撒坝猪血清酯酶多态性与繁殖性能关系的研究

采用垂直板聚丙烯酰胺凝胶电泳法(PAGE), 对115头撒坝猪的血清酯酶(ES)多态性进行了检测,计算了该位点的基因型频率、基因频率和位点多态杂合度(h),并用二因素有互作的最小二乘模型对血清酯酶多态性与繁殖性能的关系进行了分析。结果表明,撒坝猪血清酯酶3种基因型AA、AB和BB的频率分别为0.2696、0.5826和0.1478。两个等位基因A和B的基因频率分别为0.5609和0.4391。该位点的杂合度为0.4926。在3种基因型中,不同基因型母猪的繁殖性能在产仔数、仔猪初生窝重、20日龄窝重、断奶仔猪数和断奶窝重等性状上存在着显著差异(P<0.05);公、母猪不同基因型交配组合在产仔数、断奶仔猪数、仔猪断奶体重和断奶窝重等性状上亦有显著差异(P<0.05)。显示出猪的血清酯酶多态性可望作为繁殖性能选种的遗传标记。 Abstract：　Serum esterase polymorphisms of 115 Saba pigs were investigated by using the method of vertical polyacrylamide gel electrophoresis. The genotype frequency, gene frequency and heterozygosity of this locus were calculated. The relationship between the serum esterase polymorphism and reproductive performance was analyzed by the least square analysis of two factors (ES genotype of boar and sow) with interaction. The results demonstrated that the genotype frequency of AA, ABandBBwas 0.2696, 0.5826 and 0.1478 respectively, the gene frequency of the alleleAandBwas 0.5609 and 0.4391 respectively. The heterozygosity of this locus was 0.4926. There are significant differences (P<0.05) on litter size, litter weight at birth, litter weight at 20 days, litter size at weaning and litter weight at weaning of different genotypic sows. The significant differences (P<0.05) were showed on litter size, litter size at weaning, weaning weight and litter weight at weaning of different genotypic mating combinations. It indicated that the serum esterase polymorphism was expected to be the genetic marker of pig reproductive performance.     

Accuracy of canine parturition date prediction from the initial rise in preovulatory progesterone concentration

Accurate prediction of parturition date is useful for clinical management of canine parturition. For nearly all normal canine pregnancies, parturition occurs 64-66 days from the LH peak, the timing of which cannot be differentiated from the initial sharp rise in serum progesterone (P4) concentrations. We sought to determine by retrospective analysis if prebreeding serum progesterone concentrations could accurately predict parturition date. Serum progesterone concentrations recorded as serial samples from 63 bitches (19 breeds) were analyzed. Progesterone concentrations were measured by radioimmunoassay (RIA) or chemiluminescent immunoassay (CLIA). The CLIA method was validated for use in determining P4 concentrations in canine serum and results were comparable to those obtained with RIA. Bitches were grouped by nonpregnant body weight (BW) and litter size (LS). Day 0 (D0), the day of preovulatory rise in serum P4, was defined as the day that P4 concentration rose to > or =l.5 ng/ml and was at least twice the baseline concentration. The predicted parturition date, 65 days following the day of preovulatory rise in serum P4 (D65), was compared to actual parturition date, the day the first pup was delivered. We determined that mean P4 concentration at D0 for all BW groups was 2.02+/-0.18 ng/ml and there was significant variation in P4 concentrations between BW groups after D1. In addition, we determined that the accuracy of parturition date prediction within a +/-1, +/-2, and +/-3 day interval using prebreeding serum progesterone concentrations was 67, 90, and 100%, respectively, and that the accuracy was not affected by body weight or litter size.     

