Estradiol, progesterone and cyclic nucleotides in the fluid of persistent follicles and luteal phase follicles in the estrus cycle in cows]

The objective of the present study was to compare the concentrations of 17 beta-estradiol, progesterone, cyclic adenosine monophosphate and cyclic quinosine monophosphate in the largest follicles of cows that persist for seven days after insemination following the preceding synchronization of oestrus and superovulation and in follicles of the luteal phase of cycle (5th-10th days). Animals included in the experiment were selected on the basis of rectal examination. Synchronization of oestrus was achieved in 24 crossbreds of Slovak Pied x Lowland Black Pied breeds (SS x Nc) using two doses of cloprostenol of Czechoslovak provenience Oestrophan Spofa, 500 micrograms in each, within 11 days. Serum gonadotrophin at the amount of 2500 I. U. was administered forty-eight hours before administration of the second dose PGF2 alpha. Experimental animals were inseminated after 72 hours. On the 7th day after mating the cows were killed at a slaughterhouse. Evaluated were only the ovaries of the 14 cows in which the persistent large follicles occurred. Ovaries of the 13 control cows in the luteal phase between the 5th-10th days were obtained at the slaughterhouse by the method after Ireland et al. (1980). Correct determination of the phase of sexual cycle was substantiated by determination of progesterone concentrations in blood serum. Follicular fluid was obtained from the largest follicles by aspiration and centrifuged in a cooled centrifuge at 3000 G. The concentrations of 17 beta-estradiol and progesterone in follicular fluid were determined using kits from URVJT at Kosice, designated RIA-test-ESTRA (SI-125-9) or RIA-test-Prog (SI-125-6).2+ persistent follicles (9.15 +/- 5.47 nmol.l-1).(ABSTRACT TRUNCATED AT 250 WORDS)     

Dynamics of ovarian follicular development in cattle following hysterectomy and during early pregnancy

Three experiments were conducted to evaluate ovarian follicular dynamics and functional activity during pregnancy in cattle. In 11 pregnant Charolais cows of Experiment I, size of largest follicle, number of follicles and accumulated follicle size were reduced by day 27 of pregnancy on the ovary bearing the corpus luteum (CL) but not on the non-CL bearing ovary. In experiment II, local attenuation of ovarian follicular development on the CL bearing ovary of seven pregnant heifers was evident compared to the contralateral ovary without the CL. However, in four hysterectomized heifers, follicular development was sustained on both the CL- and non-CL bearing ovaries when CL maintenance was achieved without presence of the uterus or conceptus. In Experiment III, steroidogenic characteristics of the largest and second largest follicles at 17 d postestrus were evaluated for seven pregnant and six cyclic cattle. Follicle by physiological status interactions were detected for both aromatase activity of the follicle and follicular fluid concentrations of estradiol and progesterone. In cyclic cows, the largest follicle had appreciably more aromatase activity than did the second largest follicle; whereas, aromatase activity of the largest follicle from pregnant cows was less than that of cyclic cows. However, in pregnant cows the second largest follicle became the estrogen-active follicle, and this follicle occurred with a higher frequency on the ovary contralateral to the CL-bearing ovary. These changes in aromatase activity were reflected by parallel changes in estrogen concentrations of follicular fluid. The higher progesterone concentration in follicular fluid of the largest follicle in pregnant cows provided further confirmation of their atretic status. In conclusion, during early pregnancy the conceptus and/or uterus ipsilateral to the conceptus appear to secrete compounds which alter local follicular steroidogenic activity and attenuate subsequent follicular growth between 17 to 34 d of pregnancy on the CL-bearing ovary. This local mechanism acting within the ovary may contribute to the antiluteolytic effects of early pregnancy in cattle.     

