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J F Roche

Publications and source records attributed to J F Roche.

At least 55 records · Page 3Linked to original sources

Superovulation in cattle: effect of FSH type and method of administration on follicular growth, ovulatory response and endocrine patterns.

Although different FSH preparations and injection regimens are used to superovulate cattle, the optimum treatment regimen and blood concentrations of FSH to induce effective superovulatory responses are currently not known. The current objective was to evaluate the pattern of follicular growth, oestradiol-17 beta(E2) concentrations and yield of embryos in heifers following superovulation with two different pFSH preparations reportedly differing in LH content. In experiment 1, 90 synchronised heifers were superovulated at mid-cycle using a 2 x 2 factorial design comparing Folltropin (Vetrepharm; low LH) with Pluset (Serovet; FSH:LH ratio 1:1) administered either as a single or multiple (8 for Folltropin, MF and 10 for Pluset, MP) injections. Animals were inseminated during oestrus which was induced with prostaglandin F2 alpha analogue and embryos were recovered 7 days later. Overall, Pluset treatments compared with Folltropin resulted in more ovulations and unfertilized or degenerate embryos (P < 0.05). Multiple injections resulted in more (P < 0.05) freezable (MF = 55 +/- 1.2; MP = 3.8 +/- 1.0) and transferable embryos (MF = 2.68 +/- 0.9; MP = 2.71 +/- 0.9) than single injections (SF = 2.2 +/- 0.5 and 1.0 +/- 0.3 respectively; SP = 2.6 +/- 0.8 and 1.3 +/- 0.4 respectively); there was also a higher (P < 0.05) percentage embryo recovery rate. In two subsequent experiments, animals (n = 17) were superovulated with either single or multiple injections of Folltropin or Pluset as described and blood samples were collected and analysed for E2 concentrations. Ovarian scanning was carried out until 72 h after the first FSH injection, to count medium (5-9 mm) and large (> or = 10 mm) follicles. Heifers treated with SP had higher E2 concentrations in comparison with heifers treated with SF at 18, 36-48 and 84-96 h after the FSH injection. There were no differences in E2 concentrations in heifers treated with MF or MP treatments. Heifers treated with SP had greater numbers of follicles compared to SF treated heifers (21.0 +/- 3.1 vs 13.9 +/- 2.2; P = 0.089) on the third day after FSH injection. There were no differences between the numbers of medium and large follicles in heifers treated with MF or MP at any time throughout the experimental period. These data indicate that a single injection of Folltropin or Pluset can result in multiple ovulations and that the E2 profiles are different following single injections of either Folltropin or Pluset.

Animals↗

Validation of a sensitive radioimmunoassay to measure serum follicle-stimulating hormone in cattle: correlation with biological activity.

Meaningful biological interpretation of the role of follicle-stimulating hormone (FSH) requires use of a validated radioimmunoassay (RIA) that closely estimates biologically active FSH, which was the objective of this work. Three FSH antibodies [NIDDK anti ovine FSH (oFSH); JAD anti oFSH; USDA anti bFSH] were screened against three tracer preparations [USDA oFSH-19-SIAFP-I2(USDA oFSH I2); LER1976a oFSH; USDA bFSH I2] in a RIA using USDA bFSH B1 or I2 as the assay standard. Sera obtained from three heifers at 4- to 8-h intervals for 5 days after injection of PGF2 alpha during the luteal phase were assayed for both FSH immunoactivity using each of the three optimized assay formats (NIDDK anti oFSH and JAD anti oFSH with USDA oFSH I2 as tracer; USDA anti bFSH with USDA bFSH I2 as tracer), and FSH bioactivity, using a rat Sertoli cell bioassay. Cross reactivity of bLH (NIH bLH B9) in all three assay formats was minimal (0.7, 0.9 and < 0.4% at 50% binding for the NIDDK, JAD and USDA antibodies, respectively). There was parallel displacement of tracer between bovine serum dilutions of 10 to 500 microliters and the two FSH standards. Correlations between JAD and USDA RIA data and bioassay results were not significant (P > or = 0.10), but were significant (r = 0.78; P = 0.0001) for the NIDDK RIA FSH and the bioactive FSH measurements. The assay sensitivity of the NIDDK RIA was 0.55 ng USDA bFSH B1 (0.013 ng USDA bFSH I2)/ml. The inter- and intra-assay CV were between 5.8 and 7.9 %. This RIA detected a pre-ovulatory FSH surge coincident with the LH surge, in all heifers studied. Furthermore, the emergence of each wave of follicle growth (up to day 12 of the cycle), was preceded by a transient increase (P < 0.02; days 0.5 to 1.5 and 8 to 10.5 of the cycle) in serum FSH, while LH concentrations remained unchanged. In conclusion, the RIA utilising NIDDK anti oFSH and USDA oFSH I2 as tracer provides a good estimate of bioactive FSH in cattle, and detects physiologically relevant increases in serum FSH related to emergency of each new wave of follicle growth.