The relationship of conceptus number and fetal sex to maternal serum testosterone concentration in the rat

On Day 8 of pregnancy, the number of implantation sites in pregnant rats was adjusted to 1, 2, 4, 6, or greater than 10. Blood was collected on Days 11, 12, 15, 18, and 20 for the determination of serum testosterone, progesterone, and androstenedione. Serum testosterone levels exhibited a direct linear relationship with implantation number, increasing from 1 through greater than 10 implants. This linear relationship was particularly evident at Days 12, 15, and 18 of pregnancy. Serum progesterone levels increased from one to four conceptuses and plateaued above this number. There was no apparent relationship between the number of conceptuses and serum androstenedione levels, which may reflect the multiple origins of this steroid in the pregnant rat. In a separate group of rats in which the number of conceptuses was adjusted to three on Day 8 of pregnancy, blood was collected on Days 11, 12, 15, 18, and 20. Fetal sex was determined between the last bleeding and the day of parturition. Serum testosterone was determined and results were examined with regard to the number of male/female fetuses in the litter of three. There was no relationship between maternal serum testosterone levels and the number of male fetuses, indicating that the fetal testis does not make a significant contribution to circulating maternal testosterone levels.     

犬卵透明带3 DNA疫苗在小鼠模型中的避孕效果评价

为了评价犬卵透明带3(CZP3)DNA疫苗的避孕效果,采用6只雌性小鼠Mus musculus肌肉注射50μg pcDNA3-CZP3质粒后30 V电压刺激6次的方法建立实验组,以6只同等注射pcDNA3质粒的雌性小鼠为阴性对照组;初免后每周采血并用ELISA方法检测小鼠的CZP3抗体水平及第6周抗体滴度;第6周与雄鼠合笼,计算产仔数;第24周收集卵巢制成HE染色切片进行观察;最后用小鼠血清进行小鼠及犬卵透明带的间接免疫荧光实验。结果显示,1)实验组小鼠从初免第2周就产生了抗CZP3的抗体,并且与阴性对照组之间的差异有统计学意义(P<0.05);第24周实验组的抗体水平与阴性对照组之间的差异有高度统计学意义(P<0.01)。2)第6周血清中的抗体1∶4 000倍稀释时,实验组与阴性对照组之间的差异有高度统计学意义(P<0.01)。3)小鼠生育率由阴性对照组的100%降低至实验组的50%,平均产仔数由阴性对照组的(6.667±0.422)只降至实验组的(2.500±1.455)只,且实验组的抗体水平与产仔数呈高度负相关。4)卵巢HE染色切片结果显示,CZP3 DNA疫苗...     

Effects of the antiandrogen hydroxyflutamide on progesterone secretion by preovulatory rat follicles in vivo and in vitro.

Serum and ovarian progesterone levels and in vitro production of progesterone by preovulatory follicles were measured on proestrus in pregnant mare's serum gonadotropin (PMSG) primed immature rats in which the luteinizing hormone (LH) surge and ovulation were blocked by administration of the antiandrogen hydroxyflutamide. Serum progesterone levels observed at 12:00 on proestrus were significantly elevated, twofold above those observed in vehicle-treated controls, by in vivo administration of 5 mg hydroxyflutamide 4 h earlier. In control rats, proestrous progesterone did not increase until 16:00, in parallel with rising LH levels of the LH surge. No LH surge occurred in the hydroxyflutamide-treated rats, ovulation was blocked, and serum progesterone declined throughout the afternoon of proestrus, from the elevated levels present at 12:00. Administration of human chorionic gonadotropin (hCG) at 11:00 advanced the elevation of serum progesterone by 2 h in vehicle-treated controls and prevented the decline in progesterone levels in hydroxyflutamide-treated rats. The patterns of change in ovarian tissue concentrations with time and treatment were essentially similar to those observed for serum progesterone. In in vitro experiments, progesterone secretion during 24 h culture of preovulatory follicles obtained on PMSG-induced proestrus was significantly increased, sixfold, by addition to the culture media of 370 microM but not of 37 microM hydroxyflutamide. Testosterone (50 nM) and hCG (20 mIU/mL) caused 26- and 14-fold increases, respectively, in progesterone secretion by cultured follicles. Hydroxyflutamide significantly reduced the stimulatory effect of testosterone but not of hCG on progesterone secretion in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)     

Identification of genetic regions of importance for reproductive performance in female mice