Relationship between endometritis and oxidative stress in the follicular fluid and luteal function in the buffalo

In this study, alteration in the follicular fluid composition and luteal function was investigated in the buffalo with endometritis. Genitalia were classified into cytological and purulent endometritis on the basis of polymorphonuclear cell cut off while non‐endometritis served as control (n = 10/group). In the follicular phase, the number of surface follicles was counted, diameter of the largest follicle was measured and the follicular fluid was assayed for total protein, cholesterol, malondialdehyde (MDA), total antioxidant capacity (TAC), oestradiol (E₂) and progesterone (P₄). The P₄ content of corpus luteum during mid‐luteal phase was estimated by radioimmunoassay. Ovaries from the follicular phase of oestrous cycle showed no significant difference in the total number of surface follicles, size of the largest follicle and volume of follicular fluid in the buffaloes with and without endometritis (p > .05). However, the antral fluid of the largest follicle from the genitalia of buffalo with cytological and purulent endometritis showed a significant decrease in the concentration of total protein, cholesterol, TAC and E₂ and a significant increase in the concentration of MDA and P₄ (p < .05). The results indicated that there is an association between endometritis and decreased ovarian function.     

Ovine follicular fluid inhibits aromatase activity

“Within follicle” regulations may be important for the fine tuning of gonadotrophin action in ovarian follicles. While numerous growth factors, steroids or proteins which are present in follicular fluid have been shown to have the ability of positively or negatively affecting follicle function, the net effet of follicular fluid of the dominant follicle on its function is unclear.

A bioassay measuring aromatase activity of follicular walls was used (1) to check whether follicular fluid from dominant follicles can alter aromatase activity (2), to check how follicle size, atresia and specific gonadotrophins alter the effects of follicular fluid (3), to identify the nature (steroid or protein) of the active compound(s), and (4) to check whether the inhibition is specific of aromatase. Dominant follicular fluid had the ability to reduce aromatase activity. This effect was dose dependent and was obvious whether or not a protease inhibitor was added to the incubation medium. There was no difference in the magnitude of the inhibitory effect of follicular fluid when FSH (2 ng/ml) or no FSH was added to the incubation medium. LH, however, could potentialise the inhibitory effects of follicular fluid. Dominant follicular fluid was more potent to inhibit aromatase than follicular fluid from atretic follicles. Medium conditioned by granulosa cells, but not by theca cells could inhibit aromatase activity when added to the incubation medium. Charcoal treatment of dominant follicular fluid did not remove its inhibitory potential. Fractionation of dominant follicular fluid by a desalting column demonstrated that the inhibition was related to a compound(s) > 10 kDa. Finally, the effect of dominant follicular fluid on aromatase appears specific of this enzyme as follicular fluid does not affect androgen output by thecal shells or progesterone output by luteal cells.

Further research is required to check whether the activity observed in dominant follicular fluid is related to compounds known to affect aromatase activity (inhibin, mullerian inhibiting substance, heat shock protein 90, superoxyde dismutase) or to another peptide/protein.     

Effects of progestagens on follicular growth and oocyte developmental competence in FSH-treated ewes

Previous research has reported evidence for negative effects of progestagens on follicular growth and oocyte competence. In the present study, negative effects of progestagens on follicular growth and oocyte developmental competence were assessed. During the breeding season, 20 Sarda ewes were treated with two doses of cloprostenol, 10 days apart, to assure the presence of a corpus luteum (CL). On day 5 after the second cloprostenol dose, 10 ewes were treated with a progestagen sponge while 10 females remained untreated. Starting on day 7 after the second cloprostenol dose, all the ewes were treated with 6 equal doses of 24 I.U. of FSH (Ovagen™, ICP, NZ), every 12 h. The number of follicles ≥2 mm in diameter increased (P < 0.0005) in all the ewes from 24 h before to 60 h after the first FSH dose (from 12.8 ± 1.1 to 23.4 ± 1.3 in treated and from 12 ± 0.6 to 22 ± 1.2 in untreated ewes, n.s.). There were no significant differences in follicle dynamics between groups, but concentrations of estradiol in control ewes were higher than in the progestagen group (P < 0.05). Twelve hours after the last FSH dose, oocytes were collected by ovum pick-up. Recovery rates were lower for progestagen-treated ewes (71.1 versus 83%; P < 0.001). After IVP procedure, cleavage rate was also lower in the progestagen group (39.1 versus 82.6%; P < 0.001). Furthermore, blastocysts output revealed that oocyte developmental competence was lower in progestagen group (17.3 versus 30.4%; P = 0.245), although differences were not significant. These results suggest deleterious effects from progestagen on oocyte developmental competence and set the basis for new protocols for in vitro embryo production.     

