外激素促绵羊同期发情作用的研究

本文研究了外激素对新疆细毛羊性周期的影响,探讨应用外激素促进绵羊同期发情的可能性。 将391头母羊分为两群放牧,一群定期给以外激素喷雾,为试验组,另一群作为对照,定期喷以清水。结果两群羊的性周期显现不同的变化。试验组母羊平均发情7.0±0.85次;对照组为4.3±0.63次,p<0.05,相差显著。其中成畜对外激素的刺激作用反应较强,未成年羊的感受能力较弱)。 在外激素的作用下,发情羊显然增多,于7—10天内,大部分母畜发情。而对照组中发情羊数明显低予试验组,而且其发情母畜高峰期的出现,亦迟于试验组。 配种结果得出,经外激素处理组,羊配种时间较为集中,在配种开始的一周内,多数母畜发情并施行人工授精。而对照组的母羊,则表现陆续发情、没有集中的趋势。根据配种开始14日内的资料统计,试验组有87.9％的母羊发情、配种,对照组则只有37.5％母羊发情、输精。在其它未给外激素刺激的羊群中,要达到与试验组相似的配种百分数,配种时间平均需要35.8日,较给予外激素刺激的试验组延长了21.8日之多。说明外激素能产生促进绵羊同期发情、缩短配种时间的效应。 统计试验群与对照群40头母羊的产羔情况,试验组计有80％母羊怀孕并产羔,对照组母羊则只有30％母畜妊娠并产仔。 据试验羊的性周期、配种、产羔的结果,都说明     

不同品种绵羊发情周期LEPRb mRNA表达分析

为了探讨LEPR在绵羊季节性发情中的表达特点,以季节性发情的中国美利奴和常年发情的多浪羊为研究对象,利用实时荧光定量PCR技术检测繁殖季节不同发情阶段LEPRb m RNA在下丘脑、卵巢和子宫组织中的表达变化。结果显示,在发情间期、前期和发情期中国美利奴卵巢和子宫组织LEPRb的相对表达水平都低于下丘脑,但在发情后期卵巢和子宫LEPRb的相对表达水平显著高于下丘脑。不同发情时期,两品种绵羊同一组织的LEPRb的表达变化幅度有所差异,但趋势相似。且中国美利奴各组织LEPRb的表达水平都高于常年发情的多浪羊。LEPRb的表达变化与绵羊发情存在相关关系。     

绵羊的卷唇行为及其生物学意义

本文研究了绵羊对不同品种公母羊的尿液、母羊粪便、阴道分泌物、身体的气味等刺激物诱导卷唇行为的发生,持续约60-120秒。实验得出:母羊尿液是诱导公羊释放卷唇行为最强的刺激物,尤以发情母羊尿液更甚,公羊、幼羊以及其它的尿液诱导卷唇行为发生的频次低。卷唇行为发生的频次有季节变化,10月最高。卷唇行为与犁鼻器的功能紧密联系,是绵羊繁殖行为的一个构成部分,通常只有公羊发生卷唇行为,但个别母羊亦有表现。     

绵羊发情周期不同组织Cry1mRNA 转录水平相对定量研究

研究表明,时钟基因Cry1在哺乳动物季节性繁殖内分泌过程中可能发挥了重要作用。文章以多浪羊(非季节性繁殖)和中国美利奴羊(季节性繁殖)为研究对象,采用实时荧光定量PCR技术对下丘脑-垂体-性腺轴主要组织Cry1在发情周期不同阶段的表达变化情况进行了跟踪检测。结果表明:绵羊Cry1在所检测组织中均有表达,其中松果体和甲状腺Cry1的表达水平较高;不同绵羊品种Cry1在发情周期不同阶段的组织表达谱基本一致,除下丘脑外,卵巢、子宫、松果体、垂体和甲状腺中Cry1的表达水平均是在发情前期达到峰值;卵巢、子宫、垂体和松果体Cry1在发情前期和发情期的表达变化幅度存在显著的品种间差异。研究结果表明:Cry1可能具有启动发情和季节性繁殖的重要作用。     

