Hyperactivation of Stallion Sperm is Required for Successful In Vitro Fertilization of Equine Oocytes.

McPartlin, L.A.; Suárez, Susan S.; Czaya, Catherine A.; Hinrichs, Katrin; Bedford, Sylvia J.

doi:10.1093/biolreprod/78.s1.190b

Cited by 36 publications

(109 citation statements)

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“…Based on our previously published review on the beneficial effect of oviductal cells and secretions on equine IVF [47], we anticipated a beneficial effect of preincubation of jennies oocytes with oviductal fluid. Jennies oocytes were coincubated with donkey sperm treated with procaine, as previously described for equine gametes [32]. Equine oocytes were used as control.…”

Section: Discussionmentioning

confidence: 99%

“…After 27-hour IVM, COCs were partially denuded by flushing and washed in modified Whitten's Medium (MW; 100-mM NaCl, 4.7-mM KCl, 1.2-mM MgCl 2 , 22-mM Hepes, 4.8-mM lactic acid hemicalcium salt, and 1-mM pyruvic acid) supplemented with 5.5-mM glucose (anhydrous), 25-mM NaHCO 3 , and 7-mg/mL BSA, pH 7.25 [32]. Cumulus-oocyte complexes were then preincubated in droplets of 30 ml of porcine oviductal fluid for 30 minutes in an atmosphere of 5% CO 2 in air at 38.5 C in 100% humidity, as previously described [36].…”

Section: In Vitro Fertilization Procedures 251 Preincubation Of Oocmentioning

confidence: 99%

“…Then, 2 mL of semen was diluted in 2 mL of prewarmed MW supplemented with 5.5 mM glucose (anhydrous), pH 7.25 [32]. Diluted sperm was transported to the laboratory within a few minutes at 37 C and centrifuged in 15-mL conical tubes at 100 Â g for 1 minute at 37 C to remove particulate matter and dead sperm.…”

Section: Preparation Of Donkey Jack Sperm and Ivfmentioning

confidence: 99%

“…The supernatant was then transferred to a 14-mL round-bottom tube and centrifuged at 600 Â g for 5 minutes at 37 C. The pellet was resuspended in 1.5-mL of prewarmed MW supplemented with glucose, and the concentration was determined by counting on a Thoma chamber under a microscope (Olympus, IMT-2; Paris, France). Spermatozoa were then diluted at 10 Â 10 6 /mL in prewarmed MW supplemented with 5.5-mM glucose, 25-mM NaHCO 3 , and 7-mg/mL BSA, pH 7.25 (capacitating MW) [32]. Spermatozoa were incubated in 500-ml aliquots in polyvinyl alcohol-coated 5-mL round-bottom tubes at 37 C in a humidified atmosphere for 6 hours.…”

Section: Preparation Of Donkey Jack Sperm and Ivfmentioning

confidence: 99%

“…However, ICSI requires expensive equipment and expertise in micromanipulation. In 2009, a 60% rate of equine IVF was reported after treatment of fresh spermatozoa with procaine [32]. However, procaine was reported to induce cytokinesis in horse oocytes [33].…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Establishment of conditions for ovum pick up and IVM of jennies oocytes toward the setting up of efficient IVF and in vitro embryos culture procedures in donkey (Equus asinus)

et al. 2016

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Section: Discussionmentioning

confidence: 99%

Section: In Vitro Fertilization Procedures 251 Preincubation Of Oocmentioning

confidence: 99%

Section: Preparation Of Donkey Jack Sperm and Ivfmentioning

confidence: 99%

Section: Preparation Of Donkey Jack Sperm and Ivfmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Establishment of conditions for ovum pick up and IVM of jennies oocytes toward the setting up of efficient IVF and in vitro embryos culture procedures in donkey (Equus asinus)

et al. 2016

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The equine oocyte: Factors affecting meiotic and developmental competence

