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Ovary and Testis: Molecular Biological Insights

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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: closed (15 April 2024) | Viewed by 10571

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Dr. Volodimir Isachenko

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Guest Editor

1. Professor of Medical Faculty, University Temuco, Temuco, Cautín 4801057, Chile
2. Department of Obstetrics and Gynaecology, Medical Faculty, Cologne University, 50931 Cologne, Germany
Interests: cancer; cells; cryopreservation; embryo; proliferation; reproductive; spermatozoa; oocyte; ovarian tissue
Special Issues, Collections and Topics in MDPI journals

Prof. Dr. Frank Nawroth

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Guest Editor

Gynecological Endocrinology and Reproductive Medicine Center for Fertility, Prenatal Medicine, Endocrinology and Osteology, 20095 Hamburg, Germany
Interests: gynecological endocrinology; reproductive medicine

Special Issue Information

Dear Colleagues,

This Special Issue, “Ovary and testis: Molecular Biological Insights”, will cover a selection of recent research and current review articles in the field. This Special Issue is devoted not only to research directly related to the cryopreservation of cells, but also to the study of the characteristics of spermatogenesis and oogenesis theoretically associated with the cell cryo-resistance. The cryopreservation of ovarian and testicular tissues leads to cell damage. Intracellular and extracellular ice-crystal formation and osmotic damage of cells can further deteriorate their viability. The production of reactive oxygen species leads to an increase in lipid peroxidation after the cryopreservation of tissue, which is associated with a loss of viability. Cryopreservation also leads to diminished antioxidant defense activity during cooling and (or) structural damage to the cytoskeleton and (or) antioxidant enzymes. It can also lead to chromatin damage, which is extremely important because chromatin abnormalities have repercussions on cell quality. Additionally, cell DNA damage is strongly correlated with the mutagenic effects of cryopreservation. The study of such cryo-changes using molecular biological methods can aid the development of more efficient cryopreservation technologies. It is also known that the cryo-biological properties of cells can change during the process of oogenesis and spermatogenesis.

Dr. Pradeep Kumar (drpradeepkrvet@gmail.com, Central Institute for Research of Buffaloes, Hisar, India) and Dr. Wensheng Liu (liuws@gdszjk.org.cn, Center of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical University, Guangzhou, China), whose central area of research interest are mammalian testis and ovarian tissues are serving as co-Guest Editor and will assist Dr. (SU) Volodimir Isachenko and Prof. Dr. med. Frank Nawroth in managing this Issue.

Dr. Volodimir Isachenko
Prof. Dr. Frank Nawroth
Guest Editors

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Keywords

mammalian
human
ovary
testis
molecular
biology
physiology
fertility
sterility
follicles
oogenesis
spermatogenesis
gene
molecular genetics
genome
expression
DNA
RNA
cooling
cryopreservation
freezing
transplantation
proteomics
genomics
transcriptomics
metabolomics
apoptosis
cancer

Published Papers (9 papers)

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Research

Jump to: Review

16 pages, 9922 KiB

Open AccessArticle

RNA Interference-Mediated Suppression of Ecdysone Signaling Inhibits Choriogenesis in Two Coleoptera Species

by Xiao-Qing Zhang, Lin Jin, Wen-Chao Guo, Kai-Yun Fu and Guo-Qing Li

Int. J. Mol. Sci. 2024, 25(8), 4555; https://doi.org/10.3390/ijms25084555 - 22 Apr 2024

Viewed by 220

Abstract

During choriogenesis in insects, chorion (eggshell) is formed by surrounding follicular epithelial cells in ovarioles. However, the regulatory endocrine factor(s) activating choriogenesis and the effect of chemical components on eggshell deserve further exploration. In two representative coleopterans, a coccinellid Henosepilachna vigintioctopunctata and a chrysomelid Leptinotarsa decemlineata, genes encoding the 20-hydroxyecdysone (20E) receptor heterodimer, ecdysone receptor (EcR) and ultraspiracle (USP), and two chitin biosynthesis enzymes UDP-N-acetylglucosamine pyrophosphorylase (UAP) and chitin synthase (ChS1), were highly expressed in ovaries of the young females. RNA interference (RNAi)-aided knockdown of either HvEcR or Hvusp in H. vigintioctopunctata inhibited oviposition, suppressed the expression of HvChS1, and lessened the positive signal of Calcofluor staining on the chorions, which suggests the reduction of a chitin-like substance (CLS) deposited on eggshells. Similarly, RNAi of LdEcR or Ldusp in L. decemlineata constrained oviposition, decreased the expression of LdUAP1 and LdChS1, and reduced CLS contents in the resultant ovaries. Knockdown of LdUAP1 or LdChS1 caused similar defective phenotypes, i.e., reduced oviposition and CLS contents in the L. decemlineata ovaries. These results, for the first time, indicate that 20E signaling activates choriogenesis in two coleopteran species. Moreover, our findings suggest the deposition of a CLS on the chorions. Full article

