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Advances in Spermatogenesis and Male Infertility

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Biology".

Deadline for manuscript submissions: 20 September 2025 | Viewed by 13272

Special Issue Editor


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Guest Editor
The Sperm Laboratory, College of Life Sciences, Zhejiang University, Hangzhou 310058, China
Interests: cell cycle; spermatogenesis; kinesin; testis; motor protein; cytoskeleton

Special Issue Information

Dear Colleagues,

Sperm provides the zygote with genetic material from the father and is essential for reproduction. Throughout biological evolution, spermatogenesis has been a precise and conserved process which occurs in the male testis but is regulated by the hypothalamic–pituitary–gonadal axis. The blood–testis barrier, as the histological barrier of the testis, prevents the stimulation of spermatogenesis and ensures the process runs smoothly. A variety of signaling molecules are involved in various stages of and at different points in spermatogenesis, and studying its related functions may provide us with a new therapeutic direction for male reproductive diseases. However, at present, more and more environmental factors impair this process, impeding spermatogenesis and as a result decreasing male fertility rates.

The purpose of this Special Issue is to summarize and expand on the molecular mechanisms that ensure successful spermatogenesis and to explore the key processes in male reproduction, providing an effective platform on which new insights into male reproductive diseases to be described.

We welcome submissions of original research articles and comprehensive reviews.

The specific topics include, but are not limited to, the following:

  • The role of hormone regulation in spermatogenesis;
  • How somatic cells maintain spermatogenesis in the testis;
  • The maintenance mechanism ensuring the integrity of the blood–testis barrier and its role in spermatogenesis;
  • The multiple signaling pathways that synergistically regulate spermatogenesis;
  • New discoveries of molecules involved in spermatogenesis and fertility;
  • How cell fate is involved in spermatogenesis (e.g. cell self-renewal and repair, apoptosis, proliferation, autophagy, etc.);
  • Molecular mechanisms that disrupt spermatogenesis via environmental toxicology and stimulation.

Prof. Dr. Wanxi Yang
Guest Editor

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Keywords

  • male reproduction
  • hormone regulation
  • blood–testis barrier
  • male reproductive diseases
  • male infertility

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Published Papers (8 papers)

