Bovine cumulus-oocyte disconnection in vitro

Summary Cumulus-oocyte complexes were obtained from cows by aspiration of small (1–6 mm in diameter) antral follicles after slaughter. Complexes with a compact multilayered cumulus investment were cultured and processed for transmission electron microscopy after different periods of culture including a 0 h control group. In 0 h control oocytes the cumulus cells had numerous projections which penetrated the zona pellucida and established gap junctions with the oolemma. A partial loss of these junctions was noticed as an early event of oocyte maturation occurring within the first 3 h of culture. A low frequency of gap junctions was maintained until 12–18 h of culture where the junctional contact was completely disrupted. This decrease in intercellular communication was parallelled by resumption of oocyte meiosis.     

Fertility in patients with multiple sclerosis: current knowledge and future perspectives

Abstract The issue of fertility in patients with multiple sclerosis (MS) has not been exhaustively studied. Epidemiological data have suggested that spontaneous fecundity might be reduced; several endocrine and sexual disturbances potentially interfering with reproduction have been evidenced in MS patients of both sexes. Moreover, some medical treatments used in MS (e. g., mitoxantrone, cyclophosphamide) may exert detrimental effects on spermatozoa as well as on oocytes, leading to early impairment of fertility. This review illustrates the factors potentially interfering with fertility in MS and discusses the therapeutic tools that may be used to promote fertility in these patients. The safety of hormonal therapies in MS is also examined. The current applications of assisted reproductive technology (ART) are discussed, including in vitro fertilisation (IVF) techniques. Currently available methods to preserve fertility in patients that undergo cytotoxic treatments by means of sperm/oocyte cryostorage or by ovarian fragment cryopreservation and autografting are considered.     

降调节对卵子质量的影响

近10余年来，在体外受精一胚胎移植（IVF-ET）周期中应用促性腺激素释放激素激动剂（GnRH-a）进行控制性卵巢刺激（COS）已成为普遍采用的方法，其主要原因是GnRH-a可以促进卵巢内多个卵泡同步发育和成熟，抑制内源性黄体生成素（LH）峰，阻遏卵泡过早黄素化及卵子早熟。由于GnRH-a在IVF-ET进行降调节COS中的应用不同，可将COS方案分为长方案和短方案。[第一段]     

Mechanism of action of the epileptogenic drug pentylenetetrazol on a cloned neuronal potassium channel

The action of the epileptogenic agent pentylenetetrazol (PTZ) on a cloned potassium channel of the rat brain was studied. The Kv1.1 channel was expressed in oocytes ofXenopus laevis and potassium currents were investigated in outside-out and inside-out membrane patches. The results show that PTZ increased the multi-channel potassium currents at strongly negative potentials and decreased them at potentials positive to −35 mV both in outside-out and inside-out membrane patches. The extent and manner of PTZ action, the concentration dependence as well as the onset and time course of the PTZ effect were the same both in outside-out and inside-out membrane patches. The single-channel potassium currents showed an increase in open probability and frequency of opening and a decrease in close time at −50 mV and vice versa at 0 mV with application of PTZ. The amplitude of single-channel current, the open time and the latency to the first channel opening remained almost unchanged under PTZ. The results indicate that PTZ acts via the cell membrane and influences the membrane-associated part of the potassium channel. Thereby, PTZ accelerates the transition from the inactivated to the open state of the channel at strongly negative potentials and reduces it at slightly negative and positive potentials. This mechanism may be the basis for a gate function which is in favour of the development of epileptic discharges.     

异丙酚麻醉不同阶段对卵细胞受精和对早期胚胎质量的影响

目的探讨异丙酚静脉麻醉下取卵手术中卵泡抽吸液中异丙酚的浓度变化及各阶段对卵细胞受精和早期胚胎质量的影响.方法 17例实施超促排卵下体外受精-胚胎移植术的妇女预测卵泡至少有15个,按取卵的先后分3组(早、中、晚)分别培养.记录从静脉给异丙酚到每个卵泡取出时间.检测卵泡抽取液中异丙酚的浓度及观察卵细胞的受精率及卵裂率等.结果取出第G一个卵泡的平均时间是200秒,取出卵泡的平均速率是17.6s/ 个.所有患者的最后一个卵泡抽吸液中异丙酚的浓度明显高于第一个卵泡抽吸液中异丙酚的浓度(P<0.01).除未成熟的卵细胞占总卵泡百分比在3组间有明显的区别外,受精率和卵裂率无明显区别.结论随着麻醉的进展,卵泡抽吸液中异丙酚的浓度逐渐增加,但与所给剂量或持续的时间之间无必然的关联.我们不能显示卵泡抽吸液中升高的异丙酚浓度对卵细胞质量的不利影响.     

Kinetic mode switch of rat brain IIA Na channels in Xenopus oocytes excised macropatches

Na currents recorded from inside-out macropatches excised from Xenopus oocytes expressing the subunit of the rat brain Na channel IIA show at least two distinguishable components in their inactivation time course, with time constants differing about tenfold ( _h1 = approx. 150 s and _h2 = approx. 2 ms). In excised patches, the inactivation properties of Na currents changed with time, favoring the faster inactivation kinetics. Analysis of the fast and slow current kinetics shows that only the relative magnitudes of _h1 and _h2 components are altered without significant changes in the time constants of activation or inactivation. In addition, voltage dependence of both activation and steady-state inactivation of Na currents are shifted to more negative potentials in patches with predominantly fast inactivation, although reversal potentials and valences remained unaltered. We conclude that the two inactivation modes discerned in this study are conferred by two states of Na channel the interconversion of which are regulated by an as yet unknown mechanism that seems to involve cytosolic factors.     

