Germ Cell Transplantation Using Sexually Competent Fish: An Approach for Rapid Propagation of Endangered and Valuable Germlines

Sullip K Majhi,Carlos A Strüssmann,Masashi Yokota,Ricardo S Hattori,Seiichi Watanabe,Laszlo Orban

doi:10.1371/journal.pone.0006132

Sullip K Majhi, Carlos A Strüssmann + Show 4 more

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https://doi.org/10.1371/journal.pone.0006132

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Journal: PLoS ONE	Publication Date: Jul 2, 2009
Citations: 94	License type: CC BY 4.0

Affiliation: Tokyo University of Marine Science and Technology

Abstract

The transplantation of germ cells into adult recipient gonads is a tool with wide applications in animal breeding and conservation of valuable and/or endangered species; it also provides a means for basic studies involving germ cell (GC) proliferation and differentiation. Here we describe the establishment of a working model for xenogeneic germ cell transplantation (GCT) in sexually competent fish. Spermatogonial cells isolated from juveniles of one species, the pejerrey Odontesthes bonariensis (Atherinopsidae), were surgically transplanted into the gonads of sexually mature Patagonian pejerrey O. hatcheri, which have been partially depleted of endogenous GCs by a combination of Busulfan (40 mg/kg) and high water temperature (25°C) treatments. The observation of the donor cells' behavior showed that transplanted spermatogonial cells were able to recolonize the recipients' gonads and resume spermatogenesis within 6 months from the GCT. The presence of donor-derived gametes was confirmed by PCR in 20% of the surrogate O. hatcheri fathers at 6 months and crosses with O. bonariensis mothers produced hybrids and pure O. bonariensis, with donor-derived germline transmission rates of 1.2–13.3%. These findings indicate that transplantation of spermatogonial cells into sexually competent fish can shorten considerably the production time of donor-derived gametes and offspring and could play a vital role in germline conservation and propagation of valued and/or endangered fish species.

Highlights

Germ cell (GC) transplantation (GCT) is a powerful reproductive technique pioneered by Brinster and colleagues in 1994 [1]
Donor germ cells harvested from sexually immature O. bonariensis testes were successfully transplanted into partially sterilized testes of sexually competent O. hatcheri by surgical intervention
We conclude 1) that repeated administration of Busulfan and rearing at a high temperature cause pronounced germ cell loss, 2) that testes treated in this way remain sexually competent and able to support gametogenesis, as already noted previously [8,12,18,19], and 3) that germ cell transplantation (GCT) into these gonads using simple injection techniques results in recolonization of the recipient gonads with the transplanted germ cell

Summary

Introduction

Germ cell (GC) transplantation (GCT) is a powerful reproductive technique pioneered by Brinster and colleagues in 1994 [1]. Lacerda et al [8] and Takeuchi et al [9,10] demonstrated that GCT in fish, as originally devised in mammals [1,3], does not need to be performed with PGCs as transplantation of syngeneic or xenogeneic spermatogonia resulted in colonization of the recipient gonads by the transplanted cells [8,11] When these cells were transplanted to rainbow trout hatchlings, they resulted in donor-derived functional gametes that were capable of fertilization and generation of viable offspring [11]. This possibility has been experimentally demonstrated in mammals [1,13], but it has never been tested in adult fish beyond cell transplantation and demonstration of colonization of the recipient gonads [8]

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