Mitochondrial Dynamics Markers in Spermatozoa as Possible New Prognostic/Diagnostic Tool to Assess Men (In/Sub)Fertility – Fantasy or Reality?

Abstract Authors: Silvana A Andric, Isidora M Keselj, Tamara D Bogdanovic, Tamara M Tomanic, Sava M Radovic Pletikosic, Tatjana S Kostic

Laboratory for Reproductive Endocrinology and Signaling, Laboratory for Chronobiology and Aging, CeRES, DBE, Faculty of Sciences, University of Novi Sad, Novi Sad, Serbia

Abstract Text: Mitochondria are important for spermatozoa development, homeostasis and functionality as well as for the process of fertilization.

Our study investigated the transcriptional profiles of human spermatozoal mitochondrial dynamics markers and seminal plasma hormone levels in relation to spermiogram types.

The samples were collected from men participating in the national assisted-reproductive-technology-program. The study design and procedures were approved by the Ethical Committee. The normozoospermic (N), teratozoospermic (T), asthenoteratozoospermic (AT), and oligoasthenoteratozoospermic (OAT) spermatozoa were separated from seminal plasma. Testosterone, cortisol, estradiol and nitrites were measured in seminal plasma. Real-time PCR analyses were conducted to obtain transcriptional profiles of mitochondrial dynamics markers in human spermatozoa.

The results showed that the levels of hormones and nitrites in seminal plasma, as well as transcriptional patterns of mitochondrial dynamics markers in spermatozoa displayed high individual variations. A significant correlation between nitrite levels in seminal plasma and mitochondrial dynamics markers in spermatozoa from normozoospermic and teratozoospermic samples were observed. In teratozoospermic samples, correlation was also detected between gelatinous clamps and transcripts for PPARGC1A and MFN1 in spermatozoa. A significant increase in the levels of PPARGC1A, TFAM, MFN2 and OPA1 transcripts were observed in teratozoospermic spermatozoa comparing to normozoospermic. In the same samples, the levels of NFE2L2 and MFN1 transcripts decreased. MFN1/MFN2 increased in asthenoteratozoospermic spermatozoa. In oligoasthenoteratozoospermic spermatozoa, a significant increase of PPARGC1B, NRF2, MFN1/MFN2 and OPA1 transcripts were detected, while the level of TFAM transcripts decreased. Receiver-operating characteristic (ROC) curve analyses confirmed that TFAM, NFE2L2, MFN1, MFN2 and OPA1 have potentially good diagnostic parameters for teratozoospermia:  TFAM (AUC = 1.00, p = 0.0112);  NFE2L2 (AUC = 1.00, p = 0.0112);  MFN1 (AUC = 0.9524, p = 0.0304), MFN2 (AUC = 1.00, p = 0.0143), OPA1 (AUC = 1.00, p = 0.0201).

Accordingly, the above-mentioned markers could provide a solid foundation for the development of a Mito-Sperm-Signature - a potential new prognostic/diagnostic tool to assess male infertility, as semen quality and fertility are important fundamental markers of reproductive and overall health.

Acknowledgements. This study was supported by the Science Fund of the Republic of Serbia (Grant. No. 14977 Mito-Sperm-Signature) the Ministry of Science, Technological Development and Innovation of the Republic of Serbia (Grants No. ‪451-03-137/2025-03/200125, 451-03-136/2025-03/200125 and CIV-CeRES-2024 grant), the Autonomic Province of Vojvodina (grant APV9418) and the ANDRONET action