中华绒螯蟹(Eriocheir sinensis)雄性生殖系统的组织学研究

中华绒螯蟹雄性生殖系统的组织学研究表明:生精小管一侧为管壁上皮,另一侧为生发区,生殖细胞由生发区基底部同管腔增殖。输精管分为输精细管和贮精囊,管壁上皮具分泌功能,贮精囊有肌肉层。射精管壁肌肉层较厚,粘膜形成纵行皱襞。副性腺内壁为单层立方上皮。生殖系统发育有明显的季节性,8月开始发育加速,10月进入高峰,4月开始发育停滞。     

饥饿对中华绒螯蟹(Eriocheir sinensis)幼体发育的影响

对刚孵化的中华绒螯蟹第一期的蚤状幼体经不同时间的饥饿后再投喂,发现饥饿可以明显降低幼体的存活率和延长幼体的发育期。实验表明:对中华绒螯蟹第一期的蚤状幼体的饥饿时间(t)和发育期长(D)呈线性关系(D=4.6303+1.3226t r=0.970p<0.01)。对于中华绒螯蟹第一期的蚤状幼体,当起始饥饿时间超过了4d,再予以投饵,幼体均不能恢复正常的发育和蜕皮功能,得出中华绒螯蟹的不可恢复点(the point of no-return,PNR)大约为4d。通常以产生50%的幼体死亡的饥饿期即PNR₅₀,来表明幼体对饥饿的抵抗能力,实验得出中华绒螯蟹第一期的蚤状幼体的PNR₅₀大约为48h。     

中华绒螯蟹(Eriocheir japonica sinensis)线粒体ND5基因序列变异分析

不同水系河蟹已发现严重混杂,水产业中急需一种快速鉴别不同水系的分子标记。基于研究13条长约853bp的线粒体ND5基因序列分析得出,其平均碱基组成为40.2%A,30.9%T,9.2%G,19.7%C,共定义了8个单元型,单元型多样性(h)为0.885;序列间存在16个变异位点,核苷酸多样性(π)为0.004;平均遗传距离为0.004。其中单元型EJSⅠ和EJSⅡ占据了53.8%的个体数量,推测可能是较为原始类型;两单元型在NJ和UPG-MA系统发生树中分居于不同的分支,可能蕴涵着各自进化历史。通过研究表明,ND5序列的变异水平将为采用PCR-RFLP或位点特异性PCR等分子标记分辨不同水系河蟹起着重要作用。     

中华绒螯蟹Eriocheir sinensis H.Milne-Edwards的幼体发育

1.本文所报道毛蟹的幼体发育试验,全部是在实验室内进行的。 2.毛蟹的幼体发育共经五个(氵蚤)状幼体期和一个大眼幼体期。卵孵化出膜的幼体即为第一(氵蚤)状幼体,而不是早期(氵蚤)伏幼体或原(氵蚤)伏幼体。这两种幼体应该是在卵膜内度过的。 3.在水温11—22℃,盐度为9‰的实验室条件下,从幼体出膜到第一期幼蟹的出现,共经39—40天。每蜕一次皮,即进入另一发育时期,其所需的时间,随着温度的升高而缩短。其发育速度见下表: 幼体名称 水温(℃) 日数 第一(氵蚤)状幼体 11—17 7—9 第二(氵蚤)状幼体 17—18 5—6 第三(氵蚤)状幼体 18—20 6—7 第四(氵蚤)状幼体 16—19.5 5—6 第五(氵蚤)状幼体 16.5—19 7—8 大眼幼体 19—22 9—10 4.饵料、盐度、水温和水质等因子对幼体有着不同程度的影响。 5.(氵蚤)状幼体以两对颚足外肢末端的羽伏刚毛的数目,胸、腹肢的大小与形状和尾叉内面中部刚毛的数目为分期的主要依据。     

