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Mitochondrial function and mitochondria-induced apoptosis in an overstimulated rat ovarian cycle.

Female rats were treated with FSH (40 IU/kg) on the first and second diestrus days (D1 and D2) and with LH (40 IU/kg) on the proestrus (P) day to synchronize and maximize ovarian changes. Follicle area increased by 50% from D1 to P, and the estrus (E) phase showed multiple corpora lutea and massive apoptosis. Increased oxygen uptakes (42-102%) were determined in ovary slices and in isolated mitochondria in active state 3 along the proliferation phase (D1-D2-P) that returned to initial values in the E phase. Mitochondrial content and the electron transfer activities of complexes I and IV were also maximal in the P phase (20-79% higher than in D1). Production of NO by mitochondrial nitric oxide synthase (mtNOS), biochemically determined, and the mtNOS functional activity in regulating state 3 oxygen uptake were also maximal at P and 79-88% higher than at D1. The moderately increased rate of NO in the proliferative phase is associated with mitochondrial biogenesis, whereas the high rate of NO generation by mtNOS at phase P appears to trigger mitochondria-dependent apoptosis. The calculated fraction of ovary mitochondria in state 3 was at a minimal value at the P phase. Mitochondrial oxidative damage, with increased thiobarbituric acid-reactive substances and protein carbonyls, indicates progressive mitochondrial dysfunction between phases P and E. The roles of mitochondria as ATP provider, as a source of NO to signal for mitochondrial proliferation and mitochondria-dependent apoptosis, and as a source of O(2)(-) and H(2)O(2) appear well adapted to serve the proliferation-apoptosis sequence of the ovarian cycle.

Animals↗

[Catecholamine blood levels in sheep during superovulation in estrus].

The effects of a serum gonadotropin (SG) superovulation hormonal preparation were investigated on catecholamine levels (norepinephrine, dopamine and epinephrine) in the blood plasma of ewes with synchronized oestrus in the oestrus period. In this trial the blood plasma of eleven ewes of the Slovak Merino breed was analyzed to detect catecholamines in the oestrus period. Superovulation was induced by an i.m. administration of 1500 IU SG as soon as oestrus synchronization with Agelin vaginal tampons finished (20 mg chlorsuperlutin) which lasted for 10 days. Catecholamines were detected in the blood plasma before synchronization, on the day of Agelin vaginal tampons application, and in 48 and 72 hours after the hormone administration--on the days of the expected ovulation. Catecholamine concentrations in the blood plasma were determined by a radioenzymatic assay using a Catechola test (Praha). The results indicate that synchronization and hormonal stimulation influence plasma catecholamine levels. The norepinephrine (NE) concentration in the blood plasma of the control samples has the value of 8.31 +/- 0.732 pmol/ml. An insignificant increase in the NE levels (13.12 +/- 0.120 pmol/ml; Fig. 1) was recorded on day 1 of the trial, after the start of synchronization. On the day of the expected ovulation the NE concentrations rose to 17.12 +/- 1.289 pmol/ml (P less than 0.001) and they remained significantly increased (P less than 0.01) at the level of 12.89 +/- 1.020 pmol/ml on the following day of the experiment. The dopamine level (DA) in the plasma of a control sample is 6.42 +/- 0.350 pmol/ml (Fig. 2).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of placement of intravaginal sponges on LH, FSH, estrus and ovarian activity in mares during the nonbreeding season.

Eight seasonally anestrous mares were administered intravaginal polyurethane sponges on December 15 and then weekly thereafter until February 1. Control mares received no sponges or genital contact. Sponge insertion caused an immediate surge in follicle-stimulating hormone (FSH) concentrations in jugular plasma in 50% of treated mares whereas no control mares had surges in FSH (P less than .05). The effect of treatment on luteinizing hormone (LH) concentrations was much less dramatic and only three treated mares appeared to have positive responses. Sponge-treated mares exhibited positive responses in FSH concentrations 11 times out of 32 mare-days and control mares zero out of 28 (P less than .05). The magnitude of the FSH response decreased rapidly with successive responses. Sponge insertion induced estrus in four of eight treated mares; no control mares exhibited estrus (P less than .05). Sponge insertion also increased ovarian size and the incidence of large follicles. When all mares were fed altrenogest for 14 d beginning February 1, there was no beneficial effect of sponge treatment on number of mares exhibiting estrus or on pregnancy rate. These data confirm earlier speculations that sponge treatment causes surges in gonadotropins and increased ovarian size in approximately 50% of anestrous mares. However, sponge treatment does not appear to provide a practical means of preparing mares for progestogen synchronization during the nonbreeding season.

