scholarly journals Glutathione S-Transferases Play a Crucial Role in Mitochondrial Function, Plasma Membrane Stability and Oxidative Regulation of Mammalian Sperm

Antioxidants ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 100 ◽  
Author(s):  
Marc Llavanera ◽  
Ariadna Delgado-Bermúdez ◽  
Samuel Olives ◽  
Yentel Mateo-Otero ◽  
Sandra Recuero ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Mitochondrial Function ◽  
Sperm Quality ◽  
Mammalian Species ◽  
Membrane Stability ◽  
Sperm Function ◽  
Cell Protection ◽  
Mammalian Sperm ◽  
Boar Sperm ◽  
Glutathione S Transferases

Glutathione S-transferases (GSTs) are essential sperm antioxidant enzymes involved in cell protection against oxidative stress and toxic chemicals, preserving sperm function and fertilising ability. Artificial insemination (AI) in pigs is commonly carried out through the use of liquid-stored semen at 17 °C, which not only reduces sperm metabolic activity but also sperm quality and AI-farrowing rates within the 72 h of storage. While one may reasonably suggest that such enzymes are implicated in the physiology and maintenance of mammalian sperm function during liquid-storage, no previous studies conducted on any species have addressed this hypothesis. Therefore, the objective of the present work was to characterise the presence and function of sperm GSTs in mammalian sperm, using the pig as a model. In this regard, inhibition of such enzymes by ethacrynic acid (EA) during semen storage at 17 °C was performed to evaluate the effects of GSTs in liquid-preserved boar sperm by flow cytometry, immunofluorescence, and immunoblotting analysis. The results of this study have shown, for the first time in mammalian species, that the inhibition of GSTs reduces sperm quality and functionality parameters during their storage at 17 °C. These findings highlight the key role of such enzymes, especially preserving mitochondrial function and maintaining plasma membrane stability. In addition, this study has identified and localised GSTM3 in the tail and equatorial subdomain of the head of boar sperm. Finally, this study has set grounds for future investigations testing supplementation of semen extenders with GSTs, as this may improve fertility outcomes of swine AIs.

scholarly journals Calpain modulates capacitation and acrosome reaction through cleavage of the spectrin cytoskeleton

Reproduction ◽  
2010 ◽  
Vol 140 (5) ◽  
pp. 673-684 ◽  
Author(s):  
Yadira Bastián ◽  
Ana L Roa-Espitia ◽  
Adela Mújica ◽  
Enrique O Hernández-González
Keyword(s):  
Plasma Membrane ◽  
Guinea Pig ◽  
Acrosome Reaction ◽  
Mammalian Species ◽  
Physiological Changes ◽  
Signal Transducer ◽  
Mammalian Sperm ◽  
Important Player ◽  
Calpain 2 ◽  
Spectrin Cytoskeleton

Research on fertilization in mammalian species has revealed that Ca2+is an important player in biochemical and physiological events enabling the sperm to penetrate the oocyte. Ca2+is a signal transducer that particularly mediates capacitation and acrosome reaction (AR). Before becoming fertilization competent, sperm must experience several molecular, biochemical, and physiological changes where Ca2+plays a pivotal role. Calpain-1 and calpain-2 are Ca2+-dependent proteases widely studied in mammalian sperm; they have been involved in capacitation and AR but little is known about their mechanism. In this work, we establish the association of calpastatin with calpain-1 and the changes undergone by this complex during capacitation in guinea pig sperm. We found that calpain-1 is relocated and translocated from cytoplasm to plasma membrane (PM) during capacitation, where it could cleave spectrin, one of the proteins of the PM-associated cytoskeleton, and facilitates AR. The aforementioned results were dependent on the calpastatin phosphorylation and the presence of extracellular Ca2+. Our findings underline the contribution of the sperm cytoskeleton in the regulation of both capacitation and AR. In addition, our findings also reveal one of the mechanisms by which calpain and calcium exert its function in sperm.

scholarly journals Exosomes derived from HEK293T cells interact in an efficient and noninvasive manner with mammalian sperm in vitro