中华绒螯蟹(Eriocheir sinensis)肌肉中不饱和脂肪酸含量与越冬死亡、产卵关系

本文报道了中华绒螯蟹越冬时和越冬后不同性别,不同生活状态粗脂肪中不饱和脂肪酸及肌肉中不饱和脂肪酸含量及变化,分析了不饱和脂肪酸含量与越冬死亡,越冬后交配,产卵和孵化关系。越冬前雌,雄河蟹粗脂肪中不饱和脂肪酸碘（I2）值为57和48。越冬（越冬时,越冬后）死亡雌,雄河蟹粗脂肪中不饱和脂肪酸I2值分别低于28和10;而其肌肉中不饱和脂肪酸的I2值分别低于1．25和0．83。越冬后能成功交配的雄河蟹粗脂肪中不饱和脂肪酸I2值高于33。越冬后交配产卵雌河蟹粗脂肪中不饱和脂肪酸I2值高于42,抱卵并使卵正常孵化的雌河蟹粗脂肪中不饱和脂肪酸I2值大于47。脂肪酸在河蟹的细胞内氧化分解,为越冬提供了能量。     

中华绒螯蟹一龄幼蟹(Eriocheir sinensis)越冬前后脂肪酸和4种类胡萝卜素含量的变化

实验以越冬前后的中华绒螯蟹一龄幼蟹作为研究对象,利用高效气相色谱法分别检测一龄幼蟹越冬前后肝胰腺及头胸甲中脂肪酸含量变化、利用液相二级质谱法分别测定一龄幼蟹越冬前后肝胰腺、头胸甲和血淋巴中4种类胡萝卜素含量变化。结果表明:肝胰腺中的饱和脂肪酸(SFA) C20:0、单不饱和脂肪酸(MUFA) C18:1n9作为主要能量脂肪酸被动用,其含量越冬后显著下降(p0.05),多不饱和脂肪酸(PUFA)中的C18:3n3、C22:6n3越冬后含量显著升高(p0.05)。头胸甲的饱和脂肪酸(SFA) C16:0、C18:0,单不饱和脂肪酸(MUFA)中的C18:1n7作为主要能量脂肪酸被动用,越冬后含量下降显著(p0.05)。多不饱和脂肪酸(PUFA) C18:2n6、C20:3n6越冬后含量显著升高(p0.05)。越冬后头胸甲中4种类胡萝卜素含量均极显著下降(p0.01)。其中,血淋巴中虾青素、叶黄素含量极显著升高(p0.05),β-胡萝卜素含量极显著下降(p0.05)。肝胰腺中虾青素与叶黄素、β-胡萝卜素变化趋势与血淋巴的一致,角黄素含量显著下降(p0.05)。越冬期间气温、水温的短暂升高可能使一龄幼蟹短暂摄食。越冬前经育肥的扣蟹越冬后能量物质无显著变化,越冬后根据肝胰腺颜色来判断一龄幼蟹质量的方法不科学。     

中华绒螯蟹（Eriocheir sinensis）体内维生素C含量与死亡和产…

本文报道了中华绒螯蟹越冬后肝胰脏、肌肉和生殖腺维生素Ｃ的含量及变化。肝胰脏为中华绒螯蟹储存维生素Ｃ的器官;越冬后的雌性中华绒螯蟹,其肌肉和生殖腺的维生素Ｃ高于雄性;正常活动雌性或雄性,其肝胰脏、肌肉和生殖腺的维生素Ｃ含量均高于死亡的个体的维生素Ｃ含量;产卵且抱卵孵化的雌体,其肝脏维生素Ｃ的含量高于那些不产卵的雌体。维生素Ｃ含量的降低是越冬时或越冬后中华绒螯蟹死亡的主要原因之一。     

中华绒螯蟹（Eriocheir sinensis）肌肉中粗脂肪含量与死亡,产 …

本文报道了中华螯蟹越冬时,越冬后不同性别、不同生活状况肌肉中粗脂肪含量及变化。越冬时、越冬后死亡的♂♀河蟹肌肉中粗脂肪的含量均不到８％,而同时越冬成活的♂♀蟹肌肉中粗脂肪含量均高于１０％。顺利越冬并能产卵的雌蟹肌肉中粗脂肪含量为４．９６％,顺利越冬并交配的♂性肌肉中粗脂肪含量为４．１８％。产卵并抱卵孵化的♀蟹肌肉中粗脂肪含量为５．８５％,产卵但♀蟹中途死亡的其肌肉中粗脂肪的含量为５．４７％。交配不     

