Effect of Timing of Postovulatory Insemination Relative to Human Chorionic Gonadotropin/Buserelin Treatment With 1 Straw of Frozen-Thawed Semen on Mare Fertility

The reproductive management of mares for frozen semen artificial insemination (AI) can be costly and labor intensive. Predicting the exact time of ovulation can be challenging even when ovulation-inducing drugs are used. The main objective of this retrospective study was to determine whether there was an effect of interval between examinations to detect ovulation on likelihood of pregnancy and early embryonic loss in mares after postovulatory breeding with a single straw of frozen/thawed semen. The second objective was to determine the efficacy of two different drugs (human chorionic gonadotropin vs. buserelin) for timely induction of ovulation. The length of the interval from penultimate check to ovulation had no significant effect on pregnancy or embryo loss rates (4 hours: 34.1% and 13.3% vs. 8 hours: 26.1% and 0% and 16 hours: 34.5% and 10%, respectively) nor did the ovulation-inducing drug used, number of the cycle, or the stallion. In conclusion, there appears to be no advantage of checking mares for ovulation during the late evening and night hours when using a postovulatory AI protocol and ovulation-inducing drugs.     

A novel method to improve sow reproductive performance: Combination of pre-weaning immunization against inhibin and post-insemination hCG treatment

The feasibility of a novel method to improve sow reproductive performance by combining inhibin immunization and hCG treatment was tested using in vivo and in vitro experiments. In the in vivo experiment, 106 sows were administered an inhibin immunogen on day 7 prior to weaning, and 56 non-treated sows served as the controls. Sows exhibiting oestrous behaviour on day 5 after weaning were artificially inseminated. On day 5 post-insemination, a subset of 50 inhibin-immunized sows received an injection of 1 000 IU human chorionic gonadotropin(hCG). Our results showed that pre-weaning immunization against inhibin marginally improved(P=0.068) total litter size and significantly increased(P=0.044) the live litter size. The overall differences in farrowing rates and live litter size tended toward significance(P=0.10) in the three groups, and the differences in total litter size were not significant(P=0.18). In the in vitro experiment, activin and hCG dose-dependently suppressed(P0.001) and stimulated(P0.001) progesterone(P4) secretion in cultured pig granulosa cells(GCs), respectively, and the suppression effect of activin was reversed(P0.001) by hCG. Activin suppressed P4 secretion mainly by downregulating(P0.001) the expression of StAR, Cyp11 a1, and 3β-HSDII, whereas hCG prevented(P0.001) the suppression effects. These results indicate that the combination of pre-weaning immunization against inhibin and postinsemination hCG treatment provides a novel method for improving sow reproductive performance.     

Single injection of triptorelin or buserelin acetate in saline solution induces ovulation in mares the same as a single injection of hCG

The aim of this study was to assess the efficacy of different doses of buserelin acetate and another GnRH agonist, triptorelin acetate, in saline solution in a single subcutaneous injection, to induce ovulation of growing pre-ovulatory follicle in mare and compare it with the classical treatment of a single injection of hCG. The study is split into 3 experiments over different breeding seasons in the same stud with a random distribution of treatment. The first one was to compare the injection of 6 mg of buserelin with 1,500 IU of hCG; the second one consisted of comparing different doses of buserelin (6 mg and 3 mg); and the third one compared three different doses of buserelin (3, 2 and 1 mg), 0.1 mg of triptorelin with 1,500 IU of hCG as a control group. The results of all experiments showed the same efficacy between all treatments with mares ovulating between 24 and 48 hr after injection: experiment 1: hCG (78% n = 41) and buserelin 6 mg (90% n = 50); experiment 2: buserelin 6 mg (78,1% n = 192) and buserelin 3 mg (78% n = 341); and experiment 3: hCG (87% n = 106), buserelin 3 mg (84,7% n = 137), buserelin 2 mg (82,7% n = 104), buserelin 1 mg (87% n = 54) and triptorelin 0.1 mg (84,7% n = 72). In conclusion, this study contributes to erasing the dogma that has been established since 1975 that a single injection in solution without any long-acting excipient of a GnRH agonist cannot induce ovulation in the mare. This study also shows that a injection of 0.1 mg of triptorelin in solution is a good alternative for ovulation induction and is comparable to small doses of buserelin acetate in solution (1 mg) and 1,500 IU of the gold standard trigger hCG, mainly in countries where human formulation of buserelin is not available.     

