免疫分析技术在圈养大熊猫繁殖研究中的应用

免疫分析技术是利用抗原与抗体的特异性结合作用来选择性对待测物中的抗体或抗原进行识别和测定的方法,其中以放射免疫技术和酶联免疫技术运用最多.文章简述了使用放射免疫分析技术对圈养大熊猫雌二醇(E)、促黄体生成素(LH)及孕酮(P)水平进行监测,结果表明尿中的黄体生成素的峰值几乎与雌二醇峰值同时出现,之后随着孕酮浓度迅速上升而下降,认为黄体生成素高峰的出现即代表有成熟的卵排出,从而为圈养大熊猫的繁殖研究提供可靠的理论依据.酶联免疫分析技术通过测定雌性大熊猫发情期尿中雌酮-3-葡萄糖醛酸(E1G)的变化以达到预测排卵的目的,所获得的结果与采用放射免疫法相一致.通过以上论述并结合免疫分析技术一些客观存在的问题,对其应用前景进行展望.     

繁殖期雌性大熊猫尿中激素变化的研究

用ELISA法对39只次发情雌性大熊猫尿中雌酮(E1G)和19只次产前大熊猫、6只次假孕大熊猫尿中孕酮(P4)的含量进行测定、统计分析,结果表明:(1)雌性成体大熊猫,发情期多在2—5月份,尿中E1G/Cr由基础值<12ng/mg开始上升,达峰值需8—15d,平均12d。E1G/Cr峰值>60ng/mg,峰值下降12h内进行首次本交、人工授精或两者交替配种,受孕率达100%;峰值下降12—24h内进行首次本交、人工授精或两者交替配种,受孕率达83 3%;峰值下降24h后进行首次配种,受孕率为0。E1G/Cr峰值<60ng/mg的发情雌性大熊猫,无论在峰前、峰值下降时或峰后进行首次配种,受孕率都为0。(2)大熊猫产仔季节一般多在7—9月份,妊娠期多为108—184d,平均150d。有妊娠行为表现时P4/Cr由基础值<12ng/mg开始上升,达峰值时平均需30d,P4/Cr峰值>60ng/mg。大熊猫产前9—18d出现P4/Cr峰值,然后急剧下降,达产仔平均需14d。(3)假孕大熊猫由配种到假孕结束所需时间与真孕无显著性差异。在整个假孕期间,P4/Cr波动性大,峰值不太明显,假孕行为表现持续时间平均为41d。     

姜曲海母猪发情周期性腺激素及其子宫受体含量的变化规律

以长白猪为对照,测定了姜曲海母猪发情周期血浆中雌二醇和孕酮及其子宫受体含量。发现发情期血浆雌二醇的峰值为58.3±3.9pg/ml,与长白猪比差异不显著。间情期孕酮的最高水平和平均水平均比长白猪高(23.25±1.36对14.28±1.42ng/ml和18.56±1.42对11.76±0.57ng/ml,P<0.01)。发情期和间情期的细胞质雌二醇受体(CER)分别为 410±30.3和367±14.2fmol/mgDNA,其kd值分别为4.8±0.65和4.0±0.8nmol;细胞核雌二醇受体(NER)分别为1390±195.3和620±28.9fmol/mgDNA,其kd值为6.6±2.1和4.4±1.3nmol;细胞质孕酮受体(CPR)分别为364±56.0和147±46.0 fmol/mgDNA;细胞核孕酮受体(NPR)分别为337±51.8和184±67.0fmol/mgDNA。发情期和间情期子宫NER含量较长白猪高(P<0.05),说明此时姜曲海母猪子宫对雌二醇具有较高的敏感性。     

雌性大熊猫的血液性激素和阴道涂片的观察

关于大熊猫生殖生物学方面的研究,国内外研究报导较为罕见。Bonney R. C. 等(1982)用雌性大熊猫尿雌激素变化予测排卵期已得到证明,李惠福等(1982)曾报导大熊猫发情期中尿排出的N-乙酰-B-D-氨基葡萄糖苷酶的特异表现与排卵的关系,而生殖生物学其他方面的检测指标尚未见报导。本文为检测大熊猫发情期与非发情期血清雌二醇和孕酮激素水平及阴道细胞涂片变化,于1983年4月和5月分别对大     