绵羊CCNG1基因克隆及表达分析

细胞周期蛋白cyclin G1(CCNG1)是一个重要的细胞周期调控因子,参与哺乳动物颗粒细胞增殖、卵母细胞成熟等繁殖生物学过程,但其在绵羊中鲜有报道。为了研究CCNG1基因对绵羊发情调控以及季节性繁殖的影响,文章对绵羊CCNG1基因进行了克隆和组织表达谱分析,利用Real-time PCR对该基因在多浪羊(常年发情)与中国美利奴羊(季节性繁殖)发情周期不同阶段性腺轴组织的表达变化进行了实时检测。获得了绵羊CCNG1基因部分cDNA序列,其中编码区全长885 bp,编码294个氨基酸。CCNG1蛋白结构经预测存在多个磷酸化位点和蛋白激酶C磷酸化位点。CCNG1基因在所检测绵羊各组织中均有表达,但在卵巢与肾脏中为高丰度表达;CCNG1在不同绵羊品种发情周期不同阶段性腺轴组织的表达变化规律基本一致,卵巢、子宫、松果体、垂体均是在发情期达到峰值。但是在发情期和发情后期卵巢CCNG1的表达量存在显著的品种间差异(P<0.01)。研究结果表明,CCNG1可能通过参与卵泡的生长发育继而达到对绵羊发情和季节性繁殖的调控。     

褪黑素与绵羊的季节性生殖

绵羊是一种短光照型生殖动物,具有明显的生殖季节性。人们普遍认为,绵羊生殖周期与环境光周期的同步变化是通过褪黑素作用于下丘脑－垂体－性腺轴系统来实现的。近年来的不少研究结果表明,绵羊对光周期的感受性还受褪黑素受体的调控。现从生态学（光周期的生殖效应）、神经内分泌学和分子生物学（褪黑素受体）等领域的研究进展出发,对褪黑素调控绵羊自然季节性生殖的作用及机制进行系统综述。     

Leptin基因多态性与绵羊季节性发情的相关分析

根据leptin基因在GenBank中的已知序列设计两对引物,采用PCR-SSCP技术在常年发情的湖羊和季节性发情的阿勒泰羊群体中进行单核苷酸多态性(SNPs)检测,对筛查到的SNP位点进行基因型与绵羊季节性发情的关联分析。结果表明,与湖羊相比,阿勒泰羊leptin基因第1内含子上有3个连续碱基TTG的插入和C/T碱基突变;第3外显子3上发生G/T碱基突变,编码氨基酸由缬氨酸变成亮氨酸。Leptin外显子2扩增片段上检测到AA、AB、BB三种基因型,BB基因型在阿勒泰羊群体中属于优势基因型;对两个群体进行基因型频率独立性χ2检验,差异极显著(P0.001),说明BB基因型是影响季节性发情的有利基因型。研究结果提示,绵羊品种中Leptin基因序列的差异性可能是造成绵羊季节性发情的原因之一,可作为常年发情绵羊品种选育的辅助标记。     

绵羊胚胎附植分子调控研究进展

胚胎附植是哺乳动物复杂的生殖生理过程,是妊娠建立的标志和首要环节。早期胚胎发育、母体妊娠识别、胚胎附植和妊娠维持都严格依赖于孕体和中间的信号联系。大量研究证明,在绵羊胚胎附植过程中,来源于胚胎、母体子宫及宫外组织的多种生殖激素、黏附分子、细胞外基质、细胞活素物质和生长因子通过极其精密的协调共同参与和维持了孕体的发育、子宫内膜的重塑、分泌功能和子宫生长。综述了近年来绵羊胚胎附植的相关分子调控机制的最新研究进展,对胚胎附植分子调控信号的掌握将有助于诊断和确定那些引起妊娠失败的原因,为提高家畜和人类妊娠率提供参考。     

动物季节性繁殖分子调控机理研究进展

动物季节性发情繁殖涉及下丘脑-垂体-性腺轴系统复杂的神经内分泌过程,并受光照周期等环境因素的影响。褪黑激素则作为光周期信号分子调控动物季节性繁殖活动。近年来研究发现,对GnRH分泌有重要影响的Kiss1/GPR54系统既受褪黑激素的调控又受到性腺类固醇激素反馈调节,Kiss1/GPR54系统很可能是调控动物季节性繁殖的关键因子;同时动物季节性繁殖很可能还存在一条涉及TSH-DIO2/DIO3系统的逆向调控通路,该系统同样显著影响GnRH合成释放并受褪黑激素调控。文章就褪黑激素中心信号,特别是Kiss1/GPR54和TSH-DIO2/DIO3系统对繁殖季节性调控的最新研究进展进行综述。     

