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The generic GnRH agonist, Fertilan (goserelin), was tested for the ability to induce an LH surge and ovulation in estrus-synchronized gilts. Three experiments were performed to 1) examine the effect of various doses of Fertilan on secretion of LH in barrows, to select doses to investigate in gilts (Exp. 1); 2) determine doses of Fertilan that would induce a preovulatory-like rise of LH in gilts (Exp. 2); and 3) determine the time of ovulation after Fertilan treatment (Exp. 3). In Exp. 1, 10 barrows were injected on d 1, 4, 7, 10, and 13 with 10, 20, or 40 [micro]g of Fertilan; 50 [micro]g of Gonavet (depherelin; GnRH control) or saline (negative control); and sequential blood samples were collected for 480 min. There was a dose-dependent stimulation (P < 0.05) of LH release. Maximal plasma concentrations of LH ([LH.sub.MAX]) were 2.1 [+ or -] 0.2, 4.1 [+ or -] 0.3, 2.6 [+ or -] 0.4, and 3.4 [+ or -] 0.3 ng/mL after 10, 20, and 40 [micro]g of Fertilan and 50 [micro]g of Gonavet, respectively, and duration of release was 78 [+ or -] 9, 177 [+ or -] 12, 138 [+ or -] 7, and 180 [+ or -] 11 min, respectively. Fertilan doses of 10 and 20 [micro]g were deemed to be the most suitable for testing in gilts. In Exp. 2, 12 gilts received (after estrus synchronization with Regumate and eCG) injections of 10 or 20 [micro]g of Fertilan or 50 [micro]g of Gonavet 80 h after eCG to stimulate a preovulatory-like LH surge and ovulation. An LH surge was induced in 3 of the 4 gilts in both of the Fertilan groups and in all of the Gonavet-treated gilts. Characteristics of induced release of LH did not differ among groups: [LH.sub.MAX], 5.0 [+ or -] 0.9 vs. 4.6 [+ or -] 1.8 vs. 6.6 [+ or -] 1.1 ng/mL; duration, 11.7 [+ or -] 2.0 vs. 12.3 [+ or -] 2.2 vs. 14.3 [+ or -] 0.5 h; interval from GnRH injection to [LH.sub.MAX], 4.0 [+ or -] 2.0 vs. 6.7 [+ or -] 1.3 vs. 5.8 [+ or -] 1.6 h. In Exp. 3, estrus-synchronized gilts were injected with 20 [micro]g of Fertilan (n = 8) or 50 [micro]g of Gonavet (n = 4), and the time of ovulation was determined by repeated endoscopic examination. Time of ovulation ranged from 34 to 42 h postGnRH; however, ovulation occurred earlier in the Gonavet compared with the other groups (P < 0.05). Results of these experiments indicate that 1) barrows are an appropriate model for determining GnRH doses that can be effective in inducing a preovulatory-like LH surge in females; 2) the generic GnRH agonist Fertilan, at doses of 10 to 20 [micro]g, can stimulate an LH surge in gilts, with subsequent ovulation; and 3) Fertilan at doses of 10 and 20 [micro]g should be examined further for use in fixed-time insemination protocols. Key words: gonadotropin-releasing hormone agonist, luteinizing hormone, ovulation, pig

Contents The aim of this investigation was to examine the chromatin configuration of the nucleus, pattern of mitochondrial aggregation and mitochondrial activity in parallel studies in the same horse oocytes. Horse oocytes recovered by ultrasound-guided follicle aspiration in vivo were classified according to two main initial cumulus morphologies as having compact or expanded cumulus. The percentage of oocytes with a diplotene meiotic configuration at the time of recovery from the follicles was highest in compact oocytes. Oocytes with expanded cumulus layers at the time of recovery matured more rapidly in vitro and reached a proportion >50% at the metaphase II stage (M 2) sooner during in vitro maturation (IVM), than did compact oocytes. The mitochondrial aggregation pattern changed from finely distributed (Type 1) through crystalline (Type 2) to an aggregated, granulated appearance (Type 3) during IVM. The pattern of mitochondrial aggregation at the time of recovery was associated with the initial cumulus morphology of the oocyte, in that compact oocytes had a higher proportion of Type 1 aggregation, whereas expanded oocytes had a higher proportion of Type 3. The fluorescence intensity of metabolic active mitochondria, measured by fluorescence intensity (Em 570) per oocyte after MitoTracker CMTM Ros orange labelling, increased in the oocytes during IVM and depended on initial cumulus investment. Oocytes with the granulated type of aggregated mitochondria Type 3 had the highest level of metabolic activity and were in more progressed stages of meiosis (A 1–M 2). Oocytes initially having expanded layers of cumulus reached significantly higher levels of mitochondrial activity after IVM than did oocytes initially having compact cumuli. During resumption of meiosis the mitochondrial activity of oocytes with initially expanded cumulus increased continuously up to M 2, whereas in oocytes from compact cumulus–oocyte complex (COC), the activity declined after A 1/T 1 stages of meiosis. [ABSTRACT FROM AUTHOR]