Characteristics of oestrous cycles in Holstein cross-bred dairy heifers: An evidence of delayed post-ovulatory progesterone rise

The aims of the current study were to illustrate figures for the characteristics of oestrous cycles especially on follicular dynamics, corpus luteum and changes in progesterone and prostaglandin F2alpha, in the Holstein cross-bred dairy heifers. Twenty six healthy and sexual-mature virgin heifers were monitored for signs of oestrus. Their ovaries were sonically examined and the numbers and the sizes of the follicles as well as of the corpus luteum were documented. In our study, no difference in ratio of the 2-wave and 3-wave patterned cycle was evident. Seasons' change did not affect on characteristics oestrous cycles as well as on dynamics of follicles and corpus luteum. The heifers showed high variation in manifesting oestrus especially on a number of hours. The ‘bodily’ oestrous signs lasted longer than did ‘behavioural’ signs and connection of lowering of the back to standing oestrus was established. Certain diversities comparing to of existed dairy breeds were drawn for follicular dynamics, corpus luteum and its progesterone: 1) the 1st an-ovulatory dominant follicles showed higher growth rate and earlier exceeded dominant diameter; 2) the follicle tended to quicker ovulate but with a smaller diameter at ovulation; 3) the corpus luteum exhibited 4−16.5 mm in diameter of central cavity. Connecting to the levels of progesterone, 4) the corpus luteum turned into active, as well as mid-luteal, period quite late, and 5) the duration of the active period of the corpus luteum was shorter, but 6) at the end of the cycle –around the day of oestrus, progesterone remained certain low but significant levels. In conclusion, the Holstein cross-bred dairy heifers in our study faced a problem of delayed post-ovulatory progesterone rise of which underlying causes are needed to be further scrutinised either at endocrine or at cell levels.     

藏绵羊卵巢组织学及卵泡超微形态的观察

旨在通过观察藏绵羊卵泡、黄体的组织学特征及卵泡的超微形态,探讨其与生理功能的关系.本研究运用大体解剖、常规组织切片和H.E染色及透射电镜技术对藏绵羊卵巢卵泡和黄体的组织结构特点以及卵泡的超微形态进行观察和分析.结果发现,藏绵羊黄体期和卵泡期卵巢的宽度和厚度存在显著差异(P＜0.05),而重量和长度无显著差异(P＞0.0...     

6月龄和成年牦牛卵巢及卵泡发育状况研究

为了研究6月龄牦牛和成年牦牛卵巢及表面卵泡发育状况,试验比较了6月龄和成年牦牛卵巢长度、宽度、厚度、重量、卵泡数量以及卵母细胞体外成熟培养效果。结果表明:成年牦牛卵巢长度(2.29±0.43)cm、宽度(1.91±1.31)cm和厚度(1.60±1.90)cm均显著大于6月龄牦牛[(1.65±0.30)cm、(1.14±0.25)cm、(0.79±0.26)cm](P<0.05),成年牦牛卵巢体积(6.92±7.00)cm3和重量(3.19±1.58)g极显著大于6月龄牦牛体积(1.63±0.93)cm3和重量(0.87±0.44)g(P<0.01)。6月龄牦牛Ⅰ级卵泡数(14.47±8.74)枚和平均总卵泡数(15.17±8.87)枚极显著高于成年牦牛Ⅰ级卵泡数(7.97±3.72)枚和平均总卵泡数(8.98±3.87)枚(P<0.01),Ⅱ级卵泡数差异不显著(P>0.05),成年牦牛平均每头含(0.02±0.15)枚Ⅲ级卵泡,而6月龄牦牛无Ⅲ级卵泡。成年牦牛有黄体卵巢重量显著大于无黄体卵巢重量(P<0.05),有黄体卵巢含(0.06±0.24)枚Ⅲ级卵泡,而无黄体卵巢不含Ⅲ级卵泡。6月龄和成年牦牛A、B级卵母细胞体外培养成熟率分别为(81.39±3.53)%、(80.44±4.50)%,差异不显著(P>0.05),而6月龄牦牛卵巢的平均卵母细胞数和平均A、B级卵母细胞数均显著高于成年牦牛(P<0.05)。     

