scholarly journals HVCN1 but Not Potassium Channels Are Related to Mammalian Sperm Cryotolerance

2021 ◽  
Vol 22 (4) ◽  
pp. 1646
Author(s):  
Ariadna Delgado-Bermúdez ◽  
Yentel Mateo-Otero ◽  
Marc Llavanera ◽  
Sergi Bonet ◽  
Marc Yeste ◽  
...  
Keyword(s):  
Potassium Channels ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Physiological Role ◽  
Membrane Lipid ◽  
Proton Channels ◽  
Lipid Disorder ◽  
Mammalian Sperm

Little data exist about the physiological role of ion channels during the freeze–thaw process in mammalian sperm. Herein, we determined the relevance of potassium channels, including SLO1, and of voltage-gated proton channels (HVCN1) during mammalian sperm cryopreservation, using the pig as a model and through the addition of specific blockers (TEA: tetraethyl ammonium chloride, PAX: paxilline or 2-GBI: 2-guanidino benzimidazole) to the cryoprotective media at either 15 °C or 5 °C. Sperm quality of the control and blocked samples was performed at 30- and 240-min post-thaw, by assessing sperm motility and kinematics, plasma and acrosome membrane integrity, membrane lipid disorder, intracellular calcium levels, mitochondrial membrane potential, and intracellular O2−⁻ and H2O2 levels. General blockade of K+ channels by TEA and specific blockade of SLO1 channels by PAX did not result in alterations in sperm quality after thawing as compared to control samples. In contrast, HVCN1-blocking with 2-GBI led to a significant decrease in post-thaw sperm quality as compared to the control, despite intracellular O2−⁻ and H2O2 levels in 2-GBI blocked samples being lower than in the control and in TEA- and PAX-blocked samples. We can thus conclude that HVCN1 channels are related to mammalian sperm cryotolerance and have an essential role during cryopreservation. In contrast, potassium channels do not seem to play such an instrumental role.

scholarly journals Aquaglyceroporins but not orthodox aquaporins are involved in the cryotolerance of pig spermatozoa

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Ariadna Delgado-Bermúdez ◽  
Marc Llavanera ◽  
Leira Fernández-Bastit ◽  
Sandra Recuero ◽  
Yentel Mateo-Otero ◽  
...  
Keyword(s):  
Lipid Membrane ◽  
Sperm Quality ◽  
Membrane Lipid ◽  
Lipid Disorder ◽  
Long Term Storage ◽  
Boar Sperm ◽  
Functional Relevance ◽  
Positive Effect

Abstract Background Aquaporins (AQPs) are a family of transmembrane water channels that includes orthodox AQPs, aquaglyceroporins (GLPs) and superAQPs. AQP3, AQP7, AQP9 and AQP11 have been identified in boar sperm, and they are crucial for sperm maturation and osmoregulation. Water exchange is an important event in cryopreservation, which is the most efficient method for long-term storage of sperm. However, the freeze-thaw process leads to sperm damage and a loss of fertilizing potential. Assuming that the quality of frozen-thawed sperm partially depends on the regulation of osmolality variations during this process, AQPs might play a crucial role in boar semen freezability. In this context, the aim of this study was to unravel the functional relevance of the different groups of AQPs for boar sperm cryotolerance through three different inhibitors. Results Inhibition of different groups of AQPs was found to have different effects on boar sperm cryotolerance. Whereas the use of 1,3-propanediol (PDO), an inhibitor of orthodox AQPs and GLPs, decreased total motility (P < 0.05), it increased post-thaw sperm viability, lowered membrane lipid disorder and increased mitochondrial membrane potential (MMP) (P < 0.05). When acetazolamide (AC) was used as an inhibitor of orthodox AQPs, the effects on post-thaw sperm quality were restricted to a mild increase in MMP in the presence of the intermediate concentration at 30 min post-thaw and an increase in superoxide levels (P < 0.05). Finally, the addition of phloretin (PHL), a GLP inhibitor, had detrimental effects on post-thaw total and progressive sperm motilities, viability and lipid membrane disorder (P < 0.05). Conclusions The effects of the different inhibitors suggest that GLPs rather than orthodox AQPs are relevant for boar sperm freezability. Moreover, the positive effect of PDO on sperm quality suggests a cryoprotective role for this molecule.