Animals↗

Reproductive performance of dairy cows with ovarian cysts after different GnRH and cloprostenol treatments.

Cystic ovarian disease is an important cause of reproductive failure and economic loss for the dairy industry. This report describes two consecutive studies. The objective of the first was to evaluate the response of cows with ovarian cysts to two therapeutic treatments. In the second study, we compared the effectiveness of the best treatment established in Study 1 with that of the Ovsynch protocol. For Study 1, cows were considered to have an ovarian cyst if it was possible to observe a single follicular structure with a follicular antrum diameter > 25 min in the absence of a corpus luteum in three ultrasonographic examinations performed at 7 days intervals. At diagnosis (Day 0), cows were assigned to one of two treatment groups. Cows in Group GnRH/CLP (n = 31) were treated with 100 microg GnRH i.m. and 500 microg cloprostenol (CLP) i.m. on Day 14. Cows in Group GnRH-CLP/CLP(n = 32) were treated with 100 microg GnRH i.m. plus 500 microg CLP i.m. on Day 0, and 500 microg CLP i.m. on Day 14. The animals were inseminated at observed estrus and monitored weekly by ultrasonography for 4 weeks or until Al. Cows in the GnRH-CLP/CLP group showed a lower cystic persistence rate (15.6% < 45.2%; P = 0.01); a higher estrus detection rate (84.4% > 41.9%; P < 0.0001); a higher ovulation rate (75% versus 32.3%; P < 0.0001) and a higher early response rate (31% > 3%; P = 0.02) than those in the GnRH/CLP group. For the second study, 128 cows with ovarian cysts were randomly assigned to one of two treatment groups: cows in Group Ovsynch (n = 64) were treated with 100 microg GnRH i.m. on Day 0, 500 microg CLP on Day 7, and 100 microm GnRH i.m. 36 h later. Cows in this group were inseminated 24 h after the second GnRH dose (Ovsynch protocol). Cows in Group GnRH-CLP/CLP/GnRH (n = 64)were treated as those in the GnRH-CLP/CLP group of Study 1 but received GnRH 32 h after the second CLP treatment and were inseminated 24 h after this. A further group of cows without ovarian cysts inseminated at natural estrus served as the Group Control (n = 64). Cows in the GnRH-CLP/CLP/ GnRH group showed a lower cystic persistence rate (10.9% < 46.9%; P < 0.0001); higher ovulation rate (79.7% > 17.2%; P < 0.0001); higher return to estrus rate (34.3% > 12.5%; P < 0.01) and higher pregnancy rate (28.1% > 3.1%; P < 0.01) than those in Ovsynch; and a similar pregnancy rate (28.1% versus 35.9%) to Control cows. These findings indicate that lactating cows with ovarian cysts can be successfully synchronized and time inseminated using a protocol that combines GnRH and CLP, starting treatment by simultaneously administering both products. This protocol also allows the insemination of cows showing estrus within the first week of treatment. Ovarian cysts were less responsive when treatment was started with GnRH alone.

Animals↗

Role of corticosteroids in female reproduction.

Corticosteroids, ACTH, and stress can exert inhibitory and facilitory effects on reproduction. The purpose of this review is to reconcile the divergent effects of corticosteroids on gonadotropin secretion based on recent work in the area. Whether stimulation or inhibition of gonadotropin secretion is observed appears to depend on two important variables: 1) length of exposure, and 2) background of estrogen priming. The acute administration of ACTH and certain corticosteroids to estrogen-primed animals brings about the release of LH and FSH. Corticosteroids have also been shown by some investigators to cause selective release of follicle-stimulating hormone (FSH) both in vitro and in vivo. This selective facilitation of FSH release by corticosteroids may explain many deleterious effects on reproduction observed after adrenalectomy, and it may have relevance in explaining the beneficial effects of corticosteroids in inducing ovulation in anovulatory patients suffering from polycystic ovarian syndrome. Finally, evidence is presented which suggests that adrenal steroids may participate in initiation and synchronization of the preovulatory LH and FSH surge, as well as the secondary FSH surge seen on estrus in the rat.