Abstract: The aim of the study was to investigate the effect of the GnRH agonist Buserelin given on day 10 after ovulation on pregnancy rate and concentrations of progesterone and LH. Altogether 191 warmblood mares were used for two trials. Fresh or frozen/thawed semen from 27 stallions was used for A.I. In trial A 171 mares received either Buserelin (Receptal®, Hoechst, Germany, 40μg/animal) or 10ml 0.9% NaCl (placebo). On day 16 after A.I. pregnancy diagnosis was performed by ultrasound scanning of the uterus. For statistical analysis, data were analyzed by a mixed model, with four fixed factors (treatment, type of spermatozoa, A.I. number, reproductive status of the mare) and a random factor (stallion). Least Square Means (LSM) for pregnancy rate were 46.0% in GnRH agonist treated mares and 36.4% in the control group (P =0.22). In trial B 20 lactating and cycling mares were used for endocrine studies. Blood samples were recovered for analyses of progesterone and LH from days 0 to 11. The mean progesterone concentrations increased continuously from days 0 to 8 after ovulation in both groups (GnRH group: from 0.81±0.48 to 5.47±0.48ng/ml, control group: from 0.63±0.68 to 5.83±0.68ng/ml). Moreover, the progesterone concentrations from days 9 to 11 were not different between the GnRH and the control group. In contrast to this LH concentrations were markedly influenced by the GnRH agonist. On day 10 LH concentrations were significantly higher in GnRH agonist treated than in placebo treated animals. From the data obtained from individual animals it can be concluded that GnRH agonist, given during luteal phase may have different effect on luteal function. [Copyright &y& Elsevier]

Contents In many species, large numbers of macromolecules are accumulated during oocyte growth. The messenger and ribosomal RNAs produced in these cells are far in excess of those necessary to support protein synthesis. Thus, the aim of this study was to elucidate the processes of translational regulation during meiotic maturation. The relationship between transcription, translation and polyadenylation of mRNA during in-vitro maturation (IVM) of bovine oocytes was investigated. The results presented here show that overall protein synthesis is stimulated during meiotic maturation (approximately three times) concomitantly with the onset of germinal vesicle breakdown after 6 to 10 h of IVM. However, in metaphase II, the incorporation of [35 S]methionine into proteins showed only basal levels, as in the germinal vesicle (GV) stage. Furthermore, in the course of IVM, de-novo transcription strongly declines as determined by measuring the incorporation of [3 H]uridine into RNA. In contrast to this finding, the incorporation of [3 H]adenosine increased and showed a peak during the time interval from 6 to 10 h of IVM, parallel with the onset of germinal vesicle breakdown (GVBD) and translation. In the further course of maturation, only a moderate decrease of [3 H]adenosine incorporation was observed. These results indicate that (i) translation increased at the time of GVBD; (ii) these processes were accompanied by polyadenylation of mRNA; and (iii) although transcription declines, polyadenylated mRNA is accumulated until metaphase II (as shown by poly(U)-hybridization), in which protein synthesis is low. The correlation of these processes is discussed here. A detailed knowledge of the biochemical and molecular processes which occur during oocyte maturation can be useful for the improvement of IVM conditions. [ABSTRACT FROM AUTHOR]

Summary The object of this study was to compare 4 different methods of oocyte recovery from mares; 1) transvaginal follicle aspiration in vivo; 2) follicle aspiration in vitro; 3) oocyte recovery by isolation of follicles in vitro and 4) follicle scraping in vitro. Oocyte recovery was highest after follicle scraping (71.1%) and follicle isolation and rupture (61.3%). Follicle aspiration in vitro and in vivo yielded oocytes on 31.2% and 19.3% of occasions, respectively. The output of different types of cumulus-oocyte-complexes was different among the methods; the portion of compact cumulus-oocyte-complexes was significantly higher with follicle scraping (50.7%) and follicle isolation (44.5%) than with aspiration in vivo (31.9%) and in vitro (23.7%). The recovery rate of oocytes from small follicles (<15 mm) was significantly higher than from larger follicles (P < 0.05) using transvaginal follicle aspiration. The proportion of oocytes that were degenerate (exhibited shrunken, dense or visibly damaged ooplasm) ranged from 1.2% after follicle scraping, to 17.2% after aspiration in vivo. These results indicate that, for the recovery of horse oocytes in vitro, follicle scraping and follicle isolation give the highest recoveries of cumulus-intact oocytes. [ABSTRACT FROM AUTHOR]

Contents The mechanism of implantation is species specific (pig: epitheliochorial, bovine: synepitheliochorial, mouse: hemochorial). Recently, we have shown that proteolytical cleavage of the prototypical 25 kDa mRNA cap-binding protein eIF4E (eukaryotic initiation factor 4E) produces a stable variant with a molecular mass of approximately 23 kDa in porcine endometrium at the time of implantation. Here, we investigate if an eIF4E truncation also takes place in the endometrium of species with other implantation forms. Thus, eIF4E and its repressor protein 4E-BP1 were investigated in porcine, murine and bovine endometrium during the time of implantation. Our results show that eIF4E truncation is specific for the porcine implantation. In bovine and mouse uterine tissue, no cleavage of eIF4E was observed. Whereas no difference of bovine 4E-BP1 was found, in murine samples, increased phosphorylation during implantation was observed. However, porcine samples exhibit an opposite behaviour, the abundance and mainly the phosphorylation of 4E-BP1 decrease. We propose that the translation initiation in the endometrium is differently regulated by the two eIF4E forms with regard to different 4E-BP1 abundance and phosphorylation as well as different eIF4E/4E-BP1 binding dynamic depending on the type of implantation. [ABSTRACT FROM AUTHOR]

