scholarly journals Perfluorooctane Sulfonate (PFOS) and Perfluorohexane Sulfonate (PFHxS) Alters Protein Phosphorylation, Increase ROS Levels and DNA Fragmentation during In Vitro Capacitation of Boar Spermatozoa

Animals ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1934
Author(s):  
Iván Oseguera-López ◽  
Serafín Pérez-Cerezales ◽  
Paola Berenice Ortiz-Sánchez ◽  
Oscar Mondragon-Payne ◽  
Raúl Sánchez-Sánchez ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Acrosome Reaction ◽  
Perfluorooctane Sulfonate ◽  
Toxic Effects ◽  
Toxic Compounds ◽  
Worldwide Distribution ◽  
Lack Of Information ◽  
Boar Spermatozoa ◽  
Vital Parameters

Perfluorooctane sulfonate (PFOS) and perfluorohexane sulfonate (PFHxS) are toxic and bioaccumulative, included in the Stockholm Convention’s list as persistent organic pollutants. Due to their toxicity, worldwide distribution, and lack of information in spermatozoa physiology during pre-fertilization processes, the present study seeks to analyze the toxic effects and possible alterations caused by the presence of these compounds in boar sperm during the in vitro capacitation. The spermatozoa capacitation was performed in supplemented TALP-Hepes media and mean lethal concentration values of 460.55 μM for PFOS, and 1930.60 μM for PFHxS were obtained. Results by chlortetracycline staining showed that intracellular Ca2+ patterns bound to membrane proteins were scarcely affected by PFOS. The spontaneous acrosome reaction determined by FITC-PNA was significantly reduced by PFOS and slightly increased by PFHxS. Both toxic compounds significantly alter the normal capacitation process from 30 min of exposure. An increase in ROS production was observed by flow cytometry and considerable DNA fragmentation by the comet assay. The immunocytochemistry showed a decrease of tyrosine phosphorylation in proteins of the equatorial and acrosomal zone of the spermatozoa head. In conclusion, PFOS and PFHxS have toxic effects on the sperm, causing mortality and altering vital parameters for proper sperm capacitation.

scholarly journals Sperm binding to ZP2-coated beads improve the efficiency of porcine in vitro fertilisation

Reproduction ◽  
2020 ◽  
Vol 160 (5) ◽  
pp. 725-735
Author(s):  
Julieta Gabriela Hamze ◽  
María Jiménez-Movilla ◽  
Raquel Romar
Keyword(s):  
Acrosome Reaction ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
In Vitro Fertilisation ◽  
Sperm Binding ◽  
Fertilisation Rate ◽  
Gamete Recognition ◽  
Boar Spermatozoa

The role of specific zona pellucida (ZP) glycoproteins in gamete interaction has not yet been elucidated in many species. A recently developed 3D model based on magnetic sepharose beads (B) conjugated to recombinant ZP glycoproteins (BZP) and cumulus cells (CBZP) allows the study of isolated ZP proteins in gamete recognition studies. The objective of this work was to study the role of porcine ZP2, ZP3 and ZP4 proteins in sperm binding, cumulus cell adhesion and acrosome reaction triggering. ZP protein-bound beads were incubated with fresh ejaculated boar spermatozoa and isolated cumulus cells for 24 h. The number of sperm bound to the beads, the acrosomal shrouds (presence of acrosomal content) on the bead’s surface, and the acrosome integrity (by means of PNA-FITC lectin) in bound and unbound sperm were studied. Finally, in vitro matured porcine oocytes mixed with BZP2 were inseminated in vitro using fresh sperm and fertilisation results evaluated. Over 60% of beads had at least one sperm bound after 2 h of coincubation. ZP2-beads (BZP2) and cumulus-ZP2-bead complexes (CBZP2) reached the highest number of sperm per bead, whereas BZP3 and BZP4 models showed the highest number of unbound reacted sperm cells and acrosomal shrouds. Fertilisation efficiency and monospermy rate increased when oocytes were fertilised in the presence of BZP2. We, therefore, conclude that in pigs, it is mainly ZP2 that is involved in sperm-ZP binding whereas ZP3 and ZP4 induce acrosome reaction. Using magnetic sepharose ZP2-bound beads might be a valuable tool to improve the fertilisation rate in pigs.

