Stefan S. du Plessis

Effect of Oxidative Stress on Male Reproduction

Infertility affects approximately 15% of couples trying to conceive, and a male factor contributes to roughly half of these cases. Oxidative stress (OS) has been identified as one of the many mediators of male infertility by causing sperm dysfunction. OS is a state related to increased cellular damage triggered by oxygen and oxygen-derived free radicals known as reactive oxygen species (ROS). During this process, augmented production of ROS overwhelms the bodys antioxidant defenses. While small amounts of ROS are required for normal sperm functioning, disproportionate levels can negatively impact the quality of spermatozoa and impair their overall fertilizing capacity. OS has been identified as an area of great attention because ROS and their metabolites can attack DNA, lipids, and proteins; alter enzymatic systems; produce irreparable alterations; cause cell death; and ultimately, lead to a decline in the semen parameters associated with male infertility. This review highlights the mechanisms of ROS production, the physiological and pathophysiological roles of ROS in relation to the male reproductive system, and recent advances in diagnostic methods; it also explores the benefits of using antioxidants in a clinical setting.

The effect of obesity on sperm disorders and male infertility

The results of several studies point to an increased likelihood of abnormal semen parameters among overweight men, and an elevated risk for subfertility among couples in which the male partner is obese. Obesity is, therefore, associated with a higher incidence of male factor infertility. Several mechanisms might account for the effect of obesity on male infertility, both directly and indirectly, by inducing sleep apnea, alterations in hormonal profiles (reduced inhibin B and androgen levels accompanied by elevated estrogen levels) and increased scrotal temperatures, ultimately manifesting as impaired semen parameters (decreased total sperm count, concentration and motility; increased DNA fragmentation index). Neither the reversibility of obesity-associated male infertility with weight loss nor effective therapeutic interventions have been studied in-depth. The increasing prevalence of obesity calls for greater clinical awareness of its effects on fertility, better understanding of underlying mechanisms, and exploration into avenues of treatment.

Obesity: modern man's fertility nemesis

The obesity pandemic has grown to concerning proportions in recent years, not only in the Western World, but in developing countries as well. The corresponding decrease in male fertility and fecundity may be explained in parallel to obesity, and obesity should be considered as an etiology of male fertility. Studies show that obesity contributes to infertility by reducing semen quality, changing sperm proteomes, contributing to erectile dysfunction, and inducing other physical problems related to obesity. Mechanisms for explaining the effect of obesity on male infertility include abnormal reproductive hormone levels, an increased release of adipose-derived hormones and adipokines associated with obesity, and other physical problems including sleep apnea and increased scrotal temperatures. Recently, genetic factors and markers for an obesity-related infertility have been discovered and may explain the difference between fertile obese and infertile obese men. Treatments are available for not only infertility related to obesity, but also as a treatment for the other comorbidities arising from obesity. Natural weight loss, as well as bariatric surgery are options for obese patients and have shown promising results in restoring fertility and normal hormonal profiles. Therapeutic interventions including aromatase inhibitors, exogenous testosterone replacement therapy and maintenance and regulation of adipose-derived hormones, particularly leptin, may also be able to restore fertility in obese males. Because of the relative unawareness and lack of research in this area, controlled studies should be undertaken and more focus should be given to obesity as an etiolgy of male infertility.

Sperm viability, apoptosis, and intracellular reactive oxygen species levels in human spermatozoa before and after induction of oxidative stress

OBJECTIVE To investigate sperm viability, incidence of apoptosis, and intracellular basal and induced reactive oxygen species (ROS) in sperm fractions. DESIGN Prospective controlled study. SETTING Center for Reproductive Medicine at a tertiary care hospital. METHOD(S) Liquefied seminal ejaculates (n = 12) prepared by density gradient centrifugation were reconstituted to 2 mL with phosphate-buffered saline. Oxidative stress was induced by hydrogen peroxide (H(2)O(2), 100 muM). Sperm viability, intracellular ROS, and incidence of apoptosis/necrosis in neat, immature, and mature sperm fractions were assessed. RESULT(S) Before H(2)O(2) exposure, mature spermatozoa fractions showed a significantly lower incidence of apoptotic sperm and intracellular O(2)(-*) levels but higher amounts of intracellular H(2)O(2) compared with neat semen. Higher levels of intracellular H(2)O(2) were demonstrated in immature sperm fractions compared with neat or mature fractions. In all sperm fractions, intracellular H(2)O(2) levels correlated with the intracellular concentration of O(2)(-*). After H(2)O(2) exposure, neat semen showed a significantly higher percentage of apoptosis compared with the prepared mature spermatozoa. However, no differences were observed in the incidence of apoptosis between immature and mature sperm fractions. CONCLUSION(S) There is a differential shift of both intracellular H(2)O(2) and O(2)(-*) in each sperm fraction that may affect sperm quality. Sperm apoptosis is related to intracellular H(2)O(2) levels, which in turn are affected by intracellular O(-*) levels. Oxidative stress was not associated with an increased incidence of apoptosis in immature or mature sperm fractions.