Use of milk progesterone enzyme immunoassay for differential diagnosis of follicular cyst, luteal cyst, and cystic corpus luteum in cows

In 160 cows with ovarian cysts as determined by rectal palpation, differentiation was made of follicular cyst, luteal cyst, and cystic corpus luteum on the basis of milk progesterone concentrations estimated by an enzyme immunoassay before and at 10 days after cows were treated with gonadotropin-releasing hormone. Cows having a progesterone concentration in skim milk less than 1.0 ng/ml were considered to have follicular cysts and those with concentrations of 1.0 ng/ml or higher were regarded as the cases of luteal cyst or cystic corpus luteum. Luteal cyst was characterized by progesterone values remaining high in the cows for 10 days after treatment, and cystic corpus luteum was characterized by a decrease in progesterone concentration after cows were treated. By the rectal palpation procedure it was impossible to differentiate luteal cyst and cystic corpus luteum from follicular cyst. The frequencies of follicular cyst, luteal cyst, and cystic corpus luteum were 65%, 19%, and 16%, respectively. Of 104 cows with follicular cysts as defined by milk progesterone assay result, 73 (70%) responded to the treatment with gonadotropin-releasing hormone, the milk progesterone concentration increasing from 0.7 +/- 0.2 ng/ml (mean +/- SD) to 1.8 +/- 1.1 ng/ml. The accuracy of rectal palpation 10 days after treatment for judgment of luteinization of follicular cyst confirmed by milk progesterone analysis was only 30% (48 cows of 160).     

Submucosal vestibulo-vaginal administration of cloprostenol (Oestrophan-Spofa) in cows in relation to estrus activity, conception, and the level of progesterone in milk

Trials were performed to examine the effectiveness of 250 micrograms (1 ml) of cloprostenol (Oestrophan Spofa) implantation under the mucous membrane of the vaginal vestibule of 128 cows with a clinically pronounced corpus luteum on ovaries. Within 72 hours from administration, oestrus was observed in 112 animals (85.5%). Out of the 97 cows inseminated, 63 cows (64.94%) got in calf. The effectiveness of the luteolytic action was examined on the basis of progesterone check in milk in 56 treated cows. The submucous implantation of cloprostenol rapidly degraded the function of the corpus luteum since from the original level of 14.78 ng/ml progesterone decreased to 0.87 ng/ml within 72 hours. However, luteolysis did not affect all the corpora lutea. Hence the submucous administration of cloprostenol was found to be effective, and at the same time, highly economical, owing to a substantial reduction in the costs of reproduction control as well as the costs of production.     

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.     

Elevated temperature (heat stress) in vitro reduces androstenedione and estradiol and increases progesterone secretion by follicular cells from bovine dominant follicles

Dairy cattle are susceptible to heat stress-induced reductions in fertility; however, direct effects of hyperthermia on specific reproductive functions are difficult to determine in vivo. The objective of this experiment was to examine the effect of elevated temperature in vitro on follicular steroidogenesis, to gain insight into specific follicular responses associated with heat stress. Dominant follicles were obtained from Holstein heifers on day 6 post-estrus (luteal phase; n = 4) or day 8, 36 h after an injection with 25 mg PGF(2alpha) to induce regression of the corpus luteum (follicular phase; n = 4). Pieces of follicle wall were isolated from dominant follicles and cultured for 96 h with 0, 2 or 100 ng/ml LH or FSH at 37, 39 or 41 degrees C. Concentrations of androstenedione, estradiol and progesterone were determined in culture media collected every 24h. During the last 48 h of culture, basal secretion of androstenedione and estradiol by pieces of follicle wall was lower at 41 degrees C than at 37 or 39 degrees C (P < 0.05). In contrast, cumulative secretion of progesterone by pieces of follicle wall in medium alone was higher at 41 degrees C than at 37 or 39 degrees C (P < 0.05). Pieces of follicle wall responded to treatment with both low (2 ng/ml) and high (100 ng/ml) doses of gonadotropins at all temperatures. However, gonadotropin-induced secretion of androstenedione and estradiol was generally lower, whereas gonadotropin-induced secretion of progesterone was higher at 41 degrees C and sometimes at 39 than at 37 degrees C. The changes in basal steroidogenesis and in responses to gonadotropins suggest that follicular cells begin to luteinize at elevated temperatures in vitro. Premature luteinization of follicular cells in vivo has been associated with reduced fertility in cattle with persistent follicles, suggesting that the premature differentiation of follicular cells observed in the current study may be responsible, in part, for the reduced fertility of dairy cattle under heat-stressed conditions.     

