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Expression of the chicken GDNF family receptor alpha-1 as a marker of spermatogonial stem cells

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MUCKSOVA J KALINA J BAKST M YAN H BRILLARD JP BENESOVA B FAFÍLEK Bohumil HEJNAR J TREFIL P

Rok publikování 2013
Druh Článek v odborném periodiku
Časopis / Zdroj ANIMAL REPRODUCTION SCIENCE
Citace
Doi http://dx.doi.org/10.1016/j.anireprosci.2013.08.006
Klíčová slova GFR alpha 1; Chicken spermatogonial stem cell; Male germ line transplantation
Popis The identification, enrichment and subsequent isolation of spermatogonial stem cells (SSCs) are integral to the success of SCC transplants between fertile donor and sterilized recipient males. In birds generally and particularly in chicken, SSC-specific has yet to be identified. The receptor for glial cell-derived neurotrophic factor (GDNF), i.e. GDNF family receptor alpha-1 (GFR alpha 1), has been identified as a potential marker for different mouse spermatogonial subtypes. In the present study, we characterized the chicken cGFR alpha 1 receptor and compared its predicted amino-acid sequence with mouse, rat and human GFR alpha 1 proteins. Using specific polyclonal mouse anti-cGFR alpha 1 serum, a total of 2.8% cells were recognized as cGFR alpha 1 positive among isolated testicular cells recovered from sexually mature cockerels. The percentages of cGFR alpha 1-positive testicular cells with haploid, diploid, tetraploid and SP DNA content were 1.6%, 2.5%, 393% and 76.8%, respectively. The presence of cGFR alpha 1 protein on the surfaces of all cells of the seminiferous epithelium was confirmed by immunocytochemical and immunohistochemical analyses. Tissue specificity of cGFR alpha 1 mRNA expression was significantly higher in adult testes compared to brain tissue which itself was several times higher than tissues prepared from the spleen, liver and heart. No expression was observed in muscular tissue. At last, we demonstrated the successful repopulation of sterilized recipient's testes with transplanted cGFR alpha 1-positive donor testicular cells. Recipient males subsequently produced functional heterologous spermatozoa capable of fertilizing an ovum and obtaining chicks with donor cell genotypes. (C) 2013 Elsevier B.V. All rights reserved.