In Vitro Fertilization and Development of In Vitro-Matured Oocytes Aspirated from Pregnant Mares1

Li, L.Y.; Meintjes, M.; Graff, K.J.; Paul, J.B.; Denniston, R.S.; Godke, R.A.

doi:10.1093/biolreprod/52.monograph_series1.309

Cited by 46 publications

(24 citation statements)

References 26 publications

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“…This finding is not surprising in light of previous disappointing results on culture of in vivo-recovered equine embryos (Bezard et al, 1989;Ball et al, 1993). Blastocysts have been produced only sporadically from in vitro-derived embryos after co-coculture on Vero cell or oviductal monolayers, or trophoblast-conditioned medium (Li et al, 1995;Dell'Aquila et al, 1997b;Guignot et al, 1998;Choi et al, 1999b). Li et al (2001) reported 30% blastocyst development (10 of 33) when two-cell embryos, resulting from oocytes matured in co-culture, were co-cultured with cumulus cells after ICSI.…”

Section: Discussionmentioning

confidence: 99%

“…This low rate of fertilization appears to be related to the inability of spermatozoa to penetrate the equine zona pellucida in vitro, as fertilization rates are greatly improved by partial removal or by drilling of the zona pellucida (Choi et al, 1994;Li et al, 1995). Only two foals have been produced by conventional IVF, both resulting from insemination of an oocyte matured in vivo (Palmer et al, 1991;Bezard et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

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Developmental competence in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection with fresh or frozen-thawed spermatozoa

Choi¹,

Love²,

Love³

et al. 2002

Reproduction

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This study was undertaken to evaluate the development of equine oocytes in vitro and in vivo after intracytoplasmic sperm injection (ICSI) with either fresh or frozen-thawed spermatozoa, without the use of additional activation treatments. Oocytes were collected from ovaries obtained from an abattoir and oocytes classified as having expanded cumulus cells were matured in M199 with 10% fetal bovine serum and 5 microU FSH ml(-1). After 24-26 h of in vitro maturation, oocytes with a first polar body were selected for manipulation. Fresh ejaculated stallion spermatozoa were used for the experiment after swim-up for 20 min in sperm-Tyrode's albumen lactate pyruvate. Frozen-thawed spermatozoa from the same stallion were treated in a similar way. Spermatozoa were immobilized and injected into the oocytes using a Piezo drill. Presumptive zygotes were cultured in G1.2 medium for 20 or 96 h after the injection was administered, or were transferred to the oviducts of recipient mares and recovered 96 h later. In addition, bovine oocytes with first polar bodies were injected with the two types of stallion spermatozoa and fixed 20 h after injection to examine pronuclear formation. Fertilization rate (pronucleus formation and cleavage) at 20 h after injection of spermatozoa was not significantly different between fresh and frozen-thawed sperm groups in either equine or bovine oocytes. Pronucleus formation after injection of spermatozoa into bovine oocytes was significantly higher than that for equine oocytes (P < 0.05). There were no significant differences in cleavage rate or average number of nuclei at 96 h between equine oocytes injected with fresh or frozen-thawed spermatozoa. However, embryos developed in vivo for 96 h had a significantly higher number of nuclei in both sperm treatments compared with those cultured in vitro. These results indicate that good activation rates may be obtained after injection of either fresh or frozen-thawed equine spermatozoa without additional activation treatment. Injection of frozen-thawed equine spermatozoa results in similar embryo development to that obtained with fresh equine spermatozoa. In vitro culture of equine zygotes in G1.2 medium results in a similar cleavage rate but reduced number of cells compared with in vivo culture within the oviduct. Bovine oocytes may be useful as models for assessing sperm function in horses.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Developmental competence in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection with fresh or frozen-thawed spermatozoa

Choi¹,

Love²,

Love³

et al. 2002

Reproduction

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“…While in vitro capacitation and fertilization are considered to be routine procedures in many species, and despite promising results in equine intracellular sperm injection (ICSI) programs [29][30][31], there are still no reliably successful conventional in vitro fertilization (IVF) protocols for equids [32][33][34]. Most likely, this is because functional stallion sperm capacitation is not induced in vitro using routine capacitation media (i.e., media containing bovine serum albumin [BSA], bicarbonate, and Ca 2þ ) to promote sperm capacitation as described for various other mammalian species, including the mouse [19,20], pig [14], and human [35].…”

Section: Introductionmentioning

confidence: 99%

Oviduct Binding and Elevated Environmental pH Induce Protein Tyrosine Phosphorylation in Stallion Spermatozoa1

Leemans

Gadella

Šoštarić

et al. 2014

Biology of Reproduction

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Sperm-oviduct binding is an essential step in the capacitation process preparing the sperm for fertilization in several mammalian species. In many species, capacitation can be induced in vitro by exposing spermatozoa to bicarbonate, Ca 2+ , and albumin; however, these conditions are insufficient in the horse. We hypothesized that binding to the oviduct epithelium is an essential requirement for the induction of capacitation in stallion spermatozoa. Sperm-oviduct binding was established by coincubating equine oviduct explants for 2 h with stallion spermatozoa (2 3 10 6 spermatozoa/ml), during which it transpired that the highest density (per mm 2 ) of oviduct-bound spermatozoa was achieved under noncapacitating conditions. In subsequent experiments, sperm-oviduct incubations were performed for 6 h under noncapacitating versus capacitating conditions. The oviduct-bound spermatozoa showed a timedependent protein tyrosine phosphorylation response, which was not observed in unbound spermatozoa or spermatozoa incubated in oviduct explant conditioned medium. Both oviductbound and unbound sperm remained motile with intact plasma membrane and acrosome. Since protein tyrosine phosphorylation can be induced in equine spermatozoa by media with high pH, the intracellular pH (pH i ) of oviduct explant cells and bound spermatozoa was monitored fluorometrically after staining with BCECF-AM dye. The epithelial secretory cells contained large, alkaline vesicles. Moreover, oviduct-bound spermatozoa showed a gradual increase in pH i , presumably due to an alkaline local microenvironment created by the secretory epithelial cells, given that unbound spermatozoa did not show pH i changes. Thus, sperm-oviduct interaction appears to facilitate equine sperm capacitation by creating an alkaline local environment that triggers intracellular protein tyrosine phosphorylation in bound sperm.

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“…We also noticed that calcium ionophore decreased sperm motility within 30 min of incubation and this may have impaired the development of embryos produced in calcium ionophore containing medium. Treatment of stallion sperm with concentrations of calcium ionophore over 3 mM has been reported to greatly reduce stallion sperm motility (Li et al, 1995).…”

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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In Vitro Fertilization and Development of In Vitro-Matured Oocytes Aspirated from Pregnant Mares1

Cited by 46 publications

References 26 publications

Developmental competence in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection with fresh or frozen-thawed spermatozoa

Developmental competence in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection with fresh or frozen-thawed spermatozoa

Oviduct Binding and Elevated Environmental pH Induce Protein Tyrosine Phosphorylation in Stallion Spermatozoa1

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

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