女性年龄与卵细胞染色体非整倍体异常的相关关系

目的 :直接从卵细胞水平研究女性年龄与其卵细胞染色体非整倍体间的相关关系。方法 :取试管婴儿助孕技术后未能受精成功的卵细胞 ,采用多色荧光原位杂交方法检测卵细胞 13,16 ,18,2 1和 2 2号染色体的情况。结果 :卵细胞的非整倍体率为 2 7.1% ,随着女性年龄的增加 ,卵细胞的非整倍体率也随之上升 ,两者呈较强的正相关关系 ( r=0 .88,P<0 .0 0 1)。结论 :女性年龄与其染色体非整倍体异常间存在相关性 ,对于 35岁以上行试管婴儿助孕术的妇女应同时进行遗传咨询     

左右侧卵巢卵泡液中白血病抑制因子与卵子质量关系的研究

目的:对比性分析左右侧卵巢卵泡液中白血病抑制因子(LIF)含量及其与卵子和胚胎质量的关系。方法:选择首次实施体外受精/卵胞浆内单精子注射(IVF/ICSI)不孕症妇女37例,采卵日采集血清及左右侧卵巢卵泡液,按左右标记后编号。ELISA法测定LIF含量。结果:右侧卵巢卵泡液中LIF含量明显高于左侧(P0.05);右侧卵巢减数分裂中期(MII期)卵数、正常受精的双原核(2PN)数、可移植胚胎数明显多于左侧(P0.05);双侧卵泡液中LIF浓度与可移植胚胎数呈明显正相关(P0.05)。结论:右侧卵巢内卵子的数量和质量优于左侧,卵泡液中LIF含量有可能作为评估卵子正常受精及早期胚胎质量的参考指标之一。     

线粒体能量代谢和DNA复制对小鼠卵子体外成熟、受精及发育的影响

目的 探讨胞质线粒体的氧化磷酸化（OXPHOS）活性或线粒体DNA复制在卵子成熟、受精和胚胎发育过程中的作用。方法 通过在小鼠体外成熟培养液中引入不同浓度的羰基氰4-(三氟甲氧基)苯腙 (FCCP, 10 nmol/L和100nmol/L)或2′,3′-双脱氧胞苷(ddC,10μmol/L和100μmol/L),抑制线粒体OXPHOS活性或线粒体DNA复制,统计分析各组卵子的体外生发泡破裂（GVBD）率、核成熟率、受精率及囊胚形成率,以分析线粒体功能抑制对卵子成熟、受精和胚胎发育的影响。 结果 线粒体OXPHOS活性和DNA复制功能在卵子和胚胎中所发挥的作用并不完全相同。FCCP抑制线粒体OXPHOS活性可显著降低卵子的核成熟率和囊胚形成率;但对卵子的GVBD的发生率和受精率无显著影响。而ddC抑制线粒体DNA复制不影响卵子的体外成熟和受精,但可显著抑制囊胚的形成。 结论 OXPHOS活性主要影响卵子成熟及胚胎发育;线粒体DNA复制则主要影响胚胎发育;而线粒体功能抑制不影响卵子的成熟启动和体外受精。     

Seasonal changes in plasma steroid levels in Indian major carp Labeo rohita: influence of homologous pituitary extract on steroid production and development of oocyte maturational competence

Circannual variations in plasma levels of testosterone (T), 17beta-estradiol (E(2)), and 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (DHP) as well as seasonal fluctuations in ovarian steroid synthetic potential were observed in Indian major carp, Labeo rohita. A study was also conducted to examine the mechanism of the development of gonadotropin-induced maturational competence in oocytes of this fish. The present study recorded the lowest values of plasma E(2) and T in L. rohita during the period from October to January. A mild increase in the levels of these steroids observed in February was followed by their rapid rise reaching peak values in April, when the ovary contained mostly the vitellogenic follicles. In the month of May, as the postvitellogenic follicles predominated in the ovary, there was a decline in plasma concentrations of both T and E(2). Low levels of these steroids in plasma remained until January, except a small elevation detectable during June and July (spawning stage). DHP was not detected in the plasma of this fish collected during the period from August to March. Existence of DHP was first recorded in blood in the month of April (vitellogenic stage) and it quickly reached the peak value in May (postvitellogenic stage), followed by a sudden decline in the month of June. Under stimulation of fish pituitary extract (FPE), as a source of gonadotropin, in vitro production of E(2) and T by the vitellogenic follicles was shown to be highest compared to their production rate in other stages, while the postvitellogenic follicles recorded the highest rate of DHP synthesis. Acquisition of oocyte maturational competence (OMC) was shown to develop either by priming the vitellogenic stage fish with a single dose of FPE or by in vitro addition of FPE in culture. In vitro treatment of trilostane, an inhibitor of 3beta-HSD, blocked the FPE-stimulated steroid production but not the development of OMC. Presence of cycloheximide and actinomycin D in the incubation was shown to inhibit FPE-induced development of OMC, indicating the requirement of de novo protein synthesis for this process.