银杏和苏铁游动精子发现记

银杏和苏铁游动精子发现记张仲鸣（北京大学生命科学学院１００８７１）１００年前,即１８９６年,日本的两位植物学家平徽作五郎（英文名：ＳａｋｕｇｏｒｏＨｉｒａｓｅ）和池野（英文名：Ｓｅｔ－ｉｃｈｉｒｏ．Ｉｋｅｎｏ）分别发现了银杏和苏铁的具鞭毛游动精子,从...     

蕨类植物精细胞运动器和细胞骨架的研究

介绍了近年来蕨类植物游动精子运动器和细胞骨架的研究进展。游动精子由配子体精子器中的非运动细胞发育形成,其分化过程包括了运动器官和细胞骨架的合成和组装。精子发生过程中形成的运动器的各部分结构包括鞭毛、基体、多层结构及附属结构;基体是细胞中新形成的结构,在不同类群的蕨类植物中分别由双中心粒、分支生毛体和生毛体产生。鞭毛、基体和多层结构中的微管带形成了游动精子三个独特的微管列阵,由于微管蛋白的后修饰作用这些微管列阵十分稳定;centrin是运动器中的重要成分, 但功能尚不清楚,可能和细胞骨架及运动器的构建有关。     

蕨类植物精细胞运动器和细胞骨架的研究

介绍了近年来蕨类植物游动精子运动器和细胞骨架的研究进展.游动精子由配子体精子器中的非运动细胞发育形成,其分化过程包括了运动器官和细胞骨架的合成和组装.精子发生过程中形成的运动器的各部分结构包括鞭毛、基体、多层结构及附属结构;基体是细胞中新形成的结构,在不同类群的蕨类植物中分别由双中心粒、分支生毛体和生毛体产生.鞭毛、基体和多层结构中的微管带形成了游动精子三个独特的微管列阵,由于微管蛋白的后修饰作用这些微管列阵十分稳定;centrin是运动器中的重要成分,但功能尚不清楚,可能和细胞骨架及运动器的构建有关.     

绵马鳞毛蕨精母细胞和游动精子超微结构特征的研究

应用电镜技术对蕨类植物绵马鳞毛蕨(RYOPTERIS CRASSIRHIZOMA Nakai)精母细胞和游动精子的超微结构特征进行了研究。精母细胞为多边形,细胞质内含有丰富的线粒体、质体、内质网、高尔基体等常见的细胞器．在细胞质中还可见到一些同心圆膜状结构,位于质膜的附近或精母细胞的角偶。同心圆膜状结构由双层膜环绕构成,外被l层单位膜。精母细胞与精子器的璧细胞之间形成了分离腔。在精母细胞质膜外形成了嗜锇层,这些结构的形成说明精母细胞已经开始与雄配子体逐渐分离,进入独立发育的阶段。尽管精母细胞之间也有嗜锇层的形成,但嗜锇层是不连续的,其上有一些空隙,精母细胞之间可通过空隙进行物质和信息的交流。成熟的精子细胞外被l层透明的薄膜,里面为游动精子。螺旋状。由环状细胞器环绕3～4圈构成．这些环状细胞器包括多层结卡构、微管带、巨大线粒体、鞭毛带和1个长形浓缩的细胞核。游动精子的后端为一些泡囊化的细胞质．其中包括一些残存的线粒体、造粉质体及大的囊泡等。当成熟的精子细胞排出精子器后。其内的游动精子挣脱透明质膜的束缚,摆脱后端的囊泡,成为1条游动精子。本文还对绵马鳞毛蕨和其它蕨类植物精子的超微结构特征进行了比较。     

磁场对牲畜精子和植物发芽过程的影响--日本生物磁学研究简介之一

前言在１９９０年的时候，就见到了关于动物精子磁场效应的研究报告。其后，菅大助等又发表了一系列文章，报道了关于稳恒磁场（１０００mT）对牛精子存活率、游动率以及平均速率的影响以及弱磁场（６０mT）和含磁粒空间对牛精子的存活率、游动率、平均速率和人工授精的影响结果犤１，２犦。龟田裕孟等，在１９９０发表了人的精子垂直于磁场方向的取向研究犤３犦。芝田信之等，在１９９６年发表了牛的精子垂直于磁场方向取向的研究犤４犦。结果表明，人的精子，只在５０．５mT磁场下垂直取向率就达到７０％；牛的精子，在１００mT下开始受…     