Contents Three experiments were conducted to determine whether follicular fluid (FF) enters the oviduct and plays any role in the transport of oocytes into the oviduct. Experiment 1: Oestrus and ovulation were synchronized in cycling gilts (n = 21) over a 15 day period of feeding Regumate® and injections of 1000 IU pregnant mare’s serum gonadotrophin (PMSG) 24 h after the last Regumate® feed and 500 IU human chorionic gonadotrophin (hCG) 80 h after PMSG. Ipsi-lateral aspiration of FF and salpingectomy (group 1, n = 7), aspiration of FF without salpingectomy (group 2, n = 7) or ligation of the oviduct between the ampulla and infundibulum (group 3, n = 7) was performed endoscopically prior to ovulation (34–36 h after hCG). Ipsi-lateral (group 2 and 3) and contra-lateral salpingectomy was carried out in all gilts post ovulation, 42–44 h after hCG. The oviducts were flushed with 1 ml saline and the samples as well as the aspirated FF were analysed for progesterone and estradiol by RIA methods. In group 1 both progesterone and estradiol concentrations did not differ before and after ovulation. Withdrawal of FF from the ipsi-lateral ovary by aspiration (group 2) or ligation of the oviduct (group 3) did not influence the steroid content within the oviducts. Similarly low progesterone concentrations were measured in ipsi- and contra-lateral oviducts after ovulation (group 2: 0.29 ± 0.17 versus 0.24 ± 0.35 ng/ml and group 3: 0.22 ± 0.19 versus 0.21 ± 0.22 ng/ml). The high content of progesterone of FF (269.7 ± 67.9 and 389.6 ± 226.5 ng/ml in group 1 and 2, respectively) was not reflected in the oviductal fluid. Experiment 2: In five gilts 0.06 ml 3H-progesterone (30 000 dpm) were applied via a fine 27 G injection needle into the largest three follicles of the ipsi-lateral ovary prior to ovulation (34–36 h after hCG). The oviducts were flushed following... [ABSTRACT FROM AUTHOR]

This study was conducted to investigate the potential for increased oxidative stress of high- vs. average-producing dairy cows. Two experiments were performed using 11 and 13 Holstein cows (53 ± 2 d postpartum). Lipohydroperoxides (LHP) were determined in serum lipids (experiment 1) and low-density lipoprotein (experiment 2) via oxidation of ferrous to ferric ions through LHP using thiocyanate as chromogen. In experiment 1, differing milk yield and milk energy output corresponded to different concentrations of LHP. In experiment 2, analysis of regression resulted in a significant relationship between milk yield and LHP. Phospholipids isolated from lipids with 6.5 µM of LHP evoked in monocytic cells a transient increase in super-oxide formation, indicating inflammatory potential. The results show that high milk productivity can associate with oxidative stress indicated by oxidative modifications of circulating lipids and their changed bioactivity. [ABSTRACT FROM AUTHOR]

Abstract: The present study was conducted to assess effects of the gonadotropin-releasing hormone agonist (GnRHa) triptorelin in dairy heifers. The peptide was released from a commercial 4-week depot formulation (Decapeptyl®Depot) administered at animals’ estrus (day 0). First experiment (EXP I, n =5), which was aimed to explore the availability of peptide, detected a maximum of triptorelin concentration between day 2 and 5 after depot injection, and the peptide remained detectable by RIA in peripheral blood for about 3 weeks. In further experiments, the peptide release was terminated on day 9 (EXP II, n =16) or day 21 (EXP III, n =47). Treatment effects were studied on follicular development, the characteristics of cumulus-oocyte complexes (COCs) (EXP II; EXP IIIa) and secretions of LH and progesterone (EXP IIIb). Results showed that the occurrence of the pre-ovulatory LH surge was more uniform in treated heifers than that in controls. The duration of ovulation periods was similar amongst the heifers of EXP II, but more compact amongst those of EXP III each compared with the respective controls. Post-ovulatory, the number of LH pulses was significantly reduced by treatment, whereas both basal LH and progesterone concentrations were elevated on a few days. Follicular growth was reduced only by the prolonged influence of the GnRHa. There were increased proportions of both degenerated COCs and immature oocytes from small follicles (<3mm in diameter), and meiotic configuration and quality of oocytes isolated from follicles 3–5mm were changed after the prolonged, 21-day treatment. These results indicate that a continuous influence of a GnRHa over more than 1 week may increasingly impair the development of bovine follicles and oocytes. This may have some significance for the development of novel GnRH-based techniques in regulating the reproductive function in cattle. [Copyright &y& Elsevier]