绵羊犁鼻器在繁殖中的作用

许多动物的尿液中含有重要的外激素信息,其生理意义多种多样。雌性尿液中多有表示本身性别及性状况的化学信息存在,行为学调查表明,犁鼻器的主要功能在于识别尿液气味。公羊正是依据这些化学信息识别并选择配偶的。通常公羊嗅到母羊,尤其是发情母羊尿液时,表现伸颈、抬头、卷缩上唇的特有行为型式,称之为卷唇行为或性嗅反射。这个行为是与犁鼻器的功能相联系的。它的作用在于关闭外鼻孔,堵闭会厌,使吸进的空气进入犁鼻器内,犁鼻器两侧的肌肉运动,腔内静脉窦的膨胀和收缩,又促进空气的流入和排出,如此使携带化学信息的载体不断进入犁鼻器官,成为绵羊感受化学信息的方式之一。 犁鼻器的作用可为公羊及早地、准确地选择发情的配偶提供信息;同时又能唤起公羊本身的性行为,刺激雌、雄发情及性活动的同步,对保证繁殖的成功具有一定的意义。说明绵羊犁鼻器作为化学感受器在化学通讯中特别是对繁殖行为具有重要作用。     

Effect of breeding season on fertility of sheep following estrus synchronization and fixed-time artificial insemination under field conditions in semi-arid tropical region

Accelerated lambing system is heavily reliant on reproductive technologies meant to enable off the season breeding in sheep. Therefore, the present study was programmed to assess the effect of breeding season (BS) on fertility of sheep following estrus synchronization (ES) and fixed time artificial insemination (FTAI). A total of 248 and 365 ewes were synchronized and inseminated during the BS (Febuary–March and July–September) and non-breeding season (NBS) respectively, during 2012–14. Synchronization of estrus was done by AVIKESIL-S, intra-vaginal progesterone sponges kept in situ in vagina for 12 days. 200 IU eCG was administered intramuscularly on 12th day after sponge withdrawal. FTAI was performed twice in ewes exhibiting signs of estrus at 48 and 56h after sponge removal, using liquid chilled semen of Patanwadi/Malpura rams containing 100 million sperm. Breeding season had no significant (p<0.05) effect on estrus synchronization. The estrous responses during the BS and NBS were 84.68% and 83.29% respectively. The lambing percentage during BS was 66.67%, which is significantly (p<0.05) higher than the lambing percentage of NBS (57.57%). Although, the lambing percentage in NBS maneuvered ewes were lower than the BS ewes but still the technology can be used to offset the effect of anestrus and to augment production in sheep.     

Effect of season, month of parturition and lactation on estrus behavior and ovarian activity in Barki x Rahmani crossbred ewes under subtropical conditions

The correlations between some meteorological parameters and fertility data were evaluated in Barki x Rahmani crossbred ewes using the records of five consecutive years (2003-2007). Additionally, estrus detection and ultrasonic evaluation were applied on eighteen mature dry ewes during breeding and non-breeding seasons. The effect of lactation was evaluated by monitoring estrus behavior in ninety four lactating ewes from 40 to 120 d after parturition. Moreover, ultrasonography was used to identify ovarian activity in six cyclic and six acyclic non-lactating ewes. Results revealed that relative estrus occurrence and fertile mating were positively correlated (P <0.05) with high temperature and long photoperiod (conditions of summer season), and were negatively correlated (P < 0.01) with rainfall (condition of winter season). During breeding season, estrus rate, serum progesterone concentration, and diameter of largest follicle were significantly (P < 0.05) higher than those observed during the non-breeding season. Furthermore, month of parturition had a significant effect (P < 0.05) on estrus rate of lactating ewes where ewes that lambed in August, September, and October recorded higher estrus rate than those lambed in November and December. However, the lactational strength did not exert any deleterious effect on the reproductive performance of lactating ewes.In conclusion, in Egypt under subtropical conditions, Barki x Rahmani crossbred ewes exerted optimum estrus behavior and fertile mating during summer season. The reduction in estrus activity during lactation was due to the seasonal effect rather than lactational stress.     