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.

Elucidating The Physiological Role of Slo1 and Hvcn1 Channels In Mammalian Sperm Cryopreservation

Cryobiology ◽  
2021 ◽  
Vol 103 ◽  
pp. 181-182
Author(s):  
Ariadna Delgado-Bermúdez ◽  
Yentel Mateo-Otero ◽  
Marc Llavanera ◽  
Sergi Bonet ◽  
Marc Yeste ◽  
...  
Keyword(s):  
Physiological Role ◽  
Sperm Cryopreservation ◽  
Mammalian Sperm

scholarly journals Effect of N-Methylacetamide Concentration and Thawing Rate on Chicken Sperm Quality after Cryopreservation

Animals ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 824
Author(s):  
Fabio Mosca ◽  
Luisa Zaniboni ◽  
Ahmad Abdel Sayed ◽  
Nicolaia Iaffaldano ◽  
Dominga Soglia ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Kinetic Parameters ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Motile Sperm ◽  
Cell Membrane Integrity ◽  
Before And After ◽  
Freezing Procedure ◽  
Thawing Rate

In seeking alternative cryoprotectants to glycerol for a reference chicken semen freezing procedure, the aim of the present study was to compare the effect of two concentrations of N-Methylacetamide (MA) and two thawing rates on the quality of frozen-thawed semen. Semen samples were diluted in Lake pre-freezing extender, including 0.1 M trehalose in presence of 6% or 9% MA, loaded into straws, frozen in nitrogen vapors, and stored in liquid nitrogen. The following thawing treatments were used: 5 °C for 100 s and 38 °C for 30 s. Sperm quality (cell membrane integrity, motility and kinetic parameters) was assessed before and after cryopreservation. The decrease of MA concentration from 9 to 6% improved sperm quality after freezing/thawing and this effect was dependent on thawing temperature. Decreasing the MA concentration from 9 to 6% improved the proportion of undamaged membrane, motile, and progressive motile sperm recovered after thawing at 5 °C for 100 s; in contrast, no effect of the MA concentration was observed thawing at 38 °C for 30 s. Therefore, the treatment with 6% MA and thawing at 5 °C for 100 s has given the best cryoprotective action. These results contribute to improve the efficacy of the current chicken semen cryopreservation procedures.

scholarly journals The Effect of Adding Different Levels of Curcumin and Its Nanoparticles to Extender on Post-Thaw Quality of Cryopreserved Rabbit Sperm

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1508 ◽  
Author(s):  
Sameh Abdelnour ◽  
Mahmoud Hassan ◽  
Amer Mohammed ◽  
Ahmad Alhimaidi ◽  
Naif Al-Gabri ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Positive Influence ◽  
Control Group ◽  
Curcumin Nanoparticles ◽  
Total Antioxidant ◽  
Semen Extender ◽  
Apoptosis Process ◽  
Structural Levels

The cryopreservation process adversely affects sperm function and quality traits, causing some changes at biochemical and structural levels, due to mechanical, thermal, osmotic, and oxidative damage. Supplementation with curcumin nanoparticles could prevent and even revert this effect and could enhance the post/thawed sperm quality in the rabbit. The study amid to explore the effect of curcumin (CU) and curcumin nanoparticles (CUNPs) supplementation in semen extender on post/thawed rabbit sperm quality. Twelve fertile, healthy rabbit bucks were included, and the ejaculates were collected using artificial vaginas. Rabbit pooled semen was cryopreserved in tris-yolk fructose (TYF) extender without any supplement (control group) or extender supplemented with CU at levels of 0.5, 1 or 1.5 µg/mL (CU0.5, CU1.0, and CU1.5, respectively) or CUNPs at levels of 0.5, 1, 1.5 (CUNPs0.5, CUNPs1.0, and CUNPs1.5, respectively) and was packed in straws (0.25 mL) and stored in liquid nitrogen (−196 °C). Results revealed that CUNPs1.5 had a positive influence (p < 0.05) on post-thawing sperm progressive motility, viability, and membrane integrity as compared with the other groups. Percentages of dead sperm, abnormalities, early apoptotic, apoptotic, and necrotic sperm cells reduced (p < 0.05) in CUNPs1.5 as compared to other treatments. Using 1.5 µg/mL of CUNPs significantly improved total antioxidant capacity (TAC), GPx, while MDA and POC reduced (p < 0.05) in CU1.5 in comparison with other groups. SOD values were enhanced (p < 0.05) in CUNPs1.0 and CUNPs1.5 in relation with other treatments. Conclusively, the addition of curcumin and its nanoparticles to the extender can improve the post-thawed quality of rabbit sperm via redox signaling and reduce the apoptosis process.