猪卵丘卵母细胞复合体和壁层颗粒细胞上LHR/hCGR的检测及性腺激素的分泌

用取自 ３～５ ｍ ｍ 卵泡的处于减数分裂阻抑状态的猪卵丘卵母细胞复合体（ｐ Ｃ Ｅ Ｏ），培养 ４８ ｈ，用 Ｆ Ｓ Ｈ（１００ Ｉ Ｕ／ Ｌ）或 ｈ Ｃ Ｇ（０、５０、１００、２００、５００ Ｉ Ｕ／ Ｌ）刺激 ｐ Ｃ Ｅ Ｏ 分泌甾体激素；用放射受体测定法（ Ｒ Ｒ Ａ）比较 ｐ Ｃ Ｅ Ｏ 的卵丘细胞及壁层颗粒细胞（ Ｍ Ｇ Ｃ）上促黄体激素受体／人绒毛膜促性腺激素受体（ Ｌ Ｈ Ｒ／ｈ Ｃ Ｇ Ｒ）的数量； Ａ Ｍ ｅ Ｘ 石蜡切片、免疫组织化学染色，检测 Ｌ Ｈ Ｒ／ｈ Ｃ Ｇ Ｒ 在 ｐ Ｃ Ｅ Ｏ 以及 Ｍ Ｇ Ｃ 的分布情况。在 Ｆ Ｓ Ｈ 作用下，ｐ Ｃ Ｅ Ｏ 分泌的孕酮明显高于 ｈ Ｃ Ｇ 作用组和对照组 （ Ｐ ＜ ００５）； 平均每个壁层颗粒细胞上的 Ｌ Ｈ Ｒ／ｈ Ｃ Ｇ Ｒ是每个卵丘细胞的 １０５ 倍； Ｌ Ｈ Ｒ／ｈ Ｃ Ｇ Ｒ 在基膜两侧分布较多，且卵母细胞上没有 Ｌ Ｈ Ｒ／ｈ Ｃ Ｇ Ｒ，临近卵母细胞的卵丘细胞膜上较少被染色。以上结果表明，在体外试验中，可能由于 Ｃ Ｅ Ｏ 上 Ｌ Ｈ Ｒ／ｈ Ｃ Ｇ Ｒ 的数量不足或生理活性降低， Ｌ Ｈ／ｈ Ｃ Ｇ 不能促进卵母细胞恢复减数分裂。     

提高黄牛,杂种牛受胎率的研究

对２７１头黄牛、杂种牛进行提高受胎率的试验。结果表明：用ＬＲＨ－Ａ处理的母牛情期受胎率和总受胎率分别为５６．６％和６５．９３％，明显高于对照组（２４．４２％和４９．２１％）（Ｐ＜０．０５）；用ｈＣＧ处理的母牛情期受胎率和总受胎率分别为５３．３３％和７２．７２％，明显高于对照组（３３．６９％和５２．５４％）（Ｐ＜０．０５）。     

促进黄体功能提高奶牛人工授精和胚胎移植受胎率的研究

研究探讨了在人工授精后和胚胎移植过程中注射hCG以促进黄体功能从而提高奶牛的受胎率。试验一:随机选取自然发情后的4~8岁荷斯坦经产奶牛39头进行人工授精,在输精后第5天和第10天,分别肌肉注射hCG2000U/头(试验组)或生理盐水5mL/头(对照组)。在输精后的第0天(输精当天)、第7天、第14天和第21天,每组随机抽取12头牛从颈静脉采血分离血清以测定血清中孕酮浓度。结果hCG处理组牛血浆孕酮浓度在输精后第7、14和21天都显著高于对照组(P<0.05);输精后45d的受胎率在hCG处理组为63.2%,高于对照组的45.0%,但差异不显著(P>0.05)。试验二:选取16头生殖系统健康、16~17月龄的青年荷斯坦牛作为受体牛,在发情后第7天,对A、B级黄体的牛施行胚胎移植,试验组牛于胚移同时静脉注射6000UhCG,对照组牛则注射生理盐水5mL/头。结果hCG处理牛在胚移后第7和14天的血浆孕酮浓度极显著高于对照组(P<0.01);移植后第35天的受胎率试验组为62.5%,高于对照组的50%,但差异不显著(P>0.05)。试验结果表明,在人工授精和胚胎移植后特定时间,使用外源hCG能够有效促进黄体发育,提高血液中孕酮浓度,从而一定程度提高母牛受胎率。     

Induction of Out-of-Season Spawning of Pikeperch, Sander Lucioperca (L.)

