Abstract

The Egyptian or Common spiny mouse (A. cahirinus) is the first rodent species to show human-like menstruation and spontaneous decidualisation. We consider from these, and its other, human-like characteristics that this species will be a more useful and appropriate small animal model for human reproductive studies. Based on this, there is a need to develop specific laboratory-based assisted reproduction protocols including superovulation, in-vitro fertilisation, embryo cryopreservation and transfer to expand and make this model more relevant. Because standard rodent superovulation has not been successful in the spiny mouse, we have selected to test a human protocol. Female spiny mice will receive a subcutaneous GnRH agonist implant and be allowed to recover. Menstrual cycle lengths will then be allowed to stabilize prior to ovarian stimulation. After recovery, females will be injected IP once a day for 4 days with a FSH analogue, to induce follicular growth, and on day 5 will be injected IP with a hCG analogue to trigger ovulation. Females will either be culled 36hrs after trigger to collect oocytes or immediately paired with a stud male and two cell embryos collected 48hrs later. Mature oocytes will be inseminated using fresh spiny mouse spermatozoa and all in-vitro grown and in-vivo collected two cell embryos will be cryopreserved using methods developed in a close spiny mouse relative, the Mongolian gerbil. For embryo transfer, vitrified embryos will be rapidly warmed and non-surgically transferred to surrogate mice. Surrogates will be monitored until pregnancy is apparent (roughly 30 days) and then left undisturbed until birth, 38–40 days after transfer. By successfully developing robust assisted reproduction protocols in A. cahirinus we will be able to use this rodent as a more effective model for human reproduction.

Highlights

  • The Egyptian or common spiny mouse (A. cahirinus) is a small, desert-dwelling rodent native to northern Africa and the Middle East

  • Females are killed 36hrs later by isoflurane inhalation, the ovaries and oviducts removed by fine dissection and ovulated oocytes released from the oviducts into fertilisation medium (MT6) for use in in-vitro fertilisation (IVF) and, subsequently, embryo vitrification and embryo transfers (ET) (Figs 1 and 2; Experiments 2–3, 5)

  • Number of oocytes and embryos collected from control and experimental animals will be compared statistically using a one-way ANOVA and the Tukey test used for post-hoc analysis; significance is set at p

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Summary

Introduction

The Egyptian or common spiny mouse (A. cahirinus) is a small, desert-dwelling rodent native to northern Africa and the Middle East. Subsequent development of embryo vitrification in this Acomys species will streamline embryo transfer procedures, and provide the foundation for future creation of transgenic strains and for improved breeding efficiencies for captive spiny mouse colonies. Our aim with this protocol is to develop a successful agonist-based superovulation technique for A. cahirinus that will provide large numbers of oocytes for IVF, and embryos for culture, cryopreservation and transfer, and enable us to more effectively use the spiny mouse as a model for human reproduction

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