Adrenal Cortex Hormones↗

Direction of uterine contractions during estrus in ewes: a reevaluation.

In four ewes direction of propagation of uterine contractions was evaluated using an electromyographic technique during 15 entire estrous periods; 120,101 propagations were analyzed. When horns were considered separately 89.9% of all propagations were classified into three main modes: ascending (34.3%), descending (59.85%), and divergent (5.85%). When both horns were analyzed simultaneously, horn's synchronicity was observed in most instances; on this basis, eight modes of propagation were identified of which three accounted for two-thirds of all propagation: synchronous descending (24.9%), reciprocal propagation (descending on one horn and then ascending on contralateral horn) (22.1%), and isolated descending propagation (18.0%). A time-dependent pattern of propagation throughout estrus was clearly identified, the percentage of ascending propagations reaching a minimum (16.5%) and the percentage of descending propagation reaching a maximum (77%) at peak uterine motility level. By considering both direction of uterine propagation and cervical mechanical activity, a new hypothesis concerning two aspects of sperm transport (speed and mechanism) was formulated. It is hypothesized that the high prevalence of descending propagations is important to reduce sperm cell population by selecting the most vigorous spermatozoa; such selection is possible when the mechanical cervical activity is low or absent (cervix open); when cervical mechanical activity is high (cervix closed), it is suggested that both descending and ascending propagations participate in sperm transport by back and forth motion of luminal fluid within the uterine lumen.

Animals↗

Fertility of fresh and frozen-thawed goat semen during the nonbreeding season.

A total of 4109 does of a local Greek breed (Capra prisca) were synchronized with intravaginal MPA-sponges and PMSG, and 24 bucks of Alpine (n = 8), Saanen (n = 8) and Damascus (n = 8) breeds were used for studying the fertility of nonfrozen and frozen-thawed semen during the nonbreeding season (June to August). Artificial insemination (AI) was performed once (50 to 55 h after sponge withdrawal) or twice (36 and 60 h after sponge withdrawals with fresh semen (collected during the nonbreeding season, stored at 16 degrees C and inseminated within 6 h) or frozen semen (prepared from the same bucks during the preceding breeding season). The induction of estrus was successful, varying between 91.0 and 95.0%. The form of semen (fresh or frozen-thawed used for inseminating the synchronized does affected their fertility: the overall kidding rate with fresh semen (65.5%) was higher (P < 0.05) than that with frozen-thawed semen (53.4%). The fertility level was also affected by the number of inseminations performed: the overall kidding rate was significantly higher (P < 0.001) in the does inseminated twice with fresh or frozen-thawed semen (70.4 and 59. 1%, respectively) than in those inseminated only once (48.9 and 44.9%, respectively). Finally, the breed of the buck used for preparing the fresh or the frozen-thawed semen affected the fertility level of the does. The kidding rate was higher in does inseminated with fresh semen prepared from bucks of the Damascus breed than from bucks of Saanen or Alpine breed. However, when frozen-thawed semen was used the kidding rate was lower in does inseminated with semen prepared from bucks of the Damascus breed than from bucks of the Alpine or Saanen breed. It is concluded that the fresh semen of Alpine, Saanen and Damascus breed bucks, born and raised under the climate conditions prevailing in Greece (34 degrees to 41 degrees N), can be used successfully during the nonbreeding season (June to August) for inseminating does.

Journal Article↗

Relationship between size of the ovulatory follicle and pregnancy success in beef heifers.

