10 mmol/L的2-巯基乙醇和20μmol/LHgCl2混合液为激活液激活.激活后立即在显微镜下观察精子运动,发现2-巯基乙醇町逆转HgCl2对泥鳅精子的抑制作用,为探讨HgCl2对泥鳅精子运动影响的可能机制提供参考.     

肌肽棉酚对绵羊精子运动及超微结构的影响

肌肽促进绵羊精子运动增强,直线运动精子数目增多;棉酚则强烈抑制精子运动并使精子的运动方式发生改变;肌肽与棉酚混合处理精子,肌肽可不同程度抵消棉酚对精子产生的有害作用。但当棉酚完全抑制精子运动后,再加入肌肽则不能使精子恢复运动。电镜观察,肌肽对绵羊精子超微结构无任何不良影响;棉酚同造成精子严重损伤,导致生物膜系统损害、轴丝崩解、线粒体螺旋鞘紊乱等。本文通过肌肽,棉酚加入的时序不同,探讨两者对绵羊精子     

精子获能及其发展

获能是哺乳动物精子受精前必须经历的一个成熟过程,它所需时间因种而导,并具有非均一怀和可逆性,获能涉及精子质膜去获能因子（ＤＦ）的去除和膜组分的重新分布,包括膜脂成分的改变,蛋白迁移,受体暴露,钙通道激活,胞内ｃＡＭＰ产生和蛋白酪氨酸磷酸化等,获能的终点是发生顶体反应（ＡＲ）并呈现超激活运动（ＨＡＭ）受精取决于上述变化,并可用金霉素染色法,ＡＲ,ＨＡＭ和穿卵率定量检测获能。     

环境pH及渗透压对玫瑰无须鲃精子运动的影响

以玫瑰无须鲃Puntius conchonius精子为材料,应用计算机辅助精子分析系统(CASA),研究了精子在不同pH和不同渗透压的NaCl溶液中的运动百分率、运动时间和运动速率。结果表明,酸性(pH<7.0)或碱性较强(pH>9.0)的溶液均不利于精子运动,而弱碱性(pH7.5～8.5)的溶液较适合精子的运动;在渗透压较低(<75mOsm/kg)或较高(>175mOsm/kg)的NaCl溶液中,精子的运动时间和运动百分率都显著较100～150mOsm/kg渗透压溶液中的短或低(P<0.05);而运动时间最长,并且运动百分率最高的条件为pH8.0和125mOsm/kg渗透压的溶液环境。     

甘油浓度对蓝狐精子深低温冷冻保存的影响及冻融前后精子的超微结构

人工采取8只优质芬兰雄性蓝狐的精液,分别利用2％、4％、6％和8％甘油浓度的卵黄-Tris-果糖-柠檬酸钠稀释液进行稀释,制成细管冻精。在冻融后0、O．5、2、4、6h检测4种浓度组的精子运动度、质膜完整率、顶体完整率;并利用透射电镜观察冻融前后精子的超微结构变化。冻融后0h,4％甘油浓度组冻融精子的运动度、质膜完整率、顶体的完整率均最高(分别为41．8％、43．6％、48．4％),2％浓度组最低(分别为24．5％、27．6％、31．7％);随着检测时间延长,2％与4％组的精子特性差异显著,但2％、6％、8％3个组间差异不显著;6h时各组间精子的运动度均不超过10％,最高质膜完整率和顶体完整率分别为11．8％、12．7％。说明蓝狐精液稀释剂中甘油的适宜浓度应为4％,冻融后精子的活力维持时间较短。蓝狐精子冻融过程中质膜极易发生膨胀或断裂、顶体囊泡化或溃散,而质膜和顶体丢失现象较少。     