Effects of follicle stimulating hormone (FSH) on follicular development, oocyte retrieval, and in vitro fertilization (IVF) in ewes during breeding season and seasonal anestrus

Administration of FSH increases the number of developing follicles, and affects oocyte health and cleavage rate. To determine the optimal level of FSH treatment, studies were conducted during the normal breeding season and seasonal anestrus. In Experiment 1, ewes were implanted with SyncroMate-B (SMB; norgestomet) for 14 days during the breeding season. Beginning on day 12 or 13 after SMB implantation, ewes were treated with saline (control; n=10), or treated with FSH for two days (2D; n=9) or three days (3D; n=10). In Experiment 2, conducted during seasonal anestrus, ewes were implanted with SMB for 14 days (n=23) or were not implanted (n=26). The SMB-implanted and nonimplanted ewes were assigned to one of three treatments as in Experiment 1: control (n=13), 2D (n=21) or 3D (n=15). In Experiments 1 and 2, ewes were laparotomized to count the number of follicles < or = 3 mm and > 3 mm and to retrieve oocytes. Healthy oocytes from each treatment were used for IVF. In Experiment 3, ewes (n=6) were implanted twice with SMB for 14 days during seasonal anestrus. Ewes were injected with FSH for 2 days, and the oocytes were collected and fertilized as in Experiments 1 and 2. In Experiment 1, FSH-treatment increased (P < 0.05) the number of follicles > 3 mm, the number of oocytes retrieved from follicles < or = 3 mm and > 3 mm, the proportion of healthy oocytes, and the number of oocytes used for IVF. Oocytes from control and 2D ewes had greater (P < 0.01) cleavage rates than 3D ewes (68% and 71% vs. 42%). In Experiment 2, implanted and nonimplanted ewes had similar (P > 0.05) numbers of follicles, total oocytes, and healthy oocytes; therefore, data were combined. The FSH treatment increased (P < 0.01) the number of follicles > 3 mm, and the number of oocytes recovered from follicles > 3 mm. The recovery rate of oocytes and the percentage of healthy oocytes were similar for control and FSH-treated ewes. The cleavage rate in Experiment 2 ranged from 4 to 16%. In Experiment 3, the cleavage rate for ewes treated twice with SMB was 27% which tended to be greater (P < 0.07) than for the 2D ewes that received one SMB implant in Experiment 2. These data indicate that FSH increased the number of developing follicles and the number of healthy oocytes retrieved from ewes during the breeding season and seasonal anestrus. However, cleavage rates during seasonal anestrus were lower than during the normal breeding season in both FSH-treated and control ewes. Treatment of ewes for 2 days with FSH resulted in a greater cleavage rate than treatment of ewes for 3 days.     

Patterns of estrous cycles, estrous behavior, and circulating prolactin in spring and summer in ewes selected for autumn lambing and exposed to ambient or long-day photoperiods

Estrous behavior in response to ambient and long-day photoperiods was evaluated in ewes developed by 10 years of selection for ability to lamb in autumn. Following October lambing, 67 ewes were moved indoors and exposed to long-day (16L:8D) or ambient photoperiods from February 2 until July 6. Two vasectomized rams with marking harnesses were housed with each group. Estrous behavior was monitored twice weekly. Ewes from the selection line were unresponsive to long days, with no effects on estrous behavior, frequency of ovulation, or circulating prolactin. Adult ewes were anestrus for only 34±3 d, but 2- and 3-years-old ewes were anestrus for 72±7 and 57±10 d, respectively. Frequencies of ovulation based on circulating progesterone concentrations in March, May, and June were 97%, 95% and 52%, respectively, indicating that many ewes that did not exhibit estrus still ovulated. Prolactin concentrations increased from 10 ng/ml in February to 27 ng/ml in March and 173 ng/ml in June but were not affected by light treatment. Ten ewes that failed to exhibit estrus behavior for at most 24 d during the main study were then monitored for 74 additional long days. Nine of 10 ewes did not exhibit estrus for periods similar to 1 or 2 estrus cycles during this period, but eight ewes re-initiated cycles by the end of the study on September 18. Selection for ability to lamb in autumn thus resulted in ewes with an abbreviated seasonal anestrus and reduced sensitivity to long days.     