Both environmental and genetic factors can dramatically affect reproductive performance in mice. In this study we have focused on the identification of genetic regions, quantitative trait loci (QTL), which affect the breeding capacity of female mice. We have identified polymorphic microsatellite markers for the mouse strains used and performed a genomewide scan on 237 females from a gene-segregating backcross between a high breeder and a relatively poor breeder. The high-breeder mouse strain we used is the inbred NFR/N mouse (MHC haplotype H-2q), which has extraordinary good breeding properties. The moderate breeder chosen for F(1) and N2 progeny was B10.Q, which is a genetically well-characterized MHC-congenic mouse of the H-2q haplotype. Each of the 237 females of the N2 generation was allowed to mate twice with MHC-congenic B10.RIII (H-2r) males and twice with B10.Q males. A predetermined number of phenotypes related to reproductive performance were recorded, and these included litter size, neonatal growth, and pregnancy rate. Loci controlling litter size were detected on chromosomes 1 (Fecq3) and 9 (Fecq4). The neonatal growth phenotype was affected by Fecq3 and a locus on chromosome 9 (Neogq1). On chromosome 11 two loci affecting the pregnancy rate (Pregq1 and Pregq2) were identified. Furthermore, on chromosomes 13 and 17 we found loci (Pregq3 and Pregq4) influencing the outcome of allogeneic pregnancy (allogeneic by means of MHC disparity between mother and fetuses). A locus on chromosome 1 affecting maternal body weight was also identified and has been denoted Bwq7. It is well known that reproductive performance is polygenically controlled, and the identification of the major loci in this complex process opens the possibility of investigating the natural genetic control of reproduction.     

Genetic and environmental interrelations between measurements of thyroid function in a healthy Danish twin population

Serum thyrotropin (TSH), free thyroxine (T4), and free triiodothyronine (T3) levels illustrate the thyroid function set point, but the interrelations between these have never been characterized in detail. The aim of this study was to examine the associations between TSH and thyroid hormone levels in healthy euthyroid twins and to determine the extent to which the same genes influence more than one of these biochemical traits; 1,380 healthy euthyroid Danish twins (284 monozygotic, 286 dizygotic, 120 opposite-sex twin pairs) were recruited. Genetic and environmental associations between thyroid function measurements were examined using quantitative genetic modeling. In bivariate genetic models, the phenotypic relation between two measurements was divided into genetic and environmental correlations. Free T4 and free T3 levels were positively correlated (r=0.32, P<0.0001). The genetic correlation between serum free T4 and free T3 levels was rg=0.25 (95% CI 0.14-0.35), suggesting that a set of common genes affect both phenotypes (pleiotropy). The correlation between the environmental effects was re=0.41 (0.32-0.50). From this we calculated that the proportion of the correlation between free T4 and free T3 levels mediated by common genetic factors was 48%. Only 7% of the genetic component of serum free T3 levels is shared with serum free T4. Serum TSH and thyroid hormone levels did not share any genetic influences. In conclusion, thyroid hormone levels are partly genetically correlated genes that affect free T4 levels and exert pleiotropic effects on free T3 levels, although most of the genetic variance for these measurements is trait specific.     

小熊猫繁殖周期血清雌二醇和孕酮含量变化

为研究小熊猫繁殖周期血清雌二醇、孕酮含量变化规律,采用化学发光免疫分析法连续16 次测定了2只成体雌性小熊猫血清雌二醇和孕酮含量变化,历经发情间期、发情期和两次妊娠期;连续9次测定了7只小熊猫妊娠期的孕酮含量变化。结果:(1)发情间期,小熊猫血清雌二醇的水平一直维持在低水平(基础水平),进入发情前期,血清雌二醇水平明显升高,在发情期一直维持高水平,配种后迅速降至基础水平; (2)小熊猫血清孕酮含量在发情间期和发情期均维持在较低水平,直至发情期过后才出现升高,在妊娠期一直维持高水平,峰值出现在5 月;(3)发情的小熊猫不论妊娠与否,在妊娠期内血清孕酮含量均维持在高水平。研究表明:小熊猫血清雌二醇、孕酮含量变化能直接反映其繁殖规律,雌二醇对启动雌性小熊猫季节性繁殖起重要作用;在妊娠期内小熊猫血清孕酮含量升高不能作为判断小熊猫妊娠的标准;雌性小熊猫在妊娠期有假孕现象。     

