Q112444313
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Knowledge of conditions affecting sperm quality is essential for efficient culture of fish for commercial purposes and conservation of species. Two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time of flight mass spectrometry were used to characterize the proteomic profile of Acipenser dabryanus spermatozoa relative to motility and fertilization capacity. There were differential amounts of protein in 313 spots in spermatozoa of males classified to have relatively greater or lesser spermatozoa quality. The functions of 43 of 50 selected proteins were identified. The proteins in 14 spots were involved in metabolism, and of these, proteins in 11 spots were highly abundant in spermatozoa of males categorized to have spermatozoa of greater quality, including pyruvate kinase, enolase B, phosphoglycerate kinase, lactate dehydrogenase, cytosolic malate dehydrogenase, brain creatine kinase b, Ckmb protein, and nucleoside diphosphate kinase. The proteins involved in mechanics of flagellum movement were identified, including the dynein intermediate chain, radial spoke head 1 homolog; ropporin-1-like, Bardet-Biedl syndrome 5, ADP-ribosylation factor-like protein 3, tektin-4, gamma-actin, and tubulin cytoskeleton proteins to be differentially abundant in spermatozoa that were classified relatively greater or lesser quality. Heat shock proteins, copper/zinc superoxide dismutase and peroxiredoxins, which are involved in stress response were of differential abundance in spermatozoa from males with spermatozoa in the two different classification groups. Proteins were also detected that are involved in protein folding and binding, or hydrolase activity. The results are valuable for the prediction of sperm quality and for reproduction management in A. dabryanus and other threatened species.
- MeSH
- motilita spermií MeSH
- proteomika MeSH
- rybí proteiny chemie metabolismus MeSH
- ryby fyziologie MeSH
- spermie chemie metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Heat Shock Protein 90 (HSP90) is a fertility-associated protein, the expression of which positively correlates with sperm quality in many species. Geranylgeranylacetone (GGA) is reported to induce expression of HSP90. The present study aimed to investigate whether GGA induced expression of HSP90 in Acipenser sinensis sperm to exert a cryoprotective effect. Sperm from five male A. sinensis was combined with extender containing 20 mmol/L tris pH = 8.1, 10% v/v methanol, 2-5 mmol/L KCl, 15 mmol/L lactose, and 15 mmol/L trehalose, with GGA at 0, 14, 67, 135, 673, 1346, or 6731 μmol/L. After cryopreservation and thawing, the percentage of motile spermatozoa, spermatozoon curvilinear velocity (VCL), straight-line velocity (VSL), average path velocity (VAP), acrosome integrity, and membrane integrity, as well as fertility were evaluated. Sperm quality increased with the increase of GGA to 673 μmol/L, but decreased at higher concentrations. Expression levels of HSP90α were detected by Western blot in sperm frozen with GGA at 673 μmol/L (highest obtained sperm quality), 6731 μmol/L (highest GGA concentration), and a control without GGA. The expression of HSP90α increased with the increase in GGA, with lowest expression observed in the control. GGA was found to induce increase of HSP90α, and this increase was associated with higher quality cryopreserved sperm at concentrations ≤673 μmol/L. This research suggests a viable technique to increase the quality of cryopreserved A. sinensis sperm by adding GGA to induce expression of HSP90α.
- MeSH
- cytoprotekce * účinky léků genetika MeSH
- diterpeny farmakologie MeSH
- enzymová indukce účinky léků MeSH
- kryoprezervace veterinární MeSH
- kryoprotektivní látky metabolismus farmakologie MeSH
- motilita spermií genetika MeSH
- proteiny tepelného šoku HSP90 genetika metabolismus MeSH
- ryby * genetika metabolismus MeSH
- spermie MeSH
- uchování spermatu * veterinární MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Fish sperm cryopreservation is a well-established technique allowing for artificial insemination on a commercial scale. The extent of proteome alterations in seminal plasma and sperm due to cryopreservation, however, is not known. This study was conducted to evaluate the effect of cryopreservation on motility variables of sterlet Acipenser ruthenus sperm and to detect the differences in protein profiles of fresh and cryopreserved sterlet sperm and seminal plasma. Fresh sperm had 89 ± 3% motility and 160 ± 14 μm/s curvilinear velocity at 15 s post-activation. The motility rate of cryopreserved sperm (37 ± 5%) was less at 15 s post-activation. No difference (ANOVA; P > 0.05) in mean curvilinear velocity of fresh and cryopreserved sperm was detected. The protein profiles of seminal plasma and sperm were characterized using comparative proteomics to determine the influence of cryopreservation. Six altered protein spots in seminal plasma and thirteen altered spots in sperm were detected in fresh and thawed sperm. Subsequent protein characterization suggested that the proteins identified were involved in sperm metabolism, cytoskeleton, and stress response. The results broaden the understanding of the effects of cryopreservation and identify the proteins associated with cryo-injury. These data may help to determine the function of altered proteins and provide new insights into improving sperm cryopreservation.
