Capacitation status of activated bovine sperm cultured in media containing methyl-β-cyclodextrin affects the acrosome reaction and fertility

Zygote ◽  
2010 ◽  
Vol 19 (1) ◽  
pp. 21-30 ◽  
Author(s):  
Yoku Kato ◽  
Sugita Shoei ◽  
Yoshikazu Nagao
Keyword(s):  
Acrosome Reaction ◽  
Reproductive Tract ◽  
Culture Media ◽  
Female Reproductive Tract ◽  
Control Methods ◽  
Fertilization Rates ◽  
Fertility Status ◽  
Mammalian Sperm ◽  
Bovine Sperm ◽  
Pattern Characteristic

SummaryMammalian sperm undergo a series of biochemical transformations in the female reproductive tract that are collectively known as capacitation. One of the key processes involved in capacitation is the activation of sperm motility. Here, we investigated the capacitation and fertility status of activated sperm which had been cultured in media containing methyl-β-cyclodextrin (MBCD). In order to do this, single activated sperm were caught using a micropipette and stained with chlortetracycline (CTC). Firstly, we investigated the effects of preincubation upon motility, capacitation of activated sperm and fertility. Culture in preincubation media supplemented with MBCD increased the rates of activation and fertilization compared with sperm cultured by control methods (p < 0.05). Following capture, individual activated sperm mostly exhibited a pattern characteristic of capacitation.Secondly we examined the effects of culturing sperm in media with or without glucose (G) and pyruvate acid (P) upon activated motility, the capacitation of activated sperm and fertility. Supplementation of culture media with G and P resulted in higher proportions of activated sperm and increased fertilization rates compared to culture without G and P (p < 0.05). Most of the sperm activated by culture in G and P exhibited patterns characteristic of capacitation. Without G and P, individual activated sperm mostly exhibited patterns characteristic of the acrosome reaction (p < 0.05). In conclusion, activated sperm exhibited patterns characteristic of capacitation. In addition, sperm activated in media containing an energy source (glucose and pyruvate acid) appeared to exhibit acrosome reactiveness and fertility.

scholarly journals Role of spermine in mammalian sperm capacitation and acrosome reaction

1991 ◽  
Vol 278 (1) ◽  
pp. 25-28 ◽  
Author(s):  
S Rubinstein ◽  
H Breitbart
Keyword(s):  
Binding Sites ◽  
Acrosome Reaction ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Sperm Capacitation ◽  
Amino Groups ◽  
Binding Properties ◽  
Mammalian Sperm ◽  
Energy Dependent

The binding properties of seminal polyamines to ram spermatozoa and their possible role in sperm capacitation and the acrosome reaction were studied. Binding and release of [14C]spermine from ram spermatozoa occurred at a rate faster than in somatic cells and were not energy-dependent. Release of bound spermine was further facilitated by heparin, a constituent of the female reproductive tract which was reported to induce capacitation and the acrosome reaction. High- and low-affinity polyamine-binding sites were identified, of which the high-affinity site was specific to polyamines with three or more amino groups. We also found that spermine inhibited the acrosome reaction and propose that it is the major seminal decapacitating factor. Since precise timing of capacitation and the acrosome reaction are critical for successful fertilization, it is suggested that the role of seminal spermine is to prevent premature capacitation and the acrosome reaction.

scholarly journals Role of actin cytoskeleton in mammalian sperm capacitation and the acrosome reaction

Reproduction ◽  
2005 ◽  
Vol 129 (3) ◽  
pp. 263-268 ◽  
Author(s):  
Haim Breitbart ◽  
Gili Cohen ◽  
Sara Rubinstein
Keyword(s):  
Plasma Membrane ◽  
Zona Pellucida ◽  
Acrosome Reaction ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Mammalian Sperm ◽  
Acrosomal Membrane ◽  
Close Proximity ◽  
Sperm Plasma Membrane ◽  
To Come