scholarly journals O papel do dimetilsulfóxido na criopreservação de sêmen de Curimba (Prochilodus lineatus)

2015 ◽  
Vol 36 (5) ◽  
pp. 3471
Author(s):  
Antonio Sergio Varela Junior ◽  
Estela Fernandes Silva ◽  
Tainã Figueiredo Cardoso ◽  
Érica Yokoyama Namba ◽  
Rodrigo Desessards Jardim ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Fertilization Rate ◽  
Dna Integrity ◽  
Hatching Rate ◽  
Sperm Viability ◽  
Prochilodus Lineatus ◽  
Dmso Concentration ◽  
Fresh Semen

<p class="Pa7">Cryopreservation of Curimba semen (Prochilodus lineatus) is ecological and commercial importance. The objective of this study was to evaluate the effect of different concentrations (2, 5, 8 and 11%) of dimethyl sulfoxide (DMSO) diluted in Betsville Thawing Solution (BTS) on the quality of post-thaw semen Curimba. We analyzed the rate and period motility, sperm viability, membrane integrity and DNA, mitochondrial functionality, and fertilization and hatching rate. The plasma membrane and DNA integrity of a DMSO concentration of 11% obtained better results than the concentration of 5% (p &lt;0.05). However, treatment of 5% DMSO resulted in a longer latency and a higher fertilization rate and hatching, in other sperm quality equal to that of fresh semen. The results of this study indicate that 5% DMSO is ideal for cryopreservation of semen Curimba.</p>

scholarly journals Evaluation of sperm quality of Erythrolamprus poecilogyrus sublineatus (Cope, 1860) (Serpentes, Dipsadidae)

2017 ◽  
Vol 77 (3) ◽  
pp. 553-557
Author(s):  
A. C. Silva ◽  
A. S. Varela Junior ◽  
T. F. Cardoso ◽  
E. F. Silva ◽  
D. Loebmann ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Coastal Region ◽  
Rio Grande ◽  
Rio Grande Do Sul ◽  
Sperm Analysis ◽  
Pampa Region

Abstract Erythrolamprus poecilogyrus sublineatus (Cope, 1860), is a species widely distributed in the Pampa Domain, occurring in Rio Grande do Sul, Argentina and Uruguay, mainlyin the pampa region. In the coastal region of southern Brazil this is serpent is considered one of the most abundant. The purpose of the present study is to describe the techniques of sperm evaluation in vitro for E. poecilogyrus sublineatus in the coastal plain of Rio Grande do Sul, Brazil. After laparatomy the efferent vases were collected and the semen was diluted in 1ml Beltsville Thawing Solution. The characteristics of motility, membrane integrity, mitochondria, acrosome, DNA, cell viability and cellular functionality were evaluated. Fluorescent probes were used for the evaluation of sperm structure in epifluorescence microscope. With the techniques described, it was possible to identify intact and injured cells, enabling the determination of cell characteristics for the spring season (October and November). It was observed in the analyses that 80% of sperm cells were mobile and that 84.1 ± 8.0% of sperm membranes were intact. The standards found were of 48 ± 13.8% of intact acrosome, 73.6 ± 6.0 of perfect DNA and of 91.8 ± 4.0 of functional mitochondria. Thus, these values from the sperm analysis can be used as standards for the species Erythrolamprus poecilogyrus sublineatus.

scholarly journals Cell Membrane Stability and the Role of Calcium Infiltration in Postharvest Quality of Apples

