大银鱼卵膜孔结构的电镜观察

扫描电镜下，大银鱼成熟卵卵膜孔区域呈现奇异的放射沟脊状结构。在授精开始的３０秒内，测得卵膜附近约８６％的精子沿着凹沟进入精孔管；还拍摄到了通过卵孔中心的纵切面映象，通过电子计算机模拟，验证了卵膜孔的这种结构，为精子的云集和进入精孔管所提供有利条件，从而，于泥鳅之后又发现了一种真骨鱼，其精子入卵除化学因素外，还存在着不容忽视的机械动力因素。     

唇鲳精子早期入卵观察

用扫描电镜对唇鳃成熟卵子及早期精子人卵过程进行观察。结果显示,唇鲋成熟卵子在动物极中央有一深凹陷的表面光滑的精孔器,其外径2．512μm,内径2．330μm,精子直径1．567μm。混匀的精卵刚遇水时,没有精子进入精孔器。受精后1s,精孔器内出现精子。受精后5S,组织切片显示,精子已经进入卵子内,并形成具有强烈抑制多精人卵作用的受精锥。受精后10S,精子在精孔器前庭集结,尚未形成受精塞。受精后20S,在精孔器内形成受精塞。受精塞没有阻塞精孔管,经分析它不是来源于皮层反应产物。受精塞形成后,可以吸附人卵的精子,这对多精入卵有积极的抑制作用;精子尾部在入卵过程中相互缠绕,这也是减少多精入卵的重要机制。受精后30s,受精塞和吸附的精子向精孔器外移动。受精后50S,受精塞和吸附的精子堵塞精孔器。受精后60s,受精塞吸附的精子开始解体,但是由于精孔管未封闭,还有精子通过精孔管进入到质膜。在人工受精过程中,卵子的单精受精屏障会因其周围精子密度大、精子与卵子距离短、精子运动速度快而被打破,从而导致这些卵子出现多精入卵的现象。受精后80s,精孔管仍然没有封闭,精孔器附近的精子明显出现活动能力的差异：精孔器外面的精子活动能力最强,精孔管旁边的精子活动能力较弱;精孔管外堆积的精子活性消失,受精塞吸附的精子已开始解体,经初步分析,这可能是进入其内的精子耗能有所差异的结果。受精后100S,受精塞吸附的精子解体。     

日本鳗鲡精卵的超微结构以及受精过程观察

通过扫描电镜和透射电镜对经人工催产获得的日本鳗鲡(Anguilla japonica)精子、卵膜的超微结构以及受精过程进行了观察。实验观察到,除一般硬骨鱼类的精子特性外,日本鳗鲡精子有其独特的结构。精子头部为不规则的梨形,有背腹面之分。一个巨大的球形线粒体位于头部顶端。精子中段向后伸出一支根,支根位于袖套腔外精子的背侧,前端向精子头部线粒体方向延伸,支根的微管结构为"8+2"结构,并在精子入卵过程中起到切断鞭毛的作用。精子的尾部由鞭毛和鞭毛末端的结组成。鞭毛横切面呈圆形,无侧鳍,鞭毛微管结构为"9+0"结构。受精卵的整个表面密布着无规律延伸的脊、脊包围形成的窝和窝中的孔所组成的脊孔复合体,但无典型特征的受精孔。受精卵超薄切片观察发现,日本鳗鲡卵膜分为外层壳膜和内层卵黄膜。壳膜与卵黄膜间为卵周隙。壳膜只观察到放射带,未见透明带。放射带可分为三个亚层:最外层为脊孔复合体的脊,中间层为皱纹层,最内层为致密的平滑层。脊孔复合体的孔横穿整个放射带,在放射带内层形成一个乳突状结构。日本鳗鲡的卵膜不仅具有保护卵子的作用,而且还参与了受精。实验还通过扫描电镜观察了日本鳗鲡精子的入卵过程。观察结果认为:日本鳗鲡精子入卵过程可分为卵膜对精子的吸引、精子对卵膜的锚定、精核的进入和孔封闭等4个阶段。但由于研究只观察到受精过程中日本鳗鲡精子和卵膜的形态变化,因此对精子穿过卵膜的方式和特征等尚需做进一步的研究。整个受精过程为1min30s左右。此外,研究还探讨了日本鳗鲡精子结构的特殊性和受精过程的特殊性,为进一步突破日本鳗鲡人工育苗技术提供了理论依据。       