The effect of season and technique on synchronized and induced estrus and the induction of lambing in the ewe in a commercial setting

At 40 day intervals, groups of 87 to 142 commercial ewes of mixed breeding were subjected to a 5 day breeding period, following one of three estrus inducing or synchronizing treatments (intravaginal sponges containing 60 mg of medroxyprogesterone acetate) for 12 days with (progestin P.M.S.G. group) or without pregnant mares serum gonadotropin (P.M.S.G.) (Progestin group) at sponge removal, or a single injection of prostaglandin F(2alpha) (breeding season and early anestrus only) (PGF(2alpha) group)). Mean pregnancy rates (ewes lambing of those treated) and lambing percentages (lambs born per 100 ewes lambing) were 31 +/- 4%, 169 +/- 6%, 20 +/- 5%, 105 +/- 2% and 18 +/- 8%, 118 +/- 3%, respectively, for the three treatment groups above. Pregnancy rates for the progestin, P.M.S.G. group were 52% in late July, prior to the breeding season, 52% in September and declined to 14% in March (early anestrus). The time from the first of two daily injections of estradiol benzoate to lambing was 34.5 +/- 4.4 hours in ewes 142 to 146 days pregnant.     

Continuous exposure to sexually active rams extends estrous activity in ewes in spring

Sexual activity in sheep is under photoperiodic control, which is the main environmental factor responsible for the seasonality of reproduction. However, other natural environmental factors such as presence of conspecifics can slightly influence the timing of onset and offset of the breeding season. In goats, we have found that the continuous presence of bucks that were rendered sexually active out of season by previous exposure to long days, prevented goats from displaying seasonal anestrus, which suggests that the relative contribution of photoperiod in controlling seasonal anestrus should be reevaluated in small ruminant species. The aim of this study was to assess whether the presence of sexually active rams that had been stimulated by artificial photoperiod and melatonin implants, reduces seasonal anestrus in sheep, by prolonging ovulatory activity in spring. Ewes were assigned to one of two groups (n = 16 and 15), which were housed in two separate barns, and kept in contact, either with the treated or the control rams between March and July. Vasectomized rams were either exposed to 2 months of long days followed by the insertion of three subcutaneous melatonin implants (treated rams, n = 8), or exposed to natural light conditions (control rams, n = 2). Estrus was monitored daily, and weekly plasma progesterone analyses indicated ovulatory activity. Ewes that were exposed to treated rams exhibited a higher proportion of monthly estrus than ewes exposed to the control rams (P < 0.05). Thirteen of 15 ewes (one ewe was not considered because of the presence of persistent CL) exposed to stimulated rams exhibited estrous behavior in a cyclic manner. In contrast, all ewes exposed to control rams stopped estrous activity for a period of time during the study, such that this group exhibited a significantly longer anestrous season (mean ± standard error of the mean 89 ± 9 days) than did the ewes housed with treated rams (26 ± 10 days; P < 0.0001). Among 15 ewes housed with treated rams, 13 of them exhibited continuous ovulatory activity between March and July, whereas one stopped in June and two in July. All ewes kept with control rams stopped ovulating for some time; consequently, those ewes had a longer anovulation period than did the group exposed to treated rams (3 ± 3 vs. 18 ± 7 days, respectively; P < 0.05). In conclusion, continuous exposure to sexually activated rams induced by artificial photoperiod and melatonin implants in spring extended the ovarian activity of ewes in spring, which results in an increase in estrous expression.     

