AIM: The objective of this project is to evaluate the protein expression of putative spermatogonial stem cell (SCC) and meiotic markers in human testicular tissue. Marker expression was studied in testicular samples at different stages of development (fetal, prepubertal and adult) and a timeline expression was established. In addition, samples of prepuberal patients who underwent gonadotoxic treatments or were diagnosed for Klinefelter syndrome were included in order to evaluate the effect of these conditions on different marker expression. METHODS: Immunohistochemistry was performed using antibodies for TEX15, PAX7, ID4, SSEA4 and “PROTEIN A”. The samples were evaluated by light microscopy. The protein expression pattern was classified according to the intensity of the signal in three different groups (intense, faint, no signal). ANOVA and Bonferoni statistical tests were performed with the software SPSS to evaluate significant differences in the marker’s expression between groups. RESULTS: Among the different markers tried, TEX15 expression timeline was successfully established. The obtained results confirm that the protein is expressed in spermatids and in postmeiotic germ cells, as previously described in mouse. However, a faint expression was also detected in Sertoli cells. Klinefelter syndrome patients presented a lower number of spermatogonia when compared with the rest of the groups but no significant differences of expression TEX15 in spermatogonia were found among the different groups of patients. However, patients treated with hydroxyurea showed a fainter TEX15 expression in Serotoli cells, while Klinefelter syndrome patients showed a more intense expression. CONCLUSIONS: PAX7, ID4, SSEA-4 and “PROTEIN A” timeline expression could not be stablished due to technical difficulties. Differences in number of spermatogonia but no differences in TEX15 expression were observed in the different groups of prepubertal patients. Expression differences in Sertoli cells were observed according to the background of prepubertal patients.
|Date of Award||Sep 2015|
- protein expression