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Research

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27 pages, 5470 KiB  
Article
Age-Associated Proteomic Changes in Human Spermatozoa
by Mohd Amin Beg, Abrar Osama Ismail, Ayodele Alaiya, Firdous Ahmad Khan, Taha Abo-Almagd Abdel-Meguid Hamoda, Ishfaq Ahmad Sheikh, Priyanka Sharma, Omar Mohammed Baothman, Ali Hasan Alkhzaim, Zakia Shinwari, Rinad Fahad Abuzinadah, Arif Mohammed, Abdullah Mohammed Assiri, Adel Mohammad Abuzenadah, Erdogan Memili and Jean Magloire Feugang
Int. J. Mol. Sci. 2025, 26(13), 6099; https://doi.org/10.3390/ijms26136099 (registering DOI) - 25 Jun 2025
Viewed by 205
Abstract
Advancing age in men significantly contributes to declining sperm fertility. Information on age-related proteomic changes in spermatozoa is limited. This study involved normal fertile Arab men in three age groups: young adult (21–30 years; n = 6), late adult (31–40 years; n = [...] Read more.
Advancing age in men significantly contributes to declining sperm fertility. Information on age-related proteomic changes in spermatozoa is limited. This study involved normal fertile Arab men in three age groups: young adult (21–30 years; n = 6), late adult (31–40 years; n = 7), and advanced age (40–51 years; n = 5). Gradient-purified spermatozoa were analyzed using LC-MS/MS and proteomic data were processed using Progenesis QI (QIfp) v3.0 and UniProt/SwissProt. Significantly enriched annotations and clustering of proteins in the proteomic datasets were identified (2-fold change; p < 0.05). A total of 588 proteins were identified, with 93% shared across the three groups. Unique proteins were MYLK4 for the young adult group, PRSS57 for the late adult group, and HMGB4, KRT4, LPGAT1, OXCT2, and MGRN1 for the advanced age group. Furthermore, 261 (44%) proteins were differentially expressed (p < 0.05) across the three groups. Functional enrichment analysis suggested an aging-related significant increase in pathways associated with neurodegenerative diseases and protein folding, alongside decreases in glycolysis/gluconeogenesis, flagellated sperm motility, acetylation, phosphoprotein modifications, oxidation processes, and Ubl conjugation. Cluster analysis highlighted significantly upregulated proteins in young adults (e.g., H2BC1, LAP3, SQLE, LTF, PDIA4, DYNLT2) and late adults (e.g., ATP5F1B, ODF2, TUBA3C, ENO1, SPO11, TEX45, TEKT3), whereas most proteins in the advanced age group exhibited downregulation (e.g., SPESP1, RAB10, SEPTIN4, RAB15, PTPN7, USP5, ANXA1, PRDX1). In conclusion, this study revealed aging-associated proteomic changes in spermatozoa that impact critical processes, including spermatogenesis, motility, metabolism, and fertilization, potentially contributing to fertility decline. These changes provide a molecular framework for developing therapies to preserve sperm proteostasis and enhance fertility in older men. Full article
(This article belongs to the Special Issue Advances in Spermatogenesis and Male Infertility)
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16 pages, 1039 KiB  
Article
Comparison of Swim-Up and Microfluidic Sperm Sorting Methods in Selection of Sperm for Intracytoplasmic Sperm Injection
by Michal Ješeta, Adéla Doubravská, Jana Antalíková, Lenka Mekiňová, Kateřina Franzová, Kateřina Remundová, Jan Hošek, Bartosz Kempisty and Robert Hudeček
Int. J. Mol. Sci. 2025, 26(11), 5374; https://doi.org/10.3390/ijms26115374 - 4 Jun 2025
Viewed by 524
Abstract
The use of microfluidic sperm sorting (MFSS) systems in infertility treatment is increasing due to their practicality and ease of use. While often presented as highly effective, their efficacy in patients with varying sperm analysis results remains uncertain. In this study, we evaluated [...] Read more.
The use of microfluidic sperm sorting (MFSS) systems in infertility treatment is increasing due to their practicality and ease of use. While often presented as highly effective, their efficacy in patients with varying sperm analysis results remains uncertain. In this study, we evaluated the effectiveness of MFSS compared with the swim-up (SU) technique in terms of oxygen radical levels and spermiogram parameters. Samples from each patient were processed using both methods, followed by assessments of sperm concentration, motility, morphology, DNA integrity, acrosomal status, and mitochondrial membrane potential. Participants were selected based on sperm analysis and categorized as normozoospermic (n = 40) or non-normozoospermic (n = 28). An analysis of separation techniques revealed no significant differences, except for a lower percentage of DNA-fragmented sperm in the MFSS group compared with SU within the non-normozoospermic cohort (SU: 10.0% vs. MFSS: 5.69%, p = 0.027). No differences were observed between SU and MFSS in normozoospermic men. The MFSS method is a simple technique, frequently used in laboratories, that yields good results but does not offer a substantial advantage over SU. The primary benefit of MFSS appears to be a significant reduction in the proportion of sperm with DNA fragmentation compared with SU in patients with abnormal sperm analysis results. Full article
(This article belongs to the Special Issue Advances in Spermatogenesis and Male Infertility)
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31 pages, 5528 KiB  
Article
Early Zinc Supplementation Enhances Epididymal Sperm Glycosylation, Endocrine Activity, and Antioxidant Activity in Rats Exposed to Cadmium
by Sergio Marín de Jesús, Rosa María Vigueras-Villaseñor, Edith Cortés-Barberena, Joel Hernández-Rodríguez, Sonia Guadalupe Pérez-Aguirre, Sergio Montes, Leticia Carrizales-Yáñez, Isabel Arrieta-Cruz and Marcela Arteaga-Silva
