<i>In Vitro</i>Production of Equine Embryos: State of the Art

Hinrichs, Katrin

doi:10.1111/j.1439-0531.2010.01624.x

Cited by 55 publications

(47 citation statements)

References 38 publications

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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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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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“…In this latter respect, while it is common practice to culture embryos to the blastocyst stage before intrauterine transfer in most domestic species, in human medicine, premature intrauterine transfer of day 2 and 3 IVP embryos is a routine procedure that yields good results (Younis et al 2009) and circumvents the potential downsides of prolonged in vitro culture or the difficulty of transferring early embryos to the oviduct. Currently, blastocyst rates of 25-35% can be obtained after ICSI and in vitro culture of cleaved horse embryos (Hinrichs 2010). However, only few laboratories are able to obtain such good results.…”

Section: Introductionmentioning

confidence: 99%

Influence of the uterine environment on the development of in vitro-produced equine embryos

et al. 2012

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The necessity for early interaction between the embryo and the oviductal and/or uterine environment in the horse is reflected by several striking differences between equine embryos that develop in vivo and those produced in vitro. Better understanding of the salient interactions may help to improve the efficiency of in vitro equine embryo production. In an initial experiment, cleavage-stage in vitro-produced (IVP) equine embryos were transferred into the uterus of recipient mares that had ovulated recently to determine whether premature placement in this in vivo environment would improve subsequent development. In a second experiment, an important element of the uterine environment was mimicked by adding uterocalin, a major component of the endometrial secretions during early pregnancy, to the culture medium. Intrauterine transfer of cleavage-stage IVP equine embryos yielded neither ultrasonographically detectable pregnancies nor day 7 blastocysts, indicating that the uterus is not a suitable environment for precompact morula stage horse embryos. By contrast, exposure to uterocalin during IVP improved capsule formation, although it did not measurably affect the development or expression of a panel of genes known to differ between in vivo and in vitro embryos. Further studies are required to evaluate whether uterocalin serves purely as a carrier protein or more directly promotes improved capsule development.

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“…In mares, oocytes can be surrounded by both tightly packed and dispersed layers of the corona radiata, but both types of oocytes are competed for maturation (Squires et al, 2003;Betteridge, 2006;Hinrichs, 2010Hinrichs, , 2010a.…”

Section: In Vitro Maturation Of Oocytes (Ivm)mentioning

confidence: 99%

“…All of these requirements are characteristic of the respective farm animal species and have been described in detail in the literature: cattle (Hoshi, 2003;Lonergan, 2007;Lonergan and Fair, 2008), pigs (Abeydeera, 2002;Nagai et al, 2006;Lonergan and Fair, 2008;Gil et al, 2010;Dang-Nguyen et al, 2011), sheep and goats (Cognie et al, 2003;Paramio, 2010), and horses (Squires at el., 2003;Betteridge, 2006Betteridge, , 2007Hinrichs, 2010).…”

Section: Introductionmentioning

confidence: 99%

Obtaining farm animal embryos <i>in vitro</i>

Duszewska¹,

Rapala²,

Trzeciak³

et al. 2012

J. Anim. Feed Sci.

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This paper reviews the basic knowledge about obtaining farm animal embryos in vitro with special focus on differences among species and application of this procedure in the future. In vitro production of farm animal embryos consists of in vitro maturation (IVM) of oocytes, in vitro fertilization (IVF) of matured oocytes, and in vitro culture (IVC) of embryos. Oocytes can be collected from live animals (by laparotomy, laparoscopy, Ovum Pick Up) or from slaughtered ones (by puncture, sectioning). Usually immature oocytes are isolated, and during IVM they reach maturity. Matured oocytes are cultured with sperm (IVF), leading to the formation of zygotes. In the case of fertilization problems (horse, pig), intracytoplasmic sperm injection is used. The zygotes are usually cultured (IVC) to the morula and blastocyst stages. These embryos can be transferred to recipients or frozen/vitrified. Offspring have been obtained after transfer of cattle, sheep, goat, pig and horse embryos. This procedure can be used in animal breeding, biotechnology, medicine, and basic research.

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In VitroProduction of Equine Embryos: State of the Art

Cited by 55 publications

References 38 publications

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

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

Influence of the uterine environment on the development of in vitro-produced equine embryos

Obtaining farm animal embryos <i>in vitro</i>

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