Sperm motility is one of the major determinants of male fertility. Since sperm need a great deal of energy to support their fast movement by active metabolism, they are thus extremely vulnerable to oxidative damage by the reactive oxygen species (ROS) and other free radicals generated as byproducts in the electron transport chain. The present study is aimed at understanding the impact of a mitochondrial oxidizing/reducing microenvironment in the etiopathology of male infertility. We detected the mitochondrial DNA (mtDNA) 4,977 bp deletion in human sperm. We examined the gene mutation of ATP synthase 6 (ATPase6 m.T8993G) in ATP generation, the gene polymorphisms of uncoupling protein 2 (UCP2, G-866A) in the uncoupling of oxidative phosphorylation, the role of genes such as manganese superoxide dismutase (MnSOD, C47T) and catalase (CAT, C-262T) in the scavenging system in neutralizing reactive oxygen species, and the role of human 8-oxoguanine DNA glycosylase (hOGG1, C1245G) in 8-hydroxy-2'-deoxyguanosine (8-OHdG) repair. We found that the sperm with higher motility were found to have a higher mitochondrial membrane potential and mitochondrial bioenergetics. The genotype frequencies of UCP2 G-866A, MnSOD C47T, and CAT C-262T were found to be significantly different among the fertile subjects, the infertile subjects with more than 50% motility, and the infertile subjects with less than 50% motility. A higher prevalence of the mtDNA 4,977 bp deletion was found in the subjects with impaired sperm motility and fertility. Furthermore, we found that there were significant differences between the occurrences of the mtDNA 4,977 bp deletion and MnSOD (C47T) and hOGG1 (C1245G). In conclusion, the maintenance of the mitochondrial redox microenvironment and genome integrity is an important issue in sperm motility and fertility.
- MeSH
- DNA-glykosylasy genetika metabolismus MeSH
- frekvence genu MeSH
- lidé MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- mitochondriální DNA genetika metabolismus MeSH
- mitochondrie genetika metabolismus MeSH
- motilita spermií fyziologie MeSH
- mužská infertilita genetika patologie MeSH
- oxidační stres účinky léků MeSH
- peroxid vodíku farmakologie MeSH
- polymorfismus genetický MeSH
- spermie metabolismus fyziologie MeSH
- superoxiddismutasa genetika metabolismus MeSH
- uncoupling protein 2 genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Sturgeon spermatozoa are unique for their sustained motility. We investigated the relative importance of bioenergetic pathways in the energy supply of Siberian sturgeon Acipenser baerii spermatozoa during motile and immotile states. Spermatozoon motility and oxygen consumption rate (OCR) were analysed following exposure to inhibitors of oxidative phosphorylation (sodium azide, NaN3 ), glycolysis (2-deoxy-D-glucose, DOG) and β-oxidation of fatty acids (sodium fluoride, NaF), and to an uncoupler of oxidative phosphorylation (carbonyl cyanide m-chlorophenyl hydrazine, CCCP). No significant difference in curvilinear velocity was observed after addition of these reagents to activation medium (AM) or nonactivation medium (NAM) for incubation. Incubation of spermatozoa in NAM containing CCCP or NaN3 resulted in significantly decreased motility duration compared to controls. The OCR of sturgeon spermatozoa in AM (11.9 ± 1.4 nmol O2 min-1 (109 spz)-1 ) was significantly higher than in NAM (8.2 ± 1.5 nmol O2 min-1 (109 spz)-1 ). The OCR significantly declined with addition of NaN3 to AM and NAM. No significant difference in motility parameters or OCR was observed with NaF or DOG. These results suggest active oxidative phosphorylation in both immotile and motile spermatozoa. Nevertheless, mitochondrial respiration occurring during motility is not sufficient to meet the high energy demands, and the energy required for sustained motility of Siberian sturgeon spermatozoa is derived from adenosine triphosphate accumulated during the quiescent state.