电化学发光法对绒山羊孕酮水平检测效果分析

利用电化学发光法跟踪测定了绒山羊母羊血清孕酮(P4)水平。结果表明,在发情期的0~3 d P4值为(0.68±0.18)ng/m L;之后孕酮水平逐渐上升,于10~12 d间达到峰值(15.14±3.21)ng/m L,而后开始下降。母羊配种后若妊娠,在推测下一个发情期间,孕酮水平可达(12.24±3.72)ng/m L以上,妊娠中期也会保持相对较高的水平。说明辽宁绒山羊母羊在繁殖季节体内孕酮值随发情周期呈规律性变化。电化学发光法与微粒子发光法测定孕酮值符合度高。     

德国牧羊犬和拉布拉多犬发情期血清生殖激素的变化及其配种时间的选择

为研究成年雌性种德国牧羊犬及拉布拉多犬体内生殖激素促黄体生成素(LH)、雌二醇(E2)和孕酮(P)在发情期的分泌规律,寻找合理的配种时间,采用ELISA试剂盒检测2种试验犬(各5只)发情期内血清中LH、E2和P水平。结果表明:LH峰值的出现比E2峰值出现晚24h,P在E2达到峰值的时候浓度开始升高。两种犬的E2浓度存在显著性差异(P0.05),LH和P不存在差异显著性(P0.05)。E2:P与犬的发情行为有关,且有助于LH峰值的出现。当E2:P与LH:P的比值趋于稳定,即LH峰值后3~7 d时,进行配种可收到良好效果。     

大熊猫异常发情一例

行为观察、外阴变化、阴道上皮细胞角化率及雌激素测定是监测人工圈养大熊猫发情、指导配种的主要方法.本文报道了一例大熊猫发情期间这4种监测方法结果存在与正常发情不相一致的现象.该大熊猫在发情前、中期及高峰期均没有发情行为,但其外生殖器表现出开口、从小到大、从内缩到外翻;颜色也呈现了灰白、潮红、粉红、灰白的一系列正常发情母大熊猫外生殖器形态及颜色方面的变化;其E1G/Cr变化则与常见的发情期E1G/Cr变化一致,从基础水平逐步上升至峰值,峰值后迅速下降至基础值.阴道上皮细胞角化率在初期至高峰期一直保持基础水平,细胞核破裂的比例小于20％,远远小于正常发情者70％的角化率;根据雌激素含量的变化,在距离E1G/Cr峰值10h时为其实施首次人工授精,当年秋季该大熊猫顺利产下一仔.     

母犬发情期生殖激素测定及其意义

本试验从15只处于发情前期母犬中随机选取6只母犬,检测其促黄体生成素(LH)、促卵泡素(FSH)和孕酮(P4)3种激素,实现对母犬发情期连续的监测过程。采用静脉采血,抗凝低温离心后分别测定促黄体生成素(LH)、促卵泡素(FSH)、孕酮(P4)的含量,促黄体生成素采用人促黄体生成激素放射免疫分析药盒的方法测定,促卵泡素采用人促滤泡激素放射免疫分析药盒的方法测定,孕酮采用孕酮放射免疫分析药盒的方法测定。测定结果显示:母犬发情期的促黄体生成素的变化范围为5.16～6.82 ng/mL(除峰值外),其峰值平均值是116.71ng/mL;母犬发情期的促卵泡素的变化范围为3.18～4.78 ng/mL(除峰值外),其峰值平均值是43.56 ng/mL;母犬发情期的孕酮的变化范围为0.199 1～8.506 3 ng/mL,其峰值平均值是8.506 3 ng/mL。母犬在发情时,可出现LH释放波,并在LH开始释放后24 h内或开始发情的1～2 d内排卵。     

狸獭发情周期中17β-E_2和P_4分泌规律的研究

应用放射免疫分析法(RIA)首次测定了5只雌性成年狸獭发情周期中外周血浆17β-雌二醇(E_2)和孕酮(P_4)的含量。结果表明,在母狸发情盛期17β-E_2达峰值62.19pg/ml,以后开始下降,并在一定范围内上下波动,到下一个发情周期的发情期就又开始上升。而P_4含量在发悄盛期最低2.48ng/ml,到18天左右峰值达6.26ng/ml,到第二个发情周期前,又迅速下降。二者的这种变化规律与该动物发情周期的表现规律相吻合。     

银狐繁殖期血浆主要激素变化及其对繁殖功能的调控

通过放射免疫测定技术测定母银狐发情前期和发情期外周血浆雌二醇-17β[E2]、孕酮(P4)浓度以及公狐睾酮(T)浓度,并分析这些技术的变化及其对繁殖功能的调控.结果表明,银狐初配前2～3d,E2浓度达峰值(240.85±39.98)pg/mL,在母狐接受跨时E2浓度降至(146.95±38.21)pg/mL,而在发情前期和发情期,其孕酮浓度范围是0.91～77.43ng/mL,峰值是(54.77±23.66)ng/mL,在雌二酮峰值时,孕酮浓度为(28.57±21.21)ng/mL,配种时,P4浓度(33.12±22.56)ng/mL,初配后4d(排卵结束),P4达(48.28±34.84)ng/mL.公狐配种期,T浓度范围(127.2±444.4)pg/mL,,峰值是(341.2±103.2)pg/mL.     