Int. J. Mol. Sci. 2025, 26(10), 4589; https://doi.org/10.3390/ijms26104589 - 10 May 2025
Viewed by 414
Abstract
Sperm maturation involves changes in plasma membrane glycosylation for fertilization. Cadmium (Cd) exerts a negative effect by disrupting testicular and epididymal function, altering antioxidant activity. Zinc (Zn) is an essential element known for its antioxidant properties, role in testosterone synthesis, and support of [...] Read more.
Sperm maturation involves changes in plasma membrane glycosylation for fertilization. Cadmium (Cd) exerts a negative effect by disrupting testicular and epididymal function, altering antioxidant activity. Zinc (Zn) is an essential element known for its antioxidant properties, role in testosterone synthesis, and support of spermatogenesis. However, its effect on sperm membrane glycosylation, as well as endocrine and antioxidant activity, after exposure to Cd has remained unexplored. This study evaluated the impact of Zn on epididymal sperm glycosylation, endocrine activity, and antioxidant activity in Cd-exposed rats. Four groups of male Wistar rats were analyzed: control, Cd-exposed, Zn-supplemented, and Zn + Cd groups. On postnatal day 90, tissues and blood were collected for Zn and Cd quantification, testosterone levels, antioxidant activity, histological analysis, and sperm quality. The results showed that Cd concentration increased significantly, reduced testosterone levels, modified antioxidant activity, and caused structural damage in the epididymis. The Cd-exposed group showed disrupted glycosylation and distribution patterns and reduced sperm quality. The Zn + Cd group showed lower Cd accumulation, preserved testosterone levels, restored antioxidant activity, and preserved glycosylation patterns and sperm quality. This study highlights the protective role of Zn in mitigating Cd-induced reproductive toxicity, probably through the competitive inhibition of Cd uptake and antioxidant support, thereby preserving fertility. Full article
(This article belongs to the Special Issue Advances in Spermatogenesis and Male Infertility)
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27 pages, 8689 KiB  
Article
Impact of Ultraviolet C Radiation on Male Fertility in Rats: Suppression of Autophagy, Stimulation of Gonadotropin-Inhibiting Hormone, and Alteration of miRNAs
by Ahmed Mohamed Alahwany, Ahmed Hamed Arisha, Adel Abdelkhalek, Tarek Khamis, Taku Miyasho and Doaa Kirat
Int. J. Mol. Sci. 2025, 26(1), 316; https://doi.org/10.3390/ijms26010316 - 1 Jan 2025
Cited by 2 | Viewed by 3736
Abstract
While ultraviolet C (UVC) radiation has beneficial applications, it can also pose risks to living organisms. Nevertheless, a detailed assessment of UVC radiation’s effects on mammalian male reproductive physiology, including the underlying mechanisms and potential protective strategies, has not yet been accomplished. This [...] Read more.
While ultraviolet C (UVC) radiation has beneficial applications, it can also pose risks to living organisms. Nevertheless, a detailed assessment of UVC radiation’s effects on mammalian male reproductive physiology, including the underlying mechanisms and potential protective strategies, has not yet been accomplished. This study aimed to examine the critical roles of oxidative stress, autophagy, reproductive hormonal axis, and microRNAs in UVC-induced reproductive challenges in male rats. Semen, biochemical, molecular, and in silico analyses revealed significant dysregulation of testicular steroidogenesis, impaired spermatogenesis, deteriorated sperm quality, and altered reproductive hormonal profiles, which ultimately lead to a decline in fertility in male rats exposed to UVC radiation. Our data indicated that the suppression of autophagy, stimulation of gonadotropin-inhibiting hormone (GnIH), and alteration of microRNAs serve as key mediators of UVC-induced stress effects in mammalian reproduction, potentially contributing to male infertility. Targeting these pathways, particularly through pretreatment with hesperidin (HES), offers a promising strategy to counteract UVC-induced male infertility. In conclusion, the present findings emphasize the importance of understanding the molecular mechanisms behind UVC-induced male infertility and offer valuable insights into the protective mechanisms and prospective role of HES in safeguarding male reproductive health. Full article
(This article belongs to the Special Issue Advances in Spermatogenesis and Male Infertility)
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23 pages, 5912 KiB  
Article
Functional Study of PTSMAD4 in the Spermatogenesis of the Swimming Crab Portunus trituberculatus
by Yu Zhang, Qiu-Meng Xiang, Chang-Kao Mu, Chun-Lin Wang and Cong-Cong Hou
Int. J. Mol. Sci. 2024, 25(23), 13126; https://doi.org/10.3390/ijms252313126 - 6 Dec 2024
Viewed by 952
Abstract
Portunus trituberculatus holds significant economic value. The spermatogenesis is regulated by numerous signaling pathways. Among them, the TGF-β signaling pathway plays an important role in the development of testes and spermatogenesis. Smad4 is a Co-Smad protein that forms a complex with R-Smad to [...] Read more.
