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Immediate and delayed effects of heat stress on follicular development and its association with plasma FSH and inhibin concentration in cows. 热应激对奶牛卵泡发育的即时和延迟影响及其与血浆卵泡刺激素和抑制素浓度的关系
Pub Date : 2000-09-01 DOI: 10.1530/JRF.0.1200083
Z. Roth, R. Meidan, R. Braw-Tal, D. Wolfenson
The aim of this study was to characterize the immediate effects of heat stress on plasma FSH and inhibin concentrations, and its involvement in follicular dynamics during a complete oestrous cycle, and to examine a possible delayed effect of heat stress on follicular development. Holstein dairy cows were oestrous synchronized and randomly assigned to either cooled (n = 7) or heat-stressed (n = 6) treatment groups. During a complete oestrous cycle, control cows, which were cooled, maintained normothermia, whereas heat-stressed cows, which were exposed to direct solar radiation, developed hyperthermia. At the end of this oestrous cycle (treated cycle), both groups were cooled and maintained normothermia for the first 10 days of the subsequent oestrous cycle. Throughout this period, follicular development was examined by ultrasonography, and plasma samples were collected. During the second follicular wave of the treated oestrous cycle, a significantly larger cohort of medium sized follicles (6-9 mm) was found in heat-stressed cows than in cooled cows (P < 0.05). The enhanced growth of follicles in this wave in heat-stressed cows was associated with a higher plasma FSH increase which lasted 4 more days (days 8-13 of the oestrous cycle; P < 0.05), and coincided with a decrease in the plasma concentration of immunoreactive inhibin (days 5-18 of the oestrous cycle; P < 0.05). During the follicular phase (days 17-20 of the treated cycle), heat-stressed cows showed an increase in the number of large follicles (>/= 10 mm), and the preovulatory plasma FSH surge was significantly higher in heat-stressed cows than in cooled cows (P < 0.01). The effect of heat stress was also observed during the first follicular wave of the subsequent cycle: the postovulatory plasma FSH concentration was higher (P < 0.01), but fewer medium follicles developed, and the first follicular wave decreased at a slower rate in previously heat-stressed cows than in cooled cows (0.40 and 0.71 follicles per day, respectively). This study shows both immediate and delayed effects of heat stress on follicular dynamics, which were associated with high FSH and low inhibin concentrations in plasma. These alterations may have physiological significance that could be associated with low fertility of cattle during the summer and autumn.
本研究的目的是表征热应激对血浆促卵泡刺激素和抑制素浓度的直接影响,及其在完整发情周期中对卵泡动力学的影响,并研究热应激对卵泡发育的可能延迟影响。选取荷斯坦奶牛同步发情,随机分为冷应激组(n = 7)和热应激组(n = 6)。在一个完整的发情周期中,被冷却的对照奶牛保持体温正常,而暴露在太阳直接辐射下的热应激奶牛则出现体温过高。在此发情周期结束时(处理周期),两组均降温并在随后的发情周期的前10天保持常温。在此期间,通过超声检查卵泡发育,并收集血浆样本。在发情周期的第二次卵泡波中,热应激奶牛的中等大小卵泡(6-9 mm)数量显著高于冷应激奶牛(P < 0.05)。热应激奶牛在这一波中卵泡生长的增强与血浆促卵泡刺激素的增加有关,这种增加持续了4多天(发情周期的第8-13天;P < 0.05),且与血浆免疫反应抑制素浓度降低(发情周期第5 ~ 18天;P < 0.05)。在卵泡期(处理周期第17 ~ 20天),热应激奶牛的大卵泡数(>/= 10 mm)显著增加(P < 0.01),排卵前血浆FSH峰值显著高于冷应激奶牛(P < 0.01)。热应激对后续周期的第一次卵泡波也有影响:排卵后血浆FSH浓度升高(P < 0.01),但中等卵泡发育较少,第一次卵泡波下降速度慢于冷却奶牛(分别为0.40和0.71个/ d)。本研究显示热应激对卵泡动力学的即时和延迟影响,这与血浆中高FSH和低抑制素浓度有关。这些变化可能具有生理意义,可能与夏季和秋季牛的低生育率有关。
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引用次数: 244
Immediate and delayed effects of heat stress on follicular development and its association with plasma FSH and inhibin concentration in cows. 热应激对奶牛卵泡发育的即时和延迟影响及其与血浆卵泡刺激素和抑制素浓度的关系
Z Roth, R Meidan, R Braw-Tal, D Wolfenson