鮸鱼精子的生理特性及超低温冻存

通过测定精子的激活率、运动时间及寿命,研究了鮸鱼精子的生理特性,以0.5mL麦细管为冻存管、两步降温法超低温冻存鮸鱼精子。结果表明,鮸鱼精子激活与运动的适宜盐度为20~30、适宜pH值为5.5~9.0,适宜的KCl、NaCl、CaCl2溶液浓度分别为（500~600）mmol/L、（400~500）mmol/L、（300~400）mmol/L,适宜的葡萄糖溶液浓度为（800~900）mmol/L。无Ca2＋、Mg2＋及HCO3-的人工海水均能使鮸鱼精子激活,但运动时间及寿命有所下降。以Cortland溶液为稀释液,10%Gly、15%Gly、5%DMSO、10%DMSO、15%DMSO、10%EG、10%PG、15%PG及20%PG为抗冻剂,超低温冻存鮸鱼精子15d后,冻精的活力与鲜精相比无显著差异,其中,以10%Gly为抗冻剂冻存精子的效果最好,冻精的激活率、运动时间及寿命分别达（86.38±1.63）%、（8.24±1.37）min及（10.21±0.42）min。     

Movement characteristics and acrosomal status of rabbit spermatozoa recovered at the site and time of fertilization

Rabbit spermatozoa were recovered from the oviductal ampullae 11 h postcoitus by an oil microflush technique. Their movement was evaluated in the ampullar fluid, or in ampullar fluid diluted with in vitro fertilization medium, in slide preparations which were approximately 25 micron or 100 micron deep. The movement of these sperm was compared with the movement of ejaculated sperm in diluted semen. Movement parameters measured from videotapes recorded by a high-speed camera were coded according to treatment and entered into a microcomputer for statistical analysis. A total of 157 spermatozoa were recovered from the oviducts of 16 does: 152 were motile and 126 were free-swimming. Nearly all of the free-swimming sperm swam in trajectories whose average paths were circular. The flagellar beat pattern of the circular swimmers was asymmetric and nearly planar, and the sperm did not roll. Spermatozoa observed in 25-micron slide preparations produced smaller flagellar bends than sperm swimming in 100-micron preparations and tended to swim in larger circles which were oriented in the plane of the slide. Spermatozoa observed within the cumulus matrix moved in a slow, erratic, sinuous manner, but resumed rapid circling upon leaving the matrix. It was concluded that the ampullar sperm were hyperactivated, retaining this physiological condition as they entered the cumulus. The movement qualitatively resembled that of hyperactivated guinea pig and hamster spermatozoa because these species effectively swim in circles. In contrast, 80% of the ejaculated spermatozoa swam in linear trajectories, resulting from relatively symmetrical, flagellar beat patterns. The percentage of rolling spermatozoa and the rolling frequencies were less in the 25-micron than the 100-micron slide preparations. Thus, the movement parameters of both ampullar and ejaculated spermatozoa were affected by the geometry of their observation chambers. This influence should be taken into account when observing sperm motility in vitro. It could also be important in vivo, where changes in sperm movement in response to epithelial surfaces might provide an advantage for reaching the cumulus mass. Ninety-eight percent of the motile ampullar sperm were observed to have acrosomes, including all spermatozoa found within the cumulus matrix.     

Movement characteristics and power output of guinea-pig and hamster spermatozoa in relation to activation

Spermatozoa were collected from the cauda epididymidis of golden hamsters and guinea-pigs, and the acrosome reaction was induced in vitro. Movement characteristics of the spermatozoa were assessed with high-speed cinemicrography. Before the initiation of the acrosome reaction (preactivated spermatozoa), sperm movement in both species was characterized by progressive swimming by regular flagellar waves of moderate amplitude and relative high frequency. After the acrosome reaction (activated spermatozoa), sperm movement in both species was not progressive, and was characterized by whiplash-like flagellar undulations of significantly (P less than 0.05) higher amplitude and lower frequency. Calculation of the hydrodynamic power output by a new theory indicated that no significant change occurred after activation.     

Movement characteristics of bovine epididymal spermatozoa: effects of forward motility protein and epididymal maturation