Portunus trituberculatus holds significant economic value. The spermatogenesis is regulated by numerous signaling pathways. Among them, the TGF-β signaling pathway plays an important role in the development of testes and spermatogenesis. Smad4 is a Co-Smad protein that forms a complex with R-Smad to regulate the expression of target genes. The sperm structure in crustaceans differs greatly from that in mammals, with mature sperm lacking tails. Our previous studies have reported the function of R-Smad in the spermatogenesis of P. trituberculatus. In this study, we cloned the full-length cDNA sequence of PTSMAD4; immunofluorescence technology revealed that PTSMAD4 is expressed throughout all stages of spermatogenesis. We knocked down the expression of PTSMAD4 in P. trituberculatus using RNAi technology, and the immunofluorescence results show abnormal co-localization and weakened signals of PTSMAD4 and PTSMAD2. Additionally, transcriptome sequencing results enriched functional genes and pathways related to spermatogenesis. This study indicates that PTSMAD4 may participate in the spermatogenesis process through its involvement in signal transduction. This research not only lays the foundation for further study of the function of the TGF-β signaling pathway in spermatogenesis but also provides a theoretical basis for further investigation of the spermatogenesis mechanism in crustaceans. Full article
(This article belongs to the Special Issue Advances in Spermatogenesis and Male Infertility)
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16 pages, 3869 KiB  
Article
Sperm-Borne Mitochondrial Activity Influenced by Season and Age of Holstein Bulls
by Khurshaid Anwar, Georg Thaller and Mohammed Saeed-Zidane
Int. J. Mol. Sci. 2024, 25(23), 13064; https://doi.org/10.3390/ijms252313064 - 5 Dec 2024
Cited by 1 | Viewed by 1293
Abstract
Sperm mitochondria are vital organelles for energy production and pre- and post-fertilization sperm functions. The potential influence of the age of the bull and season on the sperm-borne mitochondrial copy number and the transcription activity has not yet been investigated. Therefore, the expression [...] Read more.
Sperm mitochondria are vital organelles for energy production and pre- and post-fertilization sperm functions. The potential influence of the age of the bull and season on the sperm-borne mitochondrial copy number and the transcription activity has not yet been investigated. Therefore, the expression patterns of all protein-coding mitochondrial genes were identified throughout the year along with mitochondrial copy numbers in young and old bulls’ spermatozoa. For that, high-quality semen samples (n = 32) with more than 80% quality for the morphological parameters, from young (n = 4, aged 18–24 months old) and old (n = 4, aged 40–54 months old) Holstein bulls, were collected during the four seasons (n = 4 samples each animal/season). The DNA and RNA were isolated from sperm cells and subjected to the DNA copy number and expression analyses using qPCR. Furthermore, an in silico analysis using gene ontology online tools for the abundantly expressed genes was utilized. The data were statistically analyzed using Prism10 software. There was a significant reduction in the mitochondria copy number of young bulls’ spermatozoa compared to their old counterparts during the summer (29 ± 3 vs. 51 ± 6, p < 0.001) and winter (27 ± 3 vs. 43 ± 7, p < 0.01) seasons. However, sperm-borne mitochondrial protein-coding genes were transcriptionally higher in young bulls throughout the year. Within the same group of bulls, unlike the old bulls, there was a significant (p < 0.05) induction in the transcription activity accompanied by a significant (p < 0.05) reduction in the mitochondrial copy numbers in the summer (29 ± 3) and winter (27 ± 3) compared to the spring (42 ± 9) and autumn (36 ± 5) seasons in young bulls. Additionally, the pathway enrichment of the top six expressed genes differed between age groups and seasons. In conclusion, under the same quality of semen, the early stages of age are associated with mitochondrial biogenesis and transcription activity dysregulation in a season-dependent manner. Full article
(This article belongs to the Special Issue Advances in Spermatogenesis and Male Infertility)
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29 pages, 19432 KiB  
Article
ERK/CREB and p38 MAPK/MMP14 Signaling Pathway Influences Spermatogenesis through Regulating the Expression of Junctional Proteins in Eriocheir sinensis Testis
by Hong-Yu Qi, Zhan Zhao, Bang-Hong Wei, Zhen-Fang Li, Fu-Qing Tan and Wan-Xi Yang
Int. J. Mol. Sci. 2024, 25(13), 7361; https://doi.org/10.3390/ijms25137361 - 4 Jul 2024
Viewed by 1735
Abstract
The hemolymph–testis barrier (HTB) is a reproduction barrier in Crustacea, guaranteeing the safe and smooth process of spermatogenesis, which is similar to the blood–testis barrier (BTB) in mammals. The MAPK signaling pathway plays an essential role in spermatogenesis and maintenance of the BTB. [...] Read more.
The hemolymph–testis barrier (HTB) is a reproduction barrier in Crustacea, guaranteeing the safe and smooth process of spermatogenesis, which is similar to the blood–testis barrier (BTB) in mammals. The MAPK signaling pathway plays an essential role in spermatogenesis and maintenance of the BTB. However, only a few studies have focused on the influence of MAPK on crustacean reproduction. In the present study, we knocked down and inhibited MAPK in Eriocheir sinensis. Increased defects in spermatogenesis were observed, concurrently with a damaged HTB. Further research revealed that es-MMP14 functions downstream of ERK and p38 MAPK and degrades junctional proteins (Pinin and ZO-1); es-CREB functions in the ERK cascade as a transcription factor of ZO-1. In addition, when es-MMP14 and es-CREB were deleted, the defects in HTB and spermatogenesis aligned with abnormalities in the MAPK. However, JNK impacts the integrity of the HTB by changing the distribution of intercellular junctions. In summary, the MAPK signaling pathway maintains HTB integrity and spermatogenesis through es-MMP14 and es-CREB, which provides insights into the evolution of gene function during barrier evolution. Full article
(This article belongs to the Special Issue Advances in Spermatogenesis and Male Infertility)
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Review