In order to fertilize, the mammalian spermatozoa should reside in the female reproductive tract for several hours, during which they undergo a series of biochemical modifications collectively called capacitation. Only capacitated sperm can undergo the acrosome reaction after binding to the egg zona pellucida, a process which enables sperm to penetrate into the egg and fertilize it. Polymerization of globular (G)-actin to filamentous (F)-actin occurs during capacitation, depending on protein kinase A activation, protein tyrosine phosphorylation, and phospholipase D activation. F-actin formation is important for the translocation of phospholipase C from the cytosol to the sperm plasma membrane during capacitation. Prior to the occurrence of the acrosome reaction, the F-actin should undergo depolymerization, a necessary process which enables the outer acrosomal membrane and the overlying plasma membrane to come into close proximity and fuse. The binding of the capacitated sperm to the zona pellucida induces a fast increase in sperm intracellular calcium, activation of actin severing proteins which break down the actin fibers, and allows the acrosome reaction to take place.

scholarly journals Which Low-Abundance Proteins are Present in the Human Milieu of Gamete/Embryo Maternal Interaction?

2019 ◽  
Vol 20 (21) ◽  
pp. 5305 ◽  
Author(s):  
Canha-Gouveia ◽  
Paradela ◽  
Ramos-Fernández ◽  
Prieto-Sánchez ◽  
Sánchez-Ferrer ◽  
...  
Keyword(s):  
Embryo Development ◽  
Reproductive Tract ◽  
Culture Media ◽  
Adult Life ◽  
Early Embryo ◽  
Female Reproductive Tract ◽  
High Throughput Analysis ◽  
Maternal Interaction ◽  
Oviductal Fluid ◽  
Abundance Proteins

The improvement of the embryo culture media is of high relevance due to its influence on successful implantation rates, pregnancy, neonatal outcomes, and potential effects in adult life. The ideal conditions for embryo development are those naturally occurring in the female reproductive tract, i.e., the oviductal and uterine fluids. To shed light on the differences between chemical and natural media, we performed the first comparative study of the low abundance proteins in plasma, uterine, and oviductal fluid collected, simultaneously, from healthy and fertile women that underwent a salpingectomy. The rationale for this design derives from the fact that high-abundant proteins in these fluids are usually those coming from blood serum and frequently mask the detection of low abundant proteins with a potentially significant role in specific processes related to the embryo–maternal interaction. The proteomic analysis by 1D-nano LC ESI-MSMS detected several proteins in higher amounts in oviductal fluid when compared to uterine and plasma samples (RL3, GSTA1, EZRI, DPYSL3, GARS, HSP90A). Such oviductal fluid proteins could be a target to improve fertilization rates and early embryo development if used in the culture media. In conclusion, this study presents a high-throughput analysis of female reproductive tract fluids and contributes to the knowledge of oviductal and uterine secretome.

301 THE EFFECT OF EQUINE FOLICULLAR FLUID ON IN VITRO INDUCTION OF THE ACROSOME REACTION IN FROZEN - THAWED STALLION SPERMATOZOA

2010 ◽  
Vol 22 (1) ◽  
pp. 307
Author(s):  
D. S. Silva ◽  
P. Rodriguez ◽  
N. S. Arruda ◽  
R. Rodrigues ◽  
J. L. Rodrigues
Keyword(s):  
Contact Area ◽  
Follicular Fluid ◽  
Acrosome Reaction ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Heparin Group ◽  
Fluorescent Stain ◽  
In Vitro Induction

The capacitation process occurs in vivo upon exposure of the spermatozoa through the female reproductive tract, but can be induced in vitro in the presence of several compounds. This study was conducted to assess the effect of heparin or equine follicular fluid on hyperactivated motility and in vitro induction acrosome reaction swim-up method with frozen-thawed stallion semen. Two hundred microliters of frozen-thawed equine semen was placed in a tube (45°C) to increase contact area and incubated at 37°C for 1 h. After incubation 800 μL of the supernatant was collected by centrifugation (500 × g, 10 min) to collect spermatozoa. The resulting pellet was resuspended in capacitation medium Fert-TALP supplemented with 5.0 μg mL-1 heparin or 100% follicular fluid and incubated for different times (1, 2, 3, 4, and 5 h) at 37°C. After incubation the hyperactivated motility and acrosome-reacted spermatozoa were evaluated. Hoechst stain was used to differentiate live and dead spermatozoa, and chlortetracycline (CTC) fluorescent stain was used to assess the capacitation response of sperm; data were analyzed by ANOVA. The effect of equine follicular fluid resulted in improved percentage of spermatozoa with acrosome reaction at all times of incubation (60, 63, 57, 52, and 58%) but immediately after 3 h of incubation, the hyperactivated motility decreased in heparin group and follicular fluid (42 and 30%, respectively).