Animals↗

Influence of previous photoperiodic exposure on the reproductive response to a specific photoperiod signal in ewes.

Two experiments were carried out to determine whether the reproductive response of ewes to a specific photoperiodic signal depends on the time of year that the signal is given, and, if so, whether this dependence can be attributed to the photoperiodic history of the ewes. The aim of experiment 1 was to expand upon previous findings that the reproductive response to a specific photoperiodic challenge in ewes previously maintained on natural photoperiod varies with time of year. Ewes were transferred at one of three times of year from natural photoperiod to photochambers and were immediately exposed to 35 long days (18L:6D) followed by continuous exposure to short days (8.5L: 15.5D); this treatment is referred to as LD-->SD. The three times of year when long days started corresponded to the beginning of the breeding season, the mid-breeding season, and early anestrus (September 21, December 21, March 21, respectively). In ewes exposed to LD-->SD beginning in September, the breeding season and subsequent anestrous season was not altered. In ewes exposed to LD-->SD beginning in December, the transition to anestrus was advanced (p < 0.05) relative to that in controls maintained in simulated natural photoperiod. Subsequently, half of these ewes resumed reproductive activity within 180 days; this occurred 131 +/- 8 days after transfer to short days. In contrast, all ewes exposed to LD-->SD beginning in March resumed reproductive activity; this began 100 +/- 3 days after transfer to short days (p < 0.05 versus December group). The purpose of experiment 2 was to assess the extent to which the difference in response to a photoperiodic challenge can be attributed to photoperiodic history. Ewes were maintained on short days from the winter solstice interrupted with 35 long days from March 21, June 21, September 21, or December 21. The majority of ewes exhibited an onset of reproductive activity after exposure to LD-->SD at the different times of year, and there was no group difference in latency to onset of reproductive activity. The duration of reproductive activity, however, was longer (p < 0.05) in ewes exposed to LD-->SD beginning in June than in the other groups. Thus we conclude that the seasonal difference in the ability of the photoperiodic challenge of long followed by short days to induce reproductive activity in ewes previously maintained outdoors can be attributed, in large measure, to photoperiodic history. Other factors, such as phase of the endogenous rhythm, however, may influence the duration of reproductive activity resulting from this photoperiodic challenge.

Analysis of Variance↗

Decline in serum follicle-stimulating hormone concentrations alters key intrafollicular growth factors involved in selection of the dominant follicle in heifers.

Declining FSH after a transient rise coincides with selection of a dominant follicle (DF) and atresia of the remaining cohort follicles (subordinates) in cattle. The objectives of this study were to determine 1) whether intrafollicular amounts of inhibins, activin-A, insulin-like growth factor I (IGF-I), and IGF-I-binding proteins (IGFBP) are altered during selection of the first-wave dominant follicle (DF1) and 2) whether these biochemical markers are FSH dependent. Beef heifers received six or eight 6-h injections of saline (controls) or eight 6-h injections of recombinant bovine FSH (1 mg/injection) at 38 to 42 h after estrus (Day 0). Daily ultrasound scanning was used to define selection of DF1. Controls (n = 6 per group) were ovariectomized 1) on Day 3 of the estrous cycle before DF1 selection (preselection follicles) and 2) after DF1 selection on Day 4.8 +/- 0.5. In controls, FSH declined between Days 2 and 3 and selection of DF1 occurred between Days 3 and 5. During this interval, intrafollicular estradiol concentrations increased > 5-fold in DF1, yet declined 4-fold in subordinates (p < 0.05). In DF1, total IGF-I increased 1.3-fold (p < 0.05), whereas the amounts of the 40- to 47-kDa and the 35-kDa IGFBP (ligand hybridization) decreased 2.4- and 2.5-fold, respectively (p < 0.05), compared to values in preselection follicles on Day 3; total dimeric inhibin-A decreased 1.8-fold (p < 0.05). In contrast, amounts of the 30- to 32-kDa IGFBP increased 12.4-fold (p < 0.05) in subordinates on Day 4.8 compared with preselection follicles on Day 3, while the amount of inhibins > 34 kDa decreased 4- to 9-fold (p < 0.05). In FSH-treated heifers, both selection of DF1 and atresia of subordinates were delayed by 2.2 days. Preselection follicles recovered on Day 4.9 +/- 0.1 from FSH-treated heifers were similar (p > 0.05) in almost all biochemical parameters to preselection follicles from control heifers; however, they differed markedly from both DF1 and subordinate follicles recovered from control heifers on Day 4.8 +/- 0.5. In conclusion, the decline in FSH beginning after Day 2 of the heifer estrous cycle causes differential alterations in FSH-dependent growth factors and hormones within the cohort of preselection follicles, simultaneously inducing growth and enhanced estradiol-producing capacity of the DF and atresia of subordinate follicles.

