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Surgical Techniques for the Generation of Mutant Mice

  • Anne Plück
  • Christian Klasen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 561)

Summary

The previous two chapters have described the generation of chimeric embryos by blastocyst microinjection and morula aggregation. This chapter describes the reimplantation of these embryos into pseudopregnant female recipient mice (foster mice) in order for the embryos to develop to term. Four different surgical techniques will be described in detail.

The transfer of blastocyst-stage embryos back into the uterus is most commonly used for chimeric embryos generated by microinjection or aggregation, or for rederivation purposes when strains are transferred to a clean environment. Fertilized oocytes manipulated by pronuclear microinjection or two-cell-stage embryos are reintroduced to the infundibulum of the oviduct of pseudopregnant female mice.

To generate pseudopregnant females, they need to be mated to sterile male mice. These males can be either generated by vasectomy of stud males or naturally sterile males can be used. Two different techniques for the vasectomy are shown: the vas deferens is accessed through an incision in the scrotal sac or through an abdominal incision.

Key words

Uterus transfer Oviduct transfer Vasectomy Abdominal incision Incision in the scrotal sac Rederivation Aseptic conditions 

References

  1. 1.
    Agca, Y., Bauer, B.A., Johnson, D.K., Critser, J.K., and Riley, L.K. (2007). Detection of mouse Parvovirus in Mus musculus gametes, embryos, and ovarian tissues by polymerase chain reaction assay. Comp. Med. 57, 51–56.PubMedGoogle Scholar
  2. 2.
    Whitten, W.K. (1958). Modification of the oestrous cycle of the mouse by external stimuli associated with the male. Changes in the oestrous cycle determined by vaginal smears. J. Endocrinol. 17, 307–313.PubMedCrossRefGoogle Scholar
  3. 3.
    Whitten, W.K., Bronson, F.H., and Greenstein, J.A. (1968). Estrus-inducing pheromone of male mice: transport by movement of air. Science 161, 584–585.PubMedCrossRefGoogle Scholar
  4. 4.
    Erhardt, W., Hebestedt, A., Aschenbrenner, G., Pichotka, B., and Blümel, G. (1984). A comparative study with various anesthetics in mice (pentobarbitone, ketamine-xylazine, carfentanyl-etomidate). Res. Exp. Med. 184, 159–169.CrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Centre for Mouse Genetics, Institute for GeneticsUniversity of CologneKoelnGermany

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