桫椤科笔筒树游动精子行为及形态观察

中国桫椤科植物近年出现了以幼苗减少为特征的生殖障碍新趋势,该研究首次观察了桫椤科笔筒树有性生殖关键环节即游动精子的行为及形态,以明确中国桫椤科植物生殖障碍发生的发育环节及其解剖特征。结果表明:(1)笔筒树的游动精子可分为初游期、平稳期、衰亡期。精子开始游动后经1~3s的初游期,即可达到约200r·min~(-1)的自转频率、约130μm·s~(-1)的位移速度;在17~22min的平稳期内,精子自转频率和位移速度基本稳定,对水体质量即颈卵器释放的化学诱导物质较为敏感;衰亡期为5~7min,精子的自转频率、位移速度和敏感性逐渐降低,直至消失。(2)笔筒树的精子为长条状螺旋形,长10~12μm,螺旋2.5~3.5圈,自上而下由顶脊、鞭毛带、鞭毛、精核、细胞质等5部分构成。(3)笔筒树精子的鞭毛带沿精核呈螺旋状分布在精核的前1/3处,着生有鞭毛32~48条,细胞质呈泡状,随精子游动时间的延长而逐渐萎缩,精核不均质。     

蟹类精子超微结构的比较研究

应用光镜和电镜,比较研究了三疣梭子蟹,中华绒螯蟹和长江华溪蟹的成熟精子。揭示３种蟹精子都是不能游动的无鞭毛精子,呈球形,前后略扁,精子前端出现一光滑的小圆面,圆面四周有内陷的沟环。沟环之后,精子表面凹凸不平,并伸出多数辐射臂。３种蟹精子均为高度特化的细胞,外被质膜,内含细胞核,顶体及退化的细胞质。     

中国特产2种濒危水韭雄性发育的比较研究

采用透射电镜和光镜比较观察了中华水韭和云贵水韭雄配子体及其精子的发育特征。结果显示:(1)2种水韭的雄配子体的寿命只有15~30d,终生都在小孢子壁内发育。(2)雄配子体只含有1个原叶体细胞、1个精子器壁细胞和4个精细胞,前2个细胞内含有丰富的营养物质。(3)精子由精核、微管带、鞭毛、细胞质等4部分构成。(4)中华水韭雄配子体发生率为4.5%,平均每个雄配子体产生0.46个精子,精子游动速度约53μm/s,寿命8min;云贵水韭雄配子体发生率和产精量略高于中华水韭,但精子游动速度和寿命略低于中华水韭。研究认为,中国水韭濒危的主要原因之一是雄配子体产精率低、受到生殖生态限制、水污染对精子的危害等;雄性特征表明水韭在石松类中占有较高的进化地位;结合前人的研究成果绘出了水韭雄配子体的发育模式。     

铅与镉胁迫对桫椤科笔筒树精子行为和形态的影响

中国桫椤科植物部分居群近年来出现了较大面积的病害和幼苗不足等问题,为探究幼苗不足与土壤重金属污染的关系,该研究在观察笔筒树[Sphaeropteris lepifera(Hook.)R.M.Tryon]精子正常行为形态的基础上,首次研究了铅与镉重金属胁迫下笔筒树精子受害的行为和形态变化规律。结果表明:(1)在铅浓度为1.25、5.00mg·kg^-1,镉浓度为0.25、1.00mg·kg^-1的胁迫培养条件下,笔筒树精子的寿命、游动位移速度、自转频率与对照组相比均显著下降。(2)铅离子会影响笔筒树精子的运动能力,镉离子影响精子对雌性物质浓度的辨别能力而使精子迷失方向。(3)重金属胁迫后精子形态和行为出现的变化包括精子伸展受限,鞭毛收缩、脱落,精核畸形、肿胀、降解,精子相互粘连、逆向运动及自溶解体等,且铅与镉胁迫后精子的畸形类型具有差异性。研究证明,轻污染浓度的铅与镉单一胁迫会显著影响笔筒树精子的行为和形态,严重阻断桫椤科植物的有性生殖。     