Frontostriatal activity and connectivity increase during proactive inhibition across adolescence and early adulthood

During adolescence, functional and structural changes in the brain facilitate the transition from childhood to adulthood. Because the cortex and the striatum mature at different rates, temporary imbalances in the frontostriatal network occur. Here, we investigate the development of the subcortical and cortical components of the frontostriatal network from early adolescence to early adulthood in 60 subjects in a cross‐sectional design, using functional MRI and a stop‐signal task measuring two forms of inhibitory control: reactive inhibition (outright stopping) and proactive inhibition (anticipation of stopping). During development, reactive inhibition improved: older subjects were faster in reactive inhibition. In the brain, this was paralleled by an increase in motor cortex suppression. The level of proactive inhibition increased, with older subjects slowing down responding more than younger subjects when anticipating a stop‐signal. Activation increased in the right striatum, right ventral and dorsal inferior frontal gyrus, and supplementary motor area. Moreover, functional connectivity during proactive inhibition increased between striatum and frontal regions with age. In conclusion, we demonstrate that developmental improvements in proactive inhibition are paralleled by increases in activation and functional connectivity of the frontostriatal network. These data serve as a stepping stone to investigate abnormal development of the frontostriatal network in disorders such as schizophrenia and attention‐deficit hyperactivity disorder. Hum Brain Mapp 35:4415–4427, 2014.

Insulin and leptin enhance human sperm motility, acrosome reaction and nitric oxide production

AIM To investigate the in vitro effects of insulin and leptin on human sperm motility, viability, acrosome reaction and nitric oxide (NO) production. METHODS Washed human spermatozoa from normozoospermic donors were treated with insulin (10 microIU) and leptin (10 nmol). Insulin and leptin effects were blocked by inhibition of their intracellular effector, phosphotidylinositol 3-kinase (PI3K), by wortmannin (10 micromol) 30 min prior to insulin and leptin being given. Computer-assisted semen analysis was used to assess motility after 1, 2 and 3 h of incubation. Viability was assessed by fluorescence-activated cell sorting using propidium iodide as a fluorescent probe. Acrosome-reacted cells were observed under a fluorescent microscope using fluorescein-isothiocyanate-Pisum sativum agglutinin as a probe. NO was measured after treating the sperm with 4,5-diaminofluorescein-2/diacetate (DAF-2/DA) and analyzed by fluorescence-activated cell sorting. RESULTS Insulin and leptin significantly increased total motility, progressive motility and acrosome reaction, as well as NO production. CONCLUSION This study showed the in vitro beneficial effects of insulin and leptin on human sperm function. These hormones could play a role in enhancing the fertilization capacity of human spermatozoa.

An investigation of excess residual cytoplasm in human spermatozoa and its distinction from the cytoplasmic droplet

Recent studies have shown cytoplasmic droplets to be normal morphological occurrences in human male spermatozoa. When the cytoplasm around the sperm midpiece is present in large amounts, however, pathological effects may transpire. The cytoplasmic droplet then becomes known as excess residual cytoplasm, which can impair overall sperm function and produce higher levels of reactive oxygen species, potentially leading to male infertility. Though the distinction between cytoplasmic droplets and excess residual cytoplasm has been made, some studies fail to recognize the difference and incorrectly label the latter as a cytoplasmic droplet. This review attempts to clarify excess residual cytoplasm’s effect on fertility, examine the enzymes responsible, and suggest tests and possible treatment options for those affected by this defect.