Nanomedicine ◽  
2020 ◽  
Vol 15 (20) ◽  
pp. 1965-1980
Author(s):  
Teresa Vilanova-Perez ◽  
Celine Jones ◽  
Stefan Balint ◽  
Rebecca Dragovic ◽  
Michael L Dustin ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Mitochondrial Membrane ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Sperm Function ◽  
Sperm Analysis ◽  
Mammalian Sperm ◽  
Boar Sperm ◽  
Computer Assisted Sperm Analysis

Aim: To investigate exosomes as a noninvasive delivery tool for mammalian sperm. Materials & Methods: Exosomes were isolated from HEK293T cells and co-incubated with boar sperm in vitro. Results: Internalized exosomes were detected within 10 min of co-incubation. Computer-assisted sperm analysis and flow cytometry demonstrated that even after 5-h of exposure to exosomes, there were no significant deleterious effects with regard to sperm motility, viability, membrane integrity and mitochondrial membrane potential (p > 0.05), thus indicating that exosomes did not interfere with basic sperm function. Conclusion: HEK293T-derived exosomes interacted with boar sperm without affecting sperm function. Exosomes represent a versatile and promising research tool for studying sperm biology and provide new options for the diagnosis and treatment of male infertility.

Effect of platelet activating factor (PAF) supplementation in semen extender on viability and ATP content of cryopreserved canine spermatozoa

2010 ◽  
Vol 13 (4) ◽  
pp. 571-579 ◽  
Author(s):  
W. Kordan ◽  
M. Lecewicz ◽  
R. Strzeżek ◽  
A. Dziekońska ◽  
L. Fraser
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Mitochondrial Function ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Platelet Activating Factor ◽  
Computer Assisted ◽  
Atp Content ◽  
Plasma Membrane Integrity ◽  
Semen Extender

Effect of platelet activating factor (PAF) supplementation in semen extender on viability and ATP content of cryopreserved canine spermatozoa The aim of this study was to investigate the effect of platelet activating factor (PAF) on the quality characteristics of cryopreserved canine spermatozoa. Cryopreserved semen of 5 mixed-breed dogs was treated with different concentrations of exogenous PAF (1 × 10-3M, 1 × 10-4M, 1 × 10-5M and 1 × 10-6M) and examined at different time intervals (0, 30, 60 and 120 min). Cryopreserved semen treated without PAF was used as the control. Sperm quality was evaluated for motility (computer-assisted semen analysis, CASA), mitochondrial function (JC-1/PI assay) and plasma membrane integrity (SYBR-14/PI assay and Hoechst 33258). Also, ATP content of spermatozoa was determined using a bioluminescence assay. Treatment of cryopreserved semen with 1 × 10-3 M PAF at 120 min of incubation resulted in significantly higher total sperm motility compared with the control. It was observed that PAF-improved total sperm motility was concurrent with enhanced sperm motility patterns after treatment of cryopreserved semen. Treatment of cryopreserved semen with PAF did not improve either sperm mitochondrial function or plasma membrane integrity, as monitored by different fluorescent membrane markers. Furthermore, ATP content of cryopreserved spermatozoa was significantly higher when PAF was used at a concentration of 1 × 10-3 M compared with the control and other PAF treatments, regardless of the incubation time. The findings of this study indicated that treatment with 1 × 10-3 M PAF at 120 min of incubation rendered better quality of cryopreserved canine semen, which was associated with improved sperm motility parameters and ATP content. It can be suggested that exogenous PAF addition is beneficial as a supplement for canine semen extender used for.

scholarly journals Butaphosphan and Cyanocobalamin Supplementation in Semen Extender on Chilled Boar Sperm Quality and Life Span

2020 ◽  
Vol 7 ◽  
Author(s):  
J. Suwimonteerabutr ◽  
S. Chumsri ◽  
P. Tummaruk ◽  
Morakot Nuntapaitoon
Keyword(s):  
Plasma Membrane ◽  
Life Span ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Control Group ◽  
Plasma Membrane Integrity ◽  
Progressive Motility ◽  
Boar Sperm ◽  
Acrosome Integrity ◽  
Semen Extender