蕨类植物桂皮紫萁颈卵器和精子器形态和发育的研究

利用扫描电镜技术和树脂切片技术对蕨类植物桂皮紫萁（Osmunda cinnamomeaL.var.asiatica Fernald)的颈卵器和精子器的形态和发育进行了细致的研究。颈卵器发生于雌配子体的腹面,颈部由4列壁细胞构成,6-7个细胞高,内部含有颈沟细胞,腹沟细胞和卵细胞,卵细胞在整个发育过程中,造粉体和囊泡最为显著,颈卵器内的卵细胞成熟时产生卵膜和分离腔。精子器发生于雄配子体的边缘及腹面,由7-8个壁细胞螺旋状围绕而成,壁细胞内为产精组织,精子成熟时精子器盖细胞开裂释放出游动精子。     

精胺抑制人精子的体外受精能力

以精子穿透去透明带仓鼠卵试验(SPA)为模型,评价了精胺对人精子体外受精能力的影响。精胺(0.25—8.0mmol/L)可抑制人精子体外获能和受精,其抑制作用与精胺浓度呈正相关,此种抑制作用是可逆的。用 HPLC 测定精子精胺含量表明,精子获能后精胺含量明显下降。dbcAMP(0.5—1.0mmol/L)或咖啡因(10mmol/L)可拮抗精胺抑制人精子体外获能。其拮抗作用随 dbcAMP 浓度而增强。钙离子载体 A 23187 2/μmol/L 或胰蛋白酶0.05%均可拮抗精胺抑制人精子穿卵率。上述结果提示,精胺可能通过降低精子 cAMP 含量和抑制钙内流或顶体酶活性,从而阻止人精子体外获能和受精。     

THE EFFECTS OF SERUM ON SPERMATOZOA

A spermicidal factor was found in fresh human, bovine, rabbit, guinea pig, and rat sera. It kills the spermatozoa of its own species (except in the case of human serum) and the sperms of other species. It was unstable, thermolabile, and of large molecular size. It was present in limited quantity in the fresh serum and could be used up by a definite number of spermatozoa. It could be destroyed by sodium citrate, by Seitz filtration, by trypsin, and by snake venom. This factor was not present in tissue extracts and various plasma protein fractions. The strength or concentration of this factor varies in different individuals and in different species. This factor has several characteristics similar to those of complement.     

CHEMICAL DISSECTION OF MAMMALIAN SPERMATOZOA

Spermatozoa from several mammalian species have been dissected by chemical methods to yield free heads, tails with attached midpieces, and tails from which the mitochondrial components of the midpiece were removed. Mouse and rat spermatozoa were cleaved by brief treatment with trypsin to yield free heads and tails, while human, guinea pig, and rabbit spermatozoa were cleaved by trypsin only after incubation with 2-mercaptoethanol or dithiothreitol. Spermatozoa were also cleaved at the junction of the head and the tail by treatment with acid and base. Mitochondria were removed from intact spermatozoa or isolated tails by mechanical shear after treatment with 2-mercaptoethanol or dithiothreitol. The dissected components of spermatozoa were fractionated with good yield and high purity by density gradient centrifugation. Ultrastructural analysis indicates that proteolytic cleavage to yield separated heads and tails occurs at a specific location in the neck of the spermatozoon, leaving the basal plate attached to the head of the cell. In contrast, after acid cleavage the basal plate remains with the midpiece. Proteolytic treatment has no apparent effect on any other spermatozoan structures, whereas acid or base treatment results in damage to the plasma membrane, the acrosome, and other structures. The specificity of the proteolytic cleavage suggests that a particular protein or group of proteins may be responsible for the linkage between the sperm head and tail.     