Previous research indicated that the size of the ovulatory follicle at the time of insemination significantly influenced pregnancy rates and embryonic/fetal mortality after fixed-timed AI in postpartum cows, but no effect on pregnancy rates was detected when cows ovulated spontaneously. Our objective was to evaluate relationships of fertility and embryonic/fetal mortality with preovulatory follicle size and circulating concentrations of estradiol after induced or spontaneous ovulation in beef heifers. Heifers were inseminated in 1 of 2 breeding groups: (1) timed insemination after an estrous synchronization and induced ovulation protocol (TAI n = 98); or (2) AI approximately 12 h after detection in standing estrus by electronic mount detectors during a 23-d breeding season (spontaneous ovulation; n = 110). Ovulatory follicle size at time of AI and pregnancy status 27, 41, 55, and 68 d after timed AI (d 0) were determined by transrectal ultrasonography. Only 6 heifers experienced late embryonic or early fetal mortality. Interactions between breeding groups and follicle size did not affect pregnancy rate (P = 0.13). Pooled across breeding groups, logistic regression of pregnancy rate on follicle size was curvilinear (P < 0.01) and indicated a predicted maximum pregnancy rate of 68.0 +/- 4.9% at a follicle size of 12.8 mm. Ovulation of follicles < 10.7 mm or > 15.7 mm was less likely (P < 0.05) to support pregnancy than follicles that were 12.8 mm. Ovulatory follicles < 10.7 mm were more prevalent (28% of heifers) than ovulatory follicles > 15.7 mm (4%). Heifers exhibiting standing estrus within 24 h of timed AI had greater (P < 0.01) follicle diameter (12.2 +/- 0.2 mm vs. 11.1 +/- 0.3 mm) and concentrations of estradiol (9.9 +/- 0.6 vs. 6.6 +/- 0.7) and pregnancy rates (63% vs. 20%) than contemporaries that did not exhibit behavioral estrus. However, when differences in ovulatory follicle size were accounted for, pregnancy rates were independent of expression of behavioral estrus or circulating concentration of estradiol. Therefore, the effects of serum concentrations of estradiol and behavioral estrus on pregnancy rate appear to be mediated through ovulatory follicle size, and management practices that optimize ovulatory follicle size may improve fertility.

Animals↗

Induction of estrus in cows by a new analogue of PGF2 alpha (alfaprostol).

A new analogue of PGF2 alpha, alfaprostol, was used to induce estrus in normally cycling and anestrous cows. Five groups of six animals were treated with single injections of different doses of the drug (4, 5, 6, 7 or 8 mg/animal) on day 10-11 of the estrous cycle. Six cows in anestrus, with palpable corpora lutea, were treated i.m. with 8 mg/animal. The luteolytic properties of alfaprostol were assessed by measuring milk progesterone by RIA for 96 h after treatment and by checking for the subsequent signs of estrus. The dose of 4 mg caused a slight fall in progesterone but did not induce heat; the 5 and 6 mg doses induced estrus in 3/6 animals and the 7 mg dose was effective in 4/6 animals. The 8 mg dose produced a sharp reduction in milk progesterone within 24 h, followed by estrus in both the normally cycling and the anestrous groups in 6/6 animals. No clinical signs of drug intolerance were seen. The present results show that alfaprostol might be of great use for both cycle synchronization in normally cycling cows and for treatment of anestrus caused by a persistent corpus luteum.

Anestrus↗

Reproductive response in supplemented heifers in the humid tropics of Costa Rica.

To evaluate the effect of two supplementary diets to determine the consequence on productive and reproductive performance in heifers (Bos indicus x Bos taurus) averaging between 24 and 36 months of age and grazing tropical pastures, two trials were conducted. Thirty animals (initial BW 325.1+/-33.6 kg) were divided in two groups in the initial study: supplemented (SG) and control (CG); SG received a concentrate (5.5% CP and 2.85 Mcal/kg of DE dry matter basis) at 1% of body weight (BW). In the second study, 45 heifers (initial BW 332.6+/-29.3 kg) were assigned in two treatments, with the same amount of supplement (1% BW) but with a greater nutrient content (13% CP and 3.15 Mcal/kg of DE). The proportion of animals with a corpus luteum at the end of each study was greater in the supplemented groups (P<0.05). Ovarian follicular dynamics was similar between groups in the first study, but in the second study there were more heifers in the SG group with follicles larger than 9 mm in diameter (P<0.05). Pregnancy rate was similar for SG and CG (P>0.05). The response to a regimen of estrous synchronization in both trials was numerically superior in the SG group. No differences were observed in the length of estrus. Daily gain and body condition score were similar for supplemented and control groups (0.27 compared with -0.06 in the first study and 0.90 kg compared with 0.60 in the second study, respectively). Dietary supplementation improved the number of animals initiating estrous cycles and the expression of estrus when compared with unsupplemented control heifers. The dietary regimens imposed in these studies appear to be an adequate for the management of growing heifers destined to a reproductive program.

Animal Nutritional Physiological Phenomena↗