The objective of the present study was to determine the effect of butaphosphan and cyanocobalamin supplementation in semen extender on chilled boar sperm quality and life span. A total of 35 ejaculates of boar semen were included. The semen was diluted with Beltsville thawing solution extender supplemented with different concentrations of butaphosphan and cyanocobalamin [0 (control), 0.1, 0.2, 0.3, 0.4, and 0.5%] in the diluted semen. The semen samples were evaluated using a computer-assisted sperm analysis system to determine sperm motility and sperm kinetic parameters (i.e., the curvilinear velocity, VCL; straight line velocity, VSL; average path velocity, VAP; linearity, LIN; straightness, STR; amplitude of lateral head, ALH; wobble, WOB; and beat cross frequency, BCF). Additionally, sperm viability, acrosome integrity, mitochondrial activity, and plasma membrane integrity were evaluated after 4 (day 0), 72 (day 3), 120 (day 5), and 168 (day 7) h of storage using SYBR-14–ethidium homodimer-1 (EthD-1), EthD-1, JC-1, and the short hypo-osmotic swelling test, respectively. The analyses were carried out by using the general linear mixed model (MIXED) procedure of SAS. The statistical models for each data set included group, day after storage, and interaction between group and day after storage. The boar was included as a random effect. On day 0 after storage, progressive motility, VCL, VSL, VAP, and plasma membrane integrity of boar sperm in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the 0.4 and 0.5% groups (P < 0.05). On day 3 after storage, total motility and progressive motility, VCL, VSL, VAP, LIN, WOB, BCF, and plasma membrane integrity in 0.3% of butaphosphan and cyanocobalamin supplementation were significantly greater than those in the control group (P < 0.05). The total motility and progressive motility, VAP, and WOB in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the control group on day 5 after storage (P < 0.05). No effects of butaphosphan and cyanocobalamin supplementation on acrosome integrity and mitochondria activity were found on days 3, 5, and 7 after storage. However, the motility and progressive motility and the values for all sperm kinetic parameters except ALH in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the control group on day 7 after storage (P < 0.05). In conclusion, 0.3% of butaphosphan and cyanocobalamin supplementation in semen extender improved sperm motility, sperm activity, morphology, and life span in chilled boar sperm.

scholarly journals 9 DOES STRAW CONCENTRATION AFFECT POST-THAW SPERM QUALITY IN AI BULL SIRES?

2005 ◽  
Vol 17 (2) ◽  
pp. 155
Author(s):  
J. Ballester ◽  
A. Johannisson ◽  
M. Håård ◽  
H. Gustafsson ◽  
H. Rodriguez-Martinez
Keyword(s):  
Plasma Membrane ◽  
Sperm Quality ◽  
Chromatin Condensation ◽  
Membrane Integrity ◽  
Annexin V ◽  
Membrane Stability ◽  
Semen Parameters ◽  
Sperm Chromatin ◽  
Sperm Viability ◽  
Low Concentrations