The target of this study was to evaluate the effect of extract of the European mistletoe - Viscum album quercus L. on spermatozoa motility and viability in vitro. The CASA system was used to determine the spermatozoa motility parameters at different time intervals (0, 1, 2 and 3 h) and spermatozoa viability was determined in five different doses of Viscum album quercus L [10 (QA), 6.6 (QB), 3.3 (QC), 2.5 (QD) and 2 (QE) mg/ml]. Results in experimental groups detected a significant deterioration on rabbit spermatozoa after 1, 2 and 3 hours, compared to the control. The initial total spermatozoa motility showed increased value for all doses of Viscum album quercus in comparison to control. After in vitro culture a dose-dependent decrease (QA: reduction of 69.7 %, QB: reduction of 40.9 %) was found. For the progressive spermatozoa most significant decrease (86.8 % for QA vs. 48.5 % for QB) was detected compared to the control after 3 hours of culture. Spermatozoa viability (MTT test) was decreased in all experiment groups at the end of experiment, but the differences were not significant. Significant alterations of membrane integrity were found in groups with the highest Viscum album quercus concentration (QA, QB), but acrosome integrity showed no significant changes. Results suggest negative dose- and time-dependent effect of Viscum album quercus at higher doses on spermatozoa motility and viability parameters in vitro.
- MeSH
- časové faktory MeSH
- dub (rod) * MeSH
- králíci MeSH
- motilita spermií účinky léků fyziologie MeSH
- rostlinné extrakty izolace a purifikace farmakologie MeSH
- spermie účinky léků fyziologie MeSH
- viabilita buněk účinky léků fyziologie MeSH
- Viscum album * MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Swim-up is a sperm purification method that is being used daily in andrology labs around the world as a simple step for in vitro sperm selection. This method accumulates the most motile sperm in the upper fraction and leaves sperm with low or no motility in the lower fraction. However, the underlying reasons are not fully understood. In this article, we compare metabolic rate, motility and sperm tail length of bovine sperm cells of the upper and lower fraction. The metabolic assay platform reveals oxygen consumption rates and extracellular acidification rates simultaneously and thereby delivers the metabolic rates in real time. Our study confirms that the upper fraction of bull sperm has not only improved motility compared to the cells in the lower fraction but also shows higher metabolic rates and longer flagella. This pattern was consistent across media of two different levels of viscosity. We conclude that the motility-based separation of the swim-up technique is also reflected in underlying metabolic differences. Metabolic assays could serve as additional or alternative, label-free method to evaluate sperm quality.
- MeSH
- adenosintrifosfát biosyntéza MeSH
- bazální metabolismus * MeSH
- bičík spermie metabolismus MeSH
- chov MeSH
- flagella metabolismus MeSH
- kinetika MeSH
- motilita spermií fyziologie MeSH
- oxidativní fosforylace MeSH
- skot MeSH
- spermie metabolismus MeSH
- spotřeba kyslíku MeSH
- viskozita MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Several factors regulating activation of spermatozoon motility in Eurasian burbot, Lota lota, including osmolality, calcium (Ca2+ ) ions, and temperature were investigated. Spermatozoon motility in Eurasian burbot, Lota lota was assessed at 4 and 30°C in seminal fluid, isotonic media (with and without Ca2+ ) and hypotonic media (with and without Ca2+ ). Spermatozoa were spontaneously activated in seminal fluid at 20°C and the maximum motility was recorded at 30°C, which is out of the spawning temperature range, indicating that no risk of activation occurs during routine semen handling in artificial insemination. Initiation of spermatozoon motility in L. lota is mediated by Ca2+ and sensitivity to Ca2+ is dependent on temperature.