Ultrasonic imaging of equine ovarian follicles and corpora lutea

One of the most profound theriogenology applications of transrectal diagnostic ultrasonography in mares involves the imaging of ovarian follicles and corpora lutea. The resolving capabilities (frequency) and quality of the scanner directly affect the minimal size of a structure that can be imaged and the quality of the image. High-frequency scanners (5 or 7.5 MHz) of good quality can image a 2-mm follicle and the corpus luteum throughout its functional life. A low-frequency scanner (3 or 3.5 MHz) can image a 6-mm follicle and the corpus luteum for several days after ovulation. Equine follicles are excellent subjects for transrectal imaging because they are large, filled with fluid, and readily accessible. Event the small follicles (less than 10 mm) can be diagnostically important in evaluating whether ovarian infertility has occurred and whether the follicles are responding to treatment for follicular stimulation. The large, preovulatory follicles are of special interest. Averaged over a group of 79 periods, the following significant changes were found in the preovulatory follicle: increasing diameter, shape change from spherical to pear-shaped or conical, and increasing thickness of the follicular wall. No significant changes were found in the echogenicity (gray-scale value) of the wall or fluid. In retrospect, the diameter of the follicle seemed as useful for predicting impending ovulation as any of the other ultrasound criteria. The occurrence of ovulation is readily detected by the disappearance of a large follicle that was present at a recent previous examination. In addition, the ovulation site on the day of ovulation is detectable. In one study, the site was correctly identified in 24 of 24 mares. A small amount of residual follicular fluid can sometimes (7 of 10 in one study) be detected at the site of ovulation. The residual fluid usually disappears over a period of 0.5 to 20 hours. Subsequently, the developing corpus luteum may form a central nonechogenic area with peripheral luteinization or may remain uniformly luteinized. The central areas are of apparently vascular origin (blood or a component of blood) and become clotted and organized. In one study, approximately 50 per cent of the glands developed central areas exceeding 10 per cent of the size of the gland. The central areas began to develop on Day 0 or 1 and continued to enlarge until Day 2 or 3. The relative proportion of the gland containing a central clot decreases after Day 3, but the central area usually remains visible throughout diestrus.(ABSTRACT TRUNCATED AT 400 WORDS)     

Biochemical analysis of bovine follicular fluid: albumin, total protein, lysosomal enzymes, ions, steroids and ascorbic acid content in relation to follicular size, rank, atresia classification and day of estrous cycle

To investigate some biochemical changes during bovine follicle development, ovaries were obtained from cyclic heifers (7 to 11 heifers/d on each day of the 21-d estrous cycle; N = 152). Follicular fluid from the two largest follicles from both ovaries and a pool from small follicles (N = 30/cow) were collected from each animal and analyzed for ionic, enzymatic and endocrine changes in relation to day of the estrous cycle, follicle size, rank and atretic or growing status. Follicular fluid alkaline phosphatase activity and ascorbate concentrations were highest in all follicular sizes during the earlier portion of the estrous cycle (d 1 to 12; P less than .05), then decreased to the lowest levels (d 13 to 21). As follicular size (diameter) increased lactate dehydrogenase (LDH), acid and alkaline phosphatase activity was reduced in follicular fluid (P less than .05). Alkaline phosphatase and LDH activity tended to be increased in atretic follicles (P less than .10), and was correlated with increased progesterone and androgen concentrations of follicular fluid (r = .4, P less than .05). Both albumin and total protein concentrations decreased as follicular diameter increased (P less than .05). Sodium concentrations in follicular fluid were greater in growing-antral than atretic follicles, and increased with follicular enlargement (P less than .05). Follicular potassium concentrations increased as the estrous cycle progressed (P less than .05), and tended to be elevated in atretic follicles (nonsignificant). Both Ca and Mg concentrations increased with follicular enlargement (P less than .05). Dehydroepiandrosterone and testosterone were the predominant androgens in follicular fluid (androstenedione, the lowest concentration); their concentration decreased with follicle development (P less than .05), but were quite variable. Estradiol was increased in growing follicles (P less than .01). Estrone and estradiol concentrations increased as ovulation approached, particularly in small follicles (less than or equal to 4 mm diameter). Changes of biochemical components found in follicular fluid that relate to the growth and atresia process may provide a more sensitive and accurate method to classify follicle status, and thus aid in understanding the complexity of events associated with maturation of the bovine follicle and oocyte.     

