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B. J. McLeod and J. Craigon

Summary. Time series analysis was used to detect LH and FSH episodes in untreated seasonally anoestrous ewes and prepubertal heifers, and in these animals when treated with low doses of GnRH. For comparison, these profiles were also assessed for episodic secretion by subjective, visual appraisal methods and by cycle detection—an objective threshold method. In untreated animals, time series analysis detected recurring events in the LH and FSH profiles, the period lengths of which varied between individual animals. When GnRH was injected at 2-h intervals, cycles in LH secretion with period lengths of 120 min were recorded in all animals, of 60 min in all ewes and 11/12 heifers, and of 40·5 min in 22/24 ewes and 10/12 heifers. The cycles with period lengths of 60 and 40·5 min are probably artefacts of this method of analysis. No consistent cycles in FSH release were detected in GnRH-injected anoestrous ewes, but 120-min cycles were recorded in 8/12 GnRH-injected heifers. When GnRH was administered to seasonally anoestrous ewes by continuous infusion, recurring cycles in both LH and FSH secretion were evident. However, there was no consistency in their period lengths and the mean number and frequency of cycles were similar to pretreatment values. The number of episodes detected by visual appraisal was influenced by the choice of episode definition. Both methods identified LH, but not FSH, episodes in response to each injection in all GnRH-injected animals. Cycle detection, which does not identify individual episodes, recorded LH and FSH episode frequencies similar to those detected by the more stringent method of visual appraisal.

Time series analysis detected an FSH response to GnRH injections in prepubertal heifers that was not identified by the other methods of analysis. However, because of the asymmetric nature of LH episodes, it also detected cycles in LH profiles that were probably spurious. Subjective decisions influenced the frequencies of LH and FSH episodes recorded by visual appraisal, and the variation in episode amplitude in these profiles made cycle detection inappropriate. Each of these methods can contribute to the interpretation of hormone profiles, but their constraints and limitations must be recognized.

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C. M. Howles, J. Craigon and N. B. Haynes

Summary. Two groups of 6 rams were kept under constant photoperiod consisting of short days (8 h light (L): 16 h dark (D); Group S) and long days (16 h L:8 h D; Group L) from 4 to 38 months of age. Five other rams were reared under a photoperiod representative of that occurring naturally (Group N). Testis size and plasma prolactin concentrations were obtained weekly. These data were subjected to time series analysis. The results indicated that there were persistent periodic excursions in both parameters measured. In Group N, the average cycle length for both testis volume and plasma prolactin was about 1 year and the peaks in plasma prolactin preceded those in testis volume by about 18 weeks. Rams from Group L also showed rhythmical changes in these parameters with periodicities of around 35 weeks and it is suggested that these cyclic changes may constitute true endogenous circannual rhythms; again the prolactin peaks preceded those of testis volume by about 18 weeks. Overall, rams from Group S had excursions of testis growth of a similar magnitude to those of Group L but the changes were less regular than those of Group L. Plasma prolactin was significantly lower in Group S than in Group L and there was little evidence for rhythmicity. It is proposed on the basis of the temporal relationship between peaks of prolactin and testis volume in Groups N and L, that prolactin may play a role in the timing of the reproductive cycle in the ram.

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J Ye, J Coleman, M G Hunter, J Craigon, K H S Campbell and M R Luck

Ovarian follicles in vivo are cooler than surrounding abdominal and ovarian tissues. This study investigated whether typical follicular temperatures influence the maturation and developmental potential of pig oocytes in vitro. Oocytes were synchronised at the germinal vesicle (GV) stage and incubated at 39, 37 or 35.5 °C. When compared with 39 °C, which is often used for in vitro studies, lower temperatures delayed spontaneous progression to the metaphase I and II (MI and MII) stages of meiosis. The MII was delayed by about 12 h per °C. All oocytes had normal morphology. Oocytes reaching GV breakdown (GVBD) at 39 °C were subsequently unaffected by cooling, demonstrating thermal sensitivity during the pre-GVBD stage only. Simultaneous assay of maturation-controlling kinases (maturation promoting factor (MPF) and MAPK) showed that cooling delayed kinase activation, provided it was applied prior to GVBD. Activity profiles remained coupled to the stage of meiosis. Neither enzyme was directly thermally sensitive over this temperature range. Following in vitro fertilisation, fewer blastocysts developed from embryos derived from 35.5 or 37 °C oocytes as compared with those from 39 °C oocytes. Manipulation of fertilisation timings to allow for delayed maturation showed that over-maturing or aging at lower temperatures compromises subsequent embryo development, despite normal nuclear maturation; the GV stage was again the thermally sensitive period. Cleavage rates were improved by the culture of oocytes with follicle-stimulating hormone (FSH) at 37 but not at 35.5 °C. Inclusion of 20% follicular fluid in the oocyte medium restored the blastocyst rate to that seen at higher temperatures. Thus, FSH and follicular fluid may allow oocytes to achieve normal developmental potential at in vivo temperatures.

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C. Biggs, J. E. Tilton, J. Craigon, G. R. Foxcroft, C. J. Ashworth and M. G. Hunter

Comparisons were made between characteristics of pre-ovulatory follicles recovered from prolific Chinese Meishan gilts (n = 12) and from European Large-White hybrid gilts (n = 13) in the late follicular phase preceding their fifth oestrous cycle, to determine whether there is an ovarian basis for the enhanced prolificacy in the Meishan. A total of 177 follicles per breed was classified as pre-ovulatory, based on follicular fluid oestradiol concentrations. Results obtained demonstrated high variability in all follicular characteristics in both breeds and no decrease in heterogeneity was evident in the Meishan. The Meishan follicles tended to be smaller (P < 0.06) and had less follicular fluid (P < 0.005), but total oestradiol content per follicle was similar (P > 0.1) with the result that the concentration of oestradiol in follicular fluid tended to be higher (P < 0.06) in Meishan than Large-White hybrid pigs. There were no differences between breeds in terms of testosterone concentration in follicular fluid, hCG binding to granulosa cells or total DNA content of granulosa cells. Concentrations of inhibin in follicular fluid were similar in both breeds (P > 0.1) which resulted in a trend towards less total inhibin content in Meishan than Large-White hybrid follicles (P = 0.065). Corpora lutea were recovered from both breeds (n = 12 per breed) on days 27–31 of pregnancy after mating at first, second and third oestrus:corpora lutea were smaller (P < 0.001) and contained less progesterone per corpus luteum in the Meishan (P < 0.05) than in Large White hybrid pigs. However, since ovulation rate was higher in these particular Meishan pigs (P < 0.005), total ovarian progesterone content per animal was similar in both breeds. These results demonstrate that there was no decrease in the variability in follicular characteristics from Meishan pigs. However, both follicles and corpora lutea were smaller in Meishan than in Large-White hybrid pigs, and progesterone per corpora lutea was also lower in the Meishan. In spite of their smaller size, Meishan follicular oestradiol content was similar to that of Large-White hybrid pigs, such that the oestradiol concentration in follicular fluid tended to be higher in Meishan follicles.