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The methods of assisted reproduction commonly used in domestic animals are currently being applied to non-domestic species. This is because of the limitation of maintaining the genetic variability in the wild, as it occurs in wild ruminants of the Mediterranean Basin. Despite the apparent progress of the technology, success of the offspring to grow and become healthy adult individuals has remained low in wild species. Difficulties usually arise from insufficient information about basic reproductive biology, such as the seasonal changes in ovarian and testicular activity. Directional adaptive evolution involves development of speciespecific physiological reproductive patterns to cope with various environmental factors. Thus, species originating and living at the same latitude display different reproductive strategies to entrain the breeding activity at an optimal time of the year. The aim of this paper was to present current knowledge on reproductive physiology of Mediterranean wild ruminants as a basic prerequisite for the successful use of assisted reproduction methods. Special emphasis is given to seasonal endocrine changes, ovarian cycles and testicular activity of Iberian wild ruminants, together with the role of social interactions on the regulation of these events.

The actions of prolactin (PRL) on target cells depend on the type of prolactin receptor (PRLr) predominantly expressed, particularly whether the long PRLr isoform is expressed. The aims of this study were to determine the cellular localization and the changes in expression of long and short PRLr isoforms in sheep ovary throughout the estrous cycle. Long and short PRLrs were localized mostly in the same ovarian cells. Maximum signal intensity, particularly for long PRLrs, was found in stromal cells surrounding primordial and primary follicles, and, for both PRLrs, in granulosa cells of preantral follicles and in luteal cells. Moderate signal intensity for PRLrs was found in theca cells of preantral to ovulatory follicles, and in granulosa cells of antral follicles up to the gonadotropin-dependent stage. Decreasing immunoreactivity to PRLrs was found in granulosa cells of gonadotropin-dependent to ovulatory follicles. For long PRLrs in particular, no signal was found in mural granulosa cells of gonadotropin-dependent follicles; for both isoforms, no signal was found in most granulosa cells of ovulatory follicles. In primordial to gonadotropin-dependent follicles, cellular localization of PRLr was similar on days 0, 10 and 15 of the cycle. Oocytes consistently showed positive immunostaining for PRLrs. Comparative RT-PCR analysis of long and short PRLr expression showed that the short isoform is evenly expressed throughout the estrous cycle, whereas the expression of the long form increases at the time of estrus and decreases at mid-luteal phase and at the onset of the follicular phase. Expression of long PRLrs was greater than that of short PRLrs on day 0 of cycle; expression of both isoforms was similar on day 10 and on day 15, long PRLrs expression was lower than that of short PRLrs. Our results indicate that in sheep ovary, the maximum responsiveness to PRL might occur during the preovulatory phase of the estrous cycle.