Activins↗

Variation in the ability of a long day followed by a short day photoperiod signal to initiate reproductive activity in ewes at different times of the year.

Three experiments examined the importance of the time and duration of exposure to a long day followed by a short day photoperiod signal in initiating reproductive activity in ewes. In Expt 1, ewes were maintained on short days (8.5 h light:15.5 h dark) from 21 December interrupted with either 105 long days (18 h light:6 h dark; LD) from 9 February or 35 LD from 9 February, 16 March or 20 April. Exposure to long days followed by short days advanced the onset of reproductive activity in comparison to control ewes maintained on simulated natural photoperiod. Exposure to long days for 105 days delayed the onset of reproductive activity (August 2 +/- 3 days; P < 0.05) compared with 35 days beginning on the same date (July 13 +/- 5 days). The interval from the end of the long day signal to the onset of reproductive activity was shorter (P < 0.001) however, after 105 LD than after 35 LD. In Expt 2, control ewes were moved from natural photoperiod to simulated natural photoperiod on 1 November and subsequently exposed to short days from 21 December. Four other groups were also exposed to this basic photoperiodic signal sequence but it was interrupted with either 70 LD from 16 November, or 35 LD from 16 November, 21 December or 20 April. More ewes (P < 0.05) initiated reproductive activity after exposure to 70 LD from 16 November and 35 LD from 21 December or 20 April compared with control ewes maintained on short days or ewes given 35 LD from 16 November. The interval from the end of long days to the onset of reproductive activity was less (P < 0.01) in ewes given 70 LD than in ewes given 35 LD. In Expt 3, ewes on natural photoperiod were given either 90 LD from 21 September, 35 LD from 21 September, 26 October, 30 November, 4 January or 8 February followed by short days. The majority of ewes that received long followed by short days after the winter solstice resumed reproductive activity. However, all photoperiod signals given between the autumn equinox and the winter solstice failed to initiate reproductive activity in ewes during the experiment. Thus we conclude that, in ewes, the reproductive neuroendocrine axis is insensitive to long days followed by short days between the autumn equinox and the winter solstice. The reproductive axis of ewes regains sensitivity to the inductive effects of long days followed by short days at a time close to the winter solstice. Between the winter and summer solstices, long days followed by short days maintain the anoestrous state and provide the cue for initiation of reproductive activity.

Animals↗

Effect of immunization of rams against bovine inhibin alpha 1-26 on semen characteristics, scrotal size, FSH, LH and testosterone concentrations.

The effects of inhibin immunization on inhibin antibody titres, semen characteristics, scrotal size, fertility, FSH, LH and testosterone concentrations were determined by immunizing adult rams against bovine inhibin alpha 1-26-Gly-Tyr conjugated to human serum albumin (n = 16) in non-ulcerative Freund's adjuvant and DEAE:dextran (1:1) or adjuvant alone (n = 16) on days 0 (29 June), 30, 60, 191, 303 and 394. Blood samples were collected and bovine inhibin alpha 1-26-Gly-Tyr antibody titres and serum testosterone concentrations were determined. Each month, between days 174 and 417, semen was collected every 30 min to a maximum of 15 ejaculates over 7 h and scrotal circumference was measured. Ram fertility was recorded during natural service. FSH, LH and testosterone concentrations and GnRH-induced FSH and LH release were measured in a subgroup of immunized (n = 5) and control (n = 5) rams at frequent intervals. Antibody titres were variable among immunized rams (0-46% 125I-labelled bovine inhibin alpha 1-26-Gly-Tyr at 1:1600 serum dilution) but mean titres were consistently higher than in control rams (P < or = 0.001). Immunization did not alter the semen volume, output or quality of spermatozoa or ram fertility, but increased the mean scrotal circumference (37.6 +/- 0.8 cm versus 34.4 +/- 0.7 cm, P < 0.001). Mean FSH concentrations were higher in immunized rams during two intensive blood sampling periods (in June and August) (5.8 +/- 0.7 ng ml-1 versus 3.0 +/- 0.3 ng ml-1, P < 0.001 in June; and 4.8 +/- 0.9 ng ml-1 versus 2.0 +/- 0.3 ng ml-1, P < 0.02 in August), and were correlated with antibody titres (r2 = 0.3, P < 0.05 in June; and r2 = 0.8, P < 0.001 in August). Discrete FSH pulses were not detected. Immunization did not alter mean or basal testosterone or LH concentrations, or LH pulse frequency; LH pulse amplitude was increased (1.6 +/- 0.2 ng ml-1 versus 0.8 +/- 0.2 ng ml-1, P < 0.02) and was correlated with antibody titres (r2 = 0.6, P < 0.01). Immunization enhanced GnRH-induced FSH (P < 0.05) but not LH release. In conclusion, immunization of adult rams against bovine inhibin alpha 1-26 Gly-Tyr increased scrotal circumference, mean FSH concentrations and LH pulse amplitude, without altering semen characteristics, fertility, mean LH concentrations, LH pulse frequency or mean testosterone concentrations.