饥饿胁迫对中华绒螯蟹(Eriocheir sinensis)仔蟹的影响

在26.2 ～28.4℃水温条件下,研究了饥饿对中华绒螯蟹(Eriocheir sinensis)仔蟹的形态、行为、存活及体重损失的影响,同时确定了仔Ⅱ的营养饱和储存点(PRS)和不可恢复点(PNR).结果表明:饥饿胁迫下中华绒螯蟹仔Ⅰ、仔Ⅱ、仔Ⅲ的初次死亡时间(T1)分别为8.0、14.0和20.3 d,50％死亡时间(Ts0)分别为11.4、16.0和25.5 d,100％死亡时间(T100)分别为15.0、22.0和32.3 d,耐饥饿能力为仔Ⅲ＞仔Ⅱ＞仔Ⅰ;饥饿期间中华绒螯蟹体内水分含量持续升高,干重量降低显著,干重损失速率也随时间延长逐渐减小;先饱食后饥饿的给饵模式中仔Ⅱ蜕皮率随初始饱食时间延长而升高,50％个体完成蜕皮所需饱食时间(PRS50)为2.10d,各处理组仔Ⅱ的蜕皮周期与持续饱食组均无显著差异(P＞0.05),但只有饱食3d以上才能达到持续饱食饲养蟹的增重率;先饥饿后饱食的给饵模式中,仔Ⅱ的蜕皮率随着初始饥饿时间的延长而下降,其中仔Ⅱ50％不能蜕皮的初始饥饿时间(PNR50)和100％不能蜕皮的初始饥饿时间(PNR100)分别为10 d和14 d,并且蜕皮周期相对延长,延长时间约等于初始饥饿时间,不存在额外的摄食时间来弥补饥饿期间损失的能量,各处理组蜕皮后仔蟹与对照组体重无显著差异(P＞0.05).     

锯缘青蟹精子顶体反应的研究

采用正交实验法研究了诱导锯缘青蟹精子产生顶体反应的最佳条件。结果表明：从纳精囊中获得的精子,用离子载体Ａ23187(64μg/ml）诱导,在ＣaCl2浓度为0.25％,pH为9.0的人工海水中４０分钟,可以得到最大的顶体反应率（82.5%）。在此基础上采用光镜和电镜观察顶体反应过程中显微和超微结构的变化。顶体反应过程可大致分为两个时相,第一时相为顶体囊外翻;第二时相为顶体管前伸。本文还对顶体反应的作用进行了初步探讨。     

中华绒螯蟹染色体的研究

中华绒螯蟹(Eriocheir sinensis H.Milne Edwards)属甲壳纲(Crustacea)、软甲亚纲(Malacostraca)、十足目(Decapoda)、方蟹科(Grapsidae),俗称河蟹,是我国重要的经济蟹类,其形态解剖、生态习性及生理等方面的研究已有报道(Panning,1939;Koch,1952;堵南山,1954,1957,1958;Bauchau,1960;Leersnyder,1966,1967;Dbainaut et al.,1976;Chevigue,1976;Leeranyder et al.,1977,1978;Pequenx et al.,1982;Chapelle et al.,1982;谈奇坤等,1984.),但至今未见有关细胞遗传学方     

三疣梭子蟹精子顶体反应过程中的形态和结构变化

用离子载体A2 3187和卵水人工诱导三疣梭子蟹精子的顶体反应 ,分别获得 75 33%和 84 83%的顶体反应率。应用光镜和电镜技术观察了顶体反应前后精子形态和结构的变化。未处理精子呈陀螺形 ,由顶体、核杯和 5 - 10条核辐射臂组成。顶体包括顶体囊和顶体管。顶体囊的伞形头帽拥有约 70条辐射肋。连续发生的精子顶体反应过程被人为地分为四个阶段 :(1)头帽鼓起 ;(2 )顶体囊外翻 ;(3)穿孔器前伸 ,顶体囊膜翻转 ;(4 )顶体囊膜脱落 ,顶体丝形成。直到第四阶段才观察到钉状精子的辐射臂开始收缩。探讨了辐射臂和穿孔器前冲在精子入卵中的功能     