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 815G-815
Author(s):  
G.A. Picchioni ◽  
A.E. Watada ◽  
W.S. Conway ◽  
B.D. Whitaker ◽  
C.E. Sams
Keyword(s):  
Membrane Lipid ◽  
Cellular Level ◽  
Postharvest Quality ◽  
Storage Quality ◽  
Pressure Infiltration ◽  
Cell Membrane Stability ◽  
Golden Delicious ◽  
And Storage

Postharvest CaCl2 pressure infiltration improves firmness and storage quality of apples but is still in the experimental stages. Its effectiveness could be increased if we had a better understanding of how Ca affects the tissue at the cellular level. `Golden Delicious' fruit were harvested from a commercial orchard and were pressure-infiltrated with CaCl2 (0%, 2%, or 4% w/v), stored for 6 months at 0C, and then for 7 days at 20C. Between harvest and the end of storage at 20C, the net breakdown of galactolipids and phospholipids decreased with increasing CaCl2 in infiltration solutions. During 0C storage, CaCl2-infiltrated fruit maintained greater concentrations of conjugated sterol lipids, and these lipid classes are thought to be closely associated with the plasma membrane. As membrane lipid alterations are viewed as a central factor in the senescence of fruits, Ca (from postharvest infiltration) may serve a major role in regulating fruit quality losses through its interactions with cell membranes.

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

2021 ◽  
Vol 9 ◽  
Author(s):  
Marc Llavanera ◽  
Yentel Mateo-Otero ◽  
Ariadna Delgado-Bermúdez ◽  
Sandra Recuero ◽  
Samuel Olives ◽  
...  
Keyword(s):  
Membrane Integrity ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Sperm Function ◽  
Jnk Pathway ◽  
Male Subfertility ◽  
Mammalian Sperm ◽  
Genetic Abnormalities ◽  
Protein Dysfunction

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.

scholarly journals Effect of refrigeration at -1°C on spermatozoa quality of domestic cats

2020 ◽  
Vol 40 (4) ◽  
pp. 306-314
Author(s):  
Anne K. Souza ◽  
Luiz Guilherme C. Trautwein ◽  
Cristiane S. Paranzini ◽  
Josiana F. Schnitzer ◽  
Felipe M. Perencin ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Quality Parameter ◽  
Domestic Cats ◽  
R Software ◽  
Sperm Sample ◽  
Morphological Defects ◽  
Harvesting Techniques ◽  
Adult Cats

ABSTRACT: The objective of this study was to evaluate the sperm quality obtained of domestic cats by electroejaculation and recovery of the tail of the epididymis after cooling at -1°C and 4°C for 24 and 48 hours. Twenty-nine adult cats (2 to 6kg) were used. Sperm collection was performed by electroejaculation (EEJ), and after 48 hours, the cats were orchiectomized, and sperm sample was obtained from the vas deferens and epididymis tail (EPD). The samples were diluted in ACP-117® extender, and the sperm characteristics were evaluated at three different moments: when still fresh, 24 and 48 hours after cooling. In order to compare the two refrigeration temperatures, the first stage was to analyze if there was a difference between the harvesting techniques. After this, two experiments were conducted: in the first, sperm sample from 14 cats were used and the cooling was performed at -1°C; and in the second, sample from 15 cats were used and the sperm were refrigerated at 4°C. Sperm kinetics were evaluated by computerized analysis (CASA) and concentration by Neubauer chamber, spermatic morphology was evaluated by modified Karras staining, and membrane integrity was evaluated by eosin nigrosine. The results obtained were analyzed in R software, version 3.2.5 using the Mann-Whitney test for variables with abnormal distributions, considering significance at the level of 5%. In ejaculate samples, higher values of total morphological defects were observed after 24 and 48 hours of refrigeration at 4°C (P<0.022) compared to refrigeration at -1°C, using Friedman test. To quantify the decrease in sperm quality, parameter reductions were calculated among time points (F-24h/F-48h/24h-48h). In EPD samples, a greater reduction in sperm quality was detected after 24 hours of refrigeration at 4°C, both in motility and sperm kinetics and in the movement and velocity indices, compared to refrigeration at -1°C. Based on the results, it can be concluded that cooling of feline spermatozoa at -1°C for up to 48 hours was efficient in maintaining spermatic quality collected by EEJ and EPD, and it could be an alternative to spermatozoa cryopreservation in domestic felines.

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