79 Progesterone-induced acrosome reaction and fertilization rates with bovine intracytoplasmic sperm injection

2021 ◽  
Vol 33 (2) ◽  
pp. 147
Author(s):  
L. Gatenby ◽  
K. R. Bondioli
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Acrosome Reaction ◽  
Fluorescent Microscopy ◽  
Flow Analysis ◽  
Fluorescein Isothiocyanate ◽  
Cumulus Cells ◽  
Male Factor ◽  
Fertilization Rates ◽  
Bovine Sperm ◽  
Marginal Improvement

Intracytoplasmic sperm injection (ICSI) has been a valuable tool in many species because of its ability to overcome male factor infertility problems and eliminate risk of polyspermy; more recently, it been used to improve genome editing technologies. However, limited success with bovine ICSI has hindered these applications in cattle. Numerous treatments have been used to increase the success rate, with marginal improvement. Replicating events synonymous with fertilization, such as the acrosome reaction, may improve fertilization with bovine ICSI. Progesterone (P4) is naturally found in both the cumulus cells surrounding the oocyte and follicular fluid released at ovulation and activates a physiological pathway within sperm to induce an acrosome reaction, a crucial process in fertilization. Progesterone induction of the acrosome reaction as a sperm pretreatment for ICSI has not yet been evaluated in cattle. In this study, frozen–thawed bovine sperm was used. Sperm were first thawed, washed, and separated via gradient to obtain the motile population before capacitation with heparin over 4h before treatment with or without P4 (10 μM) for 15min. To measure the acrosome reacting population, sperm were stained with fluorescein isothiocyanate-conjugated peanut agglutinin (FITC-PNA) and observed over 2h using cytometric flow analysis and fluorescent microscopy to measure the population undergoing and completing an acrosome reaction. For ICSI, a total of 220 oocytes were used, and both treated and nontreated sperm were incubated for 1h before injection to allow for completion of an acrosome reaction. Fertilization rates were measured by observing pronuclear formation and an absence of sperm 18h after ICSI. Means of acrosome reacting population gathered by flow cytometry and microscopy were analysed by ANOVA. Differences in fertilization rates between groups post ICSI were analysed using a Yates’ corrected chi-squared test. A significantly higher proportion of P4 treated versus control sperm initiated an acrosome reaction at hour 1 (80.2%±4.2 vs. 19%±9.1), which increased to (89.3%±3.9 vs. 29%±8.3) after 2h. P4 also increased the percentage of sperm that completed an acrosome reaction, from 50% (±5.1) at hour 1 and 62.5% (±7.4) at hour 2. Only 14.2% (±3.6) completed acrosome reactions in sperm not treated with P4 by hour 2. Motile sperm in both groups did not decrease over the 2-h incubation time period (P&lt;0.05). Progesterone treatment increased the percentage of fertilized embryos after ICSI, with 38.1% fertilized compared with 10.1% with heparin-treated control injections (P&lt;0.001). These results show that P4 has effects on bovine sperm that allow for higher rates of fertilization after ICSI by utilising the sperm’s physiological response to progesterone. Further embryo development using ICSI with P4-treated sperm, or additional physiologically similar treatments, should continue to be assessed.

scholarly journals KSper, a pH-sensitive K+ current that controls sperm membrane potential

2007 ◽  
Vol 104 (18) ◽  
pp. 7688-7692 ◽  
Author(s):  
Betsy Navarro ◽  
Yuriy Kirichok ◽  
David E. Clapham
Keyword(s):  
Membrane Potential ◽  
Acrosome Reaction ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Male Reproductive Tract ◽  
Sperm Flagellum ◽  
K Current ◽  
Sperm Membrane ◽  
Principal Piece ◽  
Mammalian Spermatozoa