Effect of frontal hypothalamic deafferentation on duration of breeding season and melatonin secretion in the ewe

The objectives of this study were to determine if ewes subjected to frontal hypothalamic deafferentation (FHD) during anestrus remained anestrus or began to have estrous cycles, and if melatonin secretion was disrupted by FHD. Ovary-intact ewes in Group 1 were subjected to either FHD (n = 10) or sham FHD (n = 5) in early July 1983. Estrous cycles were monitored by measuring circulating progesterone concentrations from before FHD until September 1985. Group 2 ewes (n = 4) were subjected to FHD in October 1984. In late April 1985, blood samples were taken from all ewes at 1- to 4-h intervals from 1100 h to 0700 h of the following day to monitor diurnal changes of melatonin. Hypothalami were collected for histological evaluation of lesions. All Group 1 ewes (sham FHD and FHD) initiated normal estrous cycles in August and September 1983, and all ceased cycles by mid-February 1984. All sham FHD and 4 FHD ewes remained anestrus until August or September of 1984 and then resumed normal cycles. In contrast, 5 FHD ewes resumed cycles as early as April 1984 and then cycled intermittently or almost continuously. Two Group 2 ewes cycled continuously after FHD and 2 cycled infrequently. FHD ewes that showed prolonged breeding seasons had cuts that damaged the suprachiasmatic nucleus (SCN) and adjacent structures. Mean nocturnal (2000 h-0500 h) melatonin concentrations did not differ (p greater than 0.05) between sham FHD, FHD "normal season," and FHD "continuous cycle" ewes. In summary, damage to the SCN region by FHD during anestrus had no detectable effect on either onset or cessation of the next breeding season but greatly prolonged subsequent breeding seasons. Thus, the environmental signals that both initiated and terminated the 1983 breeding season apparently had been given before FHD was performed in midsummer. Damage to the SCN region during the breeding season caused some ewes to cycle continuously. The effects of FHD apparently were not due to disruption of melatonin secretion. FHD ewes that showed prolonged breeding seasons had normal seasonal changes of plasma prolactin concentrations. This suggests that different neural structures control seasonal patterns of gonadotropin and prolactin secretion.     

The interaction between photoperiod and nutrition and its effects on seasonal rhythms of reproduction in the ewe

In sheep, the seasonal patterns of reproductive activity are driven primarily by the annual photoperiodic cycle, but can also respond to other environmental factors, such as nutrition, yet little is known about the mechanisms underlying this interaction. This study was designed to define the interaction between photoperiodic and nutritional cues on seasonal patterns of ovarian activity, and to determine if there is a central interaction between these cues. Groups of Ile-de-France ewes were maintained in two nutritional states (restricted and well fed) under a simulated annual photoperiod of 8-16 h of light per day over two breeding seasons. At the end of the first breeding season, half of the animals of each group were ovariectomized (OVX) and fitted subcutaneously with estradiol implants. Low nutritional status shortened the season of ovarian activity, determined from the pattern of progesterone concentrations, by modifying the timing of seasonal transitions between periods of ovarian activity and anestrus. The same results were observed for the seasonal rhythm of neuroendocrine activity, assessed in the OVX ewes, from the pattern of luteinizing hormone concentrations. These results were then confirmed for neuroendocrine activity induced by a photoperiodic treatment. We conclude that nutrition centrally modulates the interpretation of photoperiod to affect seasonal reproductive transitions. The mechanisms of this interaction are discussed in the paper.     

Annual variations in estrual behavior, rate and possibilities for ovulation in Peulh ewes from Niger

Annual variations in estrous and ovulation behavior were studied in 23 bicolored Peulh ewes for a period of 30 months. The percentages of estrus and ovulation shown over a year of observation were 76 and 86%, respectively. The average rate of ovulation was 1.3 +/- 0.04 (X +/- SEM). Reproductive activity was minimal from January to April, as the percentages of estrus and ovulation varied between 43.8 and 61% and between 53.1 and 86%, respectively. This period was marked by the advent of anestrus, interrupted frequently by silent and irregular ovulations. Anestrus lasted an average of 81.9 +/- 9.8 days (X +/- SEM). Reproductive activity was maximal from May to December, as the percentages of estrus and ovulation varied between 77 and 97% and between 87.5 and 100%, respectively. During this period ovarian activity was interrupted by prolonged diestrus which lasted an average of 24.9 +/- 3.3 days (X +/- SEM). The ovulation rate did not differ significantly (P greater than 0.05) between periods of minimal and maximal sexual activity. These results suggest that the reproductive potentialities of Peulh sheep on a good diet are comparable to those of certain ovine breeds in temperate zones. The results also suggest that anestrus in bicolored Peulh sheep is probably a different physiological process than the one observed in some breeds of sheep in which anestrus is marked by total ovarian inactivity.