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22 pages, 5926 KiB  
Review
Structure and Composition of Spermatozoa Fibrous Sheath in Diverse Groups of Metazoa
by Ekaterina A. Guseva, Vitaly S. Buev, Sabina E. Mirzaeva, Philipp I. Pletnev, Olga A. Dontsova and Petr V. Sergiev
Int. J. Mol. Sci. 2024, 25(14), 7663; https://doi.org/10.3390/ijms25147663 - 12 Jul 2024
Cited by 1 | Viewed by 3002
Abstract
The proper functioning and assembly of the sperm flagella structures contribute significantly to spermatozoa motility and overall male fertility. However, the fine mechanisms of assembly steps are poorly studied due to the high diversity of cell types, low solubility of the corresponding protein [...] Read more.
The proper functioning and assembly of the sperm flagella structures contribute significantly to spermatozoa motility and overall male fertility. However, the fine mechanisms of assembly steps are poorly studied due to the high diversity of cell types, low solubility of the corresponding protein structures, and high tissue and cell specificity. One of the open questions for investigation is the attachment of longitudinal columns to the doublets 3 and 8 of axonemal microtubules through the outer dense fibers. A number of mutations affecting the assembly of flagella in model organisms are known. Additionally, evolutionary genomics data and comparative analysis of flagella morphology are available for a set of non-model species. This review is devoted to the analysis of diverse ultrastructures of sperm flagellum of Metazoa combined with an overview of the evolutionary distribution and function of the mammalian fibrous sheath proteins. Full article
(This article belongs to the Special Issue Advances in Spermatogenesis and Male Infertility)
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