(This article belongs to the Special Issue Ovary and Testis: Molecular Biological Insights)

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15 pages, 2861 KiB

Open AccessArticle

Ultra-Fast Vitrification: Minimizing the Toxicity of Cryoprotective Agents and Osmotic Stress in Mouse Oocyte Cryopreservation

by Jung-Ran Cho, Eun-Hee Yu, Hyun-Joo Lee, In-Hye Kim, Ji-Hye Jeong, Dan-Bi Lee, Seong-Keun Cho and Jong-Kil Joo

Int. J. Mol. Sci. 2024, 25(3), 1884; https://doi.org/10.3390/ijms25031884 - 04 Feb 2024

Viewed by 838

Abstract

Globally, women have been adopting oocyte cryopreservation (OC) for fertility preservation for various reasons, such as inevitable gonadotoxic treatment for specific pathologic states and social preferences. While conventional vitrification (C-VIT) has improved the success rate of OC, challenges of possible toxicities of high-concentration cryoprotective agents and osmotic stress persist. To overcome these challenges, we evaluated the ultra-fast vitrification (UF-VIT) method, which reduces the equilibration solution stage exposure time compared to C-VIT by observing mouse oocyte intracellular organelles and embryonic development. Consequently, compared to fresh mouse oocytes, UF-VIT presented significant differences only in endoplasmic reticulum (ER) intensity and mitochondrial (MT) distribution. Meanwhile, C-VIT showed substantial differences in the survival rate, key ER and MT parameters, and embryonic development rate. UF-VIT exhibited considerably fewer negative effects on key MT parameters and resulted in a notably higher blastocyst formation rate than C-VIT. Meiotic spindle (spindle and chromosomes) morphology showed no significant changes between the groups during vitrification/warming (VW), suggesting that VW did not negatively affect the meiotic spindle of the oocytes. In conclusion, UF-VIT seems more effective in OC owing to efficient cytoplasmic water molecule extraction, osmotic stress reduction, and minimization of cell contraction and expansion amplitude, thus compensating for the drawbacks of C-VIT. Full article

(This article belongs to the Special Issue Ovary and Testis: Molecular Biological Insights)

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16 pages, 3074 KiB

Open AccessArticle

Comparative Transcriptomic Analyses for the Optimization of Thawing Regimes during Conventional Cryopreservation of Mature and Immature Human Testicular Tissue

by Cheng Pei, Plamen Todorov, Mengyang Cao, Qingduo Kong, Evgenia Isachenko, Gohar Rahimi, Nina Mallmann-Gottschalk, Pamela Uribe, Raul Sanchez and Volodimir Isachenko

Int. J. Mol. Sci. 2024, 25(1), 214; https://doi.org/10.3390/ijms25010214 - 22 Dec 2023