The aim of this study was to characterize the immediate effects of heat stress on plasma FSH and inhibin concentrations, and its involvement in follicular dynamics during a complete oestrous cycle, and to examine a possible delayed effect of heat stress on follicular development. Holstein dairy cows were oestrous synchronized and randomly assigned to either cooled (n = 7) or heat-stressed (n = 6) treatment groups. During a complete oestrous cycle, control cows, which were cooled, maintained normothermia, whereas heat-stressed cows, which were exposed to direct solar radiation, developed hyperthermia. At the end of this oestrous cycle (treated cycle), both groups were cooled and maintained normothermia for the first 10 days of the subsequent oestrous cycle. Throughout this period, follicular development was examined by ultrasonography, and plasma samples were collected. During the second follicular wave of the treated oestrous cycle, a significantly larger cohort of medium sized follicles (6-9 mm) was found in heat-stressed cows than in cooled cows (P < 0.05). The enhanced growth of follicles in this wave in heat-stressed cows was associated with a higher plasma FSH increase which lasted 4 more days (days 8-13 of the oestrous cycle; P < 0.05), and coincided with a decrease in the plasma concentration of immunoreactive inhibin (days 5-18 of the oestrous cycle; P < 0.05). During the follicular phase (days 17-20 of the treated cycle), heat-stressed cows showed an increase in the number of large follicles (>/= 10 mm), and the preovulatory plasma FSH surge was significantly higher in heat-stressed cows than in cooled cows (P < 0.01). The effect of heat stress was also observed during the first follicular wave of the subsequent cycle: the postovulatory plasma FSH concentration was higher (P < 0.01), but fewer medium follicles developed, and the first follicular wave decreased at a slower rate in previously heat-stressed cows than in cooled cows (0.40 and 0.71 follicles per day, respectively). This study shows both immediate and delayed effects of heat stress on follicular dynamics, which were associated with high FSH and low inhibin concentrations in plasma. These alterations may have physiological significance that could be associated with low fertility of cattle during the summer and autumn.

本研究的目的是表征热应激对血浆促卵泡刺激素和抑制素浓度的直接影响,及其在完整发情周期中对卵泡动力学的影响,并研究热应激对卵泡发育的可能延迟影响。选取荷斯坦奶牛同步发情,随机分为冷应激组(n = 7)和热应激组(n = 6)。在一个完整的发情周期中,被冷却的对照奶牛保持体温正常,而暴露在太阳直接辐射下的热应激奶牛则出现体温过高。在此发情周期结束时(处理周期),两组均降温并在随后的发情周期的前10天保持常温。在此期间,通过超声检查卵泡发育,并收集血浆样本。在发情周期的第二次卵泡波中,热应激奶牛的中等大小卵泡(6-9 mm)数量显著高于冷应激奶牛(P < 0.05)。热应激奶牛在这一波中卵泡生长的增强与血浆促卵泡刺激素的增加有关,这种增加持续了4多天(发情周期的第8-13天;P < 0.05),且与血浆免疫反应抑制素浓度降低(发情周期第5 ~ 18天;P < 0.05)。在卵泡期(处理周期第17 ~ 20天),热应激奶牛的大卵泡数(>/= 10 mm)显著增加(P < 0.01),排卵前血浆FSH峰值显著高于冷应激奶牛(P < 0.01)。热应激对后续周期的第一次卵泡波也有影响:排卵后血浆FSH浓度升高(P < 0.01),但中等卵泡发育较少,第一次卵泡波下降速度慢于冷却奶牛(分别为0.40和0.71个/ d)。本研究显示热应激对卵泡动力学的即时和延迟影响,这与血浆中高FSH和低抑制素浓度有关。这些变化可能具有生理意义,可能与夏季和秋季牛的低生育率有关。
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引用次数: 0
A staging scheme for assessing development in vitro of organogenesis stage embryos of the stripe-faced dunnart, Sminthopsis macroura (Marsupialia: dasyuridae). 一种评估条纹面蕨体外器官发生阶段胚胎发育的分期方案(有袋目:有袋目)。
Pub Date : 2000-09-01 DOI: 10.1530/jrf.0.1200099
Y P Cruz, D Hickford, L Selwood

The inaccessibility of mammalian organogenesis stage embryos has precluded their widespread use in embryological and teratological studies. As organogenesis occurs during the last 1.5 days of the 10. 7 days of gestation in the stripe-faced dunnart (Sminthopsis macroura), the aim of the present study was to investigate whether day 9 and day 10 embryos and fetuses could be grown to term in vitro. High glucose Dulbecco's modified Eagle's medium with 10% fetal calf serum (FCS) supported embryonic growth for various periods of time, some to within 5 h of the predicted time of parturition. A roller culture system maintained at 35 degrees C was used to incubate organogenesis stage embryos (n = 43). Nine unincubated (control) embryos were either fixed for microscopic analysis or frozen for microprotein determination. The results of the present study indicate that with some optimization of the culture conditions (increasing oxygen in the gas phase in the culture tubes, replacing FCS with rat serum), it might be possible for organogenesis stage S. macroura embryos to be grown to term. A scoring scheme for assessing morphological development was devised for use as a standard in staging organogenesis stage embryos. This scheme reflects the highly compressed schedule of developmental events that occurs mainly during day 9 of gestation in S. macroura embryos. In comparison, during embryogenesis in Didelphis virginiana these developmental events occur from day 8 to day 10.5 of gestation, and birth occurs on day 13.