Inositol tri-phosphate in human and ascidian spermatozoa

Using a specific protein binding assay we have shown that a spermatozoon of the ascidian Ciona intestinalis contains 1.58 ± 0.74 × 10^?19 moles of inositol 1,4,5-tri-phosphate (InsP₃), while a human spermatozoon contains 6.4 ± 0.14 × 10^?19 moles. Induction of the acrosome reaction (AR) in both species, by exposure to the calcium ionophore A23187, does not significantly alter levels of InsP₃, suggesting that phosphatidylinositol (PI) turnover is not necessary for the calcium ionophore induced AR. Furthermore, PI turnover in ascidian spermatozoa appears to be insensitive to lithium and phorbol ester. The high intracellular concentration of InsP₃ in spermatozoa, corresponding to 50–200 μM, suggests it may play a role in egg activation. © 1993 Wiley-Liss, Inc.     

Video microscopic analysis of ionophore induced acrosome reactions of lobster (Homarus americanus) sperm

Sperm from the American lobster (Homarus americanus) are normally nonmotile. However, during fertilization, the sperm undergo a calcium-dependent acrosome reaction that propels them forward about 18 μMm. The reaction occurs in two phases, eversion and ejection, which take place too quickly to permit analysis by direct observation. The purposes of this study were to examine the structural changes occurring in sperm during the normal acrosome reaction and to determine the rate of the reaction using video microscopy. The reaction was induced in vitro by ionophore A23187 and recorded using a video system attached to a Nikon Nomarski interference microscope. Videotapes were played back frame by frame (30 frames/sec), and images of reactions from 10 sperm were analyzed. The acrosome reaction, including the eversion of the acrosomal vesicle and ejection of the subacrosomal material and nucleus, can be divided into 4 steps: (1) expansion of the apical cap followed by expansion of the remainder of the acrosomal cylinder; expansion of the cylinder begins at its apical end and proceeds toward its base, (2) eversion of the apical half of the acrosomal vesicle and initial contraction of the apical cap, (3) eversion of the basal half of the acrosomal vesicle, continued contraction of the apical cap, and ejection of the subacrosomal material and nucleus, and (4) final contraction of the apical cap and ejection of the acrosomal filament. During steps 2, 3, and 4, the mean forward movement of sperm is 12.7, 3.9, and 1.1 μMm, respectively. Although the time required to complete the reaction ranged from 0.66 to 5.16 s, most sperm reacted in less than 3. s, and these sperm were considered to have typical rates. For sperm that reacted in less than 3 s, both step 1 and step 4 take about 0.2 s and show little variation among sperm. the time required to complete steps 2 and 3 averaged 0.63 and 0.37 s, respectively. Forward movement of the sperm during the acrosome reaction is caused by eversion of the inner and outer acrosomal material and contraction of the apical cap. The protein(s) responsible for this contraction is not yet known. © 1993 Wiley-Liss, Inc.     

Does the zona pellucida select spermatozoa from the medium with higher fertilizing potential?