高体重布氏田鼠产仔数较多但繁殖输出及氧化损伤不受影响

体重可影响动物几乎所有的生物学变量。高体重双亲通常产生较大胎仔数。繁殖过程代价很高,可能伴随着氧化损伤。为比较双亲体重对繁殖期能量学特征、繁殖输出和氧化损伤的影响,检测了高体重组(雌性:51. 5 g ± 1. 6 g;雄性:60. 4 g ±2. 5 g)和低体重组(雌性:35. 5 g ± 1. 2 g;雄性:49. 6 g ± 2. 8 g)布氏田鼠繁殖过程中母体的体重和摄食量,分娩时的胎仔数和胎仔总重,及后代断乳时母体的身体成分、激素含量(血清瘦素和催乳素)和氧化损伤(血清丙二醛、蛋白质羰基和肝脏丙二醛含量)。结果显示:(1)高体重组妊娠末期的母体体重增量显著高于低体重组;(2)高体重组母体的出生胎仔数显著高于低体重组,但出生及断乳时的平均胎仔重、胎仔总重和母体繁殖期间的能量摄入无显著差异; (3)血清激素含量、身体成分、身体脂肪重量及氧化损伤指标均无组间差异。结果表明,高体重布氏田鼠可产生较多的出生胎仔数,但母体并不通过摄入更多的能量来保证其繁殖输出,对氧化损伤也没有影响。这些结果对于理解不同体重的动物在繁殖期间的能量策略以及繁殖与生存之间的权衡等生活史理论具有重要意义。     

Progesterone withdrawal induced by ICI 81008 in pregnant rats.

Of a total of 343 pregnant rats treated with the prostaglandin F2alpha analogue ICI 81008, 137 aborted, while 83 had reduced and 123 intact litter. These biological variations depended primarily on the gestational timing of treatment and on the dose and route of administration of this synthetic PG. In comparison with the 107 controls, all experimental rats which aborted had a drastic reduction in plasma progesterone levels which was highly significant (P is less than 0.001) until day 18. In contrast, those animals which escaped suboptimal ICI 81001 treatment with a partly resorbed litter, only had a moderate reduction in progesterone which was statistically significant (P is less than 0.01) until day 16 when levels of this steroid normally begin to decrease. Ineffective treatment did not affect progesterone levels and intact pregnancy. In contrast to progesterone, there was no correlation between plasma estradiol-17beta levels and the consequences of ICI 81008 treatment.     

Pituitary-Specific Overexpression of Porcine Follicle-Stimulating Hormone Leads to Improvement of Female Fecundity in BAC Transgenic Mice

Follicle-stimulating hormone (FSH) is a pituitary glycoprotein that, together with luteinizing hormone, plays a crucial role in ovarian folliculogenesis and female fertility. We previously found that FSH beta is a major gene controlling high prolificacy of Chinese Erhualian pigs. To directly study the biological effects on reproductive function of porcine FSH (pFSH) for polyovulatory species, we generated a novel gain-of-function mouse model using a bacterial artificial chromosome (BAC) system to jointly introduce 92 kb and 165 kb genomic fragments comprising the pFSH α- and β-subunit genes. These directed the physiological expression of pFSH with the same temporal and spatial pattern as endogenous FSH in female transgenic (TG) mice. Serum levels of biologically active pFSH heterodimers in independent TG lines ranged from 6.36 to 19.83 IU/L. High basal pFSH activity led to a significant reduction of serum LH and testosterone levels in TG females compared to wild-type (WT) littermates, yet endogenous FSH and estradiol levels were significantly elevated. Interestingly, ovarian histology showed that the number of corpora lutea was significantly higher at 14 and 28 weeks of age in TG females and breeding curves revealed that mean litter sizes of TG females were obviously larger than for WT littermates before 52 weeks of age. These findings indicate that pituitary-specific overexpression of pFSH within physiological boundaries can increase ovulation rate and litter size, but it does not cause reproductive defects. Therefore, our TG mouse model provides exciting insights for investigating the actions of pFSH in vivo.     

