The roe deer blastocyst is in diapause between August and December, after which time it expands and elongates rapidly before implantation. Blood samples were taken from 30 animals to define temporal changes in reproductively important hormones to investigate the physiological cues present at embryo reactivation. In 15 of these animals, changes in uterine and conceptus protein synthesis and secretion, and luteal progesterone release during diapause and reactivation, were assessed after culture of these tissues in vitro. Oestradiol concentrations remained low during diapause (1.07 +/- 0.4 pg ml(-1)) and expansion (1.2 +/- 0.4 pg ml(-1)) but increased by 30 times at trophoblast elongation (49.17 +/- 0.37 pg ml(-1)). Prolactin remained at basal concentrations (4.69 +/- 0.86 ng ml(-1)) and increased after implantation (12.34 +/- 2.71 ng ml(-1)). Peripheral progesterone concentrations and luteal progesterone release remained constant throughout diapause, reactivation and implantation (peripheral progesterone: 3.82 +/- 1.97 ng ml(-1); luteal progesterone: 6.72 +/- 0.81 ng mg(-1) protein). Incorporation of a radiolabel into conceptus secretory proteins increased by four times at expansion compared with diapause, whereas incorporation into endometrial secretions remained constant. At elongation, incorporation into endometrial secretions increased two times and conceptus secretions increased 32 times. Two-dimensional electrophoresis and fluorography showed that the profile of endometrial secretory proteins was constant until implantation when qualitative changes were evident. Although a role for an endocrine maternal trigger of reactivation from diapause cannot be dismissed, these data provide no supporting evidence and indicate that the conceptus itself may drive reactivation.

Abstract: Roe deer blastocysts exhibit obligate embryonic diapause between early August and late December. The blastocyst then expands and elongates rapidly before implantation. The objective of this study was to ascertain the cues for reactivation of the diapausing blastocyst. Blood samples and reproductive tracts were collected from roe does during diapause, blastocyst expansion and subsequent implantation. Peripheral concentrations of oestradiol-17/3, progesterone and prolactin were measured by radioimmunoassay. Luteal progesterone release was determined following in vitro incubation. Conceptuses and endometrial tissue were cultured with 3H-leucine for 24 hours to measure de novo synthesis of secretory proteins. Endometrial secretory proteins were separated by two-dimensional electrophoresis. Results showed that peripheral progesterone concentrations declined by 55% just prior to expansion and did not rise until a 3-fold increase after implantation. Luteal progesterone release remained constant until expansion when it declined by 50% before increasing 2-fold at elongation and implantation. Concentrations of oestradiol-17/3 remained at a consistently low level during diapause and expansion until a 30-fold increase at elongation with concentrations remaining elevated after implantation. Plasma prolactin levels remained at basal concentrations during late diapause and then increased marginally at reactivation before decreasing again at elongation and implantation. Incorporation of radiolabel into both conceptus and endometrial secretory proteins was low during diapause, but incorporation in the conceptus increased 4¬fold at expansion and by 24-fold at the expanded trophoblast stage. Incorporation into endometrial secretoty proteins remained constant until the expanded trophoblast stage and implantation when a 2-fold increase was recorded. Furthermore, the profile of endometrial secretory proteins was constant during diapause and expansion but changed qualitatively following implantation. These data indicate that both endometrial protein synthesis and sectetion did not change during late diapause and early expansion. The increase in conceptus protein synthesis not only precedes that of the endometrium but consistently low luteal progesterone release, peripheral progesterone and oestradioi-17/J concentrations at early expansion suggests that reactivation is not in response to a maternal uterine trigger.

Four ♀ Roe deer Capreolus capreolus were hand‐reared and released into a 10 ha enclosed natural habitat. This paper describes the hand‐rearing procedure, nutrition and development, which we compare with other documented cases. We illustrate the handling techniques that enabled us to maintain a close relationship with the hand‐reared Roe deer in their adulthood and study them under semi‐free‐ranging conditions. The benefits of conducting research with hand‐reared Roe deer are described, together with possible biases that must be taken into consideration when planning a research programme. The Roe deer were hand‐reared in order to conduct a detailed feasibility study for the Roe deer reintroduction programme in Israel, where they have been locally extinct for 100 years.

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