口服醋酸棉酚前后人精子体外受精能力及精核转变的细胞学观察

本文报道口服醋酸棉酚对人精子体外受精能力、原核及染色体形成的影响。结果表明,在口服醋酸棉酚前的9名男性精子对金黄地鼠卵的穿透率平均为62%。每日口服20mg醋酸棉酚15天后,精子对卵的穿透率下降至47%,30天后下降至24%,50天后下降至8%。即口服醋酸棉酚50天后达到了不再具有生殖能力的精子穿透率阈值(10%)以下。原核及染色体形成的观察表明,即使口服醋酸棉酚50天后,仍可见有完整原核的形成,并未见有明显的染色体畸变。综述上列结果,似乎表明口服醋酸棉酚虽可影响人精子对去除透明带金黄地鼠卵的穿透率,而进入卵内的精子仍可形成原核及染色体。因此,仅从成熟精子的生理功能而论,口服醋酸棉酚期间它似乎并不产生可察觉的致畸效应,这与醋酸棉酚对体细胞并无致畸作用的报道相一致。     

Failure of human spermatozoa to penetrate zona free mouse and rat ova in vitro

When incubated for 8 to 26 hours with zona-free mouse or rat ova, human spermatozoa failed to attach to or penetrate any of the ova. The ova were capable of being fertilized since both intra- and inter-species penetration of spermatozoa and formation of pronuclei occurred between rat and mouse gametes. When mouse spermatozoa were incubated for three to eight hours with rat ova, a high proportion of the ova were penetrated, formation of pronuclei occurred and in 9 out of 36 ova incubated for 40 hours after insemination, regular cleavage and formation of morphologically normal 2-cell embryos occurred. Human spermatozoa retained their morphological integrity and motility only when the culture medium contained purified bovine serum albumin (3 mg/ml) or human serum (5% v/v) and not when unpurified BSA from several different commercial sources was used as a protein source. In this latter medium, the ova of both rats and mice degenerated after 8-hour incubation in the presence of human spermatozoa but not when human spermatozoa were absent or in the presence of either rat or mouse spermatozoa. Electron microscopy indicated that the human spermatozoa incubated for eight hours in medium containing purified BSA had undergone an acrosome reaction. These spermatozoa also attached to and penetrated human oocytes which had been matured in vitro.     

Binding of zona binding inhibitory factor-1 (ZIF-1) from human follicular fluid on spermatozoa

Previous studies showed that zona binding inhibitory factor-1 (ZIF-1) was the glycoprotein mainly responsible for the spermatozoa zona binding inhibitory activity of human follicular fluid. ZIF-1 has a number of properties similar to glycodelin-A. A binding kinetics experiment in the present study demonstrated the presence of two binding sites of ZIF-1 on human spermatozoa. These binding sites were saturable, reversible, and bound to (125)I-ZIF-1 in a time-, concentration-, and temperature-dependent manner. Glycodelin-A shared one common binding site with ZIF-1 on spermatozoa, and it could displace only 70% of the (125)I-ZIF-1 bound on human spermatozoa. ZIF-1 and glycodelin-A formed complexes with the soluble extract of human spermatozoa. Coincubation of solubilized zona pellucida proteins reduced the binding of ZIF-1 to two complexes of the extract, suggesting that the ZIF-1 binding sites and zona pellucida protein receptors on human spermatozoa were closely related. ZIF-1, but not glycodelin-A, significantly suppressed progesterone-induced acrosome reaction of human spermatozoa. The carbohydrate moieties derived from ZIF-1 reduced the binding of native ZIF-1 on human spermatozoa as well as the zona binding inhibitory activity of the glycoprotein, although the intensity of the effects are lower when compared with the native protein. These effects are not due to the action of the molecules on the motility, viability, and acrosomal status of the treated spermatozoa. Deglycosylated ZIF-1 had no inhibitory effect on both ZIF-1 binding and zona binding capacity of spermatozoa. We concluded that the carbohydrate part of ZIF-1 was critical for the functioning of the glycoprotein.     