Changes in the ovary and the peripheral concentrations of sex hormones after the aspiration of follicular fluid from the spontaneous follicular cysts of dairy cows

The aim of the present study was to clarify the ovarian and hormonal dynamics after the aspiration of follicular fluid in cows with follicular cysts. Follicular fluid was aspirated from the follicular cysts and follicles that were fated to become cystic follicles and other coexisting normal follicles, respectively, in lactating cows (n = 3). After the aspiration procedure, new follicles developed and reached a diameter of 25 mm without ovulation within 13–19 days. The plasma concentrations of inhibin decreased and follicle-stimulating hormone increased rapidly after the aspiration procedure, and subsequently increased and decreased, respectively, as a new follicle grew. No luteal structures developed after the aspiration procedure, and the animals’ plasma progesterone levels remained low. The present study indicates that the cystic follicles are never luteinized by the aspiration of follicular fluid, and consequently, new follicular cysts are observed to repeatedly develop.     

犬不同生殖周期阶段卵巢的组织结构观察

为了探讨犬卵巢组织结构和生殖周期阶段的相关性,试验对犬不同生殖周期阶段卵巢的外观形态和组织结构进行观察。结果表明,犬卵泡期、黄体期和乏情期卵巢体积分别为812.63、1081.80和446.03 mm³,黄体期高于卵泡期和乏情期(P＜0.05),卵泡期高于乏情期(P＜0.05);卵泡期、黄体期和乏情期卵巢质量分别为0.89、1.14和0.71 g,卵泡期低于黄体期且高于乏情期,但3者之间不存在显著性差异(P＞0.05);卵泡期卵巢中可见较多次级卵泡和少量成熟卵泡,黄体期卵巢中可见部分次级卵泡和闭锁卵泡,并有大量黄体存在,乏情期卵巢中卵泡类型主要以原始卵泡为主。可见,犬卵巢形态及组织结构与所处生殖周期阶段有关。     

湖羊不同发育阶段卵泡内代谢产物和激素含量的比较研究

本试验旨在研究代谢产物、代谢激素和生殖激素在湖羊黄体期不同发育卵泡内的变化。选用体质量40kg左右的湖羊11头,同期发情结束后第12天屠宰,按不同大小卵泡分离卵泡液。试验结果表明,与≤2.5mm卵泡相比,>2.5mm卵泡内的葡萄糖浓度显著提高(P<0.05),胰高血糖素浓度显著降低(P<0.05),乳酸脱氢酶(LDH)活性和睾酮浓度极显著降低(P<0.01),雌二醇浓度极显著提高(P<0.01),而血氨、游离脂肪酸、尿素、胰岛素和孕酮浓度差异不显著。雌二醇浓度与LDH活性呈极显著负相关(P<0.01),与葡萄糖浓度呈显著正相关(P<0.05),与胰高血糖素浓度呈显著负相关(P<0.05),与睾酮浓度呈极显著负相关(P<0.01),与孕酮浓度接近正相关(P=0.051)。试验结果表明代谢产物和激素共同参与调节卵泡发育。     

Changes in surface follicles and ovarian weight in testing for responsiveness in dairy cows after induction of superovulation using PMSG and cloprostenol

