Abstract
Spermatogonial stem cells (SSCs) continuously undergo self-renewal and differentiation to sustain spermatogenesis throughout adulthood in males. In stallions, SSCs may be used for the production of progeny from geldings after cryopreservation and therapy for infertile and subfertile stallions. Undifferentiated cell transcription factor 1 (UTF1) is a putative marker for undifferentiated spermatogonia in humans and rats. The main purposes of this study are to determine the following: 1) changes in the expression pattern of UTF1 at various reproductive stages of stallions, 2) subpopulations of spermatogonia that express UTF1. Testicular samples were collected and categorized based on the age of the horses as follows: pre-pubertal (<1 yr), pubertal (1–1.5 yr), post-pubertal (2–3 yr), and adult (4–8 yr). Western blot analysis was utilized to determine the cross-activity of the UTF1 antibody to horse testes tissues. Immunohistochemistry was conducted to investigate the UTF1 expression pattern in germ cells at different reproductive stages. Whole mount staining was applied to determine the subpopulation of UTF1-positive spermatogonia. Immunohistological analysis showed that most germ cells in the pre-pubertal and pubertal stages were immunolabeled with UTF1, whereas only a few germ cells in the basal compartment of the seminiferous tubule cross-sections of post-pubertal and adult tissues were UTF1-positive. No staining was observed in the Sertoli or Leydig cells at any reproductive stages. Whole mount staining showed that As, Apr, and chains of 4, 8, 16 Aal spermatogonia were immunolabeled with UTF1 in the post-pubertal stallion tubule. Isolated single germ cells were also immunolabeled with UTF1. In conclusion, UTF1 is expressed in undifferentiated spermatogonia, and its antibody can be used as a putative marker for SSCs in stallions.
Highlights
Spermatogonial stem cells (SSCs) undergo self-renewal and differentiation for continuous sperm production throughout adult life
Finding a putative marker for undifferentiated spermatogonia will contribute to the development of a new assistant reproductive technique using SSCs, as well as to better understanding the biological activity of spermatogonia for stallion sperm production
The present study was conducted to identify the subpopulations of spermatogonia that expressed UTF1 and to determine the changes in the pattern of UTF1 expression at different reproductive stages
Summary
Spermatogonial stem cells (SSCs) undergo self-renewal and differentiation for continuous sperm production throughout adult life. It has become increasingly evident in mice that Asingle (As), Apaired (Apr), and Aaligned (Aal) spermatogonia are undifferentiated spermatogonia [1,2], which later express the c-kit tyrosine receptor as a commitment step to undergo differentiation into spermatozoa [2]. The putative SSC markers have been used in the localization of SSCs in fixed testicular tissues and SSC colonies of seminiferous tubules using immunohistochemical and whole mount staining techniques, respectively [2,7]. Expression pattern of these molecules at early reproductive stage, the precise size of spermatogonia colony expressing these molecules by whole mount staining, and possible use of these molecules for immunocytochemistry have remained unclear
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