Toxic effects of endosulfan based insecticides on mammalian sperm: Evaluation of in vitro chromatin decondensation and DNA fragmentation

2015 ◽  
Vol 238 (2) ◽  
pp. S107
Author(s):  
M.C. Sanchez ◽  
C. Alvarez Sedo ◽  
M. Romanato ◽  
F. Lo Nostro ◽  
L. Calvo ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Toxic Effects ◽  
Chromatin Decondensation ◽  
Mammalian Sperm

Bimodal redistribution of surface transmembrane glycoproteins during Ca2+-dependent secretion (acrosome reaction) in boar spermatozoa

1989 ◽  
Vol 93 (3) ◽  
pp. 467-479
Author(s):  
A.P. Aguas ◽  
P.P. da Silva
Keyword(s):  
Plasma Membrane ◽  
Acrosome Reaction ◽  
Freeze Fracture ◽  
Secretory Process ◽  
Membrane Domains ◽  
Cytoplasmic Vesicle ◽  
Acrosomal Membrane ◽  
Freeze Fracture Electron Microscopy ◽  
Boar Spermatozoa

We used the acrosome reaction of boar sperm cells to study the dynamics of surface transmembrane glycoproteins (TMG) during a secretory process. The acrosome reaction is the Ca2+-dependent fusion of a large cytoplasmic vesicle (the acrosome) with the overlying segment of the plasma membrane (acrosomal cap) that leads to the release of the acrosomal enzymes. After triggering the acrosome reaction in vitro (2 mM-CaCl2 in the presence of 10 microM-A23187), we used freeze-fracture electron microscopy to follow the topographical rearrangement of a population of acrosomal-cap large intramembrane particles that correspond to transmembrane proteins that bind wheat germ agglutinin. We found that these TMG move in the direction of either one of two opposite poles, proximal and distal, of the acrosomal cap. This bimodal movement of the TMG reorganizes the acrosomal cap into three extensive domains. The first two, on the apical rim and on the equator, are membrane domains to which the TMG are directed and where they accumulate. The third, a large in-between area of protein clearing, corresponds to the region from which TMG were preferentially located before displacement induced by the Ca2+ effect. The topography of these new membrane domains of the acrosomal cap becomes coincident with that of the structural domains of the subjacent acrosomal membrane. Mirroring of the acrosomal membrane by the plasma membrane is followed by fusion between the two membranes, formation of an exquisite labyrinth of hybrid-membrane tubules, followed by fission and release of the acrosomal contents through intertubular fenestrae.

Seminal plasma proteins inhibit in vitro- and cooling-induced capacitation in boar spermatozoa

2010 ◽  
Vol 22 (6) ◽  
pp. 893 ◽  
Author(s):  
Melissa L. Vadnais ◽  
Kenneth P. Roberts
Keyword(s):  
Mass Spectrometry ◽  
Seminal Plasma ◽  
Plasma Proteins ◽  
Acrosome Reaction ◽  
Heparin Binding ◽  
Total Proteins ◽  
Binding Properties ◽  
Seminal Plasma Proteins ◽  
Boar Spermatozoa

Dilute boar seminal plasma (SP) has been shown to inhibit in vitro capacitation and cooling-induced capacitation-like changes in boar spermatozoa, as assessed by the ability of the spermatozoa to undergo an ionophore-induced acrosome reaction. We hypothesised that the protein component of SP is responsible for this effect. To test this hypothesis, varying concentrations of total SP protein or SP proteins fractionated by heparin binding were assayed for their ability to inhibit in vitro capacitation, as well as cooling- and cryopreservation-induced capacitation-like changes. In vitro capacitation and cooling-induced capacitation-like changes were prevented by 10% whole SP, as well as by total proteins extracted from SP at concentrations greater than 500 μg mL−1. No amount of SP protein was able to prevent cryopreservation-induced capacitation-like changes. Total SP proteins were fractionated based on their heparin-binding properties and the heparin-binding fraction was shown to possess capacitation inhibitory activity at concentrations as low as 250 µg mL−1. The proteins in the heparin-binding fraction were subjected to mass spectrometry and identified. The predominant proteins were three members of the spermadhesin families, namely AQN-3, AQN-1 and AWN, and SP protein pB1. We conclude that one or more of these heparin-binding SP proteins is able to inhibit in vitro capacitation and cooling-induced capacitation-like changes, but not cryopreservation-induced capacitation-like changes, in boar spermatozoa.