哺乳动物器官发生阶段胚胎的不可获得性阻碍了它们在胚胎学和致畸学研究中的广泛应用。器官发生在10天的最后1.5天。本研究的目的是研究第9天和第10天的胚胎和胎儿能否在体外生长至足月。含有10%胎牛血清(FCS)的高糖Dulbecco改良Eagle培养基在不同时期支持胚胎生长,有些在预测分娩时间的5小时内。采用维持在35℃的滚轮培养系统培养器官发生期胚胎(n = 43)。9个未孵育的(对照)胚胎被固定用于显微镜分析或冷冻用于微量蛋白测定。本研究结果表明,通过优化培养条件(增加培养管中气相的氧气,用大鼠血清代替FCS),有可能使器官发生期大鼠棘球蚴胚胎发育至足月。设计了一种评估形态发育的评分方案,作为器官发生阶段胚胎分期的标准。这一模式反映了大花蛇胚胎主要发生在妊娠第9天的高度压缩的发育事件时间表。相比之下,在弗吉尼亚州Didelphis的胚胎发生过程中,这些发育事件发生在妊娠第8天至10.5天,出生发生在第13天。
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引用次数: 11
Immunohistochemical localization of prostaglandin G/H synthase 1 and 2 in sheep placenta after glucocorticoid-induced and spontaneous labour. 糖皮质激素诱导和自然分娩后绵羊胎盘中前列腺素G/H合成酶1和2的免疫组织化学定位。
Pub Date : 2000-09-01 DOI: 10.1530/REPROD/120.1.33
W. McLaren, I. R. Young, G. Rice
Enhanced prostaglandin production and release by the placenta is an essential element in the normal transition to labour in many animal species. In sheep, expression of prostaglandin G/H synthase (PGHS) is the central enzyme regulating this process. In this study immunohistochemistry was used to examine the distribution of cells expressing PGHS-1 and PGHS-2 in ovine placenta in association with spontaneous parturition (n = 6) and glucocorticoid-induced labour (n = 5). Labour was induced in ewes after the intrafetal injection of betamethasone on day 131 of gestation. Animals administered an intrafetal injection of isotonic saline (n = 5) acted as non-labour controls. In placentomes collected from all groups, immunoreactive PGHS-1 was present in the mononuclear trophoblast cells of the fetal placenta. Cells in the maternal mesenchyme and epithelial syncytium were weakly immunopositive for this enzyme. PGHS-1 immunoreactivity was also demonstrated in the endothelial cells of the chorionic vessels. The PGHS-2 isozyme was localized exclusively to the trophoblast epithelial cells. Immunoreactive PGHS-2 was not detectable in the maternal epithelial syncytium or the stroma of the cotyledons. The binucleate cells of the fetal placenta were consistently immunonegative for both PGHS isozymes. These results indicate that the cellular localization of PGHS-1 and PGHS-2 in ovine placenta does not change during the last 15 days of pregnancy. Co-localization of these isozymes indicates that the source of arachidonic acid and the site of prostanoid formation are the same. Quantitation of the percentage area of positive staining for PGHS-1 and PGHS-2 using image analysis software demonstrated a significant increase in PGHS-2 in the fetal trophoblast after glucocorticoid-induced labour and spontaneous parturition. This finding indicates that increased formation of the PGHS-2 isozyme is responsible for the large increase in prostaglandin production by the ovine placenta at term labour.