Mature mammalian spermatozoa are quiescent in the male reproductive tract. Upon ejaculation and during their transit through the female reproductive tract, they undergo changes that enable them to fertilize the egg. During this process of capacitation, they acquire progressive motility, develop hyperactivated motility, and are readied for the acrosome reaction. All of these processes are regulated by intracellular pH. In the female reproductive tract, the spermatozoan cytoplasm alkalinizes, which in turn activates a Ca2+-selective current (ICatSper) required for hyperactivated motility. Here, we show that alkalinization also has a dramatic effect on membrane potential, producing a rapid hyperpolarization. This hyperpolarization is primarily mediated by a weakly outwardly rectifying K+ current (IKSper) originating from the principal piece of the sperm flagellum. Alkalinization activates the pHi-sensitive IKSper, setting the membrane potential to negative potentials where Ca2+ entry via ICatSper is maximized. IKSper is one of two dominant ion currents of capacitated sperm cells.

scholarly journals Integration of Sperm Motility and Chemotaxis Screening with a Microchannel-Based Device

2010 ◽  
Vol 56 (8) ◽  
pp. 1270-1278 ◽  
Author(s):  
Lan Xie ◽  
Rui Ma ◽  
Chao Han ◽  
Kai Su ◽  
Qiufang Zhang ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Reproductive Tract ◽  
Cumulus Cells ◽  
Female Reproductive Tract ◽  
Straight Channel ◽  
Vitro Fertilization ◽  
Mammalian Sperm ◽  
Reaction Capability ◽  
Mammalian Female

BACKGROUND Sperm screening is an essential step in in vitro fertilization (IVF) procedures. The swim-up method, an assay for sperm motility, is used clinically to select the ideal sperm for subsequent manipulation. However, additional parameters, including acrosome reaction capability, chemotaxis, and thermotaxis, are also important indicators of mammalian sperm health. To monitor both sperm motility and chemotaxis simultaneously during sperm screening, we designed and constructed a microdevice comprising a straight channel connected with a bibranch channel that mimics the mammalian female reproductive tract. METHODS The width and length of the straight channel were optimized to select the motile sperms. We selectively cultured cumulus cells in the bibranch channel to generate a chemoattractant-forming chemical gradient. Sperm chemotaxis was represented by the ratio of the sperm swimming toward different branches. RESULTS The percentage of motile sperms improved from 58.5% (3.8%) to 82.6% (2.9%) by a straight channel 7 mm in length and 1 mm in width. About 10% of sperms were found to be chemotactically responsive in our experiment, which is consistent with previous studies. CONCLUSIONS For the first time, we achieved the combined evaluation of both sperm motility and chemotaxis. The motile and chemotactically responsive sperms can easily be enriched on a lab-on-a-chip device to improve IVF outcome.

scholarly journals Inhibition of mouse fertilization in vivo by intra-oviductal injection of an anti-equatorin monoclonal antibody

Reproduction ◽  
2001 ◽  
pp. 649-655 ◽  
Author(s):  
K Yoshinaga ◽  
DK Saxena ◽  
T Oh-oka ◽  
I Tanii ◽  
K Toshimori
Keyword(s):  
Monoclonal Antibody ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Pathological Changes ◽  
Histological Studies ◽  
Control Side ◽  
Mammalian Sperm ◽  
The Right ◽  
And Control

The monoclonal antibody mMN9 recognizes an antigenic molecule, equatorin, which is localized at the equatorial segment of the mammalian sperm acrosome. Our previous results using an IVF system indicated that mMN9 blocked sperm-oocyte fusion. Antibody-containing and control solutions were injected directly into the right and left oviductal ampullae, respectively, of anaesthetized female mice to assess the effect of mMN9 on fertilization in vivo. After hCG treatment, the females were mated, and their oviductal eggs and implanted embryos were examined. mMN9 was retained in the oviductal lumen at 20 h after injection. The rates of fertilization and concomitant pregnancy were significantly lower than in the control side (P < 0.05). In addition, histological studies showed no evidence of pathological changes in the female reproductive tract after the injections. These results indicate that mMN9 inhibits mouse fertilization significantly under in vivo conditions and that this injection method should be useful for studying the effects of antibodies and agents on fertilization in vivo.

scholarly journals Sperm preparedness and adaptation to osmotic and pH stressors relate to functional competence of sperm in Bos taurus