Hinrichs

2010

Molecular Reproduction Devel

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SUMMARYThere is currently much interest in assisted reproduction techniques in the horse, however, many aspects of oocyte maturation, fertilization, and embryo development in the horse differ from those in other species. Because of the close attachment of the equine oocyte to the follicle wall, scraping of the follicle is the most effective method for oocyte recovery. A notable feature of equine oocytes is that those with expanded cumuli (Ex oocytes), which originate from atretic follicles, have higher meiotic competence (ability to mature to metaphase II in vitro) than do oocytes with compact cumuli (Cp oocytes). Cp oocytes originate in viable follicles but are largely juvenile. Recovery and culture of equine oocytes immediately after slaughter yields a higher maturation rate than that obtained from oocytes after ovary storage; this is related to damage to chromatin in Cp oocytes during storage. In contrast, developmental competence (rate of blastocyst development in vitro) is higher in oocytes recovered from the ovary after a delay. The optimum duration of maturation varies based on cumulus morphology and time of recovery from the ovary, but there is no difference in developmental competence between Ex and Cp oocytes. Because standard in vitro fertilization is not repeatable in the horse, oocyte transfer (surgical transfer of oocytes to the oviducts of inseminated mares) has been developed to allow fertilization of isolated oocytes. Fertilization in vitro may be achieved using intracytoplasmic sperm injection; culture of injected oocytes in a medium with high glucose can yield over 30% blastocyst development.Mol. Reprod. Dev. 77: 651-661,

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Relationship between cyclic AMP‐dependent protein tyrosine phosphorylation and extracellular calcium during hyperactivation of boar spermatozoa

Harayama¹,

Noda²,

Ishikawa³

et al. 2012

Molecular Reproduction Devel

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In mammalian spermatozoa, the state of protein tyrosine phosphorylation is modulated by protein tyrosine kinases and protein tyrosine phosphatases that are controlled via cyclic AMP (cAMP)-protein kinase A (PKA) signaling cascades. The aims of this study were to examine the involvement of cAMP-induced protein tyrosine phosphorylation in response to extracellular calcium and to characterize effects of pharmacological modulation of the cAMP-induced protein phosphorylation state and calmodulin activity during hyperactivation in boar spermatozoa. Ejaculated spermatozoa were incubated with cBiMPS (a cell-permeable cAMP analog) and CaCl(2) at 38.5°C to induce hyperactivation, and then used for Western blotting and indirect immunofluorescence of phosphorylated proteins and for the assessment of motility. Both cBiMPS and CaCl(2) were necessary for hyperactivation. The increase in hyperactivated spermatozoa exhibited a dependence on the state of cBiMPS-induced protein tyrosine phosphorylation in the connecting and principal pieces. The addition of calyculin A (an inhibitor for protein phosphatases 1/2A (PP1/PP2A), 50-100 nM) coincidently promoted hyperactivation and cAMP-induced protein tyrosine phosphorylation in the presence of cBiMPS and CaCl(2). Moreover, the addition of W-7 (a calmodulin antagonist, 2-4 µM) enhanced the percentages of hyperactivated spermatozoa after incubation with cBiMPS and CaCl(2), independently of protein tyrosine phosphorylation. These findings indicate that cAMP-induced protein tyrosine phosphorylation in the connecting and principal pieces is involved in hyperactivation in response to extracellular calcium, and that calmodulin may suppress hyperactivation via the signaling cascades that are independent of cAMP-induced protein tyrosine phosphorylation.

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Hyperactivation of Stallion Sperm is Required for Successful In Vitro Fertilization of Equine Oocytes.

Cited by 36 publications

References 0 publications

Establishment of conditions for ovum pick up and IVM of jennies oocytes toward the setting up of efficient IVF and in vitro embryos culture procedures in donkey (Equus asinus)

Establishment of conditions for ovum pick up and IVM of jennies oocytes toward the setting up of efficient IVF and in vitro embryos culture procedures in donkey (Equus asinus)

The equine oocyte: Factors affecting meiotic and developmental competence

Relationship between cyclic AMP‐dependent protein tyrosine phosphorylation and extracellular calcium during hyperactivation of boar spermatozoa

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