Animals↗

Circulating FSH isoform patterns during recurrent increases in FSH throughout the oestrous cycle of heifers.

Blood samples were collected from heifers (n = 6; 450 +/- 7.7 kg) 2-4 times a day during the first and second follicular waves, and during the gonadotrophin surge to determine whether changes in circulating FSH isoforms occur during cyclic quantitative changes in FSH throughout the oestrous cycle. Serum was assayed for FSH, LH, oestradiol and progesterone. Selected samples collected during the first (samples 1-3) and second (samples 4-6) postovulatory recurrent FSH increase and during the subsequent gonadotrophin surge (samples 7 and 8) were analysed for FSH isoforms by chromatofocusing. No change (P > 0.05) in isoform profile occurred during the first or second recurrent FSH increase, when oestradiol concentrations were 0.6 +/- 0.07 and 0.6 +/- 0.09 pg ml-1 and progesterone concentrations were 0.03 +/- 0.01 and 2.4 +/- 0.19 ng ml-1, respectively. The percentage of FSH eluting in the pH range 7.4-7.0 increased (P < 0.05) from 14.2 +/- 2.2 during the luteal phase (samples 1-6) to 20.2 +/- 2.3 (sample 7) and 31.4 +/- 3.4% (sample 8) during the preovulatory gonadotrophin surge, while oestradiol concentrations were higher (P < 0.05; 4.9 +/- 0.39 pg ml-1) than in the luteal phase of the cycle. In summary, FSH isoform patterns did not change during the cyclic quantitative changes in FSH associated with emergence of the first or second follicular wave. However, during the gonadotrophin surge, in association with increased oestradiol concentrations, an increase in the amount of less acidic isoforms of FSH was observed. Therefore, qualitative changes in FSH are not important in the physiological regulation of follicle turnover during the luteal phase of the oestrous cycle of heifers.

Animals↗

Normal or induced secretory patterns of luteinising hormone and follicle-stimulating hormone in anoestrous gonadotrophin-releasing hormone-immunised and cyclic control heifers.

The objective was to determine the effect of gonadotrophin-releasing hormone (GnRH), GnRH analogue (GnRH-A) or oestradiol administration on luteinising hormone (LH) and follicle-stimulating hormone (FSH) release in GnRH-immunised anoestrous and control cyclic heifers. Thirty-two heifers (477 +/- 7.1 kg) were immunised against either human serum albumin (HSA; controls; n = 8), or a HSA-GnRH conjugate. On day 70 after primary immunisation, control heifers (n = 4 per treatment; day 3 of cycle) received either (a) 2.5 micrograms GnRH or (b) 2.5 micrograms of GnRH-A (Buserelin) and GnRH-immunised heifers (blocked by GnRH antibody titre; n = 6 per treatment) received either (c) saline, (d) 2.5 micrograms GnRH, (e) 25 micrograms GnRH or (f) 2.5 micrograms GnRH-A, intravenously. On day 105, 1 mg oestradiol was injected (intramuscularly) into control (n = 6) and GnRH-immunised anoestrous heifers with either low (13.4 +/- 1.9% binding at 1:640; n = 6) or high GnRH antibody titres (33.4 +/- 4.8% binding; n = 6). Data were analysed by ANOVA. Mean plasma LH and FSH concentrations on day 69 were higher (P < 0.05) in control than in GnRH-immunised heifers (3.1 +/- 0.16 vs. 2.5 +/- 0.12 ng LH ml-1 and 22.5 +/- 0.73 vs. 17.1 +/- 0.64 ng FSH ml-1, respectively). The number of LH pulses was higher (P < 0.05) in control than in GnRH-immunised heifers on day 69 (3.4 +/- 0.45 and 1.0 +/- 0.26 pulses per 6 h, respectively). On day 70, 2.5 micrograms GnRH increased (P < 0.05) LH concentrations in control but not in GnRH-immunised heifers, while both 25 micrograms GnRH and 2.5 micrograms GnRH-A increased (P < 0.05) LH concentrations in GnRH-immunised heifers, and 2.5 micrograms GnRH-A increased LH in controls. FSH was increased (P < 0.05) in GnRH-immunised heifers following 25 micrograms GnRH and 2.5 micrograms GnRH-A. Oestradiol challenge increased (P < 0.05) LH concentrations during the 13-24 h period after challenge with a greater (P < 0.05) increase in control than in GnRH-immunised heifers. FSH concentrations were decreased (P < 0.05) for at least 30 h after oestradiol challenge. In conclusion, GnRH immunisation decreased LH pulsatility and mean LH and FSH concentrations. GnRH antibodies neutralised low doses of GnRH (2.5 micrograms), but not high doses of GnRH (25 micrograms) and GnRH-A (2.5 micrograms). GnRH immunisation decreased the rise in LH concentrations following oestradiol challenge.