Viewed by 653

Abstract

Cryopreservation of human testicular tissue, as a key element of anticancer therapy, includes the following stages: saturation with cryoprotectants, freezing, thawing, and removal of cryoprotectants. According to the point of view existing in “classical” cryobiology, the thawing mode is the most important consideration in the entire process of cryopreservation of any type of cells, including cells of testicular tissue. The existing postulate in cryobiology states that any frozen types of cells must be thawed as quickly as possible. The technologically maximum possible thawing temperature is 100 °C, which is used in our technology for the cryopreservation of testicular tissue. However, there are other points of view on the rate of cell thawing, according to how thawing should be carried out at physiological temperatures. In fact, there are morphological and functional differences between immature (from prepubertal patients) and mature testicular tissue. Accordingly, the question of the influence of thawing temperature on both types of tissues is relevant. The purpose of this study is to explore the transcriptomic differences of cryopreserved mature and immature testicular tissue subjected to different thawing methods by RNA sequencing. Collected and frozen testicular tissue samples were divided into four groups: quickly (in boiling water at 100 °C) thawed cryopreserved mature testicular tissue (group 1), slowly (by a physiological temperature of 37 °C) thawed mature testicular tissue (group 2), quickly thawed immature testicular tissue (group 3), and slowly thawed immature testicular tissue (group 4). Transcriptomic differences were assessed using differentially expressed genes (DEG), the Kyoto Encyclopedia of Genes and Genomes (KEGG), gene ontology (GO), and protein–protein interaction (PPI) analyses. No fundamental differences in the quality of cells of mature and immature testicular tissue after cryopreservation were found. Generally, thawing of mature and immature testicular tissue was more effective at 100 °C. The greatest difference in the intensity of gene expression was observed in ribosomes of cells thawed at 100 °C in comparison with cells thawed at 37 °C. In conclusion, an elevated speed of thawing is beneficial for frozen testicular tissue. Full article

(This article belongs to the Special Issue Ovary and Testis: Molecular Biological Insights)

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17 pages, 6076 KiB

Open AccessArticle

miR-29a Is Downregulated in Progenies Derived from Chronically Stressed Males

by Marta F. Riesco, David G. Valcarce, Alba Sellés-Egea, Anna Esteve-Codina, María Paz Herráez and Vanesa Robles

Int. J. Mol. Sci. 2023, 24(18), 14107; https://doi.org/10.3390/ijms241814107 - 14 Sep 2023

Viewed by 878

Abstract

Recent research has provided compelling evidence demonstrating that paternal exposure to different stressors can influence their offspring’s phenotypes. We hypothesized that paternal stress can negatively impact the progeny, altering different miRs and triggering different physiological alterations that could compromise offspring development. To investigate this, we exposed zebrafish male siblings to a chronic stress protocol for 21 days. We performed RNA-sequencing (RNA-seq) analyses to identify differentially expressed small noncoding RNAs in 7-day postfertilization (dpf) larvae derived from paternally stressed males crossed with control females compared with the control progeny. We found a single miRNA differentially expressed—miR-29a—which was validated in larva and was also tested in the sperm, testicles, and brain of the stressed progenitors. We observed a vertical transmission of chronic stress to the unexposed larvae, reporting novel consequences of paternally inherited chronic stress at a molecular level. The deregulation of mi-R29a in those larvae could affect relevant biological processes affecting development, morphogenesis, or neurogenesis, among others. Additionally, these disruptions were associated with reduced rates of survival and hatching in the affected offspring. Full article

(This article belongs to the Special Issue Ovary and Testis: Molecular Biological Insights)

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15 pages, 7801 KiB

Open AccessCommunication

RNA Transcripts in Human Ovarian Cells: Two-Time Cryopreservation Does Not Affect Developmental Potential

by Yang Zhou, Wanxue Wang, Plamen Todorov, Cheng Pei, Evgenia Isachenko, Gohar Rahimi, Peter Mallmann, Frank Nawroth and Volodimir Isachenko

Int. J. Mol. Sci. 2023, 24(8), 6880; https://doi.org/10.3390/ijms24086880 - 07 Apr 2023

