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In the 1960s, sperm cryopreservation was developed as a method to preserve fertility. Currently, techniques for the cryopreservation of human spermatozoa have been widely used in assisted reproduction. However, although sperm cryobiology has made notable achievements, the optimal method for the recovery of viable spermatozoa after cryopreservation remains elusive. Postthawing sperm quality can be affected by cryoprotectants, ice formation, storage conditions, and osmotic stress during the freezing process. This review discusses recent advances in different cryopreservation techniques, cryoprotectants, and freezing and thawing methods during cryopreservation and new indications for the use of cryopreserved spermatozoa.

Infertility is a global health problem involving about 15% of couples. Approximately half of the infertility cases are related to male factors. The oxidative stress, which refers to an imbalance in levels of reactive oxygen species (ROS) and antioxidants, is one of the main causes of infertility in men. A small amount of ROS is necessary for the physiological function of sperm including the capacitation, hyperactivation and acrosomal reaction. However, high levels of ROS can cause infertility through not only by lipid peroxidation or DNA damage but inactivation of enzymes and oxidation of proteins in spermatozoa. Oxidative stress (OS) is mainly caused by factors associated with lifestyle. Besides, immature spermatozoa, inflammatory factors, genetic mutations and altering levels of sex hormones are other main source of ROS. Since OS occurs due to the lack of antioxidants and its side effects in semen, lifestyle changes and antioxidant regimens can be helpful therapeutic approaches to overcome this problem. The present study aimed to describe physiological ROS production, roles of genetic and epigenetic factors on the OS and male infertility with various mechanisms such as lipid peroxidation, DNA damage, and disorder of male hormone profile, inflammation, and varicocele. Finally, the roles of oral antioxidants and herbs were explained in coping with OS in male infertility.

In this study, we present an analysis of the male reproductive system and spermatozoa of Anopheles darlingi Root, 1926, the primary malaria vector in Brazil. The reproductive system consists of a pair of unifollicular testes, deferent ducts, a muscular ejaculatory duct, and a pair of accessory glands. The average spermatozoa length was 188 [micro]m, with a continuous variation from 92 to 246 [micro]m.This significant variation may be associated with the mosquito's copulatory behavior, in which females are monandrous. This scenario may reduce the selective pressure for uniformity of male gametes in this species.

PurposeIt is known that sperm preparation techniques in in vitro fertilisation (IVF) are intended to select the best-quality sperm. The aim of this study is to compare sperm the density gradient method and microfluidic chip (Fertile Plus) method in infertile patients by analysing fertilisation rates, pregnancy rates, and sperm morphology and DNA fragmentation rates posed by these two methods.MethodsUsing semen samples obtained from the patients, sperms were prepared with gradient (n = 312) and microfluidic chip methods (n = 116). Fertilisation and pregnancy rates were compared in the first time and in the recurrent IVF trial patients. In addition, the morphology and DNA fragmentation comparison of sperm samples were evaluated by Toluidine blue in situ chemical staining method.ResultsThere was no statistically significant difference between fertilisation and pregnancy rates when compared with study groups in first-time IVF treatment patients. However, in recurrent IVF failure patients, there was a significant difference in fertilisation rates but no statistically significant difference was found in pregnancy rates. The microfluidic chip method significantly decreased sperm DNA fragmentation index according to density gradient method.ConclusionsMicrofluidic chip method may be recommended in patients with recurrent unsuccessful in vitro trials. The sperm DNA fragmentation test prior to the treatment will be helpful in selecting the appropriate sperm-washing method.

Successful reproduction is critical to the persistence of at-risk species; however, reproductive characteristics are understudied in many wild species. New Zealand's endemic tuatara (Sphenodon punctatus), the sole surviving member of the reptile order Rhynchocephalia, is restricted to 10% of its historic range. To complement ongoing conservation efforts, we collected and characterized mature sperm from male tuatara for the first time. Semen collected both during mating and from urine after courting contained motile sperm and had the potential for a very high percentage of viable sperm cells (98%). Scanning electron microscopy revealed a filiform sperm cell with distinct divisions: head, midpiece, tail, and reduced end piece. Finally, our initial curvilinear velocity estimates for tuatara sperm are 2-4 times faster than any previously studied reptile. Further work is needed to examine these trends at a larger scale; however, this research provides valuable information regarding reproduction in this basal reptile.

[This corrects the article DOI: 10.1371/journal.pone.0338399.].