Anestrus↗

Factors affecting superovulation in heifers treated with PMSG.

In this study we determined 1) if the immunoneutralization of PMSG affected the ovulatory response, the number of large follicles and embryo yield compared with that of PMSG alone or pFSH, and 2) whether the stage of the estrous cycle at which PMSG was injected affected the ovulatory response and yield of embryos in superovulated heifers. Estrus was synchronized in 99 (Experiment 1) and 71 (Experiment 2) heifers using prostaglandin F2alpha (PG) analogue, cloprostenol, given 11 d apart in replicate experiments over 2 yr. In Experiments 1 and 2, heifers were randomly allocated to 1 of 3 treatments (initiated at mid-cycle): Treatment 1--24 mg of pFSH (Folltropin) given twice daily for 4 d; Treatment 2--a single injection of 2000 IU PMSG; Treatment 3--2000 IU PMSG followed by 2000 IU of Neutra-PMSG at the time of first insemination. In Experiment 3, 116 heifers were given 2000 IU PMSG on Day 2 (n = 28), Day 3 (n = 27), Day 10 (n = 41) or Day 16 (n = 20) of the estrous cycle. The PG was given at 48 h (500 microg cloprostenol) and 60 h (250 microg cloprostenol) after the first gonadotropin treatment. Heifers were inseminated twice during estrus, and embryos were recovered on Day 7, following slaughter and graded for quality. The numbers of ovulations and large follicles (> or =10 mm) were also counted. There was no effect of treatment on ovulation rate in Experiment 1, but in Experiment 2 it was greater (P < 0.002) in heifers given PMSG (14.7 +/- 1.5) than pFSH (7.5 +/- 1.4) or PMSG-neutra-PMSG (8.7 +/- 1.5). The number of large follicles was higher following PMSG than pFSH treatment in Experiment 1, and it was higher (P < 0.004) in heifers given PMSG (5.5 +/- 0.8) than pFSH (1.12 +/- 0.7) or PMSG-neutra-PMSG (2.7 +/- 0.8) in Experiment 2. The use of Neutra-PMSG did not affect the numbers of embryos recovered or numbers of Grade 1 or 2 embryos, but it did decrease the number of Grade 3 embryos in both experiments. In Experiment 3, the ovulation rate decreased (P < 0.004) when PMSG was given on Day 3 (5.7 +/- 1.46) of the cycle rather than on Day 2 (12.3 +/- 1.64), Day 10 (13.4 +/- 1.45) or Day 16 (12.5 +/- 1.87). There was no effect of day of treatment on the numbers of large follicles. The mean numbers of embryos recovered were lower (P < 0.01) in heifers treated on Day 3 (2.1 +/- 0.67) than on Day 2 (6.8 +/- 1.0), Day 10 (6.4 +/- 0.86) or Day 16 (7.8 +/- 1.87). It is concluded that Neutra-PMSG given to heifers treated with PMSG did not improve embryo yield or quality and that treatment with PMSG early in the cycle can result in acceptable embryo yields provided sufficient time elapses between treatment and luteolysis.

Journal Article↗

Alterations in intrafollicular levels of different molecular mass forms of inhibin during development of follicular- and luteal-phase dominant follicles in heifers.