The present study was carried out to test whether the zona pellucida selects spermatozoa with higher fertilization potential. Fertilization rates of mouse oocytes after sperm microinjection under the zona pellucida (SMUZ) of zona-bound spermatozoa and of spermatozoa incubated in the absence of oocytes and treated (acid-treated group) or not (control group) with acid Tyrode's solution were compared. SMUZ was performed at 15, 30, 60, and 90 min after the insemination of fresh oocytes required for selecting spermatozoa bound to the zona pellucida. At these times, the percentages of acrosome-reacted spermatozoa (PARS) were evaluated using the chlortetracycline fluorescence method. Fertilization rate in the three groups analysed increased from 25.9% to 47.3% in the control group, from 29.3% to 44.0% in the acid-treated group, and from 19.5% to 40.0% in the zona-bound group from 15 to 90 min after insemination, respectively. The global fertilization potential was significantly lower in the zona-bound group compared to the other two groups. The PARS in the zona-bound group at 15 (11.48 ± 3.02); 30 (16.74 ± 3.71), and 90 (19.68 ± 3.68) min after insemination were significantly (P < 0.05) lower than those found in the acid-treated group (39.26 ± 6.69, 38.20 ± 6.24, and 42.83 ± 5.39, respectively). At 90 min after insemination, the PARS in the zona-bound group was also significantly (P 0.05) lower than the control group (36.72 ± 4.51). No significant correlation between either time and PARS or PARS and fertilization rate was observed. It appears that the zona pellucida does not select from the medium spermatozoa with higher fertilization potential. © 1993 Wiley-Liss, Inc.     

Capacity of rete testicular and cauda epididymal boar spermatozoa to undergo the acrosome reaction and subsequent fusion with egg plasma membrane

The capacity to undergo the acrosome reaction and subsequent fusion with egg plasma membrane was examined in rete testicular and cauda epididymal spermatozoa from boars. Sperm penetration assay using zona-free hamster eggs demonstrated that the penetration rates for rete testicular spermatozoa preincubated for induction of the acrosome reaction for 2 and 3 h were 55% and 97%, respectively. However, most of the eggs (93%) were penetrated with polyspermy by cauda epididymal cells preincubated for 2 h. Results obtained by the triple-stain technique revealed the percentages of acrosome-reacted spermatozoa in the rete testicular and cauda epididymal samples preincubated for 3 h to be 61% and 74%, respectively. These results indicate that many rete testicular spermatozoa possess the capacity to undergo the acrosome reaction and subsequent fusion with egg plasma membrane in vitro, which appears to be completely established only after sperm transit through at least the proximal part of the epididymis. © 1993 Wiley-Liss, Inc.     

Subzonal insemination with a single spermatozoon using manipulation assisted sperm adhesion onto the ooplasmic membrane in mouse ova.

A simple and successful method of microinjection of a single spermatozoon under the zona pellucida of a mouse oocyte has been developed. A characteristic of this method is that the tip of the sperm injection needle pierces the zona pellucida without touching the ooplasmic membrane. All the ova (277) used for this series of experiments had normal morphology after the injection procedure. Spermatozoa preincubated in culture medium for capacitation and those treated with ionophore A23187 for induction of acrosome reaction were used. In combination with some of these injections, a manipulation assisting the adhesion of the sperm head onto the ooplasmic membrane was employed. The fertilization rate (67.3%) of the ova injected with the ionophore-treated sperm using the sperm-adhesion treatment was significantly higher (P less than 0.005) than that obtained by the injection of the preincubated sperm without applying the adhesion treatment (23.6%). All three of the recipients that received the 24 fertilized ova became pregnant and gave birth to 11 offspring (45.8%). The inseminations performed with the sperm-adhesion treatment using the immotile sperm from the preincubated population and/or those from the ionophore-treated population did not result in fertilization in any case. These results suggest that the fertilization rate of subzonal insemination with motile ionophore-treated sperm can be improved by applying the sperm-adhesion treatment and that sperm motility might be involved in the establishment of fertilization, even after the adhesion of the sperm head with the mouse ovum membrane.     

The SLO3 sperm-specific potassium channel plays a vital role in male fertility

Here we show a unique example of male infertility conferred by a gene knockout of the sperm-specific, pH-dependent SLO3 potassium channel. In striking contrast to wild-type sperm which undergo membrane hyperpolarization during capacitation, we found that SLO3 mutant sperm undergo membrane depolarization. Several defects in SLO3 mutant sperm are evident under capacitating conditions, including impaired motility, a bent “hairpin” shape, and failure to undergo the acrosome reaction (AR). The failure of AR is rescued by valinomycin which hyperpolarizes mutant sperm. Thus SLO3 is the principal potassium channel responsible for capacitation-induced hyperpolarization, and membrane hyperpolarization is crucial to the AR.