在许多动物物种中,胎盘增强前列腺素的产生和释放是正常向分娩过渡的重要因素。在绵羊中,前列腺素G/H合成酶(PGHS)的表达是调节这一过程的中心酶。本研究采用免疫组织化学方法检测了PGHS-1和PGHS-2在绵羊胎盘中的分布与自然分娩(n = 6)和糖皮质激素诱导分娩(n = 5)的关系。母羊在妊娠第131天胎腔内注射倍他米松诱导分娩。给予胎儿内注射等渗盐水(n = 5)的动物作为非分娩对照组。在所有组的胎盘中,免疫反应性PGHS-1存在于胎儿胎盘的单核滋养细胞中。母体间质和上皮合胞体细胞对该酶呈弱免疫阳性。PGHS-1在绒毛膜血管内皮细胞中也表现出免疫反应性。PGHS-2同工酶仅局限于滋养层上皮细胞。母体上皮合胞体和子叶间质中未检测到免疫反应性PGHS-2。胎儿胎盘的双核细胞对两种PGHS同工酶均呈免疫阴性。这些结果表明,PGHS-1和PGHS-2在绵羊胎盘中的细胞定位在妊娠后期15天内没有变化。这些同工酶的共定位表明花生四烯酸的来源和前列腺素的形成部位是相同的。利用图像分析软件对PGHS-1和PGHS-2的阳性染色面积百分比进行定量分析,结果显示糖皮质激素引产和自然分娩后,胎儿滋养细胞中PGHS-2显著增加。这一发现表明,PGHS-2同工酶的形成增加是足月分娩时羊胎盘前列腺素产量大幅增加的原因。
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引用次数: 9
Effects of recent sexual experience and melatonin treatment of rams on plasma testosterone concentration, sexual behaviour and ability to induce ovulation in seasonally anoestrous ewes. 公羊近期性经验和褪黑素处理对季节性不发情母羊血浆睾酮浓度、性行为和促排卵能力的影响
Pub Date : 2000-09-01 DOI: 10.1530/REPROD/120.1.169
H. Rosa, D. Juniper, M. Bryant
The aim of this study was to determine whether advancing the seasonal changes associated with rams by treatment with exogenous melatonin and allowing the rams previous sexual experience would increase the proportion of anoestrous ewes ovulating in early July. North Country Mule ewes (n = 225) were grouped by live body weight and body condition score and allocated randomly to the following treatments: (i) isolated from rams (control; n = 25); (ii) introduced to rams (treatment 2); (iii) introduced to rams that had mated with ewes during the previous 2 days (treatment 3); (iv) introduced to rams implanted with melatonin (treatment 4); and (v) introduced to rams that were implanted with melatonin and had mated with ewes during the previous 2 days (treatment 5). Treatments 2-5 were replicated (2 x 25 ewes) and two rams were introduced to each replicate group. Introductions began on 4 July and were completed by 11 July. The rams were withdrawn from the ewes after 8 days. Melatonin was administered as a subcutaneous implant (Regulin((R))) on 22 May and again on 20 June. Blood samples were taken from all rams to determine plasma melatonin and testosterone concentrations (19 samples in 6 h). The behaviour of the sheep was videotaped continuously during the first 3 h after the ram was introduced. Ovulation was detected by an increase in plasma progesterone concentrations from < 0.5 ng ml(-1) to > 0.5 ng ml(-1). Mean +/- SE plasma melatonin concentrations were 649.7 +/- 281.4 and 18.3 +/- 2.4 pg ml(-1) in rams with and without melatonin implants, respectively (P < 0.001). Melatonin implants also increased plasma testosterone concentrations from 4.30 +/- 1.88 to 10.10 +/- 1.10 ng ml(-1) (P < 0.01), the libido of the rams and the proportion of ewes that ovulated in response to the rams (43 and 56% (treatments 4 and 5) versus 24% (treatments 2 and 3)). In conclusion, implanting rams with melatonin before introducing them to seasonally anoestrous ewes increases the proportion of ewes that ovulate in response to introduction of a ram, but previous sexual experience of rams appears to have little or no effect.