Movement characteristics of untreated bovine caudal epididymal spermatozoa were compared by high-speed cinemicrography with those of theophylline-activated caput epididymal spermatozoa with and without added forward motility protein (FMP). Comparison of individual movement characteristics clearly established the importance of FMP in converting the nonprogressive motility of theophylline-activated caput sperm into the progressive swimming of mature caudal sperm. Although the total or curvilinear distance traveled in 1 sec by theophylline-activated caput sperm was not changed by the addition of FMP, the linear progression was doubled and the percentage of progressively motile sperm was tripled by this protein. Untreated caudal sperm were 80% motile and theophylline-activated caput sperm were nearly 50% motile; the percentage of motile sperm that were progressive was the same for theophylline-activated caput sperm with FMP and for untreated caudal sperm. Caput sperm without FMP roll infrequently, if at all, but caput sperm with FMP and caudal sperm roll at 4.7 Hz. The beat frequency increases significantly with the addition of FMP and is even higher for caudal sperm. The hydrodynamic power output rises concomitantly with the beat frequency. Perhaps the most striking difference between caput sperm without FMP and those with it is in the swimming paths they follow. Caput sperm without FMP exhibit frequent reversals in direction, or yawing of the sperm heads as they loop back and cross over their tails in an apparently very flexible bending. Their average swimming paths are circles. Caput sperm with FMP and caudal sperm do not show this behavior, but swim in average paths which are linear. The minimum radius of curvature of the tail of caput sperm without FMP is much smaller than that for the other two cell types. These studies clarify the role of FMP in epididymal development of sperm motility.     

Morphological characteristics of stallion spermatozoa

A study of the morphological characteristics of stallion spermatozoa was conducted at the semen laboratory of the Department of Animal Production during four breeding seasons. A total of 590 ejaculates collected from 216 stallions aged from 2 to 26 years and including 13 breeds or colour types was examined. Overall means for the spermatozoal characteristics of these stallions and the classes of head and tail abnormalities are presented and compared with results of other works. Scanning electron micrographs are included to illustrate recognised abnormalities.     

Hydroxyflutamide alters the characteristics of live boar spermatozoa

Our previous study revealed that in vitro incubation of boar ejaculates with hydroxyflutamide (OH-Flu) causes changes in sperm plasma membrane integrity and its stability and sperm mitochondrial oxidative capability. To broaden the knowledge of cellular physiology of spermatozoa, we investigated direct effects of OH-Flu administered for 2 and 24 hours at concentrations of 5, 50, and 100 μg/mL, on sperm mitochondrial membrane potential and mitochondrial superoxide anion production using JC-1 dye and MitoSOX Red fluorescent probe, respectively. We further measured phosphatidylserine membrane translocation (PST) from the inner to the outer layer of the sperm plasma membrane using an annexin-V binding assay. To provide new information of direct effects of OH-Flu on cell signaling pathway, we measured sperm intracellular calcium ion dynamics using Fluo-3. Finally, we assessed sperm motility using a computer-assisted spermatozoa analysis system. Motile sperm were highlighted using the “C-Ruch” computer program for detailed analysis of the straight line velocity distribution. For each functional test, boar spermatozoa were examined and analyzed by flow cytometry and/or confocal microscopy. The results revealed a significant decrease (P < 0.05) in sperm mitochondrial membrane potential and a concomitant increase (P < 0.05) in mitochondrial superoxide anion production after a 2-hour incubation with 50 μg OH-Flu compared with the respective controls and other doses used (P < 0.05). The adverse effects of OH-Flu become strengthened over time (P < 0.05). Notably, 50 and 100 μg OH-Flu appeared to be effective in decreasing sperm motility. Hydroxyflutamide significantly decreased (P < 0.05) the fast sperm subpopulation percentage after 15 minutes and reduced the straight line velocity distribution (P < 0.05). An assessment of PST revealed an increase in the percentage of PST-positive spermatozoa (P < 0.05) only after exposure to OH-Flu for 24 hours. Moreover, OH-Flu at all concentrations induced a rapid increase in sperm intracellular calcium ion concentration. Altogether, the altered in vitro characteristics of live boar spermatozoa provide new insight into direct effects of OH-Flu on sperm mitochondrial membrane potential, superoxide anion production, translocation of membrane phosphatidylserine, free calcium ion dynamics, and sperm motility.     

Comparative dimensional characteristics of spermatozoa in muridae

A comparative study has been made of the dimensional characteristics in six species of Muridae (Malardia meltada, Bandicota bengalensis, Albino rat, Rattus norvegicus, Mus booduga, Rattus rattus). Characteristic differences were found in the dimensions of their spermatozoa. Head dimensions viz., acrosome length, width, area and post-acrosomal area, vary significantly between different species but the differences are significant within individuals of the same species. Flagellum dimensions viz., mid-piece length, breadth, area and main-piece length also vary significantly between different species; within individuals of the same species the differences are not significant except for mid-piece breadth, which varies significantly both between and within the species. The ratio of flagellum to head varies significantly between species but the difference is not significant within individuals of the same species. The significance of the variabilities is discussed.     