- MeSH
- kryoprezervace veterinární MeSH
- motilita spermií * MeSH
- proteom * MeSH
- ryby fyziologie MeSH
- spermie fyziologie MeSH
- transkriptom MeSH
- uchování spermatu veterinární MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Conventional sperm analysis (i.e., motility and fertility) has been used to evaluate sperm quality. Understanding the quality of sperm on the molecular level in the sturgeons, Acipenser baerii and A. schrenckii, is essential for the improvement of the conservation of genetic resources and farming performance. In this study, we used the iTRAQ proteomics approach to perform proteomic profiling of spermatozoa associated with sperm quality in sturgeons (Data are available via ProteomeXchange with identifier PXD006108). The results showed 291 and 359 differentially expressed proteins in A. baerii and A. schrenckii, respectively, of which 72 were common to both species and all were upregulated in high quality compared with low quality samples. The differentially expressed proteins were mainly categorized into the generation of precursor metabolites and energy and oxidation, and they were localized to the mitochondria. Three distinguishing pathways, Arginine and proline metabolism, Pyruvate metabolism and the Citrate cycle (TCA cycle) were found to play an important role in energy metabolism, and some substrates could be used in the sperm medium for storage and cryopreservation. The quantity levels of two proteins, CKMT1 and LDHB, were verified by western blot analysis. Moreover, other potential biomarkers involved in oxidation reduction, ubiquitin-proteasome-dependent proteolysis, chaperones and binding activity were also discussed. Our study is the first to use the iTRAQ-based proteomics approach to analyse the sturgeon spermatozoa proteome, and the results that we obtained are valuable for the prediction of sperm quality and reproduction management in these threatened species.
- MeSH
- druhová specificita MeSH
- fertilita MeSH
- motilita spermií MeSH
- proteom metabolismus MeSH
- ryby klasifikace metabolismus MeSH
- spermie metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- validační studie MeSH
- MeSH
- biodegradace MeSH
- čištění vody metody MeSH
- environmentální zdraví statistika a číselné údaje MeSH
- látky znečišťující vodu analýza toxicita MeSH
- mikrobiologie vody MeSH
- testy toxicity metody MeSH
- vodní organismy účinky léků fyziologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- úvodní články MeSH
- úvodníky MeSH
This study was conducted to investigate Brachymystax lenok tsinlingensis spermatozoa cell morphology and ultrastructure through scanning and transmission electron microscopy. Findings revealed that the spermatozoa can be differentiated into three major parts: a spherical head without an acrosome, a short mid-piece, and a long, cylindrical flagellum. The mean length of the spermatozoa was 36.11±2.84μm, with a spherical head length of 2.78±0.31μm. The mean anterior and posterior head widths were 2.20±0.42μm and 2.55±0.53μm, respectively. The nuclear fossa was positioned at the base of the nucleus that contained the anterior portion of flagellum and a centriolar complex (proximal and distal centrioles). The short mid-piece was located laterally to the nucleus and possessed just one spherical mitochondrion with a mean diameter of 0.65±0.14μm. The spermatozoa flagellum was long and cylindrical, and could be separated into two parts: a long main-piece and a short end-piece. The main piece of the flagellum had short irregular side-fins. The axoneme composed the typical '9+2' microtubular doublet structure and was enclosed by the cell membran e. This study confirmed that B. lenok tsinlingensis spermatozoa can be categorized as teleostean "Type I" spermatozoa; 'primitive' or 'ect-aquasperm type' spermatozoa. To the best of the authers knowledge, this was the first study conducted on the morphology and ultrastructure of B. lenok tsinlingensis spermatozoa.
- MeSH
- akrozom ultrastruktura MeSH
- axonema ultrastruktura MeSH
- buněčné jádro ultrastruktura MeSH
- centrioly ultrastruktura MeSH
- flagella ultrastruktura MeSH
- mikroskopie elektronová rastrovací MeSH
- mitochondrie ultrastruktura MeSH
- Salmonidae anatomie a histologie růst a vývoj MeSH
- spermie ultrastruktura MeSH
- transmisní elektronová mikroskopie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Tributyltin (TBT), as antifouling paints, is widely present in aquatic environment, but little is known regarding the toxicity of TBT on fish brain. In this study, the effects of exposure to TBT on the antioxidant defense system, Na(+) -K(+) -ATPase activity, neurological enzymes activity and Hsp 70 protein level in brain of juvenile common carp (Cyprinus carpio) were studied. Fish were exposed to sublethal concentrations of TBT (5, 10 and 20 μg/L) for 7 days. Based on the results, with increasing concentrations of TBT, oxidative stress was apparent as reflected by the significant higher levels of oxidative indices, as well as the significant inhibition of all antioxidant enzymes activities. Besides, the activities of Acetylcholinesterase (AChE), Monoamine oxidases (MAO) and Na(+) -K(+) -ATPase were significantly inhibited after exposure to TBT with higher concentrations. In addition, the levels of Hsp 70 protein were evaluated under TBT stress with dose-depended manner. These results suggest that selected physiological responses in fish brain could be used as potential biomarkers for monitoring residual organotin compounds present in aquatic environment.