本研究的目的是确定通过外源性褪黑素治疗和允许公羊之前的性经验来推进与公羊相关的季节变化是否会增加7月初排卵的无情母羊的比例。按活体重和体况评分对225只北方母羊进行分组,随机分为以下处理:(1)与公羊隔离(对照;N = 25);(ii)引入RAMS(治疗2);(iii)引入前2天内与母羊交配的公羊(处理3);(iv)对植入褪黑素的RAMS进行治疗(治疗4);(v)引入植入褪黑激素并在前2天内与母羊交配的公羊(处理5)。处理2-5重复(2 × 25只母羊),每个重复组引入2只公羊。介绍工作于7月4日开始,7月11日完成。8天后将公羊从母羊中取出。5月22日和6月20日分别皮下植入褪黑素(Regulin(R))。所有公羊均采集血样,测定血浆褪黑素和睾酮浓度(6小时内采集19个样本)。在公羊引入后的前3小时内,对羊的行为进行连续录像。通过血浆孕酮浓度从< 0.5 ng ml(-1)增加到bb0 0.5 ng ml(-1)来检测排卵。植入和未植入褪黑素的公羊血浆中+/- SE褪黑素的平均浓度分别为649.7 +/- 281.4和18.3 +/- 2.4 pg ml(-1) (P < 0.001)。褪黑激素植入也使血浆睾酮浓度从4.30 +/- 1.88提高到10.10 +/- 1.10 ng ml(-1) (P < 0.01),提高了公羊的性欲和对公羊有反应的母羊排卵比例(处理4和5分别为43%和56%,处理2和3分别为24%)。综上所述,在将公羊引入季节性发情母羊之前,给公羊注射褪黑激素可以增加母羊在引入公羊后排卵的比例,但公羊之前的性经验似乎没有什么影响。
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引用次数: 57
Effects of angiotensin II on the acrosome reaction in equine spermatozoa. 血管紧张素II对马精子顶体反应的影响。
Pub Date : 2000-09-01 DOI: 10.1530/REPROD/120.1.135
K. Sabeur, A. Vo, B. Ball
Angiotensin II is a hormone with a wide array of physiological effects that exerts its effect via interaction with two major subtypes of receptor. The results of this study show that angiotensin II (from 1 to 100 nmol l(-1)) initiates acrosomal exocytosis in equine spermatozoa that have undergone capacitation in vitro in a TALP-TEST (Tyrode's albumin lactate pyruvate; 188.7 mmol TES l(-1), 84.8 mmol Tris l(-1)) buffer with cAMP. The acrosome reaction and sperm viability were assessed with fluorescein isothiocyanate-Pisum sativum agglutinin (FITC-PSA) and Hoechst 33258, respectively. The initiation of the acrosome reaction by angiotensin II was strongly inhibited by losartan, a specific angiotensin II type 1 receptor antagonist. Although angiotensin II as well as progesterone both initiated the acrosome reaction in equine spermatozoa, there was no synergistic effect when both agonists were added simultaneously. Initiation of acrosomal exocytosis by angiotensin II was accompanied by a rapid and transient calcium influx that was assessed in capacitated spermatozoa loaded with Fura-2AM. In addition, the angiotensin II-mediated calcium influx was inhibited when spermatozoa were preincubated with losartan. Western blotting with an antibody against angiotensin II type 1 receptor detected a major sperm protein of 60 kDa. Indirect immunofluorescence of non-capacitated spermatozoa with the angiotensin II type 1 receptor antibody revealed labelling in the midpiece and tail. In capacitated spermatozoa, the angiotensin II type 1 receptor was localized mainly over the anterior region of the sperm head, the equatorial segment and occasionally on the postacrosomal region in addition to the sperm tail. In conclusion, this study demonstrated the ability of angiotensin II to stimulate the acrosome reaction in capacitated equine spermatozoa. This effect is mediated via the angiotensin II type 1 receptor and is accompanied by an increase in intracellular calcium.