Cited by 2 | Viewed by 1316

Abstract

Sometimes, for medical reasons, when a frozen tissue has already thawed, an operation by re-transplantation may be cancelled, and ovarian tissues should be re-frozen for transplantation next time. Research about the repeated cryopreservation of ovarian cells is rarely reported. It has been published that there is no difference in the follicle densities, proportions of proliferation of early preantral follicles, appearance of atretic follicles, or ultrastructural quality of frozen-thawed and re-frozen-rethawed tissue. However, the molecular mechanisms of a repeated cryopreservation effect on the developmental potential of ovarian cells are unknown. The aim of our experiments was to investigate the effect of re-freezing and re-thawing ovarian tissue on gene expression, gene function annotation, and protein–protein interactions. The morphological and biological activity of primordial, primary, and secondary follicles, aimed at using these follicles for the formation of artificial ovaries, was also detected. Second-generation mRNA sequencing technology with a high throughput and accuracy was adopted to determine the different transcriptome profiles in the cells of four groups: one-time cryopreserved (frozen and thawed) cells (Group 1), two-time cryopreserved (re-frozen and re-thawed after first cryopreservation) cells (Group 2), one-time cryopreserved (frozen and thawed) and in vitro cultured cells (Group 3), and two times cryopreserved (re-frozen and re-thawed after first cryopreservation) and in vitro cultured cells (Group 4). Some minor changes in the primordial, primary, and secondary follicles in terms of the morphology and biological activity were detected, and finally, the availability of these follicles for the formation of artificial ovaries was explored. It was established that during cryopreservation, the CEBPB/CYP19A1 pathway may be involved in regulating estrogen activity and CD44 is crucial for the development of ovarian cells. An analysis of gene expression in cryopreserved ovarian cells indicates that two-time (repeated) cryopreservation does not significantly affect the developmental potential of these cells. For medical reasons, when ovarian tissue is thawed but cannot be transplanted, it can be immediately re-frozen again. Full article

(This article belongs to the Special Issue Ovary and Testis: Molecular Biological Insights)

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14 pages, 4955 KiB

Open AccessArticle

Genome-Wide Landscape of mRNAs, lncRNAs, and circRNAs during Testicular Development of Yak

by Yongfu La, Xiaoming Ma, Pengjia Bao, Min Chu, Ping Yan, Chunnian Liang and Xian Guo

Int. J. Mol. Sci. 2023, 24(5), 4420; https://doi.org/10.3390/ijms24054420 - 23 Feb 2023

Cited by 1 | Viewed by 1013

Abstract

Testicular development is a tightly regulated process in mammals. Understanding the molecular mechanisms of yak testicular development will benefit the yak breeding industry. However, the roles of different RNAs, such as mRNA, lncRNA, and circRNA in the testicular development of yak, are still largely unclear. In this study, transcriptome analyses were performed on the expression profiles of mRNAs, lncRNAs, and circRNAs in testis tissues of Ashidan yak at different developmental stages, including 6-months-old (M6), 18-months-old (M18), and 30-months-old (M30). A total of 30, 23, and 277 common differentially expressed (DE) mRNAs, lncRNAs, and circRNAs were identified in M6, M18, and M30, respectively. Furthermore, functional enrichment analysis showed that the common DE mRNAs during the entire developmental process were mainly involved in gonadal mesoderm development, cell differentiation, and spermatogenesis processes. Additionally, co-expression network analysis identified the potential lncRNAs related to spermatogenesis, e.g., TCONS_00087394 and TCONS_00012202. Our study provides new information about changes in RNA expression during yak testicular development, which greatly improves our understanding of the molecular mechanisms regulating testicular development in yaks. Full article

(This article belongs to the Special Issue Ovary and Testis: Molecular Biological Insights)

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Review

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16 pages, 2783 KiB

Open AccessReview

Current Fertility Preservation Steps in Young Women Suffering from Cancer and Future Perspectives

by Alicia Marco, Marta Gargallo, Jesús Ciriza, Ariella Shikanov, Laura Baquedano, Javier García Pérez-Llantada and Clara Malo

Int. J. Mol. Sci. 2024, 25(8), 4360; https://doi.org/10.3390/ijms25084360 - 15 Apr 2024

Viewed by 308

Abstract

Childhood cancer incidence, especially in high-income countries, has led to a focus on preserving fertility in this vulnerable population. The common treatments, such as radiation and certain chemotherapeutic agents, though effective, pose a risk to fertility. For adult women, established techniques like embryo and egg freezing are standard, requiring ovarian stimulation. However, for prepubescent girls, ovarian tissue freezing has become the primary option, eliminating the need for hormonal preparation. This review describes the beginning, evolution, and current situation of the fertility preservation options for this young population. A total of 75 studies were included, covering the steps in the current fertility preservation protocols: (i) ovarian tissue extraction, (ii) the freezing method, and (iii) thawing and transplantation. Cryopreservation and the subsequent transplantation of ovarian tissue have resulted in successful fertility restoration, with over 200 recorded live births, including cases involving ovarian tissue cryopreserved from prepubescent girls. Despite promising results, challenges persist, such as follicular loss during transplantation, which is attributed to ischemic and oxidative damage. Optimizing ovarian tissue-freezing processes and exploring alternatives to transplantation, like in vitro systems for follicles to establish maturation, are essential to mitigating associated risks. Further research is required in fertility preservation techniques to enhance clinical outcomes in the future. Ovarian tissue cryopreservation appears to be a method with specific benefits, indications, and risks, which can be an important tool in terms of preserving fertility in younger women. Full article