scholarly journals Simultaneous Morphology, Motility and Fragmentation Analysis of Live Individual Sperm Cells for Male Fertility Evaluation

2021 ◽  
Author(s):  
Keren Ben-Yehuda ◽  
Simcha K Mirsky ◽  
Mattan Levi ◽  
Itay Barnea ◽  
Inbal Meshulach ◽  
...  
Keyword(s):  
Deep Learning ◽  
Dna Fragmentation ◽  
Male Fertility ◽  
Human Sperm ◽  
Sperm Cells ◽  
Interferometric Imaging ◽  
Vitro Fertilization ◽  
Lack Of Information ◽  
Common Clinical Practice

We present a new technique for simultaneously analyzing morphology, motility and DNA fragmentation of live human sperm cells at the single-cell level for male fertility evaluation. It relies on quantitative stain-free interferometric imaging and multiple deep-learning frameworks. In the common clinical practice, only motility evaluation is carried out on live human cells, while full morphological evaluation and DNA fragmentation assays require different staining protocols, and therefore cannot be performed simultaneously on the same cell. This results in a lack of information regarding the intersection of these scores. We use a clinic-ready interferometric module and deep learning to acquire dynamic sperm cells without chemical staining, and evaluate all three scores per each cell together with virtual staining. We show that the number of cells that pass each criterion separately does not accurately predict how many would pass all criteria, thus the triple evaluation per cell is necessary for accurate fertility grading. This stain-free evaluation is expected to decrease the uncertainty in male fertility evaluation, as well as be applied for sperm selection during in vitro fertilization.

scholarly journals Glycolipid migration from the apical to the equatorial subdomains of the sperm head plasma membrane precedes the acrosome reaction. Evidence for a primary capacitation event in boar spermatozoa

1995 ◽  
Vol 108 (3) ◽  
pp. 935-946 ◽  
Author(s):  
B.M. Gadella ◽  
M. Lopes-Cardozo ◽  
L.M. van Golde ◽  
B. Colenbrander ◽  
T.W. Gadella
Keyword(s):  
Plasma Membrane ◽  
Acrosome Reaction ◽  
Plasma Membranes ◽  
Phase Segregation ◽  
Sperm Head ◽  
Lipid Phase ◽  
Sperm Cells ◽  
Fluorescence Lifetime Imaging ◽  
Boar Spermatozoa

In order to extend the static information of immunolabelling sulphogalactolipids in fixed boar spermatozoa, a fluorescent sulphogalactolipid analogue, galactose(3-sulphate)-beta 1–1′[(N-lissamine rhodaminyl)-12-aminodode-canoyl]-sphingosine, was incorporated into plasma membranes of living spermatozoa and its lateral distribution over the sperm head was studied. The fluorescent lipid was enriched in the apical ridge subdomain of freshly ejaculated sperm cells. After sperm binding to the zona pellucida the lipid redistributed to the equatorial segment of the sperm surface. A similar shift occurred during capacitation in vitro with 2 mM CaCl2 or with 4% (w/v) bovine serum albumin. The desulphated derivative galactose-beta 1–1′[(N-lissamine rhodaminyl)-12-aminododecanoyl]-sphingosine was also incorporated into the plasma membrane of freshly ejaculated sperm cells and clearly stained the apical ridge subdomain and the (pre)-equatorial subdomains of the sperm heads. The desulphogalactolipid analogue showed a slightly faster migration to the equatorial segment of the sperm plasma membrane than did its sulphated counterpart. The measured fluorescence intensity distributions correlated linearly with the spatial probe distribution, which was checked by fluorescence lifetime imaging microscopy. The observed migration of the incorporated glycolipids precedes the acrosome reaction and is one of the underlying molecular events likely to be important in the process of sperm capacitation. The results of this study suggest that lipid phase segregation is an important driving force for the organization of the sperm head plasma membrane into subdomains.