血管紧张素II是一种具有广泛生理作用的激素,通过与两种主要受体亚型的相互作用来发挥其作用。本研究结果表明,血管紧张素II(从1到100 nmol l(-1))在体外TALP-TEST (Tyrode's白蛋白乳酸丙酮酸;188.7 mmol TES l(-1), 84.8 mmol Tris l(-1)) cAMP缓冲液。用异硫氰酸荧光素凝集素(FITC-PSA)和Hoechst 33258分别评价顶体反应和精子活力。血管紧张素II的顶体反应被氯沙坦(一种特异性血管紧张素II 1型受体拮抗剂)强烈抑制。虽然血管紧张素II和黄体酮都能引起马精子顶体反应,但同时加入这两种激动剂时没有协同作用。在装载Fura-2AM的有能力精子中,血管紧张素II引发顶体胞外分泌伴随着快速和短暂的钙内流。此外,当精子与氯沙坦预孵育时,血管紧张素ii介导的钙内流被抑制。抗血管紧张素II型1受体抗体的Western blotting检测到60 kDa的主要精子蛋白。血管紧张素II型1受体抗体的间接免疫荧光显示在中间和尾部有标记。在能态精子中,血管紧张素II型1受体主要位于精子头部前部和赤道段,除了精子尾部外,偶尔也位于顶体后区域。总之,本研究证明了血管紧张素II能够刺激马精子顶体反应。这种作用是通过血管紧张素II型1受体介导的,并伴随着细胞内钙的增加。
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引用次数: 28
Effects of bromocriptine administration during the follicular phase of the oestrous cycle on prolactin and gonadotrophin secretion and follicular dynamics in merino monovular ewes. 发情周期卵泡期给药溴隐亭对美利奴单卵母羊泌乳素和促性腺激素分泌及卵泡动力学的影响。
R A Picazo, A González De Bulnes, A Gómez Brunet, A del Campo, B Granados, J Tresguerres, A López Sebastián

Two experiments using Spanish Merino ewes were conducted to investigate whether the secretion of prolactin during the follicular phase of the sheep oestrous cycle was involved in the patterns of growth and regression of follicle populations. In both experiments, oestrus was synchronized with two cloprostenol injections which were administered 10 days apart. Concurrent with the second injection (time 0), ewes (n = 6 per group) received one of the following treatments every 12 h from time 0 to 72 h: group 1: vehicle injection (control); group 2: 0.6 mg bromocriptine (0.03 mg per kg per day); and group 3: 1.2 mg bromocriptine (0.06 mg per kg per day). In Expt 1, blood samples were collected every 3 h from 0 to 72 h, and also every 20 min from 38 to 54 h to measure prolactin, LH and FSH concentrations. In Expt 2, transrectal ultrasonography was carried out every 12 h from time 0 until oestrus, and blood samples were collected every 4 h to measure prolactin, LH and FSH concentrations. Ovulation rates were determined by laparoscopy on day 4 after oestrus. Bromocriptine markedly decreased prolactin secretion, but did not affect FSH concentrations, the mean time of the LH preovulatory surge or LH concentrations in the preovulatory surge. Both doses of bromocriptine caused a similar decrease in LH pulse frequency before the preovulatory surge. The highest bromocriptine dose led to a reduction (P < 0.01) in the number of 2-3 mm follicles detected in the ovaries at each time point. However, bromocriptine did not modify the total number or the number of newly detected 4-5 mm follicles at each time point, the number of follicles > 5 mm or the ovulation rate. In conclusion, the effects of bromocriptine on gonadotrophin and prolactin secretion and on the follicular dynamics during the follicular phase of the sheep oestrous cycle indicate that prolactin may influence the viability of gonadotrophin-responsive follicles shortly after luteolysis.