Fish spermatozoa acquire potential for motility in the sperm duct where they are immotile. Osmolality of the seminal plasma is a key factor to maintain spermatozoa in the quiescent state in either freshwater or marine fishes. However, potassium (K+) ions prevent spermatozoa motility in salmonid and sturgeon fishes, while CO2 inhibits spermatozoa motility in flatfishes. Once, spermatozoa are released at spawning, their motility is initiated in hypo-osmotic and hyper-osmotic environments in freshwater and marine fishes, respectively. Some substances produced by the testes (a progestin), or released from oocytes (peptides) induce spermatozoa hypermotility in some marine fishes including the Atlantic croaker and Pacific herrings, respectively. Duration of spermatozoa motility is short, lasting for a few seconds to few minutes in most fishes due to rapid depletion of energy required for the beating of the motility apparatus called axoneme. In the osmotic-activated spermatozoa, K+ and water effluxes occur in freshwater and marine fishes, respectively, which trigger spermatozoa motility signaling. In general, initiation of axonemal beating is associated with an increase in intracellular calcium (Ca2+) ions in spermatozoa of both freshwater and marine fishes and a post- or pre-increase in intracellular pH, while cyclic adenosine monophosphate (cAMP) remains unchanged. However, axonemal beating is cAMP-dependent in demembranated spermatozoa of salmonid and sturgeon fishes. Calcium from extracellular environment or intracellular stores supply required Ca2+ concentration for axonemal beating. Several axonemal proteins have been so far identified in fishes that are activated by Ca2+ and cAMP, directly or mediated by protein kinase C and protein kinase A, respectively. The present study reviews differences and similarities in complex regulatory signals controlling spermatozoa motility initiation in fishes, and notes physiological mechanisms that await elucidation.
- MeSH
- druhová specificita MeSH
- motilita spermií fyziologie MeSH
- ryby fyziologie MeSH
- signální transdukce fyziologie MeSH
- spermie fyziologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Though bivalve mollusks are keystone species and major species groups in aquaculture production worldwide, gamete biology is still largely unknown. This review aims to provide a synthesis of current knowledge in the field of sperm biology, including spermatozoa motility, flagellar beating, and energy metabolism; and to illustrate cellular signaling controlling spermatozoa motility initiation in bivalves. Serotonin (5-HT) induces hyper-motility in spermatozoa via a 5-HT receptor, suggesting a serotoninergic system in the male reproductive tract that might regulate sperm physiology. Acidic pH and high concentration of K+ are inhibitory factors of spermatozoa motility in the testis. Motility is initiated at spawning by a Na+-dependent alkalization of intracellular pH mediated by a Na+/H+ exchanger. Increase of 5-HT in the testis and decrease of extracellular K+ when sperm is released in seawater induce hyperpolarization of spermatozoa membrane potential mediated by K+ efflux and associated with an increase in intracellular Ca2+ via opening of voltage-dependent Ca2+ channels under alkaline conditions. These events activate dynein ATPases and Ca2+/calmodulin-dependent proteins resulting in flagellar beating. It may be possible that 5-HT is also involved in intracellular cAMP rise controlling cAMP-dependent protein kinase phosphorylation in the flagellum. Once motility is triggered, flagellum beats in asymmetric wave pattern leading to circular trajectories of spermatozoa. Three different flagellar wave characteristics are reported, including "full", "twitching", and "declining" propagation of wave, which are described and illustrated in the present review. Mitochondrial respiration, ATP content, and metabolic pathways producing ATP in bivalve spermatozoa are discussed. Energy metabolism of Pacific oyster spermatozoa differs from previously studied marine species since oxidative phosphorylation synthetizes a stable level of ATP throughout 24-h motility period and the end of movement is not explained by a low intracellular ATP content, revealing different strategy to improve oocyte fertilization success. Finally, our review highlights physiological mechanisms that require further researches and points out some advantages of bivalve spermatozoa to extend knowledge on mechanisms of motility.