The experiment reported here characterized changes in serum concentrations of FSH, follicular fluid (FF) concentrations of immunoreactive alpha inhibins and dimeric inhibins, and different molecular forms of inhibin during development of dominant follicles in both luteal and follicular phases of the estrous cycle in heifers. Follicular status was determined daily by ultrasound examination of ovaries, and blood samples were taken at 4-6-h intervals. Heifers (5-6 per treatment) were ovariectomized on Days 0 (follicular phase, pre-LH surge), 1-Ov (post-LH surge, preovulation), 1 + Ov (postovulation), 3 (selection phase), 6 (dominance phase), and 12 (atretic phase) of the estrous cycle. Follicles were classified on the basis of FF estradiol (E) to progesterone (P) ratio and by ultrasound morphology. Concentrations of E, P, immunoreactive alpha inhibin, LH, and FSH were determined by RIA, and concentrations of dimeric inhibins were determined by a two-site dimeric-specific immunoradiometric assay (IRMA); the different molecular mass forms of inhibin were determined by immunoblot analysis. Serum FSH was transiently elevated during Days 0.5-1.5 and 8-10.5, in association with emergence of each new wave of follicular growth. The follicular-phase dominant follicle of heifers before the preovulatory LH/FSH surge had less immunoreactive alpha inhibin and 29-kDa inhibin but more 110-kDa inhibin compared with concentrations after the surge (p < 0.05). During growth of the dominant follicle between Days 3 and 6, there were no changes in either proportions or amounts of any inhibin forms (p > 0.05). However, while concentrations of dimeric inhibins decreased between Days 1 and 6, the concentration of immunoreactive alpha inhibin increased (p < 0.05). Atresia of the luteal-phase dominant follicle during Days 6-12 was associated with a decrease in amounts and proportions of the 110- and > 160-kDa forms, an increase in amounts and proportions of the 34-kDa form, and an increase in dimeric inhibin concentration (p < 0.05). Alterations in FF E concentrations were positively correlated with changes in amounts of the 77- and 110-kDa precursor forms of inhibin, but negatively correlated with 34-kDa inhibin. The results of this experiment demonstrate that 1) seven different molecular mass forms of inhibin were present in bovine FF during all stages of development of follicular- and luteal-phase dominant follicles examined, 2) amounts and proportions of most inhibin forms are altered differently in follicular- and luteal-phase dominant follicles, 3) FF inhibins and E are regulated by different factors, and 4) RIA, IRMA, and immunoblot measurements of inhibin in bovine FF produced divergent results.

Animals↗

Control and regulation of folliculogenesis--a symposium in perspective.

The Intervet Symposium 'Control and Regulation of Folliculogenesis' took place at the Annual Conference of the Society for the Study of Fertility in University College, Dublin in July, 1995. Five papers were presented, of which abstracts have been published. Discussion centred on monotocous species and principally addressed the question 'How is the ovulatory follicle selected and nurtured?' Many follicles start to develop simultaneously, but most become atretic. During the luteal phase of the cycle, several waves of follicles are initiated, and although one in each wave gains dominance, all ultimately atrophy. The dominant follicle in the wave produced towards the end of the luteal phase ovulates. The biochemical characteristics of dominant and subordinate follicles were contrasted, and the endocrine environment in which they waxed and waned was analysed. Interaction of autocrine and paracrine effectors, principally with FSH, LH and their receptors, ultimately determines follicular destiny.

Animals↗

Effect of photoperiod before and after birth on puberty in ewe lambs.

Previous studies have shown that exposure to long days followed by short days after 12 wk of age advances the onset of puberty in ewe lambs. The aim of this experiment was to determine whether or not puberty would be advanced by exposure of lambs to such a photoperiodic regime when the long days were given before birth and the short days immediately after birth. Ewes kept in natural photoperiod (NP) were housed on January 15 and allocated at random to one of four treatments: 1) NP/NP, natural photoperiod before and after parturition (n = 26); 2) SD/SD, short days (SD; 9L:15D) before and after parturition (n = 27); 3) LD/NP, long days (LD; 18L:6D) before parturition and NP after parturition (n = 27); and 4) LD/SD, LD before parturition and SD after parturition (n = 26). Mean (+/- SEM) birth date for all lambs was March 20 +/- 1 day, and the number of days of exposure to long and short days prior to parturition was 64 +/- 2 and 67 +/- 2 days, respectively. From ewes on the four treatments, 24, 20, 14, and 16 female lambs, respectively, were reared. Lambs were weighed at birth and at weekly intervals after weaning. Reproductive activity in ewe lambs was assessed from serum progesterone concentrations in blood samples collected twice weekly from August 1 until puberty was reached or the experiment terminated on November 17 (34 wk after birth).(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Immunization of prepubertal beef heifers against gonadotropin-releasing hormone: immune, estrus, ovarian, and growth responses.