以西班牙美利奴母羊为试验对象,研究了绵羊发情周期卵泡期泌乳素的分泌是否与卵泡种群的生长和退化有关。在这两个实验中,两次氯前列醇注射间隔10天同步发情。在第二次注射的同时(时间0),母羊(n = 6 /组)在时间0 ~ 72 h每12 h接受以下处理中的一种:1组:车辆注射(对照);第二组:溴隐亭0.6 mg (0.03 mg / kg / d);第三组:溴隐亭1.2 mg (0.06 mg / kg / d)。在实验1中,从0到72 h每3小时采集一次血样,从38到54 h每20分钟采集一次血样,测量催乳素、LH和FSH浓度。在实验2中,从时间0到发情,每隔12 h进行经直肠超声检查,每隔4 h采集血液,测量催乳素、LH和FSH浓度。在发情后第4天腹腔镜检查排卵率。溴隐亭可显著降低催乳素分泌,但对FSH浓度、促黄体生成素平均时间及促黄体生成素浓度无明显影响。两种剂量的溴隐亭在排卵前的高潮前引起了相似的LH脉冲频率的降低。溴隐亭最高剂量组各时间点卵巢中2 ~ 3 mm卵泡数量减少(P < 0.01)。而溴隐亭对各时间点新检出4 ~ 5 mm卵泡总数、新检出4 ~ 5 mm卵泡数、> 5 mm卵泡数及排卵率均无影响。综上所述,溴隐亭对绵羊发情周期卵泡期促性腺激素和催乳素分泌以及卵泡动力学的影响表明,催乳素可能影响促性腺激素反应性卵泡在卵泡溶解后不久的活力。
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引用次数: 0
Loss of myometrial oxytocin receptors during oxytocin-induced and oxytocin-augmented labour. 催产素诱导和增强分娩时子宫肌层催产素受体的丧失。
Pub Date : 2000-09-01 DOI: 10.1530/jrf.0.1200091
S Phaneuf, B Rodríguez Liñares, R L TambyRaja, I Z MacKenzie, A López Bernal

Oxytocin is used widely for the induction and augmentation of labour, but there is little information about the dynamics of oxytocin receptors in human myometrium during parturition, and the possible effect of oxytocin infusion. This information is important because G protein-coupled receptors, such as the oxytocin receptor, undergo desensitization after prolonged or repeated stimulation. The concentration of myometrial oxytocin receptors and the steady state of its mRNA were measured in patients undergoing Caesarean sections before or during spontaneous or induced labour. The concentration of receptors before labour was 477 (175-641) fmol mg(-1) protein (median, quartile range), and decreased to 140 (72-206; P < 0.05) and 118 (69-75; P < 0.01) fmol mg(-1) protein during prolonged oxytocin-augmented and oxytocin-induced labour, respectively. The corresponding oxytocin receptor mRNA concentrations decreased by 60- and 300-fold, respectively. The decrease in receptor binding and mRNA in women receiving oxytocin infusion indicates that homologous receptor desensitization occurs in vivo.

催产素被广泛用于引产和助产,但关于分娩时子宫肌层中催产素受体的动态变化以及催产素输注可能产生的影响的信息很少。这一信息很重要,因为G蛋白偶联受体,如催产素受体,在长时间或反复刺激后会发生脱敏。对剖宫产患者在自然分娩或引产前或产程中子宫肌层催产素受体的浓度及其mRNA的稳定状态进行了测量。分娩前受体浓度为477 (175-641)fmol mg(-1)蛋白(中位数,四分位数范围),降至140 (72-206);P < 0.05)和118 (69 ~ 75;P < 0.01) fmol mg(-1)蛋白在延长催产素增强产程和催产素诱导产程中的表达。相应的催产素受体mRNA浓度分别下降了60倍和300倍。在接受催产素输注的女性中,受体结合和mRNA的减少表明体内发生了同源受体脱敏。
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引用次数: 110
Successful capacitation and homologous fertilization in vitro in Calomys musculinus and Calomys laucha (Rodentia - sigmodontinae). 肌鼠和牙鼠成功获能和体外同种受精的研究。
Pub Date : 2000-09-01 DOI: 10.1530/jrf.0.1200041
A Lasserre, E Cebral, A D Vitullo

Small South American rodents of the genus Calomys have been used extensively for virology and ecological research. Previous studies have demonstrated that Calomys musculinus and Calomys laucha have a relatively short oestrous cycle and that superovulation and parthenogenetic activation can be induced. The purpose of this study was to determine the requirements for in vitro manipulation of the male gamete and in vitro fertilization. Two culture media and different concentrations of spermatozoa were tested for their ability to support sperm motility, hyperactivation and the acrosome reaction. The ability of capacitated Calomys spermatozoa to penetrate zona-free hamster eggs was also evaluated. In vitro fertilization was assessed by examining attachment and binding to the zona pellucida, second polar body extrusion, pronucleus formation and the fertilizing sperm tail. The results of the study showed that: (i) Tyrode's albumin lactate pyruvate (TALP) medium was more effective than T6 medium for maintaining sperm motility in vitro; (ii) hyperactivation was achieved with TALP but not with T6; (iii) the acrosome reaction was easily distinguished by light microscopy and depends on time and sperm concentration; (iv) capacitated spermatozoa are able to penetrate zona-free hamster eggs; and (v) superovulated oocytes can be fertilized in vitro. This is the first report of capacitation and in vitro fertilization for Calomys sp. These results provide opportunities to use C. musculinus and C. laucha as new laboratory animals for research into reproductive biology.

