Microsatellite detection of donor-derived sperm DNA following germ cell transplantation in cattle
Sally Stockwell A C D E , Muren Herrid A B E F , Rhonda Davey A B , Alan Brownlee C , Keryn Hutton A B and Jonathan R. Hill A B DA CSIRO Food Futures National Research Flagship.
B CSIRO Livestock Industries, FD McMaster Laboratory, Armidale, NSW 2350, Australia.
C CSIRO Livestock Industries, Queensland Bioscience Precinct, Brisbane, Qld 4072, Australia.
D Present address: School of Veterinary Science, University of Queensland, St Lucia, Qld 4072, Australia.
E These authors contributed equally to the present work.
F Corresponding author. Email: muren.herrid@csiro.au
Reproduction, Fertility and Development 21(3) 462-468 https://doi.org/10.1071/RD08130
Submitted: 5 June 2008 Accepted: 14 November 2008 Published: 4 March 2009
Abstract
Although autologous and heterologous transplantation has resulted in colonisation of recipient testes in cattle, the ability of the transplanted spermatogonial stem cells to complete spermatogenesis has not yet been determined. The objective of the present study was to identify and validate microsatellite markers that can distinguish the genotype of different individuals and therefore can be used to detect the presence of donor DNA in recipient semen samples. In a previous study by this group, successful colonisation of recipient testes by heterologous transfer using a fluorescent dye was shown. In the present work, some of the same recipient animals were investigated further to monitor donor-derived sperm production. The bovine microsatellite detection method was developed specifically to test the ejaculates of the recipients and can also be used to pre-match individuals before germ cell transplantation. Semen was collected from the recipients 52–98 weeks after transfer and the presence of donor DNA in the samples was determined using microsatellite markers. In one of the recipients, all collected semen samples were shown to be positive for donor-derived cells; however, the percentage of donor spermatozoa in the recipient ejaculate declined with time. The donor DNA was also detected in both single cell suspensions and testis tissue from this recipient. These results demonstrate for the first time that testicular germ cell transplantation between different breeds of cattle is feasible and the recipients thereof are able to produce spermatozoa of donor origin. This technology has potential applications in livestock breeding systems and may provide an alternative to artificial insemination.
Additional keywords: donor sperm, spermatogenesis.
Acknowledgements
We thank Brendan Hatton and Andrew Eichorn for assistance with experimental animal management and funding from the CSIRO Food Future National Research Flagship and CSIRO Livestock Industries. The authors declare that there is no conflict of interest that would prejudice the impartiality of this scientific work.
Brinster, R. L. , and Avarbock, M. R. (1994). Germline transmission of donor haplotype following spermatogonial transplantation. Proc. Natl. Acad. Sci. USA 91, 11 303–11 307.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
Williams, J. L. , Usha, A. P. , Urquhart, B. G. , and Kilroy, M. (1997). Verification of the identity of bovine semen using DNA microsatellite markers. Vet. Rec. 140, 446–449.
| PubMed | CAS |
Zhang, Z. , Renfree, M. B. , and Short, R. V. (2003). Successful intra- and interspecific male germ cell transplantation in the rat. Biol. Reprod. 68, 961–967.
| Crossref | GoogleScholarGoogle Scholar | PubMed | CAS |