- MeSH
- druhová specificita MeSH
- energetický metabolismus MeSH
- flagella fyziologie MeSH
- mlži fyziologie MeSH
- motilita spermií fyziologie MeSH
- spermie cytologie fyziologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Motility is a characteristic function of the male gamete, which allows spermatozoa to actively reach and penetrate the female gamete in organisms with internal and external fertilization. Sperm motility is acquired under the control of many extrinsic and intrinsic factors and is based on a specialized structure of the sperm flagellum called "axoneme". An overview of how the sperm flagellum is organized, and it operates to support cell motility is presented, with special focus on the molecular mechanisms and factors involved in the development, maintenance and control of motility. Data obtained in aquatic organisms with external fertilization, such as sea urchins, ascidians or fishes are critically analyzed because they constitute model species on which most of the present day understanding of sperm motility function is based. In most animal species, sperm motility is dependent on a long appendage called flagellum. Flagella are essential organelles found in most eukaryotic cells; their basic structure is the axoneme, which consists of a scaffold of microtubules and is responsible for movement in an autonomous manner if ATP-energy is present. Flagellar beat propels the cell through the medium which surrounds sperm cells and is responsible of the translational drive of spermatozoa. The present paper includes: (1) an introduction to typical sperm morphology and ultrastructure in most aquatic species, (2) the motility apparatus or axoneme of the spermatozoa: the axoneme, (3) the structural and biochemical composition of the axoneme, (4) the axonemal motor or dynein, and its operation, (5) the regulation of motility at axoneme and cell membrane levels, including several effectors such as Ca2+ ions, (6) biophysical features of the wave propagation mechanism in motile spermatozoa, (7) the energy production and consumption, and (8) the building of a flagellum. Flagellar beating in aquatic animals is illustrated using several examples in figures and video-clips. These types of data are also used for computer simulation of various aspects of the modulation of sperm motility of marine animals.
All extant groups of Elasmobranches have internal fertilization and the structure of the male reproductive organs is very specific: sperm passes from the internal organs via the cloaca, but the male copulating organ (clasper) is distant from the cloaca. This suggests that sperm can contact the surrounding medium before fertilization. Because of this involvement with the environment, external signaling in sperm motility activation could occur in these species even though their fertilization mode is internal. In this case, spermatozoa of Elasmobranches should hypothetically possess a specific structure and membrane lipid composition which supports physiological functions of the sperm associated with environmental tonicity changes occurring at fertilization. Additionally, sperm motility properties in these taxa are poorly understood. The current study examined sperm lipid composition and motility under different environmental conditions for the ocellate river stingray, Potamotrygon motoro, an endemic South America freshwater species. Sperm samples were collected from six mature males during the natural spawning period. Sperm motility was examined in seminal fluid and fresh water by light video microscopy. Helical flagellar motion was observed in seminal fluid and resulted in spermatozoon progression; however, when diluted in fresh water, spermatozoa were immotile and had compromised structure. Lipid class and fatty acid (FA) composition of spermatozoa was analyzed by thin layer and gas chromatography. Spermatozoa FAs consisted of 33 ± 1% saturated FAs, 28 ± 1% monounsaturated FAs (MUFAs), and 41 ± 1% polyunsaturated FAs (PUFAs), and a high content of n-6 FAs (32 ± 2%) was measured. These results allowed us to conclude that sperm transfer from P. motoro male into female should occur without coming into contact with the hypotonic environment so as to preserve potent motility. In addition, this unusual reproductive strategy is associated with specific spermatozoa structure and lipid composition. Low level of docosahexaenoic acid and relatively low PUFA/MUFA ratio probably account for the relatively low fluidity of freshwater stingray membrane and can be the main reason for its low tolerance to hypotonicity.
- MeSH
- analýza spermatu veterinární MeSH
- lipidy chemie MeSH
- motilita spermií fyziologie MeSH
- rejnokovití fyziologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Motility analysis of spermatozoa relies on the investigation of either head trajectories or flagellum characteristics. Those two sets of parameters are far from being independent, the flagellum playing the role of motor, whereas the head plays a passive role of cargo. Therefore, quantitative descriptions of head trajectories represent a simplification of the complex pattern of whole sperm cell motion, resulting from the waves developed by the flagellum. The flagellum itself responds to a large variety of signals that precisely control its axoneme to allow activation, acceleration, slowing down or reorientation of the whole spermatozoon. Thus, it is obvious that analysis of flagellum characteristics provides information on the original source of movement and orientation of the sperm cell and presents additional parameters that enrich the panoply of quantitative descriptors of sperm motility. In this review, we briefly describe the methodologies used to obtain good-quality images of fish spermatozoa (head and especially flagellum) while they move fast and the methods developed for their analysis. The paper also aims to establish a link between classical analyses by computer-aided sperm analysis (CASA) and the descriptors generated by fish sperm flagellum analysis, and emphasises the information to be gained regarding motility performance from flagellum motion data.