南美小啮齿动物已被广泛用于病毒学和生态学研究。以往的研究表明,卡罗米斯肌肉和卡罗米斯laucha有一个相对较短的发情周期,可以诱导超排卵和孤雌生殖激活。本研究的目的是确定雄性配子的体外操作和体外受精的要求。试验了两种培养基和不同浓度精子对精子活力、超激活和顶体反应的支持能力。我们还评价了能化的Calomys精子穿透无带仓鼠卵的能力。通过观察与透明带的附着和结合、第二极体的挤压、原核的形成和受精精子尾巴来评估体外受精。结果表明:(1)Tyrode乳酸丙酮酸白蛋白(TALP)培养基比T6培养基更能有效地维持精子的体外活力;(ii) TALP实现了过度激活,而T6没有;(iii)顶体反应很容易在光镜下区分,并取决于时间和精子浓度;(iv)有能力的精子能够穿透无带的仓鼠卵;(5)超排卵的卵母细胞可以在体外受精。本研究为Calomys sp.获能和体外受精的首次报道,为利用C. musculinus和C. laucha作为新的实验动物进行生殖生物学研究提供了机会。
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引用次数: 8
Potential role of alphav and beta1 integrins as oocyte adhesion molecules during fertilization in pigs. 整合素和整合素在猪受精过程中作为卵母细胞粘附分子的潜在作用。
Pub Date : 2000-09-01 DOI: 10.1530/REPROD/120.1.65
J. Linfor, T. Berger
Integrin molecules are cell adhesion molecules that are thought to be involved in sperm-oocyte interaction in rodents and humans. The objective of this study was to evaluate whether integrin molecules were present on the surface of pig oocytes, consistent with involvement in sperm-oocyte interaction in this species. Immunocytochemistry and confocal microscopy were used to evaluate the presence of beta1, and alpha1, alpha2, alpha3, alpha4, alpha5, alpha6 and alphav integrin subunits on the plasma membrane of pig oocytes. The beta1 and alphav integrin subunits were present consistently at the surface of pig oocytes; however, the remaining alpha integrin subunits evaluated were not routinely detected. The antibodies to the beta1 and alphav integrin subunits recognized appropriately sized protein bands on western blots of partially purified oocyte plasma membrane. These two antibodies also recognized oocyte plasma membrane protein isolated from a sperm plasma membrane affinity column. Sperm plasma membrane proteins of 137 and 93 kDa appeared to be the ligands for the beta1 integrin subunit as revealed by a western sandwich blot. Antibody to an extracellular domain of the beta1 integrin subunit reduced pig sperm-oocyte binding (P < 0.05), also indicating an assisting role for a beta1 oocyte integrin subunit in sperm-oocyte interaction in pigs. These results are consistent with an alphavbeta1 pig oocyte integrin interacting with a ligand on the sperm plasma membrane during fertilization.
整合素分子是一种细胞粘附分子,被认为与啮齿类动物和人类的精子-卵细胞相互作用有关。本研究的目的是评估整合素分子是否存在于猪卵母细胞表面,与该物种参与精子-卵母细胞相互作用一致。采用免疫细胞化学和共聚焦显微镜检测猪卵母细胞质膜上β 1、α 1、α 2、α 3、α 4、α 5、α 6和α v整合素亚基的存在。猪卵母细胞表面一致存在β 1和α v整合素亚基;然而,剩余的α整合素亚基没有被常规检测到。针对β 1和α v整合素亚基的抗体在部分纯化的卵母细胞质膜上识别适当大小的蛋白带。这两种抗体也能识别从精子细胞膜亲和柱中分离的卵母细胞细胞膜蛋白。western sandwich blot显示,137和93 kDa的精子质膜蛋白可能是β 1整合素亚基的配体。β a1整合素亚基胞外结构域的抗体降低了猪精卵细胞结合(P < 0.05),也表明β a1整合素亚基在猪精卵相互作用中起辅助作用。这些结果与猪卵母细胞整合素在受精过程中与精子质膜上的配体相互作用是一致的。
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引用次数: 45
期刊
Journal of reproduction and fertility
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