Summary.

The effect of differing lighting schedules upon the timing of the influx of leucocytes into the vaginal smear of the guinea-pig, and hence of ovulation, was examined. The timing of this event was altered following advancement of the light—dark sequence by 12 hr, and implies that the timing of ovulation in this species is influenced by light.

High prolificacy due to a gene that has a large effect on ovulation rate has been noted in Booroola and Inverdale ewes. High prolificacy in the Belclare breed (a composite developed from stocks selected for very large litter size or high ovulation rate) may be related to the segregation of two genes. The aims of this study were (i) to compare the morphological and functional features of ovulatory follicles from carriers (which could only be heterozygous for the genes of interest) and non-carriers, and (ii) to identify markers of the Belclare genes among secreted or cellular ovarian proteins. Belclare carrier ewes had more ovulatory follicles (4.9 ± 0.4) than did non-carrier ewes (2.0 ± 0.2) (P < 0.001). Ovulatory follicles from carriers were also smaller (4.4 ± 0.1 mm versus 5.7 ± 0.2 mm, P < 0.001) and contained a significantly reduced number of granulosa cells (P < 0.001). However, the proportion of proliferating granulosa cells in ovulatory follicles was similar in both groups. The in vitro secretion of steroids per follicle was only marginally lower in follicles from Belclare carriers compared with non-carriers. Furthermore, similar concentrations of steroidogenic enzymes were present in both groups, indicating that steroidogenic potential per granulosa cell is similar between carriers and non-carriers. Possible markers of the Belclare genes were identified among cellular proteins of follicular walls by two-dimensional PAGE and image analysis. Two spots at 78 and 49 kDa were always absent in samples from non-carriers. When secreted proteins in follicles from carriers were compared with those from non-carriers, two spots at 53 and 41 kDa were restricted to samples from carriers and three spots at 97, 91 and 45 kDa were unique to samples from non-carriers. Interestingly, the spot at 91 kDa is also affected by the Booroola gene.

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.

Four experiments were carried out to determine the effect of the presence of ewes and rams on the reproductive state of ewes. In Expt 1, the breeding season of ewes kept with a vasectomized ram ended later (April 18 ± 8 days; mean ± sem) than that of ewes isolated from rams (6 March ± 7 days; P < 0.01). In Expt 2, the end of the breeding season was later (5 May ± 6 days; P < 0.05) and the onset of the next breeding season earlier (29 September ± 2 days; P < 0.001) in ewes maintained with rams, compared with ewes isolated from rams (14 April ± 7 days and 1 November ± 2 days, respectively). There was no difference in the timing of, or variation in, reproductive transitions between ewes maintained either as individuals or in groups. In Expt 3, all ewes exposed to artificial short days from the date of the winter solstice and interrupted with 35 long days in spring resumed cyclicity (median date, 7 September; range, 59 days). Most ewes (seven of nine) exposed to short days from the date of the winter solstice and isolated from other ewes did not resume cyclicity in the following 11 months. In contrast, all ewes resumed cyclicity (median date, 19 October; range, 144 days) when exposed to short days but housed in social contact with other ewes that became reproductively active in early September; however, the onset of cyclicity was later than in ewes exposed to long days (P < 0.01). In Expt 4, the number of LH pulses per 6 h in ewes exposed to rams was higher (P < 0.001) and the time of first ovulation earlier (16 August ± 5 days; P < 0.05) than it was in ewes that were isolated from rams and exposed to either oestrous or anoestrous ewes. We conclude that there was a chronic stimulus from rams to ewes that increased the duration of the breeding season and decreased anoestrus. There was no acute effect of introduction of oestrous ewes to anoestrous ewes on LH pulse frequency and time of first ovulation of the breeding season under the natural photoperiod, and the onset of the breeding season of housed anoestrous ewes exposed to a constant photoperiod was advanced by housing them with cyclic ewes. These results highlight a role for social or other animal-related stimuli in seasonal reproduction in ewes.