Effect of sperm immobilisation and demembranation on the oocyte activation rate in the mouse.

To analyse the effect of the state of the sperm plasma membrane on oocyte activation rate following intracytoplasmic sperm injection (ICSI), three types of human and mouse spermatozoa (intact, immobilised and Triton X-100 treated) were individually injected into mouse oocytes. At 30, 60 and 120 min after injection, maternal chromosomes and sperm nuclei within oocytes were examined. Following human sperm injection, the fastest and the most efficient oocyte activation and sperm head decondensation occurred when the spermatozoa were treated with Triton X-100. Intact spermatozoa were the least effective in activating oocytes. Thus, the rate of mouse oocyte activation following human sperm injection is greatly influenced by the state of the sperm plasma membrane during injection. When mouse spermatozoa were injected into mouse oocytes, the rates of oocyte activation and sperm head decondensation within activated oocytes were the same irrespective of the type of sperm treatment prior to injection. We witnessed that live human spermatozoa injected into moue oocytes often kept moving very actively within the ooplasm for more than 60 min, whereas motile mouse spermatozoa usually became immotile within 20 min after injection into the ooplasm. In 0.002% Triton X-100 solution, mouse spermatozoa are immobilised faster than human spermatozoa. These facts seem to suggest that human sperm plasma membranes are physically and biochemically more stable than those of mouse spermatozoa. Perhaps the physical and chemical properties of the sperm plasma membrane vary from species to species. For those species whose spermatozoa have 'stable' plasma membranes, prior removal or 'damage' of sperm plasma membranes would increase the success rate of ICSI.     

A high activity carbonic anhydrase isoenzyme (CA II) is present in mammalian spermatozoa

Human and rat spermatozoa were stained for different carbonic anhydrase (CA) isoenzymes using specific antisera to human CA I, II and VI in conjunction with the immunofluorescence technique. The spermatozoa of both species were found to contain only CA II, which was located principally in the postacrosomal region of the human spermatozoa and in the acrosomal cap region of the rat spermatozoa. The presence of CA II could be confirmed by immunoblotting, which revealed a 29 K polypeptide in both the human and rat spermatozoa. No CA I or VI-specific fluorescence could be detected in the spermatozoa of either species. The immunoblottings were also negative. The results show mammalian spermatozoa to contain the high activity carbonic anhydrase isoenzyme II. Its presence is probably linked to hydration of CO2 produced by active energy metabolism and thereby to the maintaining of an adequate intraspermatozoal bicarbonate concentration as required for the maintenance of sperm motility.     

Steroid metabolism by monkey and human spermatozoa

Freshly ejaculated spermatozoa from monkey and human were washed and incubated with tritium labelled androgens or estradiol to study the pattern of spermatozoa steroid metabolism. When equal concentrations of steroid substrates were used for incubation, monkey and human spermatozoa showed very similar pattern of steroid conversion. Spermatozoa from both species converted testosterone mainly to androstenedione, but reverse conversion of androstenedione to testosterone was negligible. Estradiol-17 beta was converted mainly to estrone. The close similarity between the spermatozoa of monkey and men in their steroid metabolic pattern indicates that the rhesus monkey could be an useful animal model to study the effect of drugs on the metabolic pattern of human spermatozoa.     

The occurrence of polyenoic fatty acids with greater than 22 carbon atoms in mammalian spermatozoa.

Fatty acids with carbon chain lengths greater than 22 (VLCFA) have been detected in boar, ram, bull and human spermatozoa. Saturated and mono-unsaturated fatty acids were present in all spermatozoa but, except for human spermatozoa, polyenoic fatty acids were quantitatively the most important components. Marked differences in polyenoic fatty acid composition were observed. Whereas human spermatozoa contain predominantly di-, tri- and tetraenoic fatty acids with up to 32 carbon atoms, boar, ram and bull spermatozoa also contain pentaenoic and/or hexaenoic acids with up to 34 carbon atoms. Human and boar spermatozoa differ markedly from those of the ram and bull in that only n-6 series acids are present.