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Maharajan Lavanya ◽  
Santhanahalli Siddalingappa Archana ◽  
Divakar Swathi ◽  
Laxman Ramya ◽  
Arunachalam Arangasamy ◽  
...  
Keyword(s):  
Reproductive Tract ◽  
Bos Taurus ◽  
Rejection Rate ◽  
Female Reproductive Tract ◽  
Fertilization Process ◽  
Progressive Motility ◽  
Bovine Sperm ◽  
Functional Attributes ◽  
Adaptive Ability ◽  
Head Area

AbstractThe adaptive ability of sperm in the female reproductive tract micromilieu signifies the successful fertilization process. The study aimed to analyze the preparedness of sperm to the prevailing osmotic and pH stressors in the female reproductive tract. Fresh bovine sperm were incubated in 290 (isosmotic-control), 355 (hyperosmotic-uterus and oviduct), and 420 (hyperosmotic-control) mOsm/kg and each with pH of 6.8 (uterus) and 7.4 (oviduct). During incubation, the changes in sperm functional attributes were studied. Sperm kinematics and head area decreased significantly (p < 0.05) immediately upon exposure to hyperosmotic stress at both pH. Proportion of sperm capacitated (%) in 355 mOsm/kg at 1 and 2 h of incubation were significantly (p < 0.05) higher than those in 290 mOsm media. The magnitude and duration of recovery of sperm progressive motility in 355 mOsm with pH 7.4 was correlated with the ejaculate rejection rate (R2 = 0.7). Using this information, the bulls were divided into good (n = 5) and poor (n = 5) osmo-adapters. The osmo-responsive genes such as NFAT5, HSP90AB1, SLC9C1, ADAM1B and GAPDH were upregulated (p < 0.05) in the sperm of good osmo-adapters. The study suggests that sperm are prepared for the osmotic and pH challenges in the female reproductive tract and the osmoadaptive ability is associated with ejaculate quality in bulls.

scholarly journals Effect of flagellar beating pattern on sperm rheotaxis and boundary-dependent navigation

2020 ◽  
Author(s):  
Meisam Zaferani ◽  
Farhad Javi ◽  
Amir Mokhtare ◽  
Alireza Abbaspourrad
Keyword(s):  
Reproductive Tract ◽  
Complex Structure ◽  
Clinical Importance ◽  
Microfluidic System ◽  
Female Reproductive Tract ◽  
Circular Motion ◽  
Sperm Cells ◽  
Progressive Motility ◽  
Mammalian Sperm ◽  
Correct Path

AbstractThe study of navigational mechanisms used by mammalian sperm inside a microenvironment yields better understanding of sperm locomotion during the insemination process, which aids in the design of tools for overcoming infertility. Near- and far-field hydrodynamic interactions with nearby boundaries and rheotaxis are known to be some of the steering strategies that keep sperm on the correct path toward the egg. However, it is not known how the beating patterns of sperm may influence these navigational strategies. In this study, we investigate the effect of flagellar beating pattern on navigation of sperm cells both theoretically and experimentally using a two-step approach. We first isolate bovine sperm based on their rheotactic behavior in a zone with quiescent medium using a microfluidic system. This step ensures that the swimmers are able to navigate upstream and have motilities higher than a selected value, even though they feature various flagellar beating patterns. We then explore the flagellar beating pattern of these isolated sperm and their subsequent influence on boundary-dependent navigation. Our findings indicate that rheotaxis enables sperm to navigate upstream even in the presence of circular motion in their motility, whereas boundary-dependent navigation is more sensitive to the circular motion and selects for progressive motility. This finding may explain the clinical importance of progressive motility in semen samples for fertility, as the flow of mucus may not be sufficiently strong to orient the sperm cells throughout the process of insemination.SignificanceFinding the egg and moving toward it while traversing the complex structure of the female reproductive tract is necessary for mammalian sperm. Previous studies have shown how sperm use navigational steering mechanisms that are based on swimming upstream (i.e. rheotaxis) and along the boundaries of the female reproductive tract. We demonstrate that the performance of theses navigational mechanisms is associated with the primary characteristics of sperm motility. In fact, sperm rheotaxis is more sensitive to the motility and thus average velocity of sperm while navigation via rigid boundaries is more sensitive to the flagellar beating pattern and selects for symmetric beating. Our results can be expanded to other autonomous microswimmers and their subsequent navigation mechanisms.

Sign in / Sign up

Export Citation Format

Share Document