革胡子鲶受精过程的扫描电镜观察

应用扫描电镜观察和描述了革胡子鲶成熟卵和精子的形态、卵壳膜的表面结构和形态、受精孔的位置和结构、精子入卵过程的程序和变化。讨论了精子入卵过程及精孔细胞在解体之后可能转变为一种能够吸引精子在精孔区聚集的“受精素”物质等问题。     

泥鳅精子入卵的动力作用

在扫描电镜下,泥鳅成熟卵卵膜孔外围呈现完整的左涡旋状结构,受精时精子是顺着涡旋的流线进入卵膜孔。涡旋纹理接近对数螺线。本文分析了真骨鱼类的受精因素,除已知的化学因素外,还存在物理因素。也讨论了泥鳅成熟卵卵膜孔形态形成的必然性。     

黄颡鱼受精早期精子入卵扫描电镜观察

用扫描电镜对黄颡鱼(Pseudobagrus fulvidraco)成熟精、卵及授精早期精子入卵过程进行了观察。成熟精子为鞭毛型形态,全长为11·2～12·4μm,头部直径1·1～1·3μm,鞭毛长10·0～11·3μm。成熟的黄颡鱼卵呈圆形,具单一受精孔,卵膜上以受精孔为中心分布有无数辐射状沟嵴。授精前,受精孔暴露在外面;授精2s时,受精孔被纤维状物质覆盖,之后大量精子很快黏附在覆盖物上;至授精10s,漏斗状受精孔又暴露出来。黄颡鱼在授精10s～1min内完成精子入卵过程,可观察到几乎所有样品的精孔区出现一圈环状隆起。大量精子处于隆起外侧,只有少数越过隆起到达受精孔前庭。授精1·5min,精孔区的隆起变成两圈,精子鞭毛解体。授精3min,可见迟到的精子被挡在外面。授精5min,精孔区的精子头部解体,受精孔几乎被分泌物覆盖,受精塞清晰可见。至授精20min,精子几乎全部解体。讨论了精子入卵的动力作用、精卵识别和单精受精机制。     

唇精子早期入卵观察

用扫描电镜对唇成熟卵子及早期精子入卵过程进行观察.结果 显示,唇成熟卵子在动物极中央有一深凹陷的表面光滑的精孔器,其外径2.512 μm,内径2.330 μm,精子直径1.567 μm.混匀的精卵刚遇水时,没有精子进入精孔器.受精后1 s,精孔器内出现精子.受精后5 s,组织切片显示,精子已经进入卵子内,并形成具有强烈抑制多精入卵作用的受精锥.受精后10 s,精子在精孔器前庭集结,尚未形成受精塞.受精后20 s,在精孔器内形成受精塞.受精塞没有阻塞精孔管,经分析它不是来源于皮层反应产物.受精塞形成后,可以吸附入卵的精子,这对多精入卵有积极的抑制作用;精子尾部在入卵过程中相互缠绕,这也是减少多精入卵的重要机制.受精后30 s, 受精塞和吸附的精子向精孔器外移动.受精后50 s, 受精塞和吸附的精子堵塞精孔器.受精后60 s, 受精塞吸附的精子开始解体,但是由于精孔管未封闭,还有精子通过精孔管进入到质膜.在人工受精过程中,卵子的单精受精屏障会因其周围精子密度大、精子与卵子距离短、精子运动速度快而被打破,从而导致这些卵子出现多精入卵的现象.受精后80 s, 精孔管仍然没有封闭,精孔器附近的精子明显出现活动能力的差异:精孔器外面的精子活动能力最强,精孔管旁边的精子活动能力较弱;精孔管外堆积的精子活性消失,受精塞吸附的精子已开始解体,经初步分析,这可能是进入其内的精子耗能有所差异的结果.受精后100 s,受精塞吸附的精子解体.     

