Porcine model for the study of sperm capacitation, fertilization and male fertility

Zigo, Michal; Maňásková-Postlerová, Pavla; Zuidema, Dalen; Kerns, Karl; Jonáková, Věra; Tůmová, Lucie; Bubeníčková, Filipa; Šutovský, Peter

doi:10.1007/s00441-020-03181-1

Cited by 43 publications

(34 citation statements)

References 305 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…It is suggested that SP contains decapacitation factors that bind it to the plasma membrane of sperm in order to maintain fertilization potential [ 6 , 53 ]. Most SP proteins maintain an uncapacitated sperm state, which prevents sperm agglutination and premature acrosome reaction, and further immune-suppresses the female reproductive tract [ 54 ]. In our study, plasma membrane integrity was not significantly affected after thawing by any protein fraction concentration.…”

Section: Discussionmentioning

confidence: 99%

Effect of Seminal Plasma Protein Fractions on Stallion Sperm Cryopreservation

Bubeníčková

Maňásková-Postlerová

Šimoník

et al. 2020

IJMS

View full text Add to dashboard Cite

Seminal plasma (SP) is the natural environment for spermatozoa and contains a number of components, especially proteins important for successful sperm maturation and fertilization. Nevertheless, in standard frozen stallion insemination doses production, SP is completely removed and is replaced by a semen extender. In the present study, we analyzed the effects of the selected seminal plasma protein groups that might play an important role in reducing the detrimental effects on spermatozoa during the cryopreservation process. SP proteins were separated according to their ability to bind to heparin into heparin-binding (Hep+) and heparin-non-binding (Hep−) fractions. The addition of three concentrations—125, 250, and 500 µg/mL—of each protein fraction was tested. After thawing, the following parameters were assessed: sperm motility (by CASA), plasma membrane integrity (PI staining), and acrosomal membrane integrity (PNA staining) using flow cytometry, and capacitation status (anti-phosphotyrosine antibody) using imaging-based flow cytometry. Our results showed that SP protein fractions had a significant effect on the kinematic parameters of spermatozoa and on a proportion of their subpopulations. The 125 µg/mL of Hep+ protein fraction resulted in increased linearity (LIN) and straightness (STR), moreover, with the highest values of sperm velocities (VAP, VSL), also this group contained the highest proportion of the fast sperm subpopulation. In contrast, the highest percentage of slow subpopulation was in the groups with 500 µg/mL of Hep+ fraction and 250 µg/mL of Hep− fraction. Interestingly, acrosomal membrane integrity was also highest in the groups with Hep+ fraction in concentrations of 125 µg/mL. Our results showed that the addition of protein fractions did not significantly affect the plasma membrane integrity and capacitation status of stallion spermatozoa. Moreover, our results confirmed that the effect of SP proteins on the sperm functionality is concentration-dependent, as has been reported for other species. Our study significantly contributes to the lack of studies dealing with possible use of specific stallion SP fractions in the complex puzzle of the improvement of cryopreservation protocols. It is clear that improvement in this field still needs more outputs from future studies, which should be focused on the effect of individual SP proteins on other sperm functional parameters with further implication on the success of artificial insemination in in vivo conditions.

show abstract

Section: Discussionmentioning

confidence: 99%

Effect of Seminal Plasma Protein Fractions on Stallion Sperm Cryopreservation

Bubeníčková

Maňásková-Postlerová

Šimoník

et al. 2020

IJMS

View full text Add to dashboard Cite

show abstract

“…Herein, cell biology and immunological approaches were performed through pharmacologically inhibiting the formation of the GSTP1-JNK heterocomplex, prior to analyzing sperm quality and functionality parameters, the presence and localization of GSTP1, and the activation of JNK. Therefore, the present study aimed to investigate the function of this heterocomplex in mammalian sperm physiology, using the pig as a model, which has recently been stablished as a suitable animal model for research in human reproduction (Zigo et al, 2020). Accordingly, we hypothesized that the dissociation of the GSTP1-JNK heterocomplex, known to occur under cellular stress conditions, enhances the JNK signaling pathway and disrupts sperm physiology.…”

Section: Introductionmentioning

confidence: 99%

Deactivation of the JNK Pathway by GSTP1 Is Essential to Maintain Sperm Functionality

Llavanera

Mateo‐Otero

Delgado-Bermúdez

et al. 2021

Front. Cell Dev. Biol.