To develop an effective immunization protocol against human serum albumin-Cys-Gly-GnRH (HSA-GnRH) conjugate to delay the onset of puberty in heifers, 58 heifers (8 mo of age; mean +/- SE BW = 203 +/- 1 kg) were randomly assigned to each of six treatments: 1) controls, .1 mg of HSA, with diethylaminoethyl (DEAE)-dextran as adjuvant, on d 0 and 28; 2) .1 mg of HSA-GnRH, with DEAE-dextran, on d 0; 3) as 2) and booster on d 28; 4) as 3) but boosters also on d 84, 140, 196, and 252; 5) as 2) but half the conjugate given with DEAE-dextran adjuvant and half with non-ulcerative Freund's adjuvant (NUFA), injected in two separate sites; and 6) as 2) but the conjugate given with DEAE-dextran and NUFA, emulsified and injected in two sites. The duration of the experiment was 342 d. Mean plasma GnRH antibody titers (samples every 2 wk) for heifers in Treatments 2 to 6 were 9.4 +/- 1.16, 20.6 +/- 2.21, 43.9 +/- 2.86, 27.9 +/- 2.67, and 44.5 +/- 3.75% binding at a plasma dilution of 1:640. The mean number of times estrus was observed in heifers was less (P < .05; pooled SEM = .53) in Treatments 4 (.2) and 6 (2.4) than in Treatments 1, 2, 3, and 5 (7.8, 7.0, 7.0, and 6.6, respectively). The mean interval to the onset of puberty (the first increase in plasma progesterone > or = .5 ng/mL for > or = 10 d with samples at 3- to 4-d intervals) was greater (P < .05; pooled SEM = 11.6) for heifers in treatments 4 (339 d) and 6 (276 d) than for heifers in Treatments 1, 2, 3, and 5 (164, 159, 165, and 170 d, respectively). Mean ADG of heifers was reduced (P < .05) in treatments 2, 3, 4, and 6 (.71, .72, .68, and .69 kg, respectively) compared with controls (.77). In summary, the multiple booster immunization treatment induced and maintained sufficient anti-GnRH titer to delay puberty for 175 d; a single immunization against GnRH with DEAE and NUFA increased antibody titers enough to delay puberty for 112 d. However, GnRH immunization treatments reduced ADG of heifers in Treatments 2, 3, 4, and 6.

Aging↗

Growth and estrous behavior of heifers actively immunized against prostaglandin F2 alpha.

To determine the effects of active immunization against prostaglandin F2 alpha (PGF) on estrous activity and performance traits of beef heifers, 50 14-mo-old cyclic heifers (358 +/- 3.3 kg) were assigned to two treatments (n = 25 per treatment): 1) heifers (controls) given 3.3 mg of human serum albumin (HSA) on d 0 (primary) and 27 (booster), and 2) heifers (PGF-immunized) given 3.3 mg of PGF-HSA on d 0 and 27. The adjuvant was DEAE-dextran, and the duration of the experiment was 167 d. Plasma progesterone concentrations (every 3 to 4 d) were used to monitor corpus luteum (CL) presence; PGF antibody titers were determined every 2 wk. Heifers were checked twice daily for estrous behavior and were weighed every 2 wk. Data were analyzed using ANOVA. Antibody titers for PGF-immunized heifers increased to a peak (43 +/- 2.9% binding at a plasma dilution of 1:1,250) on d 55 +/- 4.6. Antibody titers were greater (P = .02) in PGF-immunized than in control heifers by d 15 and remained elevated (P < or = .001) throughout the experiment. Twenty-four of 25 PGF-immunized heifers formed persistent CL with a mean duration of 129 +/- 6.4 d. The mean number of estrous period per heifer were less for PGF-immunized (1.5 +/- .27) than for control heifers (7.0 +/- .32). Mean daily live weight gain of the PGF-immunized heifers was decreased (P < .05; .75 +/- .024 kg) compared with that of controls (.83 +/- .014 kg), largely due to a 31.5% decrease during the 28-d period after booster.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗

Immunization of heifers against gonadotropin-releasing hormone: antibody titers, ovarian function, body growth, and carcass characteristics.