Effects of a simulated estrous cycle on sodium, volume, ACTH, and AVP in sheep

The studies were designed to test for effects of acute increases in estradiol and progesterone, similar in magnitude and duration to those in the ovine estrous cycle, on adrenocorticotropic hormone (ACTH) and plasma vasopressin (AVP) under resting conditions and in response to hypotension. Ewes (7 per group) were studied as intact, ovariectomized, ovariectomized and treated with progesterone for 7-8 days, or subsequently treated with estradiol. During progesterone treatment plasma sodium and AVP were increased significantly. However, neither plasma volume nor blood pressure was altered. Plasma AVP responses to hypotension were not altered by either progesterone or estradiol treatment. The peak plasma ACTH response to hypotension was not altered by steroid treatment; however, the duration of the response was greater in progesterone-treated ewes than in intact ewes. The results indicate that changes in gonadal steroids similar to those in the ovine estrous cycle cause a small increase in plasma sodium that stimulates AVP, but do not alter regulation of blood pressure or volume or AVP or ACTH responses to hypotension.     

Dynamics of the Equine Preovulatory Follicle and Periovulatory Hormones: What's New?

Recent studies (2005–2008) on the interrelationships among the preovulatory follicle and periovulatory circulating hormones are reviewed. Close temporal and mechanistic relationships occur between estradiol/inhibin and follicle-stimulating hormone (FSH), between estradiol and luteinizing hormone (LH), and between progesterone and LH. Estradiol from the dominant follicle forms a surge that reaches a peak 2 days before ovulation. Estradiol, as well as inhibin, has a negative effect on FSH, and estradiol has a negative effect on LH. When estradiol decreases, the negative effect diminishes and accounts for the beginning of an FSH increase and a transition from a slow to rapid increase in LH on the day of the estradiol peak. The decrease in estradiol and the reduction or cessation in the growth of the preovulatory follicle beginning 2 days before ovulation are attributable to the development of a reciprocal negative effect of LH on follicle estradiol production when LH reaches a critical concentration. The LH decrease after the peak of the LH surge on the day after ovulation is related to a negative effect of a postovulatory increase in progesterone. Measurable repeatability within mares between consecutive estrous cycles occurs during the preovulatory period in diameter of the ovulatory follicle and concentrations of LH and FSH. Hormone-laden follicular fluid passes into the peritoneal cavity at ovulation and transiently alters the circulating concentrations of LH and FSH. Double ovulations are associated with greater estradiol concentrations and reduced concentrations of FSH.     

Effect of l-arginine methyl ester (l-NAME) on hormonal profile and estrous cycle length in buffaloes (Bubalus bubalis)

The present study was undertaken to evaluate the effect of L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor on serum nitric oxide, progesterone, estradiol profiles and estrous cycle length in buffaloes. Murrah buffaloes (n?=?16) exhibiting regular estrous cycles were randomly allocated to two groups of eight animals. In the treatment group, buffaloes were administered 400?mg/h?L-NAME over 2?h (total dose?=?800?mg) via the coccygeal artery and the aorta abdominalis on day 15 of the estrous cycle. In the control group, normal saline was infused on the same day of the cycle by the same route. Blood samples were collected every 4?h on days 15 and 16, and once daily from days 17 to 21 of the estrous cycle for the assay of progesterone, estradiol and nitric oxide. L-NAME treatment significantly (p?相似文献   

Association of increased estradiol and progesterone blood values with altered bovine polymorphonuclear leukocyte function

Polymorphonuclear leukocyte (PMN) function and serum concentrations of estradiol, progesterone, and cortisol (hydrocortisone) were monitored concurrently in clinically normal cows during the estrous cycle. Five parameters were used to evaluate PMN function: (i) random migration under agarose, (ii) ingestion of 125I-labeled Staphylococcus aureus, (iii) nitroblue tetrazolium (NBT) reduction, (iv) iodination, and (v) antibody-dependent cell-mediated cytotoxicity. Increased serum estradiol concentrations were associated with enhanced random migration, but had no apparent effect on NBT reduction, iodination, or ingestion of S aureus by bovine PMN. Increased serum estradiol was also associated with increased serum cortisol. Increased serum progesterone values were associated with a depression of NBT reduction and iodination by PMN, but with enhanced random migration and antibody-dependent cell-mediated cytotoxicity. These results indicate that physiologic changes in steroid hormone values during the normal estrous cycle of the cow are associated with alterations in PMN function.     