- MeSH
- acetylcholinesterasa metabolismus MeSH
- antioxidancia metabolismus MeSH
- biologické markery metabolismus MeSH
- chemické látky znečišťující vodu toxicita MeSH
- kapři * genetika metabolismus MeSH
- mozek účinky léků metabolismus MeSH
- oxidační stres účinky léků MeSH
- sodíko-draslíková ATPasa metabolismus MeSH
- trialkylcínové sloučeniny toxicita MeSH
- věkové faktory MeSH
- vodní hospodářství MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In this study, the chronic toxic effects of tributyltin (TBT), an antifouling paints commonly present in surface and ground water, on morphological indices, reactive oxygen species (ROS) generation, and ATPase activity and heat shock protein (Hsp) 70 protein in tissues (liver, gill, and white muscle) of common carp were investigated. Fish were exposed at sublethal concentrations of TBT (75 ng/L, 0.75 μg/L, and 7.5 μg/L) for 60 days. When compared with the control, there was significant lower condition factor in fish exposed at the higher concentration of TBT. ROS levels in three tissues increased significantly at higher TBT concentrations (0.75 and 7.5 μg/L). The hepatic antioxidant enzymes (total antioxidative capacity and superoxide dismutase) activities were induced at higher concentrations (0.75 μg/L) of TBT. When compared with the hepatic antioxidant enzymes activities in fish exposed to 0.75 μg/L of TBT, there was a decreasing trend in those exposed to TBT with a concentration of 7.5 μg/L. However, all the antioxidant enzymes activities were significantly inhibited in gill and muscle of fish exposed to higher TBT concentrations (0.75 and 7.5 μg/L). Moreover, there was significant lower Na-K-ATPase in three tissues after long-term exposure to higher concentration of TBT, but a significant higher Hsp70 protein levels was observed. In short, environmental concentrations of TBT could not induce obvious impacts on fish, but long-term exposure to higher concentrations of TBT could affect seriously the health status of fish. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 937-944, 2016.
- MeSH
- antioxidancia metabolismus MeSH
- biologické markery metabolismus MeSH
- chemické látky znečišťující vodu toxicita MeSH
- játra účinky léků enzymologie MeSH
- kapři metabolismus MeSH
- orgánová specificita MeSH
- proteiny tepelného šoku HSP70 metabolismus MeSH
- rybí proteiny metabolismus MeSH
- sodíko-draslíková ATPasa metabolismus MeSH
- superoxiddismutasa metabolismus MeSH
- trialkylcínové sloučeniny toxicita MeSH
- žábry metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Tributyltin (TBT) is a ubiquitous contaminant in aquatic environment, but the detailed mechanisms underlying the toxicity of TBT have not been fully understood. In this study, the effects of TBT on behavior, energy metabolism and endoplasmic reticulum (ER) stress were investigated by using Chinese rare minnow larvae. Fish larvae were exposed at sublethal concentrations of TBT (100, 400 and 800 ng/L) for 7 days. Compared with the control, energy metabolic parameters (RNA/DNA ratio, Na(+)-K(+)-ATPase) were significantly inhibited in fish exposed at highest concentration (800 ng/L), as well as abnormal behaviors observed. Moreover, we found that the PERK (PKR-like ER kinase)-eIF2α (eukaryotic translation initiation factor 2α) pathway, as the main branch was activated by TBT exposure in fish larvae. In short, TBT-induced physiological, biochemical and molecular responses in fish larvae were reflected in parameters measured in this study, which suggest that these biomarkers could be used as potential indicators for monitoring organotin compounds present in aquatic environment.
- MeSH
- chování zvířat účinky léků MeSH
- Cyprinidae růst a vývoj MeSH
- DNA vazebné proteiny genetika metabolismus MeSH
- endoribonukleasy genetika metabolismus MeSH
- energetický metabolismus účinky léků MeSH
- kinasa eIF-2 genetika metabolismus MeSH
- larva účinky léků metabolismus MeSH
- stres endoplazmatického retikula účinky léků MeSH
- transkripční faktor ATF4 genetika metabolismus MeSH
- transkripční faktory genetika metabolismus MeSH
- trialkylcínové sloučeniny toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Čína MeSH
The aim of the present study was to investigate the effect of Tributyltin (TBT) on brain function and neurotoxicity of freshwater teleost. The effects of long-term exposure to TBT on antioxidant related indices (MDA, malondialdehyde; SOD, superoxide dismutase; CAT, catalase; GR, glutathione reductase; GPx, glutathione peroxidase), Na+-K+-ATPase and neurological parameters (AChE, acetylcholinesterase; MAO, monoamine oxidase; NO, nitric oxide) in the brain of common carp were evaluated. Fish were exposed to sublethal concentrations of TBT (75 ng/L, 0.75 μg/L and 7.5 μg/L) for 15, 30, and 60 days. Based on the results, a low level and short-term TBT-induced stress could not induce the notable responses of the fish brain, but long-term exposure (more than 15 days) to TBT could lead to obvious physiological-biochemical responses (based on the measured parameters). The results also strongly indicated that neurotoxicity of TBT to fish. Thus, the measured physiological responses in fish brain could provide useful information to better understand the mechanisms of TBT-induced bio-toxicity.