Effects of dichlorvos treatment on mouse reproduction

To test whether exposure to dichlorvos vapors for treatment of mouse ectoparasites resulted in temporary cessation of breeding, we exposed harem breeding groups of mice to varying concentrations of dichlorvos vapors and examined the effects of exposure on litter frequency and litter size. All exposure levels resulted in decreased plasma cholinesterase concentrations in treated mice for up to 10 days following the completion of exposure. Litter frequency and size were unaffected by dichlorvos exposure, and gestation times were not prolonged. Therefore, treatment with dichlorvos vapors during breeding did not affect reproduction in exposed mice.     

Pre-parturitional changes in serum prolactin, placental lactogen, growth hormone, progesterone, and corticosterone in the C3H/HeN mouse

Pre-parturitional changes in serum prolactin, placental lactogen, growth hormone, progesterone, and corticosterone in the C3H/HeN mouse are described. Serum prolactin concentrations display an apparent biphasic pre-parturitional increase. Both serum placental lactogen and growth hormone concentrations are elevated during the second half of pregnancy. Serum placental lactogen concentrations remain elevated until parturition, whereas serum growth hormone concentrations decline on the last two days of pregnancy. Serum progesterone and corticosterone concentrations are elevated during the latter half of pregnancy and decline on the day preceding parturition.     

Immunosuppression, sialic acid, and sialyltransferase of bovine serum as a function of progesterone concentration

Serum samples from progesterone-oestrogen-treated ovariectomized Holstein cows (N = 4) were compared with samples from control ovariectomized Holstein cows (N = 4) to determine the effects of physiological levels (0-6 ng/ml) of circulating progesterone. The average progesterone level in treated animals rose from 1 ng/ml (Day 0) to plateau at 5 ng/ml (Days 12 to 36). Sera from progesterone-oestrogen-treated cows during Days 4 to 10 significantly suppressed stimulation of lymphocytes by phytohaemagglutinin as compared with sera from control cows (P = 0.02), whereas no differences were detected in serum samples from Days 12 to 36. Serum samples from progesterone-oestrogen-treated or control cows did not affect the stimulation of lymphocytes by pokeweed mitogen. Sialyltransferase activity (P = 0.0002) and sialic acid content (P = 0.006) were both significantly elevated in serum from progesterone-oestrogen-treated animals compared with controls during Days 8 to 16, whereas no significant differences were observed at later times. The results suggest that suppression of phytohaemagglutinin-induced stimulation, sialic acid content, and sialyltransferase activity are sensitive not to the circulating level of progesterone but rather to increases in progesterone concentration, with maximal effects observed at Days 8, 12 and 12, respectively, after the start of progesterone treatment. The work provides a preliminary basis for further studies on the mechanism of immunosuppression by steroids and during pregnancy.     

Exogenous steroid effects on litter size and early embryonic survival in swine

These studies confirm the efficacy of 12.5 mug estrone and 25 mg progesterone, administered on Days 16 and 17 of pregnancy, on increased litter size in swine. Substitution of estradiol (with ten-fold greater biological activity) for estrone in this regimen eliminated the positive effect on litter size and estradiol given alone at dosages of 3.1 and 6.3 mug were without effect. However, when estradiol was given in combination with progesterone (25 mg) at a dosage equivalent to 10% the effective estrone dosage (1.25 mug), a 22.6% increase in total pigs born per litter over controls was observed. Estrone alone at dosages of 6.25 and 12.5 mug did not alter litter size. At a dosage of 18.75 mug, an 8.7% decrease in total pigs born was noted. The amniotic fluid from sows treated for ten days with 12.5 mug estrone and 25 mg progesterone near the time of implantation, had significantly higher protein concentration, pH and osmolality.     

Repeatability of blood serum progesterone levels in dairy heifers on day 7 of the estrous cycle

Thirty normally cycling dairy heifers were used to determine the repeatability of blood serum progesterone levels on Day 7 ± 0.25 d of the estrous cycle. The experimental group consisted of 16 Holsteins and 14 dairy crossbreds ranging in age from 18 to 24 months. Day of the estrous cycle was determined from twice daily observations for standing heat (Day 0). Serum progesterone levels for Day 7 ± 0.25 d were determined by radioimmunoassay from blood samples collected by jugular venipuncture over three to four consecutive estrous cycles. Levels of blood serum progesterone for Day 7 ± 0.25 d ranged from 0.57 to 6.03 ng/ml. Least square means for the Holstein (2.74 ng/ml) and dairy crossbred (3.38 ng/ml) groups were different (P<0.006). The repeatability for levels of blood serum progesterone on Day 7 of the estrous cycle was low (0.0115).     