Relationship between uterine secretion of prostaglandin F2 alpha induced by oxytocin and endogenous concentrations of estradiol and progesterone at three stages of the bovine estrous cycle

The purpose of this experiment was to determine whether differences among cows in the ability of oxytocin to stimulate uterine secretion of prostaglandin F2 alpha (PGF2 alpha) were related to the endogenous ovarian steroid environment. Sexually mature heifers were treated with oxytocin (.33 IU/kg BW) at three stages of the estrous cycle: early (d 3 to 5; n = 5), middle (d 10 to 11; n = 5) or late (d 16 to 17; n = 5). To assess uterine responsiveness to oxytocin, concentrations of 13,14-dihydro-15-keto-PGF2 alpha (PGFM) were quantified in jugular venous plasma samples collected at 1/2-h intervals for 8 h postinjection. The ovarian steroid environment at the time of injection was estimated by measuring the concentrations of progesterone and estradiol in jugular venous plasma samples collected at 4-h intervals for 12 h immediately prior to injection. Concentrations of PGFM increased immediately following injection of oxytocin either early or late in the estrous cycle. The response was much less during the middle of the estrous cycle. The magnitudes of response, early and late in the estrous cycle, were similar and greater than that observed during the middle of the estrous cycle (P less than .05). There was a positive relationship (R2 greater than .8; P less than .05) between magnitude of the response to oxytocin and ratio of estradiol to progesterone both early and late in the estrous cycle. Thus, individual differences in uterine secretion of PGF2 alpha in response to oxytocin were related to stage of the cycle and to differences in the endogenous ovarian steroid environment within each stage of the estrous cycle.     

Ovarian follicular growth, function and turnover in cattle: a review

Studies in cattle assessing changes in number and size of antral follicles, concentrations of estradiol, androgens and progesterone in serum and follicular fluid, and numbers of gonadotropin receptors per follicle during repetitive estrous cycles and postpartum anestrus are reviewed. The rate of growth of small follicles (1 to 3 mm) into larger follicles increases as the estrous cycle progresses from d 1 to 18 (d 0 = estrus). Size of the largest antral follicle present on the ovary also increases with advancement of the estrous cycle. Most large follicles (greater than 10 mm) persist on the ovarian surface for 5 d or more between d 3 and 13 of the bovine estrous cycle. After d 13, most of these large follicles are replaced more frequently by new growing follicles (turnover) with an increased probability for recruitment of the ovulatory follicle after d 18. More research is needed to determine the time required for growth of bovine follicles from small to large antral size and evoke recruitment of the ovulatory follicle. Factors that regulate selection of the ovulatory follicle are unknown but may involve increased frequency of LH pulses in blood, altered blood flow and(or) changes in intrafollicular steroids and proteins. Quantitative evaluation of ovarian follicles indicated occurrence of consistent short-term changes in fluid estradiol and numbers of luteinizing hormone receptors in cells of large follicles only during the pre-ovulatory period. Presumably, low concentrations of follicular estradiol found during most of the estrous cycle are not due to a lack of aromatizable precursor or follicle-stimulating hormone receptors. Follicular fluid concentrations of progesterone increase only near the time of ovulation. Little is known about changes in follicular growth, turnover and function during postpartum anestrus in cattle. However, preliminary data suggest that the steroidogenic capacity of large follicles changes markedly during the postpartum period.     

皖西白鹅雌鹅生殖激素水平研究

本文研究了皖西白鹅雌鹅几种主要生殖激素在生殖期与非生殖期的变化情况.相关激素测定的结果表明血清中垂体泌乳素(PRL)、雌二醇(E2)和孕酮(P)含量在生殖期和非生殖期呈现规律性变化.随着雌鹅进入生殖期,雌二醇(E2)和孕酮(P)含量都逐渐升高,特别是在生殖期的后期,二者都出现了峰值,然后随着雌鹅离开生殖期而逐渐下降.而垂体泌乳素(PRL)在血清中的含量变化情况却与雌二醇(E2)和孕酮(P)含量变化情况相反.在生殖后期含量下降,而离开生殖期后,含量反而升高.     