人工复合三倍体鲤鱼多精受精的细胞学基础研究(英文)

为了获取一个可育的、没有遗传分离的、快速生长的养殖品系,我们通过人工方法诱导了人工复合三倍体鲤鱼。它含有两套鲤鱼（一套是散鳞镜鲤、一套是兴国红鲤）和一套鲫鱼（红鲫）共三套完整的染色体组。在这种复合三倍体中,发现有些受精卵具有罕见于硬骨鱼类中的受精生物学现象,即多精受精。这是一个非常有趣的生物学现象。为了了解其潜在的机制,我们以普通红鲤作为父本、经过干法授精,通过扫描电子显微镜对其受精的早期过程进行了超微结构研究,并以普通二倍体鲤鱼受精的早期过程作为对照。（１）通过对三次产卵季节收集的近１０００余粒未受精的成熟人工复合三倍体卵细胞进行观察,发现：每个成熟卵子在其动物极上只有一个敞开的受精孔。它由前庭和精孔管组成。前庭对外开口的平均半径为１０～１２微米。精孔管的外孔径为３．００～３．５０微米（Ｆｉｇ．Ａ＆Ｂ）,只比精子头部直径（２．８０～３．００微米）稍大一点。排除了人工复合三倍体鲤鱼成熟卵细胞一次允许两个精子并排进入的可能性。（２）人工复合三倍体鲤鱼在受精后约１５～３０秒,可以看见大量精子聚集在受精孔的前庭区域（Ｆｉｇ．Ｃ）。而在卵子表面的其它区域,没有发现有精子附着的现象。表明：在受精孔的前庭区域有一些吸引     

泥鳅精子和卵子结构及受精过程的细胞学观察

研究旨在探讨泥鳅精子受精时限及为其人工繁殖提供基础资料。采用光镜、电镜技术对泥鳅（Misgurnus anguillicaudatus）精子、卵子和不同时间段的受精卵进行了观察。结果显示:泥鳅精子头部无顶体,主要为核占据,核凹窝较浅,中段具不对称的袖套,尾部轴丝为9+2结构,无侧鳍。卵子动物极卵膜仅有一受精孔,受精孔为深凹陷、短孔道型。在20-21℃水温条件下,授精后3s,精子开始穿过精孔管或已经进入卵子;授精后5-8s,形成精子星光;授精后70s,卵子处于第二次减数分裂后期;授精后6-8min,第二极体形成,等待排出;授精后20-25min,雌雄原核融合;授精后25-30min,受精卵进入第一次有丝分裂中期;授精后40-45min,第一次有丝分裂结束,二细胞形成。研究表明:成熟卵子精孔管内口径（2.2860.364）m,而精子头部直径与其接近,单精受精;入水后150s内可受精。     

三种短鼻蝗精子超微结构的比较研究（直翅目：蝗总科）

本文通过对癞短鼻蝗Ｆｉｌｃｈｎｅｒｅｌｌａ ｐａｍｐｈａｇｉｄｅｓ ｋａｒｎｙ、青海短鼻蝗Ｆ．ｋｕｋｕｎｏｒｉｓ Ｂ．－Ｂｉｅｎｋｏ和肃南短鼻蝗Ｆ．ｓｕｎａｎｅｎｓｉｓ Ｌｉｕ精子的比较,发现它们的精子在外形头和尾的长度、线粒体衍生体的大小及中心粒侧体超微形态结构方面存在着一定的差异,而精子发生过程中七、八两期精细胞线粒体周围微管的排数和精子的线粒体衍生体超微形态结构十分相似。     

Increased velocity and induction of chemotactic response in mouse spermatozoa by follicular and oviductal fluids.

The dynamic parameters of mouse sperm cells exposed to follicular and oviductal fluids were assessed. Spermatozoa were tracked on a chemotactic Zigmond chamber and recorded using a videomicroscopy system. The results were evaluated with computer-supported image analysis. Follicular fluid at a dilution of 10(-4) markedly increased the proportion of spermatozoa with high velocity, and stimulated chemotactic behaviour. The highest velocities were observed in sperm cells exposed to oviductal fluid, and a greater proportion of these cells had high velocity compared with those exposed to follicular fluid. Chemotaxis was induced in spermatozoa exposed to oviductal fluid at dilutions of 10(-3) and 10(-5). These results suggest the presence of temporal subpopulations of responsive spermatozoa, considering the distance travelled towards both follicular and oviductal fluids and the proportion of sperm cells migrating towards the gradient in the highest distance ranges. This is the first report on the effect of isolated follicular and oviductal fluids on dynamic parameters and chemotaxis of mouse spermatozoa. The findings support previous work showing that the motility and directionality of mouse sperm cells is increased by factors in the microenvironment of the egg. Although the significance of these factors in vivo is unknown, it is possible that there is a relay mechanism involving sequential activity of both oviductal and follicular fluids to direct the male gametes towards the egg.     

