Pikeperch (Sander lucioperca) is a highly profitable commercial species whose economic value has greatly increased in the last decade. As in other species, the quality of spermatozoa in this species is a principal feature inherent in fertilization success and efficient natural and artificial reproduction. The capacity of fish spermatozoa to be activated and tolerate environmental changes (in osmolality, ion composition, external pH, temperature, etc.) during the motility period contributes to fertilization success. In this study, we investigated the effects of environmental osmolality and ion composition on spermatozoa motility. To determine if the activation mechanism is affected by sperm quality parameters, we measured semen characteristics such as semen volume, spermatozoa concentration, seminal fluid osmolality and ion composition, and spermatozoa lipid composition. An additional parameter of sperm quality reflecting spermatozoa osmoresistance, the swelling rate, was measured by the nephelometry method. We detected that sperm samples with the highest content of palmitic (C16:0) and palmitoleic (C16:1) acids showed the lowest motility activation under the studied conditions, suggesting that these fatty acids are possible markers for the determination of spermatozoa quality in fish. Our results show that pikeperch spermatozoa can be activated under different osmotic conditions and that cell swelling always accompanies motility. However, spermatozoa sustain their volume under hypotonic conditions when motility is not initiated, suggesting that pikeperch spermatozoa activation is mainly controlled by ion composition rather than the osmolarity of the surrounding medium.

• Klíčová slova
• Fish spermatology, Pikeperch, Semen quality, Spermatozoa motility, Spermatozoa volume,
• MeSH
• analýza spermatu veterinární   MeSH
• motilita spermií fyziologie   MeSH
• okounovití * fyziologie   MeSH
• sperma * fyziologie   MeSH
• spermie fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

In most fish exhibiting external fertilization, spermatozoa become motile after release into water, triggered by differences between intracellular and extracellular conditions such as osmotic pressure, ion composition, and pH. The rapid change in osmolarity initiating spermatozoon motility induces osmotic pressure, resulting in active water movement across the cell membrane. Mechanisms of ion and water transport across the plasma membrane and cell volume regulation are important in maintaining structure and functional integrity of the cell. The capacity of the fish spermatozoon plasma membrane to adapt to dramatic environmental changes is an essential prerequisite for motility and successful fertilization. Adaptation to change in external osmolality may be the basis of spermatozoon function and an indicator of sperm quality. The involvement of specific water channels (aquaporins) in cell volume regulation and motility is highly likely. The goal of this review is to describe basic mechanisms of water transport and their role in fish spermatozoon physiology, focusing on osmoresistance, cell volume regulation, motility, and survival.

• Klíčová slova
• Aquaporins, Motility, Osmoresistance,·Water transport,
• MeSH
• akvaporiny fyziologie   MeSH
• kryoprezervace MeSH
• lidé MeSH
• lipidy fyziologie   MeSH
• osmoregulace * MeSH
• ryby fyziologie   MeSH
• spermie fyziologie   MeSH
• uchování spermatu MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• akvaporiny MeSH
• lipidy MeSH

Currently, considering cryopreservation of bull semen, there is no clear consensus over the comparability of cryoprotective efficacy of extenders with soybean lecithin and those based on egg yolk. The objective of this study was to prove the use of Low Density Lipoprotein (LDL) extracted from hen-egg yolk as an enhancing factor for soybean lecithin-based extenders. In total, 35 ejaculates of (seven bulls x five ejaculates per bull) were collected and cryopreserved at a commercial insemination centre. The effect of the LDL addition to the extenders AndroMed® and Bioxcell® was tested in a 6% (v/v) concentration on spermatozoa after thawing. Modified extender composition effects were assessed on sperm functional parameters motility, plasma membrane, mitochondrial membrane potential and acrosomal integrity after thawing by CASA, flow cytometry and fluorescent microscopy, respectively. Based on kinematic parameters determined from CASA, k-means cluster analysis was used to classify individual spermatozoon into specific subpopulations (fast, medium fast and slow). A subpopulation of fast spermatozoa was increased in the presence of LDL in both selected extenders (P < 0.05). Moreover, the positive effect of LDL on sperm motility was confirmed by decreasing the percentage of sperm in slow subpopulation (P < 0.05). The effect of LDL addition on the incidence of spermatozoa with intact plasma membrane was not demonstrated in any case of extender used (P > 0.05). The percentage of sperm with intact acrosome was improved when LDL was added to Bioxcell® extender (P < 0.05). On the other hand, addition of LDL to AndroMed® extender improved mitochondrial intactness after thawing (P < 0.05). In conclusion, our results showed that adding LDL to selected soybean lecithin-based extenders considerably ameliorated the functional parameters of spermatozoa after thawing and thus this lipoprotein could represent an improving agent for soybean lecithin-based extender for bull semen cryopreservation.

• Klíčová slova
• Low Density Lipoprotein, cryopreservation, spermatozoa,
• Publikační typ
• časopisecké články MeSH

In many fish species, sperm cryopreservation has deleterious effects and leads to a significant decrease in spermatozoa viability. However, the effect of cryopreservation on sperm cells that survive this process and are still viable is not fully understood. The objective of this study was to compare the viability and proteomes of fresh and cryopreserved sterlet (Acipenser ruthenus) sperm samples before and after live-dead cell separation using Percoll density gradient centrifugation. Both fresh and cryopreserved sperm samples were divided into two groups (with or without application of Percoll separation). At each step of the experiment, sperm quality was evaluated by video microscopy combined with integrated computer-assisted sperm analysis software and flow cytometry for live-dead sperm viability analysis. Sperm motility and the percentage of live cells were reduced in the cryopreserved group compared to the fresh group from 89% to 33% for percentage of motility and from 96% to 70% for live cells. Straight line velocity and linearity of track were significantly lower in cryopreserved samples than in those separated by Percoll before and after cryopreservation. However, the percentages of motile and live spermatozoa were higher than 90% in samples subjected to Percoll separation. Proteomic analysis of spermatozoa by two-dimensional differences in-gel electrophoresis coupled with matrix-assisted laser-desorption/ionization time-of-flight/time-of-flight mass spectrometry revealed that 20 protein spot abundances underwent significant changes in cryopreserved samples compared to fresh ones. However, only one protein spot was significantly altered when samples before and after cryopreservation followed by Percoll separation were compared. Thus, the results of this study show that cryopreservation leads to minimal proteomic changes in the spermatozoa population, retaining high motility and viability parameters. The results also suggest that global differences in protein profiles between unselected fresh and cryopreserved samples are mainly due to protein loss or changes in the lethal and sublethal damaged cell subpopulations.

• MeSH
• centrifugace - gradient hustoty metody   MeSH
• kryoprezervace metody   MeSH
• motilita spermií fyziologie   MeSH
• oxid křemičitý chemie   MeSH
• povidon chemie   MeSH
• proteomika MeSH
• ryby fyziologie   MeSH
• spermie fyziologie   MeSH
• uchování spermatu metody   MeSH
• viabilita buněk fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• oxid křemičitý MeSH
• Percoll MeSH Prohlížeč
• povidon MeSH