To investigate the effects of active immunization of cyclic beef heifers with different doses of a human serum albumin-Cys-Gly-GnRH (HSA-GnRH) conjugate on antibody titers, ovarian function, body growth, and carcass characteristics, 32 heifers (BW = 477 +/- 7.1 kg; mean +/- SE) were assigned to one of four immunization treatments: .1 mg of HSA or .01, .1, or 1.0 mg of HSA-GnRH, respectively. All heifers received a primary (d 0) and booster (d 28) immunization using DEAE-dextran as adjuvant. The duration of the experiment was 158 d. Overall antibody titers against GnRH were greater (P < .05) for heifers immunized against GnRH (13 +/- 3.3, 22 +/- 3.8, and 19 +/- 2.8% binding at a plasma dilution of 1: 640 for Treatments 2 to 4, respectively) than for controls (1 +/- .1%). The numbers of heifers that became anestrous (plasma progesterone < .5 ng/mL for > 21 d) were 1/8, 8/8, 7/8, and 8/8, respectively. The interval from primary immunization to anestrus (40.7 +/- 6 d) and the duration of anestrus (78 +/- 7 d) were not affected by dose of HSA-GnRH conjugate. The number of ovulations detected was reduced (P < .05) in GnRH-immunized (4.6 +/- .64, 4.0 +/- .70, and 3.6 +/- .60 for Treatments 2 to 4, respectively) compared with control heifers (9.4 +/- .20). During induced anestrus, follicular growth was generally arrested (< 5 mm in diameter) and plasma estradiol decreased.(ABSTRACT TRUNCATED AT 250 WORDS)

Adjuvants, Immunologic↗

Effects of single or primary plus booster prostaglandin F2 alpha immunization regimens on immune, ovarian, and growth responses of heifers.

Growth rate of heifers is reduced by prostaglandin F2 alpha (PGF) immunization following a primary and booster regimen. The objective was to attenuate the immune response with or without a booster immunization; specifically, the effects of booster interval, dose of PGF-human serum albumin (HSA) conjugate at booster, adjuvant type, or single immunization with one or two adjuvants were examined. Three experiments were conducted using 175 cyclic heifers. Plasma PGF antibody titers were measured every 2 wk and progesterone concentrations every 3 to 4 d. In Exp. 1, single immunization with one adjuvant (3.3 mg of PGF-HSA in either DEAE-dextran [DEAE] or non-ulcerative Freund's adjuvant [NUFA]; or 10.0 mg of PGF-HSA in NUFA) did not induce sufficient antibody titers to consistently induce persistent corpora lutea (CL). Booster intervals of either 14, 21, or 28 d increased titers sufficiently to induce persistent CL (34/35 heifers), but ADG of heifers was less (P < .05) than for those given a single immunization. In Exp. 2, 1.0 mg of conjugate for booster immunization induced a greater (P < .05) immune response than 3.3 mg, and both doses decreased (P < .05) ADG. Single immunization, with half the conjugate dose in DEAE and half in NUFA injected separately, induced persistent CL in 7/8 heifers without decreasing ADG compared with controls. In Exp. 3, single immunization, with half the conjugate dose in DEAE and half in NUFA injected separately, prolonged (P < .05) the intervals to peak titer compared with the booster treatment, but the incidence (13/15 vs 8/8) and duration (120 +/- 4.8 vs 111 +/- 7.9 d) of persistent CL were similar, and ADG was greater (P < .05). In conclusion, attempts to attenuate the immune response following booster immunization were unsuccessful. Single immunization, using two adjuvants separately, induced persistent CL for at least 120 d without decreasing ADG compared with the primary and booster regimen.

Animals↗

Oocyte structure and follicular steroid concentrations in superovulated versus unstimulated heifers.

A highly variable yield of viable embryos in superovulated cattle is a major hindrance to the embryo transfer industry. To trace the cause of this problem, investigations were carried out on the intrafollicular steroids and structure of oocytes originating from follicles of follicular stimulating hormone (FSH)-stimulated (superovulated) and unstimulated heifers. Unstimulated heifers were slaughtered at midcycle, or administered cloprostenol (PG) at midcycle and slaughtered after 24, 48, or 72 hr, while superovulated heifers were administered 4 injections of pFSH (2 injections per day) and slaughtered 12 hr later, or administered 6, 7, or 8 injections of FSH in combination with PG at the 5th and 6th injection, and slaughtered 24, 36, or 60 hr, respectively, after the first PG injection. The follicular fluid from the largest (presumptive dominant) follicle of the unstimulated heifers and from potentially ovulatory follicles (> or = 8 mm in diameter) of the superovulated heifers were assayed for estradiol-17 beta (E2) and progesterone (P4), while the oocyte cumulus complexes from such follicles were processed for transmission electron microscopy. The mean E2 and especially P4 concentrations of the potentially ovulatory follicles of the superovulated heifers were lower than similar follicles of the unstimulated animals (83.7 +/- 76.7 ng/ml vs. 208.1 +/- 357.0 ng/ml, P > 0.05 and 31.1 +/- 38.7 ng/ml vs. 150.3 +/- 202, P < 0.05, respectively). The unstimulated oocytes had, in general, spherical oocyte nuclei and compact nucleoli before PG administration, while after PG, undulation of the nuclear envelope and nucleolus vacuolization was characteristic.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