Fragmentation dynamics of frozen-thawed ram sperm DNA is modulated by sperm concentration

This study investigated the hypothesis that post-thaw incubation of ram sperm at high concentrations results in a faster rate of DNA fragmentation than when sperm are incubated at a lower concentration. Ejaculates from 10 rams were frozen-thawed, prepared in sperm concentrations of 100, 50, 25, 12, and 6 × 10⁶ sperm/mL, and incubated for 6 h at 37 °C. Sperm DNA fragmentation was assessed using the sperm chromatin dispersion test (Sperm-Halomax®) at 1, 3, 4, and 6 h of incubation at 37 °C. On fitting a binary logistic regression with a cubic over time and treating ram and dilution levels as factors, there were significant effects with respect to the ram, dilution and time (all P-values were very much smaller than 0.001). Therefore, DNA fragmentation dynamics of incubated frozen-thawed ram sperm were not only dependent on the inherent sperm DNA fragmentation expressed immediately after thawing, but also on the concentration of sperm incubated in the sample. Although there was evidence of individual ram variation in SDF during the incubation period, the general finding of the current study was that lower sperm concentrations resulted in a slower rate of DNA fragmentation These findings have important implications for the post-thaw manipulation of ram sperm used for AI and advanced reproductive procedures that use sperm at low concentrations. Our data also emphasised the highly dynamic nature of sperm DNA fragmentation and the importance of conducting the procedure in a standardised manner.     

Cooperation and the evolution of anisogamy

In the lights of the concept of cooperation wholes, I discuss why the differentiation of sperm and ova can occur with a mathematical model. Most of Parker's explanations for anisogamy are not completely proper, because it is proved that sperm competition is neither sufficient nor necessary for anisogamy and cooperation to deal with fertilization risks is the real key to understand the evolution of anisogamy. According to the computer simulation results, the transport of gametes between different individuals, risks of the transport, the consequent inequality of sperm and eggs and competition among different individuals were the main causes of gamete differentiation. But these factors have different roles and effects. The transport risk is the main reason for individuals of different mating types to cooperate and differentiate into sperm and egg producers. The transported gametes have an advantage to evolve into sperm to seek for a larger gamete number over the fixed gametes, because they suffer more risks as they can encounter the same fixed gamete and less sibling competition as they can be dispersed better. Gamete competition among different individuals just causes the transported gametes to become as small as possible if they have already become smaller beyond a critical state. In the final discussion, I further put the evolution of anisogamy into a broader background of levels of selection and of the evolution of cooperation, the most important existential mode of matters that makes life as life.     

Conditions for the evolution of soldier sperm classes

Abstract.— There has been wide disagreement as to whether sperm competition among animals can produce a soldier class of sperm to fight against other males. Utilizing mathematical models, we analyze the appropriate conditions for the evolution and maintenance of a soldier sperm class. We conclude that: (1) soldier sperm evolve even if one soldier sperm can kill or block less than one competing sperm; (2) soldier sperm evolve faster when there is a large variance in the number of competing sperm; (3) soldier ratio increases until reproductive sperm are too scarce to fertilize all ova or a sperm intensely refuses to become a soldier; and (4) soldier sperm are more likely to be smaller than reproductive sperm. Our models suggest that the conditions for the evolution of a soldier sperm class are not stringent.     

The dynamics of sperm cooperation in a competitive environment

Sperm cooperation has evolved in a variety of taxa and is often considered a response to sperm competition, yet the benefit of this form of collective movement remains unclear. Here, we use fine-scale imaging and a minimal mathematical model to study sperm aggregation in the rodent genus Peromyscus. We demonstrate that as the number of sperm cells in an aggregate increase, the group moves with more persistent linearity but without increasing speed. This benefit, however, is offset in larger aggregates as the geometry of the group forces sperm to swim against one another. The result is a non-monotonic relationship between aggregate size and average velocity with both a theoretically predicted and empirically observed optimum of six to seven sperm per aggregate. To understand the role of sexual selection in driving these sperm group dynamics, we compared two sister-species with divergent mating systems. We find that sperm of Peromyscus maniculatus (highly promiscuous), which have evolved under intense competition, form optimal-sized aggregates more often than sperm of Peromyscus polionotus (strictly monogamous), which lack competition. Our combined mathematical and experimental study of coordinated sperm movement reveals the importance of geometry, motion and group size on sperm velocity and suggests how these physical variables interact with evolutionary selective pressures to regulate cooperation in competitive environments.     