The influence of estradiol-17beta and progesterone on peripheral serum concentrations of luteinizing hormone and follicle stimulating hormone in the ovariectomized ewe

Serum gonadotropin concentrations were high and variable and fluctuated episodically in short and long term ovariectomized ewes. Treatment with solid silastic implants releasing progesterone (serum levels 1.81 +/- 0.16 ng/ml) had no consistent effect. Treatment with implants releasing estradiol-17beta significantly depressed mean serum gonadotropin concentrations and peak height to values usually seen in intact ewes. This occurred regardless of implant size and serum estradiol-17beta concentrations (range 11 +/- 0.3 pg/ml to 98 +/- 12.8 pg/ml). Progesterone and estradiol-17beta together significantly depressed the frequency of peaks in LH concentration. Following progesterone removal, 95% of the ewes treated with progesterone and estradiol-17beta implants experienced a transient increase in serum LH concentrations similar to the preovulatory surge in intact ewes. Eighty-four percent of the LH surges were accompanied by a surge in serum FSH concentrations. However, following progesterone removal, 5.1 +/- 2.1 FSH surges were observed over six days. Gonadotropin surges occurred regardless of estradiol-17beta implant size and with or without the influence of supplemental estradiol-17beta.     

Further investigation of the role of progesterone in the control of embryo transport in the mouse

Combined ovariectomy and adrenalectomy retarded mouse embryo transport while either operation alone did not. Serum progesterone levels were reduced after ovariectomy and after the combined operation but detectable levels were still present on Day 4 following both procedures. Embryo transport was also retarded after administration of testosterone propionate. This effect was abolished by progesterone and was not mimicked by 5 alpha-dihydrotestosterone. From these results it is concluded that progesterone influences embryo transport and that androgenic effects are probably a result of antagonism of progesterone.     

Sexual steroid levels and their clinical significance in the early neonatal age

Serum concentrations of progesterone (P) and estradiol (E2) in the umbilical cord blood were measured in healthy full term newborn infants and were related to the serum levels of indirect bilirubin at 72 hr postpartum in the same group of neonates. In those having higher concentration of bilirubin than 204 mumol/l at 72 hr after birth, cord P and E2 levels were significantly higher than in the rest of the group. Furthermore, strong correlation was found between serum levels of progesterone at 24 hr after birth and that of bilirubin at 72 hr postpartum in the same subjects. According to the authors' conclusion serum progesterone levels 24 hours after birth prognosticate the occurrence of neonatal hyperbilirubinaemia.     

Response of ovaries and serum steroid levels after treatment of cystic ovarian disease in dairy cattle I. treatment with corticosteroids and a combination of human chorionic gonadotropin and progesterone

Of 38 cows with ovarian follicular cysts (cysts), 10 were injected intramuscularly with 20 mg of betamethasone or 10 mg of dexamethasone (CC) and 28 intravenously with a combination of 3,000 IU of human chorionic gonadotropin and 125 mg of progesterone (HCG·P). Ovarian responses and changes in serum levels of sex steroids were examined after both of the treatments. Percent of the cows in which cysts luteinized 10 days after the treatment in the HCG·P-treated cows was significantly higher (P<=0.05) than in the CC-treated ones. Serum levels of progesterone and estrogens (estrone and estradiol-17β) in the cows were not affected 10 days after CC injection. On the contrary, serum progesterone increased significantly (P<=0.05) and serum estradiol-17β decreased significantly (P<=0.05) 10 days after HCG·P injection. Serum progesterone did not respond to HCG·P treatment in the cows previously treated unsuccessfully with gonadotropin, while progesterone increased significantly (P<=0.05) in response to HCG·P treatment in those given no treatment before.