View full text Add to dashboard Cite

Fifty percent of male subfertility diagnosis is idiopathic and is usually associated with genetic abnormalities or protein dysfunction, which are not detectable through the conventional spermiogram. Glutathione S-transferases (GSTs) are antioxidant enzymes essential for preserving sperm function and maintaining fertilizing ability. However, while the role of GSTP1 in cell signaling regulation via the inhibition of c-Jun N-terminal kinases (JNK) has been enlightened in somatic cells, it has never been investigated in mammalian spermatozoa. In this regard, a comprehensive approach through immunoblotting, immunofluorescence, computer-assisted sperm assessment (CASA), and flow cytometry analysis was used to characterize the molecular role of the GSTP1–JNK heterocomplex in sperm physiology, using the pig as a model. Immunological assessments confirmed the presence and localization of GSTP1 in sperm cells. The pharmacological dissociation of the GSTP1–JNK heterocomplex resulted in the activation of JNK, which led to a significant decrease in sperm viability, motility, mitochondrial activity, and plasma membrane stability, as well as to an increase of intracellular superoxides. No effects in intracellular calcium levels and acrosome membrane integrity were observed. In conclusion, the present work has demonstrated, for the first time, the essential role of GSTP1 in deactivating JNK, which is crucial to maintain sperm function and has also set the grounds to understand the relevance of the GSTP1–JNK heterocomplex for the regulation of mammalian sperm physiology.

show abstract

“…Capacitation occurs within the female reproductive tract and consists of a series of changes that prepare sperm to fertilize the oocyte [21][22][23]. While plasma membrane integrity indicates that sperm are viable [37,38], its destabilization, which involves changes in its architecture and eventually leads to death, is one of the features of sperm capacitation [39].…”

Section: Discussionmentioning

confidence: 99%

“…Upon ejaculation, mammalian spermatozoa are not yet ready to fertilize the oocyte, but rather need to undergo a series of changes that occur within the female reproductive tract and are known as sperm capacitation [21][22][23]. Elucidating the role of potassium channels during capacitation of pig spermatozoa may be conducted through the use of blocking agents, in a similar fashion to previous studies [4,24,25].…”

Section: Introductionmentioning

confidence: 97%

Inhibition of Potassium Channels Affects the Ability of Pig Spermatozoa to Elicit Capacitation and Trigger the Acrosome Exocytosis Induced by Progesterone

Noto

Recuero

Valencia

et al. 2021

IJMS

View full text Add to dashboard Cite

During capacitation, sperm undergo a myriad of changes, including remodeling of plasma membrane, modification of sperm motility and kinematic parameters, membrane hyperpolarization, increase in intracellular calcium levels, and tyrosine phosphorylation of certain sperm proteins. While potassium channels have been reported to be crucial for capacitation of mouse and human sperm, their role in pigs has not been investigated. With this purpose, sperm samples from 15 boars were incubated in capacitation medium for 300 min with quinine, a general blocker of potassium channels (including voltage-gated potassium channels, calcium-activated potassium channels, and tandem pore domain potassium channels), and paxilline (PAX), a specific inhibitor of calcium-activated potassium channels. In all samples, acrosome exocytosis was induced after 240 min of incubation with progesterone. Plasma membrane and acrosome integrity, membrane lipid disorder, intracellular calcium levels, mitochondrial membrane potential, and total and progressive sperm motility were evaluated after 0, 120, and 240 min of incubation, and after 5, 30, and 60 min of progesterone addition. Although blocking potassium channels with quinine and PAX prevented sperm to elicit in vitro capacitation by impairing motility and mitochondrial function, as well as reducing intracellular calcium levels, the extent of that inhibition was larger with quinine than with PAX. Therefore, while our data support that calcium-activated potassium channels are essential for sperm capacitation in pigs, they also suggest that other potassium channels, such as the voltage-gated, tandem pore domain, and mitochondrial ATP-regulated ones, are involved in that process. Thus, further research is needed to elucidate the specific functions of these channels and the mechanisms underlying its regulation during sperm capacitation.

show abstract

Porcine model for the study of sperm capacitation, fertilization and male fertility

Cited by 43 publications

References 305 publications

Effect of Seminal Plasma Protein Fractions on Stallion Sperm Cryopreservation

Effect of Seminal Plasma Protein Fractions on Stallion Sperm Cryopreservation

Deactivation of the JNK Pathway by GSTP1 Is Essential to Maintain Sperm Functionality

Inhibition of Potassium Channels Affects the Ability of Pig Spermatozoa to Elicit Capacitation and Trigger the Acrosome Exocytosis Induced by Progesterone

Contact Info

Product

Resources

About