Pikeperch were induced to spawn 3 months prior to the natural spawning period through photothermal and hormonal stimulation. Females (five specimens in each group) were stimulated with injection of human chorionic gonadotropin (hCG) once (200 IU kg^–1), twice (200 IU kg^–1, second dose after 48 h–400 IU kg^–1) or three times (200 IU kg^–1, after 24 h–200 IU kg^–1 and after another 24 h–200 IU kg^–1). The control group was injected once with 0.9% NaCl. The males were stimulated with a single hormone dose of 200 IU kg^–1. Eggs were obtained from all the hormonally treated fish. None of the control group females, which were only stimulated photothermally, ovulated any eggs. The time of ovulation was 66–71 h following the first injection, and the eggs viability until the eyed stage (from 71.5 to 77.5%) did not depend on the number of hormone doses (P > 0.05). The out-of-season spawning method described in this paper could be used to provide pikeperch larvae for intensive culture systems (recirculating water systems) before natural spawning season and to produce larger-sized pikeperch fingerlings for stocking.     

A comparison of human chorionic gonadotropin and luteinizing hormone releasing hormone analogue for ovulation induction in black sea bass Centropristis striata (Linnaeus, 1758)

Mature black sea bass, Centropristis striata L. (200–800 g), were captured in coastal South Carolina during the spawning season and administered hormones for ovulation induction and strip spawning. During both study years, control groups of females were incorporated into the study design and administered sham injections containing physiological saline solution. In 2004, females received a single intramuscular injection of human chorionic gonadotropin (hCG) (330 IU kg⁻¹) (n=8) or two injections of hCG at 24‐h intervals (n=8). In 2005, females received a single injection of hCG (n=10) or an analogue of luteinizing hormone releasing hormone (LHRHa) (n=10). In 2004, all fish administered a single dose of hCG ovulated at least once. Six fish ovulated on two consecutive days and one fish ovulated on 3 days consecutively. In contrast, six of eight fish receiving two doses of hCG ovulated once, five ovulated on 2 days successively and three fish ovulated 3 days in succession. Of the fish that spawned, no differences were found in any reproductive parameters. In 2005, all fish administered hCG or LHRHa ovulated at least once. Three fish administered hCG ovulated twice, four fish ovulated on three consecutive days and one fish 4 days successively. All fish administered LHRHa spawned at least twice, six fish ovulated thrice and three fish ovulated 4 days, successively. A significant difference in fertility was found between hCG (75.6±11.4%) and LHRHa (55.6±27.4%). The results of this study indicate that both hCG and LHRHa are effective for ovulation induction in prespawning black sea bass.     

hCG通过ERK1/2调节睾丸间质细胞StAR蛋白的表达

以2—3周龄的仔猪睾丸间质细胞为试验材料，研究了hCG在快速调节期对StAR蛋白表达的调节机制。结果表明：hCG、8-Br-cAMP均能使StAR蛋白、mRNA和P-ERK1／2的水平随刺激时间的延长逐渐增加，8-Br-cAMP作用的速度较hCG更快，增幅也更明显；加入PKI，StAR蛋白、mRNA和P-ERK1／2活性明显下降，但StAR mRNA仍然可以检测到；加入MAPK的抑制剂PD98059后，StAR蛋白、mRNA的水平均降低。由此可知，hCG在诱导StAR蛋白的表达过程中，首先通过cAMP—PKA直接调节蛋白前体蛋白向成熟蛋白的转化，随着时间的延长，通过cAMP-PKA-ERK1／2级联，磷酸化转录因子，促进StAR基因的表达。