(This article belongs to the Special Issue Ovary and Testis: Molecular Biological Insights)

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23 pages, 5755 KiB

Open AccessReview

Meiotic Cell Cycle Progression in Mouse Oocytes: Role of Cyclins

by Hye Min Kim, Min Kook Kang, Se Yoon Seong, Jun Hyeon Jo, Min Ju Kim, Eun Kyeong Shin, Chang Geun Lee and Seung Jin Han

Int. J. Mol. Sci. 2023, 24(17), 13659; https://doi.org/10.3390/ijms241713659 - 04 Sep 2023

Viewed by 2269

Abstract

All eukaryotic cells, including oocytes, utilize an engine called cyclin-dependent kinase (Cdk) to drive the cell cycle. Cdks are activated by a co-factor called cyclin, which regulates their activity. The key Cdk–cyclin complex that regulates the oocyte cell cycle is known as Cdk1–cyclin B1. Recent studies have elucidated the roles of other cyclins, such as B2, B3, A2, and O, in oocyte cell cycle regulation. This review aims to discuss the recently discovered roles of various cyclins in mouse oocyte cell cycle regulation in accordance with the sequential progression of the cell cycle. In addition, this review addresses the translation and degradation of cyclins to modulate the activity of Cdks. Overall, the literature indicates that each cyclin performs unique and redundant functions at various stages of the cell cycle, while their expression and degradation are tightly regulated. Taken together, this review provides new insights into the regulatory role and function of cyclins in oocyte cell cycle progression. Full article

(This article belongs to the Special Issue Ovary and Testis: Molecular Biological Insights)

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17 pages, 2246 KiB

Open AccessReview

Cryopreservation of Ovarian and Testicular Tissue and the Influence on Epigenetic Pattern

by Tom Trapphoff and Stefan Dieterle

Int. J. Mol. Sci. 2023, 24(13), 11061; https://doi.org/10.3390/ijms241311061 - 04 Jul 2023

Cited by 1 | Viewed by 1908

Abstract

Ovarian tissue cryopreservation (OTC) or testicular tissue cryopreservation (TTC) are effective and often the only options for fertility preservation in female or male patients due to oncological, medical, or social aspects. While TTC and resumption of spermatogenesis, either in vivo or in vitro, has still be considered an experimental approach in humans, OTC and autotransplantation has been applied increasingly to preserve fertility, with more than 200 live births worldwide. However, the cryopreservation of reproductive cells followed by the resumption of gametogenesis, either in vivo or in vitro, may interfere with sensitive and highly regulated cellular processes. In particular, the epigenetic profile, which includes not just reversible modifications of the DNA itself but also post-translational histone modifications, small non-coding RNAs, gene expression and availability, and storage of related proteins or transcripts, have to be considered in this context. Due to complex reprogramming and maintenance mechanisms of the epigenome in germ cells, growing embryos, and offspring, OTC and TTC are carried out at very critical moments early in the life cycle. Given this background, the safety of OTC and TTC, taking into account the epigenetic profile, has to be clarified. Cryopreservation of mature germ cells (including metaphase II oocytes and mature spermatozoa collected via ejaculation or more invasively after testicular biopsy) or embryos has been used successfully for many years in medically assisted reproduction (MAR). However, tissue freezing followed by in vitro or in vivo gametogenesis has become more attractive in the past, while few human studies have analysed the epigenetic effects, with most data deriving from animal studies. In this review, we highlight the potential influence of the cryopreservation of immature germ cells and subsequent in vivo or in vitro growth and differentiation on the epigenetic profile (including DNA methylation, post-translational histone modifications, and the abundance and availability of relevant transcripts and proteins) in humans and animals. Full article

(This article belongs to the Special Issue Ovary and Testis: Molecular Biological Insights)

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