There is increasing interest in decreasing the number of spermatozoa per AI dose, owing to the discrimination of threshold concentration and fertility, economical revenues and the use of sexed semen. This study evaluated the quality (as sperm viability, acrosome integrity, membrane stability and chromatin stability) post-thaw of semen collected from four elite AI sires and frozen at 15 × 106 (control) or 2 × 106 spermatozoa (spz) per dose to disclose eventual deleterious extension effects. Semen collected via a.v. from 4 élite AI sires was split-processed and frozen in 0.25 mL plastic straws under commercial conditions, at either 2 × 106 or 15 × 106 spz/straw (the latter as control). The semen parameters were within acceptable limits of normality. The post-thaw samples showed acceptable motility (above 50%). Sperm viability and stability were assessed post-thaw using flow cytometry of cells loaded with SYBR-14 and propidium iodide (PI) for sperm viability (membrane integrity), and with carboxy-SNARF-1 (SNARF; Invitrogen AB, Frolunda, Sweden), PI, and FITC-Pisum sativum agglutinin (PSA, triple stain) for acrosome status. Membrane stability status was measured with Annexin-V/PI, while sperm chromatin condensation and stability were assessed following in situ acid-induced DNA denaturation and staining with acridine orange. No significant differences were seen between the two concentrations regarding sperm motility, plasma membrane integrity (SYBR/PI, SNARF), or chromatin stability. The highly extended semen (2 × 106 spz/straw), however, showed a higher (P < 0.05) frequency of spermatozoa with translocated phosphatydil serine (as detected with Annexin-V), indicating their plasma membranes had become unstable. Also, there were more spermatozoa showing acrosomal damage (PSA). In conclusion, low concentrations of spermatozoa do not properly sustain conventional cryopreservation, although damages appear restricted to the sperm membrane. These changes in the stability of the plasma membrane apparently did not affect the viability of the spermatozoa, although they may negatively affect their lifespan. These findings may have an impact on the care that must be taken when cryopreserving low concentrations of spermatozoa with conventional freezing protocols. This work was supported by the Swedish Farmers’ Foundation for Research in Agriculture (SLF) and FORMAS, Stockholm.

scholarly journals Regulation and roles of Ca2+ stores in human sperm

Reproduction ◽  
2015 ◽  
Vol 150 (2) ◽  
pp. R65-R76 ◽  
Author(s):  
Joao Correia ◽  
Francesco Michelangeli ◽  
Stephen Publicover
Keyword(s):  
Plasma Membrane ◽  
Ion Channel ◽  
Regulatory Mechanism ◽  
Human Sperm ◽  
Good Evidence ◽  
Current Understanding ◽  
Sperm Function ◽  
Mammalian Sperm ◽  
Intracellular Organelles

[Ca2+]isignalling is a key regulatory mechanism in sperm function. In mammalian sperm the Ca2+-permeable plasma membrane ion channel CatSper is central to [Ca2+]isignalling, but there is good evidence that Ca2+stored in intracellular organelles is also functionally important. Here we briefly review the current understanding of the diversity of Ca2+stores and the mechanisms for the regulation of their activity. We then consider the evidence for the involvement of these stores in [Ca2+]isignalling in mammalian (primarily human) sperm, the agonists that may activate these stores and their role in control of sperm function. Finally we consider the evidence that membrane Ca2+channels and stored Ca2+may play discrete roles in the regulation of sperm activities and propose a mechanism by which these different components of the sperm Ca2+-signalling apparatus may interact to generate complex and spatially diverse [Ca2+]isignals.

scholarly journals Mitochondria functionality and sperm quality

Reproduction ◽  
2013 ◽  
Vol 146 (5) ◽  
pp. R163-R174 ◽  
Author(s):  
Alexandra Amaral ◽  
Bárbara Lourenço ◽  
Mónica Marques ◽  
João Ramalho-Santos
Keyword(s):  
Sperm Quality ◽  
Eukaryotic Cell ◽  
Genetically Engineered ◽  
Sperm Function ◽  
Cellular Functions ◽  
Apoptotic Pathway ◽  
Atp Production ◽  
Genetically Engineered Mouse Models ◽  
Mammalian Sperm ◽  
Intracellular Ca

Although mitochondria are best known for being the eukaryotic cell powerhouses, these organelles participate in various cellular functions besides ATP production, such as calcium homoeostasis, generation of reactive oxygen species (ROS), the intrinsic apoptotic pathway and steroid hormone biosynthesis. The aim of this review was to discuss the putative roles of mitochondria in mammalian sperm function and how they may relate to sperm quality and fertilisation ability, particularly in humans. Although paternal mitochondria are degraded inside the zygote, sperm mitochondrial functionality seems to be critical for fertilisation. Indeed, changes in mitochondrial integrity/functionality, namely defects in mitochondrial ultrastructure or in the mitochondrial genome, transcriptome or proteome, as well as low mitochondrial membrane potential or altered oxygen consumption, have been correlated with loss of sperm function (particularly with decreased motility). Results from genetically engineered mouse models also confirmed this trend. On the other hand, increasing evidence suggests that mitochondria derived ATP is not crucial for sperm motility and that glycolysis may be the main ATP supplier for this particular aspect of sperm function. However, there are contradictory data in the literature regarding sperm bioenergetics. The relevance of sperm mitochondria may thus be associated with their role in other physiological features, particularly with the production of ROS, which in controlled levels are needed for proper sperm function. Sperm mitochondria may also serve as intracellular Ca2+stores, although their role in signalling is still unclear.

