The Germ Cell–Specific Markers ZPBP2 and PGK2 in Testicular Biopsies Can Predict the Presence as well as the Quality of Sperm in Non-obstructive Azoospermia Patients


To assess the role of three testis-specific genes including ZPBP2, PGK2, and ACRV1 in the prediction of sperm retrieval result and quality of retrieved sperm by microdissection testicular sperm extraction (micro-TESE) in non-obstructive azoospermia (NOA) patients. This was a case-control study including 57 testicular samples of NOA patients including 32 patients with successful sperm retrieval (NOA+) and 25 patients with failed sperm retrieval (NOA-), and 9 samples of men with normal spermatogenesis in the testes as the positive control (OA). We investigated the expression of candidate genes by RT-qPCR and germ cell population patterns by DNA flow cytometry in testicular biopsy samples. The association between PGK2 expressions with the quality of retrieved spermatozoa was also evaluated. The RT-qPCR data revealed a significantly higher expression of ZPBP2 and PGK2 in the NOA+ in comparison to NOA- group (P = 0.002, and P = 0.002, respectively). Flow cytometry results revealed that the haploid cell percentage was significantly higher in NOA+ vs. NOA- group (P = 0.0001). In samples with a higher percentage of haploid cells, expression levels of ZPBP2 and PGK2 were higher (P = 0.001). The PGK2 expression was significantly associated with retrieved sperm quality (P = 0.01). Our results contribute to the search for the biomarkers for predicting the presence of testicular sperm and would be useful to avoid unnecessary multiple micro-TESE. Overall, the expression pattern of the ZPBP2 and PGK2 may be useful in predicting sperm recovery success and quality of retrieved sperm in NOA patients.

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The authors are grateful to the patients who participated in the study for their important collaboration. We also thank the andrologists from the Andrology Unit of the Royan Institute, Tehran, Iran, who helped provide samples for this study. A special acknowledgment is dedicated to the staff of the Embryology Laboratory of Tehran University of Medical Sciences for their constant support.


This work was financially supported by a grant from the Tehran University of Medical Sciences (No. 37940). This is an original article as a part of a Ph.D. student thesis.

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Mehdi Abbasi and Mohammad Ali Sadighi Gilani designed the project, supervised the study, and reviewed the manuscript; Mohammad Ali Sadighi Gilani provided samples and performed clinical assessment; Nasrin Ghanami Gashti performed sample collection and the experiments, wrote the manuscript, and performed data analysis; Mohammad Ali Sadighi Gilani and Nasrin Ghanami Gashti performed recruitment of patients and clinical data acquisition; Ayob Jabari and Maryam Qasemi supported and assisted in conducting experiments; Narjes Feizollahi performed materials support. All authors approved the final manuscript for publishing.

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Correspondence to Mehdi Abbasi.

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Ghanami Gashti, N., Sadighi Gilani, M. ., Jabari, A. et al. The Germ Cell–Specific Markers ZPBP2 and PGK2 in Testicular Biopsies Can Predict the Presence as well as the Quality of Sperm in Non-obstructive Azoospermia Patients. Reprod. Sci. (2021).

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  • Male infertility
  • Non-obstructive azoospermia
  • Micro-TESE
  • ZPBP2
  • PGK2