“In vitro” capacitation and subsequent acrosome reaction are related to changes in the expression and location of midpiece actin and mitofusin-2 in boar spermatozoa

2012 ◽  
Vol 77 (5) ◽  
pp. 979-988 ◽  
Author(s):  
L. Ramió-Lluch ◽  
J.M. Fernández-Novell ◽  
A. Peña ◽  
D. Bucci ◽  
T. Rigau ◽  
...  
Keyword(s):  
Acrosome Reaction ◽  
Mitofusin 2 ◽  
Boar Spermatozoa

Lectin histochemistry during in vitro capacitation and acrosome reaction in boar spermatozoa: new lectins for evaluating acrosomal status of boar spermatozoa

1996 ◽  
Vol 98 (1) ◽  
pp. 93-100 ◽  
Author(s):  
Juan María Vázquez ◽  
Emilio Martínez ◽  
Luis Miguel Pastor ◽  
Jordi Roca ◽  
Carmen Matas ◽  
...  
Keyword(s):  
Acrosome Reaction ◽  
Lectin Histochemistry ◽  
Boar Spermatozoa

scholarly journals Effect of acrosome reaction progress in frozen-thawed boar spermatozoa on the efficiency of in vitro oocyte fertilization

2010 ◽  
Vol 55 (No. 9) ◽  
pp. 429-437 ◽  
Author(s):  
S. Martecikova ◽  
P. Hulinska ◽  
Z. Reckova ◽  
A. Pavlik ◽  
M. Jeseta ◽  
...  
Keyword(s):  
Functional Status ◽  
Acrosome Reaction ◽  
Total Efficiency ◽  
Caffeine Concentration ◽  
Embryo Production ◽  
Reaction Progress ◽  
Oocyte Fertilization ◽  
In Vitro Embryo Production ◽  
Boar Spermatozoa

A good functional status of cryopreserved boar spermatozoa is very important for successful fertilization of porcine oocytes and in vitro embryo production. The purpose of the study was to evaluate the changes in functional status of boar spermatozoa separated from frozen-thawed semen and capacitated in vitro by caffeine. The effect of acrosome reaction development in spermatozoa on the efficiency of oocyte fertilization has been studied in boars A, B and C. Motile spermatozoa were separated by Percoll gradient, untreated (control) or treated with both 1mM and 2mM caffeine, and capacitated or co-cultured with matured oocytes. The motility, viability, chromatin and acrosome integrity, and fertilizing ability of spermatozoa were assessed. The separation significantly increased (P < 0.05) the percentage of viable spermatozoa in all tested boars and percentages of motile and acrosome intact spermatozoa in boars B and C. The capacitation significantly decreased (P < 0.05) the percentages of viable and motile spermatozoa, but after capacitation, the motility and viability were significantly higher (P < 0.05) for the caffeine-treated spermatozoa than for the untreated controls. A fall in the proportion of acrosome-intact spermatozoa was different for each caffeine concentration and each boar, but in all boars, acrosome reaction progress was faster and, similarly, monospermy and the total efficiency of fertilization were significantly higher (P < 0.01) for the spermatozoa treated with 1mM caffeine than for those treated with 2mM caffeine. It can be concluded that there is a potential relationship between the acrosome reaction progress in frozen-thawed boar spermatozoa and the efficiency of fertilization of porcine oocytes. A faster AR induced in spermatozoa by appropriate caffeine treatment resulted in a higher monospermy rate and total efficiency of fertilization. Thus, it is important to test sires before their semen is used for in vitro embryo production. The faster AR induced by 1mM caffeine was more effective in terms of monospermy and total efficiency of fertilization.

Sign in / Sign up

Export Citation Format

Share Document