TNF-α and IL-6 affect human sperm function by elevating nitric oxide production

Many studies have reported the effects of cytokines on human sperm function, even though their role and the mechanisms involved remain unclear. The effects of increasing concentrations of the cytokines TNF-α and IL-6 on human sperm motility and viability were assessed, and the possible mechanisms involved were investigated. TNF-α and IL-6 significantly reduced progressive motility at higher concentrations in a dose- and time-dependent manner. No differences were observed in cell viability. Both cytokines increased nitric oxide production in a dose-dependent manner. TNF-α and IL-6 did not statistically differ in their detrimental effects on human spermatozoa. These results indicate that TNF-α and IL-6 have an effect on sperm function. This effect is possibly mediated via an increase in nitric oxide production.

Oxidative phosphorylation versus glycolysis: what fuel do spermatozoa use?

Spermatozoa are highly specialized cells. Adenosine triphosphate (ATP), which provides the energy for supporting the key functions of the spermatozoa, is formed by 2 metabolic pathways, namely glycolysis and oxidative phosphorylation (OXPHOS). It is produced in the mitochondria through OXPHOS as well as in the head and principal piece of the flagellum through glycolysis. However, there is a great discrepancy as to which method of ATP production is primarily utilized by the spermatozoa for successful fertilization. Mitochondrial respiration is considered to be a more efficient metabolic process for ATP synthesis in comparison to glycolysis. However, studies have shown that the diffusion potential of ATP from the mitochondria to the distal end of the flagellum is not sufficient to support sperm motility, suggesting that glycolysis in the tail region is the preferred pathway for energy production. It is suggested by many investigators that although glycolysis forms the major source of ATP along the flagellum, energy required for sperm motility is mainly produced during mitochondrial respiration. Nevertheless, some studies have shown that when glycolysis is inhibited, proper functioning and motility of spermatozoa remains intact although it is unclear whether such motility can be sustained for prolonged periods of time, or is sufficiently vigorous to achieve optimal fertilization. The purpose of this article is to provide an overview of mammalian sperm energy metabolism and identify the preferred metabolic pathway for ATP generation which forms the basis of energy production in human spermatozoa during fertilization.

Proteomics: a subcellular look at spermatozoa

BackgroundMale-factor infertility presents a vexing problem for many reproductively active couples. Many studies have focused on abnormal sperm parameters. Recent advances in proteomic techniques, especially in mass spectrometry, have aided in the study of sperm and more specifically, sperm proteins. The aim of this study was to review the current literature on the various proteomic techniques, and their usefulness in diagnosing sperm dysfunction and potential applications in the clinical setting.MethodsReview of PubMed database. Key words: spermatozoa, proteomics, protein, proteome, 2D-PAGE, mass spectrometry.ResultsRecently employed proteomic methods, such as two-dimensional polyacrylamide gel electrophoresis, mass spectrometry, and differential in gel electrophoresis, have identified numerous sperm-specific proteins. They also have provided a further understanding of protein function involved in sperm processes and for the differentiation between normal and abnormal states. In addition, studies on the sperm proteome have demonstrated the importance of post-translational modifications, and their ability to bring about physiological changes in sperm function. No longer do researchers believe that in order for them to elucidate the biochemical functions of genes, mere knowledge of the human genome sequence is sufficient. Moreover, a greater understanding of the physiological function of every protein in the tissue-specific proteome is essential in order to unravel the biological display of the human genome.ConclusionRecent advances in proteomic techniques have provided insight into sperm function and dysfunction. Several multidimensional separation techniques can be utilized to identify and characterize spermatozoa. Future developments in bioinformatics can further assist researchers in understanding the vast amount of data collected in proteomic studies. Moreover, such advances in proteomics may help to decipher metabolites which can act as biomarkers in the detection of sperm impairments and to potentially develop treatment for infertile couples.Further comprehensive studies on sperm-specific proteome, mechanisms of protein function and its proteolytic regulation, biomarkers and functional pathways, such as oxidative-stress induced mechanisms, will provide better insight into physiological functions of the spermatozoa. Large-scale proteomic studies using purified protein assays will eventually lead to the development of novel biomarkers that may allow for detection of disease states, genetic abnormalities, and risk factors for male infertility. Ultimately, these biomarkers will allow for a better diagnosis of sperm dysfunction and aid in drug development.