Reproduction, development and homeostasis depend on motile cilia, whose rhythmic beating is powered by a microtubule-based molecular machine called the axoneme. Although an atomic model of the axoneme is available for the alga Chlamydomonas reinhardtii 1 , structures of mammalian axonemes are incomplete 1 , 2 , 3 , 4 – 5 . Furthermore, we do not fully understand how molecular structures of axonemes vary across motile-ciliated cell types in the body. Here we use cryoelectron microscopy, cryoelectron tomography and proteomics to resolve the 96-nm modular repeat of axonemal doublet microtubules (DMTs) from both sperm flagella and epithelial cilia of the oviduct, brain ventricles and respiratory tract. We find that sperm DMTs are the most specialized, with epithelial cilia having only minor differences across tissues. We build a model of the mammalian sperm DMT, defining the positions and interactions of 181 proteins including 34 newly identified proteins. We elucidate the composition of radial spoke 3 and uncover binding sites of kinases associated with regeneration of ATP and regulation of ciliary motility. We discover a sperm-specific, axoneme-tethered T-complex protein ring complex (TRiC) chaperone that may contribute to construction or maintenance of the long flagella of mammalian sperm. We resolve axonemal dyneins in their prestroke states, illuminating conformational changes that occur during ciliary movement. Our results illustrate how elements of chemical and mechanical regulation are embedded within the axoneme, providing valuable resources for understanding the aetiology of ciliopathy and infertility, and exemplifying the discovery power of modern structural biology. Cryoelectron microscopy, cryoelectron tomography and proteomics are used to resolve the 96-nm modular repeat of axonemal doublet microtubules from both sperm flagella and epithelial cilia of the oviduct, brain ventricles and respiratory tract.

This study investigates the proteomic profile of the fallopian tube following exposure to human sperm, with a focus on its role in sperm capacitation, final sperm maturation, successful fertilization, and early embryonic development. Twenty reproductive-age women undergoing hysterectomy during the luteal phase were divided into two groups. The control group was instructed to abstain from intercourse for at least one week prior to surgery. The case group was instructed to have intercourse 24 to 48 h before surgery, ensuring intravaginal ejaculation. A 1 cm segment of the ampullary region of the fallopian tube was collected. Proteomic analysis was performed using a multiplexed tandem mass tag (TMT)-based proteomics approach. Western blot analysis was used to validate the data. A total of 90 proteins were upregulated and 200 proteins were downregulated in the case group compared to the control group. These proteins were involved in key biological pathways, including inflammation-related pathways, angiogenesis, coagulation, metabolic processes, and positive regulation of hormone secretion (logFC > 1; p-value < 0.05). Our findings suggest that the presence of healthy sperm creates a stress-free environment within the fallopian tube, activating signaling pathways that support the selection of high-quality sperm and prepare the tube for successful fertilization.

Background Subfertility decreases the efficiency of the cattle industry because artificial insemination employs spermatozoa from a single bull to inseminate thousands of cows. Variation in bull fertility has been demonstrated even among those animals exhibiting normal sperm numbers, motility, and morphology. Despite advances in research, molecular and cellular mechanisms underlying the causes of low fertility in some bulls have not been fully elucidated. In this study, we investigated the metabolic profile of bull spermatozoa using non-targeted metabolomics. Statistical analysis and bioinformatic tools were employed to evaluate the metabolic profiles high and low fertility groups. Metabolic pathways associated with the sperm metabolome were also reported. Results A total of 22 distinct metabolites were detected in spermatozoa from bulls with high fertility (HF) or low fertility (LF) phenotype. The major metabolite classes of bovine sperm were organic acids/derivatives and fatty acids/conjugates. We demonstrated that the abundance ratios of five sperm metabolites were statistically different between HF and LF groups including gamma-aminobutyric acid (GABA), carbamate, benzoic acid, lactic acid, and palmitic acid. Metabolites with different abundances in HF and LF bulls had also VIP scores of greater than 1.5 and AUC- ROC curves of more than 80%. In addition, four metabolic pathways associated with differential metabolites namely alanine, aspartate and glutamate metabolism, β-alanine metabolism, glycolysis or gluconeogenesis, and pyruvate metabolism were also explored. Conclusions This is the first study aimed at ascertaining the metabolome of spermatozoa from bulls with different fertility phenotype using gas chromatography-mass spectrometry. We identified five metabolites in the two groups of sires and such molecules can be used, in the future, as key indicators of bull fertility.