An investigation of the fertilizing characteristics of pyriform-shaped bovine spermatozoa

Two breeding trials were done to determine the effect of pyriform-shaped bovine spermatozoa on fertility. In breeding trial (1), heifers were superstimulated by injecting follicle-stimulating hormone (FSH) and randomly allotted to an affected Bull A (n = 21) or a Control Bull, C (n = 18). Semen from Bull A contained 85% pyriform spermatozoa while that from the Control Bull C contained 91% normal sperm and 2% pyriform spermatozoa. Fertilization rate was lower (P = 0.01) for Bull A (total of 63 ova/embryos; 68.5%) than for the Control Bull, C (total of 81 ova/embryos; 84.4%). In breeding trial (2), 37 oestrus-synchronized heifers were randomly allotted to Bull A (n = 19) or Bull C (n = 18). Pregnancy rates at Day 60 (37% and 61% for Bulls A and C, respectively; P = 0.22) and rate of embryo/fetal loss between Days 22 and 60 of pregnancy (23% and 8% for Bulls A and C, respectively; P = 0.55) were not different. In vitro experiments involving the same Bull A and another Control Bull, B, were designed to explain the mechanism of infertility caused by pyriform sperm. The mean (+/- SEM) number of sperm bound to the zona pellucida was lower (P < 0.05) for Bull A (24.6+/-1.2) than for the Control Bull, B (46.6+/-1.9) and the percentage of zonae penetrated by sperm from Bull A (56.0%) and Bull B (82.8%) was also different (P < 0.05). The percentage of pyriform sperm from Bull A bound to (53%), and penetrating (49%) the zona pellucida was lower (P < 0.05) than that in the inseminate (85%). Although fertilization rates (64.1% and 72.8%) were not different (P = 0.5), cleavage rates (48.2% vs. 74.1%) and morula production rates (24.8% vs. 37.7%) were less (P < 0.05) for Bull A than for Bull B, respectively. In summary, pyriform sperm had reduced capability to bind to and penetrate the zona pellucida, and zygotes (resulting from the fertilization of oocytes by pyriform sperm) appeared to have a reduced ability to initiate cleavage.     

Movement of cynomolgus and rhesus monkey spermatozoa collected from the lower female reproductive tract

Postcoital (pc) cervical mucus was collected in 73 menstrual cycles of cynomolgus monkeys and in 43 cycles of rhesus monkeys at 2,6,10,30 hr pc. Videomicrography was used to analyze sperm numbers and movement in the mucus. Both cynomolgus and rhesus monkeys had comparable populations of motile sperm in the mucus at 2 hr pc. However, by 6 hr pc, cervical mucus from cynomolgus monkeys contained twice as many total sperm and motile sperm as mucus from rhesus monkeys (P <.05). Mean swimming speeds of the free-swimming cervical sperm were similar for the two species at this time. No motile sperm were recovered in mucus from rhesus monkeys at 30 hr pc. In cynomolgus monkeys, however, 14 of the 26 animals examined at 30 hr pc had motile sperm in their mucus. These sperm exhibited lower percent molility, percent free-swimming sperm, and swimming speed than those sperm observed at 6 hr pc. Uterine sperm were collected by transcervical or transuterine aspiration from cynomolgus monkeys. In the transcervical technique, sperm were successfully obtained in four of nine animals examined at 6 hr and in four of five animals at 30 hr pc. The percentage of motile sperm in the uterine fluid was high, 82% ± 4%, and the swimming speeds (86 ± 2μm/sec) were higher than those observed in cervical mucus. Approximately 5–10% of the uterine sperm exhibited swimming motions similar to the hyperactivated motility seen in most mammals. These findings indicate that the sperm cervical mucus interaction in vivo in cynomolgus monkeys has more similarities to the human situation than does the interaction in rhesus monkeys.