Capacitation and acrosome reaction in buffalo bull spermatozoa assessed by chlortetracycline and agglutinin fluorescence assay

Kaul, Gautam; Sharma, G. S.; Singh, Bibhuti N.; Gandhi, K. K.

doi:10.1016/s0093-691x(01)00494-0

Cited by 30 publications

(19 citation statements)

References 21 publications

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“…The cells were fixed in 2% formaldehyde in PBS for 20 minutes at room temperature. To visualize the acrosome, spermatozoa were stained with FITC labeled Pisum sativum agglutinin according to previously described methodology . The state of the acrosome was observed at ×1000 under a fluorescence microscope (Olympus, Center Valley) in 200 cells counted at random, in duplicated slides.…”

Section: Methodsmentioning

confidence: 99%

“…To visualize the acrosome, spermatozoa were stained with FITC labeled Pisum sativum agglutinin according to previously described methodology. 37 The state of the acrosome was observed at ×1000 under a fluorescence microscope (Olympus, Center Valley) in 200 cells counted at random, in duplicated slides. Spermatozoa were considered to have an intact acrosome when the whole sperm head was green, while those exhibiting a green label only in the equatorial region were considered as acrosome-reacted spermatozoa.…”

Section: Induced Acrosome Reactionmentioning

confidence: 99%

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Extracellular vesicles from oviductal isthmus and ampulla stimulate the induced acrosome reaction and signaling events associated with capacitation in bovine spermatozoa

Franchi

Moreno-Irusta

Domïnguez

et al. 2019

J of Cellular Biochemistry

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Cells can communicate with other neighboring or distant cells through the secretion of extracellular vesicles (EV), composed of a lipid bilayer and bearing surface molecules that allow them to recognize target cells. In this way, EV induce signaling via different mechanisms, modulating the physiological state of the recipient cell. EV have been identified in both male and female reproductive fluids, however, the possible role of EV isolated from female reproductive fluids has become an emerging field only recently. It is known that ejaculated mammalian spermatozoa need to undergo physiological preparation in the female reproductive tract to fertilize the egg. EV secreted by different regions of the female tract constitute signals that may have a key role in regulating sperm functions. The aims of the present study were isolating EV from different regions of the bovine oviduct and analyzing their interaction and physiological effects on spermatozoa. Here, we report the characterization of bovine oviductal fluid EV from the isthmus and ampulla region and their effect on the induced acrosome reaction and signaling events associated with sperm capacitation. EV induced an increase in sperm protein tyrosine phosphorylation, while cell survival of cryopreserved bovine spermatozoa was maintained. We also show that EV uptake regulates the sperm calcium levels by inducing an immediate increase in the intracellular calcium concentration and sperm priming, after a pre‐incubation period, of the progesterone‐induced intracellular calcium rise. Our data contribute to understand the role of EV in the communication between the female reproductive tract and the sperm physiology, information that may be used to improve the efficiency of reproductive assisted technologies.

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Section: Methodsmentioning

confidence: 99%

Section: Induced Acrosome Reactionmentioning

confidence: 99%

Extracellular vesicles from oviductal isthmus and ampulla stimulate the induced acrosome reaction and signaling events associated with capacitation in bovine spermatozoa

Franchi

Moreno-Irusta

Domïnguez

et al. 2019

J of Cellular Biochemistry

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“…A high incidence of polyspermic penetration in caffeine-containing IVF medium has been reported (Mao et al, 2005). Calcium ionophore has been successfully used to induce capacitation and AR in buffalo spermatozoa (Kitiyanant et al, 2002;Kaul et al, 2001); however, the effect of this agent on IVF and on the subsequent development of buffalo embryos has not been reported. The present study shows that calcium ionophore used alone or combined with either heparin or caffeine failed to improve cleavage rate and blastocyst development in IVF buffalo embryos compared to other treatments.…”

Section: Discussionmentioning

confidence: 99%

α-Tocopherol and l-ascorbic acid increase the in vitro development of IVM/IVF swamp buffalo (Bubalus bubalis) embryos