Effect of transportation on the estrous cycle and concentrations of hormones in mares

Effect of transportation on estrous behavior, duration of the estrous cycle, ovulation, pregnancy rates and concentrations of serum cortisol, plasma ascorbic acid (AA), LH, estradiol and progesterone in mares was investigated. Fifteen mares were transported for 792 km (12 h) during the preovulatory stage of estrus. Transported mares were bled immediately before transport (baseline), at midtrip and 0, 12, 24, 48 and 72 h post-transport and twice daily from d 1 before transport to d 1 (estrogen) or 3 (LH) post-ovulation. Blood samples also were taken for progesterone on d 0, 2, 6, 10, 15, 16, 17, 18, 19 and 20 post-ovulation. Nontransported control mares (n = 15) were bled on the same schedule as transported mares. There was no difference (P greater than .05) in number of mares ovulating, estrous behavior, duration of the estrous cycle or pregnancy rate between groups. Cortisol in transported mares increased to concentrations greater (P less than .05) than those in control mares at midtrip and 0 h post-transport. Concentrations of AA in transported mares also increased (P less than .05) at midtrip, then decreased (P less than .05) below baseline at 24 h post-transport. Concentrations of LH and estradiol increased (P less than .05) above baseline throughout the blood-sampling period. Increases apparently were due to preovulatory surges of these hormones. Increase in LH concentrations in transported mares, however, was greater (P less than .05) than that in control mares at 0 h post-transport.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effect of progesterone alone or in combination with estradiol on follicular dynamics, gonadotropin profiles, and estrus in beef cows following calf isolation and restricted suckling

The effects of calf isolation and restricted suckling on LH pulse characteristics and interval to first ovulation (postpartum interval) were studied in 52 multiparous beef cows, with or without exogenous progesterone. At 30 d postpartum, cows were randomly allocated to one of four treatments (n = 13/treatment): 1) Ad lib, ad libitum access of cows to calves; 2) CI/RS, calf isolation/restricted suckling, where suckling was restricted to once daily; 3) CI/RS+P4, same as CI/RS but cows received an intravaginal progesterone-releasing device at calf isolation for 6 d; or 4) CI/RS+P4+E2, as CI/RS+P4 but the intravaginal progesterone-releasing device had a 10-mg estradiol capsule attached. Daily ovarian scanning and twice-daily blood sampling were performed from d 25 postpartum until the day of second ovulation. A random sample of cows from each treatment (n = 31 in total) were blood-sampled at 15-min intervals for 10 h on d 29, 32, 35, and 38. Ovulatory response to treatment was regarded as ovulation of either the dominant follicle growing at d 30 or the subsequent DF. There was a treatment x day effect (P = .09) on LH pulse frequency, but neither progesterone (CI/ RS+P4) nor progesterone and estradiol (CI/RS+P4+E2) treatment suppressed the calf isolation/restricted suckling-induced increase in LH pulse frequency. The estradiol capsule (CI/RS+P4+E2) delivered sufficient estradiol to delay new follicle wave emergence (treatment x stage; P < .001) and the associated preemergence increase in concentrations of FSH (treatment, P < .05) in cows treated at the postselection stage of follicle wave development, prolonging dominance of the dominant follicle present at treatment initiation (P < .001). The number of cows that ovulated in response to treatment was greater (P < .001) in cows with calf isolation/restricted suckling than in cows suckled ad libitum. Hence, cows assigned to the Ad lib treatment had a longer postpartum interval (P < .001) than cows of the other treatments. Exogenous progesterone treatment increased the frequency of cows exhibiting clinical signs of estrus at first ovulation (P < .001) and reduced the frequency of short estrous cycles (P < .001). We conclude that, in beef cows with calves, a 6-d progesterone treatment does not suppress the calf isolation/restricted suckling-induced increase in LH pulse frequency. Hence, on progesterone withdrawal, the LH pulse frequency is sufficient to stimulate first ovulation, accompanied by overt estrous expression and elimination of a short estrous cycle in most cows.     

Changes in sexual behavior and fecal steroid hormone concentrations during the breeding season in female Hokkaido brown bears (Ursus arctos yesoensis) under captive condition

The longitudinal changes in fecal steroid hormone concentrations and sexual behavior in 2 mated/pregnant and 3 non-mated female Hokkaido brown bears were investigated during the breeding season. Behavioral estrus (standing) lasted for 14 and 32 days in the mated females and for 25 to 36 days in the non-mated females. In non-mated females, sexual behavior, such as female-female mounting and masturbation, was observed for several days before and after the estrous period. In mated females, mean fecal estradiol-17beta concentrations were higher in the estrous period than in the post-estrous period, while fecal progesterone concentrations were higher in the post-estrous period than in the estrous period. The similar trends of steroid hormone changes were observed in the non-mated females.