黑玛丽精子包特性及pH和温度对其精子活力的影响

通过研究黑玛丽Poecilia latipinna精子包破裂的过程和影响因素、精子运动的情况及影响因素,结果发现,当精液用Hank’s平衡盐溶液(HBSS)稀释约5 min后精子包开始破裂,约12 min后全部破裂。释放出的精子暂时处于休眠状态,约50 min后,处于HBSS稀释液中的精子会被激活。应用计算机辅助精子分析系统对黑玛丽精子在不同pH和不同温度下HBSS中的精子运动百分数、运动时间和平均运动速率进行观察。在pH7~8的中性或弱碱性溶液中,精子运动活力较强;而在酸性(pH<7)或碱性较强(pH>9)的溶液中,精子的运动活力都会降低。在不同温度下的HBSS中精子的活力不同,精子在低温(4℃)条件下的运动时间显著长于在室温(20℃)条件下,但运动速度较慢。本研究初步探讨了黑玛丽的精子包特性以及pH和温度对精子运动活力的影响,旨在对黑玛丽等卵胎生鱼类的人工授精,以及生殖生物学特性等的研究提供更丰富的基础资料。     

Theory of resource allocation strategy in territorial male tropical reef fishes

A mathematical model is proposed to explain energy resource allocation between sperm production and territoriality in male reef fishes (Labridae species) from the point of view of optimization. Labridae species are typically characterized by both TP (terminal phase) and IP (initial phase) males. The former are considered to release a lower amount of sperm but show aggressive territoriality. In the model, TP male reproductive success is considered as depending upon both fertilization probability (depending on sperm density) and the individual's own territorial activities. Between these factors, a trade-off exists by which the fertilization probability can be enhanced only by reducing territoriality. Therefore, the male has to decide how much of the total available energy resource should be allocated to each. The model showed that under high fertilization efficiency the male can achieve high success by spending less of the resource on sperm production and correspondingly more for territoriality. The TP male reproductive success increases with decreasing male density in the habitat. Nevertheless, when intruding males cannot be excluded completely by territorial behavior of the TP male, females prefer high male density. If females can control the number of intruding males to some degree, conflict may arise between the sexes. Received: December 8, 1999 / Accepted: May 24, 2000     

Sperm competition and maternal effects differentially influence testis and sperm size in Callosobruchus maculatus

The evolutionary factors affecting testis size are well documented, with sperm competition being of major importance. However, the factors affecting sperm length are not well understood; there are no clear theoretical predictions and the empirical evidence is inconsistent. Recently, maternal effects have been implicated in sperm length variation, a finding that may offer insights into its evolution. We investigated potential proximate and microevolutionary factors influencing testis and sperm size in the bruchid beetle Callosobruchus maculatus using a combined approach of an artificial evolution experiment over 90 generations and an environmental effects study. We found that while polyandry seems to select for larger testes, it had no detectable effect on sperm length. Furthermore, population density, a proximate indicator of sperm competition risk, was not significantly associated with sperm length or testis size variation. However, there were strong maternal effects influencing sperm length.     

New transient species of sperm whale myoglobin in photodissociation of dioxygen from oxymyoglobin

We carried out the flash photolysis of oxy complexes of sperm whale myoglobin, cobalt-substituted sperm whale myoglobin, and Aplysia myoglobin. When the optical absorption spectral changes associated with the O2 rebinding were monitored on the nanosecond to millisecond time scale, we found that the transient spectra of the O2 photoproduct of sperm whale myoglobin were significantly different from the static spectra of deoxy form. This was sharply contrasted with the observations that the spectra of the CO photoproduct of sperm whale myoglobin and of the O2 photoproducts of cobalt-substituted sperm whale myoglobin and Aplysia myoglobin are identical to the corresponding spectra of their deoxy forms. These results led us to suggest the presence of a fairly stable transient species in the O2 photodissociation from the oxy complex of sperm whale myoglobin, which has a protein structure different from the deoxy form. We denoted the O2 photo-product to be Mb*. In the time-resolved resonance Raman measurements, the nu Fe-His mode of Mb* gave the same value as that of the deoxy form, indicating that the difference in the optical absorption spectra is possibly due to the structural difference at the heme distal side rather than those of the proximal side. The structure of Mb* is discussed in relation to the dynamic motion of myoglobin in the O2 entry to or exit from the heme pocket. Comparing the structural characteristics of several myoglobins employed, we suggested that the formation of Mb* relates to the following two factors: a hydrogen bonding of O2 with the distal histidine, and the movement of iron upon the ligation of O2.