We evaluated the ovaries of 34 cows cross-bred of the Slovak Pied and Lowland Black-pied breeds which were culled and intended for slaughter during the winter type of feed rations. For superovulation treatment we used PMSG in the preparation Serum Gonadotropin (Bioveta, Nat. Ent., Ivanovice na Hané) and cloprostenol in the preparation Oestrophan inj. Spofa. We weighed the excised ovaries, fixed them in formalin 10% and made a quantitative evaluation of the surface follicles and differentiated them into recruited and selected or dominant follicles. We determined the concentration of 17 beta-estradiol and testosterone with the aid of the 3H RIA set from the firm Sorin after extraction by diethylether with separation of free and bound hormone with active charcoal. 72 hours after the giving of cloprostenol 43% reacted positively to superovulation treatment and after seven days a 50% positive response was recorded. After a dose of 2000 i. u. of PMSG (n = 6) embryo was obtained, whereas after 3000 i. u. of PMSG (n = 8) we flushed out eight embryos, of which four zygotes were suitable for transfer. After a higher dosage of PMSG there was an increase in the average weight of the ovaries, in right-hand ovaries significantly with P less than 0.05. After super-ovulation treatment the concentration of 17 beta-estradiol (E2) in the follicular fluid from follicles seven days after insemination was found to be so low as to be below the limit of detection, with the exception of four samples (mean = 8.60 nmol.l-1). The greatest concentration of E2 was from animals (n = 5) 72 hours after the giving of cloprostenol--1099.61 nmol.l-1) of follicular fluid. The concentration of testosterone was lower in the follicles of untreated cows in the follicular phase (mean = 5.92 nmol.l-1) compared with the follicles of super-ovulated animals the seventh day after insemination (mean = 14.12 nmol.l-1). The number of recruited and especially selected surface antral follicles 72 hours (n = 7) after the giving of cloprostenol and seven days (n = 8) after insemination was significantly higher in the group of brood cows reacting positively to superovulation in comparison with the animals which did not respond. It appears that the simultaneous monitoring of hormonal and morphological changes in the follicular system will help in objectivising the evaluation of the functional activity of stimulated follicles.     

Cell‐specific Distribution of Progesterone Receptors in the Bovine Ovary

This study describes the localization of progesterone receptors (PR) in the bovine ovary. Ovaries were obtained from 11 non‐pregnant and two pregnant cows. Progesterone receptors were visualized by immunohistochemistry on paraffin sections. Nuclear staining for PR was observed in cells of the follicles, corpora lutea, theca layers, surface epithelium, tunica albuginea, and in superficial and deep stroma cells. No staining was noticed in apoptotic bodies of atretic follicles. Expression of PR in follicle cells indicates an intrafollicular role of progesterone. The higher expression in thecal cells compared with follicle cells indicates that thecal cells mediate some effects of progesterone on the follicular development. Superficial stroma cells showing high expression might have a similar influence on primordial and primary follicles. In general, luteal cells had a lower expression than follicle cells, which may be explained by the down‐regulatory effect of locally produced progesterone. The lower expression in luteal cells during pregnancy can be due to the longer life span of this corpus luteum and concomitant degeneration of its PR. The high and rather constant expression of PR in cells of the surface epithelium remains to be elucidated.     

Effect of Dose of Cloprostenol on the Interval to Ovulation in the Diestrous Mare: A Retrospective Study

Although the ovulatory effects of prostaglandins are well documented in several domestic species including horses, there has been little attention paid to the use of this ovulatory effect for clinical purposes. Mares often grow large follicles during the luteal phase that may or may not ovulate before progesterone levels decline. Clinical observations of administering prostaglandins in diestrous mares with large follicles suggest that there may be a negative correlation between follicular diameter and interval from treatment to ovulation. The objectives of this study were twofold: to investigate the cloprostenol dose rate effect on interval to ovulation and to confirm the negative correlation between follicular diameter and interval to ovulation. The hypothesis tested was that high doses of cloprostenol given in diestrus to mares with larger follicles would induce ovulation more rapidly than in mares given lower doses or with smaller follicles. To test the hypothesis, a total of 1,234 estrous cycles were induced with different doses of cloprostenol (ranging from 8.75 to 625 μg). All mares had at least one follicle of 28 mm or larger. Dominant follicles were followed by transrectal ultrasound examinations every other day until ovulation was detected. There was a significant effect of dose (P < .000) and follicular diameter (P < .000) on the interval from treatment to ovulation. The shortest mean interval (2.4 days) was observed after administration of 625 μg in mares with follicles 36 mm or larger, whereas the longest (4.9 days) occurred after 8.75 μg in follicles of 28 to 31 mm.