Saikhun

Faisaikarm

Zhang

et al. 2008

Animal

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This study was conducted to investigate the effects of capacitating agents added at in vitro fertilization (IVF) and antioxidants supplemented during in vitro culture (IVC) on the development of buffalo embryos. In experiment I, in vitro embryo development of buffalo embryos was compared when the IVF medium was supplemented with heparin, caffeine and calcium ionophore A23187 either alone or in combination. There was no significant difference (P . 0.05) in the cleavage rates of oocytes among the treatment groups but the development rate to the blastocyst stage and the cell numbers of blastocyst in the heparin-treated group were significantly higher (P , 0.05) than that of other treatments. In experiment II, in vitro embryo development of buffalo embryos was compared when IVC medium was supplemented with either a-tocopherol (250 and 500 mM) or L-ascorbic acid (250 and 500 mM). The rate of development to the blastocyst stage of embryos cultured in medium supplemented with 250 mM a-tocopherol (33%, 41/123) and 250 mM L-ascorbic acid (31%, 38/123) was significantly higher (P , 0.05) than that of those cultured in medium alone (19%, 20/108) but not significantly different (P , 0.05) from medium supplemented with either 500 mM a-tocopherol (24%, 30/123) or 500 mM L-ascorbic acid (25%, 33/133). These results suggest that buffalo spermatozoa treated with heparin were suitable for IVF and that a-tocopherol and L-ascorbic acid added during IVC increased the rate of buffalo embryo development.Keywords: buffalo, embryo, development, a-tocopherol, L-ascorbic acid IntroductionSince the birth of the first buffalo calf from an in vitro fertilized (IVF) oocyte (Madan et al., 1991), a number of articles on in vitro embryo production (IVP) have been published. However, success, in terms of production of transferable stage embryos and birth of calves following embryo transfer, has been limited (Gasparrini, 2002;Nandi et al., 2002). Moreover, the efficiency of IVP in buffalo is much lower than that in cattle (Nandi et al., 2002;Gasparrini, 2002). IVP in cattle can be considered to be a mature technology and available for commercialization (van Wagtendonk-de Leeuw, 2006); on the other hand, IVP systems in buffalo are suboptimal and require substantial improvements (Nandi et al., 2002;Gasparrini, 2002). Several factors might affect the IVP in this species, such as inadequate oocyte maturation, an inappropriate timing of insemination and a non-optimal time of gamete co-incubation. Moreover, the important factors that might affect embryo development are the chemical agents used for capacitation of spermatozoa during IVF and the oxidative damage of embryos during in vitro culture (IVC). Several agents have been used to induce capacitation of buffalo spermatozoa. Heparin is structurally similar to glycosaminoglycans, which were shown to induce capacitation and acrosome reaction (AR) in bovine spermatozoa (Parrish et al., 1988) and several mammalian species including buffalo (Chauhan et al., 1997;Kitiyanant et al., 2002). Caffeine stimulate...

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“…The sperm capacitation and acrosome reaction (AR) were assessed by performing chlortetracycline (CTC) staining (Kaul, Sharma, Singh, & Gandhi, ). In brief, 100 µl semen aliquot was diluted with 1.4 ml phosphate‐buffered saline (PBS, pH 7.4) and centrifuged at 200 × g for 5 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Modulation of seminal plasma content in extended semen improves the quality attributes of ram spermatozoa following liquid preservation at 3–5°C

Paul

Balaganur

Kumar

et al. 2018

Reprod Domestic Animals

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Seminal plasma (SP) is known to induce motility and capacitation in spermatozoa curtailing their lifespan when preserved. Hence, this study was conducted to examine the effects of removal of SP from sperm surface prior to liquid preservation either by high dilution (1/15) or by washing and the poststorage treatment with SP (15% and 25%, v/v) on the quality attributes of liquid-preserved ram semen. Over the period of storage, the rapid motility (66.0% and 71.1% vs. 58.3%), straightness (87.1% and 82.1% vs. 79.4%), average path velocity (152.3 and 152.0 µm/s vs. 133.3 µm/s) and the straight-line velocity (131.3 and 127.8 µm/s vs. 108.5 µm/s) were significantly (p < 0.05) higher in both the high-dilution and wash groups as compared to the control (1/3 dilution). The functional membrane integrity (82.3% vs. 77.2%) and noncapacitated sperm count (65.0% vs. 58.7%) were also significantly (p < 0.05) higher in the high-dilution and wash groups, respectively, as compared to the control. The poststorage treatment of sperm with SP significantly (p < 0.05) increased the functional membrane integrity (70.1% vs. 53.8%) and most of the motility attributes as compared to the control (without SP). In conclusion, both the removal of SP prior to liquid preservation and poststorage treatment with SP significantly improved the quality attributes of ram spermatozoa.

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Capacitation and acrosome reaction in buffalo bull spermatozoa assessed by chlortetracycline and agglutinin fluorescence assay

Cited by 30 publications

References 21 publications

Extracellular vesicles from oviductal isthmus and ampulla stimulate the induced acrosome reaction and signaling events associated with capacitation in bovine spermatozoa

Extracellular vesicles from oviductal isthmus and ampulla stimulate the induced acrosome reaction and signaling events associated with capacitation in bovine spermatozoa

α-Tocopherol and l-ascorbic acid increase the in vitro development of IVM/IVF swamp buffalo (Bubalus bubalis) embryos

Modulation of seminal plasma content in extended semen improves the quality attributes of ram spermatozoa following liquid preservation at 3–5°C

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