scholarly journals Blocking NHE Channels Reduces the Ability of In Vitro Capacitated Mammalian Sperm to Respond to Progesterone Stimulus

2021 ◽  
Vol 22 (23) ◽  
pp. 12646
Author(s):  
Marc Yeste ◽  
Sandra Recuero ◽  
Carolina Maside ◽  
Albert Salas-Huetos ◽  
Sergi Bonet ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Mammalian Species ◽  
Physiological Role ◽  
Membrane Lipid ◽  
Equatorial Region ◽  
Lipid Disorder ◽  
Head Displacement ◽  
Mammalian Sperm ◽  
Few Data

Few data exist about the presence and physiological role of Na+/H+ exchangers (NHEs) in the plasma membrane of mammalian sperm. In addition, the involvement of these channels in the ability of sperm to undergo capacitation and acrosomal reaction has not been investigated in any mammalian species. In the present study, we addressed whether these channels are implicated in these two sperm events using the pig as a model. We also confirmed the presence of NHE1 channels in the plasma membrane of ejaculated sperm by immunofluorescence and immunoblotting. The function of NHE channels during in vitro capacitation was analyzed by incubating sperm samples in capacitating medium for 300 min in the absence or presence of a specific blocker (DMA; 5-(N,N-dimethyl)-amiloride) at different concentrations (1, 5, and 10 µM); acrosome exocytosis was triggered by adding progesterone after 240 min of incubation. Sperm motility and kinematics, integrity of plasma and acrosome membranes, membrane lipid disorder, intracellular calcium and reactive oxygen species (ROS) levels, and mitochondrial membrane potential (MMP) were evaluated after 0, 60, 120, 180, 240, 250, 270, and 300 min of incubation. NHE1 localized in the connecting and terminal pieces of the flagellum and in the equatorial region of the sperm head and was found to have a molecular weight of 75 kDa. During the first 240 min of incubation, i.e., before the addition of progesterone, blocked and control samples did not differ significantly in any of the parameters analyzed. However, from 250 min of incubation, samples treated with DMA showed significant alterations in total motility and the amplitude of lateral head displacement (ALH), acrosomal integrity, membrane lipid disorder, and MMP. In conclusion, while NHE channels are not involved in the sperm ability to undergo capacitation, they could be essential for triggering acrosome exocytosis and hypermotility after progesterone stimulus.

Ionic control of sperm function

1995 ◽  
Vol 7 (4) ◽  
pp. 905 ◽  
Author(s):  
LR Fraser
Keyword(s):  
Ionic Composition ◽  
Ion Fluxes ◽  
Sperm Function ◽  
Considerable Evidence ◽  
Functional Potential ◽  
Mammalian Sperm ◽  
Acrosomal Exocytosis ◽  
Extracellular Environment

Successful sperm function leads to fertilization. It is dependent on the extracellular environment, especially the array and concentration of various ions. Considerable evidence indicates that this is because of consequent effects on the intracellular ionic composition. Although both cations and anions undoubtedly play a role in a modulating sperm function, most of the evidence currently available concerns cations. Therefore, this review will concentrate on cations, focussing on Ca2+, Na+, K+ and H+. Their requirements for successful capacitation (mammalian sperm) and acrosomal exocytosis (both invertebrate and mammalian sperm) will be considered. In particular, the mechanisms which may control ion fluxes, leading to changes in the intracellular ionic composition and subsequently to changes in sperm functional potential, will be addressed.

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