Transvaginal Ultrasound-Guided Oocyte Retrieval from Cyclic and Pregnant Horse and Pony Mares for In Vitro Fertilization1

Meintjes, M.; Bellow,; Paul, J.B.; Broussard, J.R.; Li, L.Y.; Paccamonti, D.; Eilts, B.E.; Godke, R.A.

doi:10.1093/biolreprod/52.monograph_series1.281

Cited by 40 publications

(19 citation statements)

References 23 publications

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“…Several studies were then conducted to further develop this method and investigate the influence of various parameters (different vacuums, single-or double-lumen needles with various diameters) [22,23,35,37]. Recovery rates displayed high variability, ranging from as low as 12% [23,37] to as high as 47% [38]. The technique used in this study was established in our laboratory by Duchamp et al [22], who obtained an average of 0.29 COCs per nonpreovulatory follicle.…”

Section: Discussionmentioning

confidence: 99%

Equine Oocyte Competence for Nuclear and Cytoplasmic in Vitro Maturation: Effect of Follicle Size and Hormonal Environment1

Goudet

Bézard

Duchamp

et al. 1997

Biology of Reproduction

108

102

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Equine oocyte competence after in vitro maturation (IVM) was investigated in terms of the diameter of the follicle of origin and the stage of the estrous cycle, with three criteria of maturation: nuclear stage after DNA Hoechst staining, meiotic spindle morphology after tubulin immunocytochemical staining, and cortical granule localization after lectin labeling. Seven successive in vivo ultrasound-guided follicular punctures were performed on 10 cyclic saddle mares, alternatively at the end of the follicular phase (after induction of ovulation with a gonadotropin injection) and in midluteal phase (with or without a gonadotropin injection). Expanded cumulus-oocyte complexes (COCs) were stained at collection, and compact COCs were stained after in vitro culture. They were observed under a confocal microscope. Successive punctures on one mare provided 0.9 preovulatory COCs and 8 immature COCs per 22 days. Among the preovulatory oocytes, 55% had completed nuclear and cytoplasmic maturation, 86% of which displayed a normal meiotic spindle. Of the 262 oocytes cultured in vitro, 37% completed nuclear maturation. The nuclear and cytoplasmic maturation rate significantly increased with follicle diameter. The IVM rate tended to be higher in follicular phase and tended to increase in luteal phase with the gonadotropin injection. The meiotic spindle morphology was not significantly different between the classes of follicular diameters. This study provided the opportunity to increase the number of characterized oocytes collected per cycle and per mare. This is the first report showing the progressive acquisition of meiotic competence in the equine oocyte during antral follicle growth and is the only description of the equine meiotic spindle.

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Section: Discussionmentioning

confidence: 99%

Equine Oocyte Competence for Nuclear and Cytoplasmic in Vitro Maturation: Effect of Follicle Size and Hormonal Environment1

Goudet

Bézard

Duchamp

et al. 1997

Biology of Reproduction

108

102

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“…These rates are commonly low when immature (i.e., not preovulatory) follicles are aspirated (12–25%; Brück et al, 1992; Cook et al, 1993; Duchamp et al, 1995; Mari et al, 2005). Relatively high oocyte recovery rates from immature equine follicles (45–60%) have been reported with use of aspiration techniques that include flushing of each follicle with medium up to eight times via a double‐lumen aspiration needle with separate in‐ and outflow channels (Meintjes et al, 1995; Bøgh et al, 2002; Colleoni et al, 2007). In contrast, recovery rates of oocytes aspirated from the gonadotropin‐stimulated dominant preovulatory follicle, within 12 hr of ovulation, are typically high (65–85%; Hinrichs et al, 1990; Carnevale and Ginther, 1995; Hinrichs et al, 2000; Carnevale et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

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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“…Therefore, one cannot attribute this failure to poor oocyte maturation. In 1995, two studies on IVM and fertilization of equine oocytes reported very good cleavage rates after zona drilling [58,59]. In both, the oocytes were collected from earlypregnant mares.…”

Section: Discussionmentioning

confidence: 99%

Chorionic Gonadotropin Secretion Is Associated with an Inhibition of Follicular Growth and an Improvement in Oocyte Competence for In Vitro Maturation in the Mare1

Goudet

Leclercq

Bézard

et al. 1998

Biology of Reproduction

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This study reports the follicular growth and oocyte competence for in vitro maturation and fertilization under the influence of circulating eCG. Three to 7 successive ultrasound-guided follicular punctures were performed on 4 pregnant mares from Day 23 until Day 75 of pregnancy and on 5 control mares whose embryonic vesicle was crushed on Day 22. All follicles larger than 5 mm were punctured 24 h after the largest follicle reached 18 mm. Expanded cumulus oocyte complexes (COCs) were stained at recovery to analyze the nuclear stage. Compact COCs were cultured in vitro for 46 h and either stained or processed for in vitro fertilization (IVF) and stained 26 h after IVF. In the control group, no mares showed an increase in eCG levels, whereas all the pregnant mares had concentrations higher than 100 ng/ml from Day 37. The number of follicles flushed during each puncture attempt significantly decreased with time for 3 of 4 pregnant mares. No significant change in this number was observed for the 5 control mares. The maturation rate of the oocytes from follicles 10-14 mm was significantly higher in the pregnant vs. the control group (14 of 17, 82%, vs. 13 of 30, 43%). The difference was not significant for the oocytes from follicles smaller than 9 mm or larger than 15 mm. After IVF, no oocyte was fertilized. The results led us to conclude that eCG is associated with an inhibition of follicular growth and an improvement in oocyte competence for in vitro maturation.

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Transvaginal Ultrasound-Guided Oocyte Retrieval from Cyclic and Pregnant Horse and Pony Mares for In Vitro Fertilization1

Cited by 40 publications

References 23 publications

Equine Oocyte Competence for Nuclear and Cytoplasmic in Vitro Maturation: Effect of Follicle Size and Hormonal Environment1

Equine Oocyte Competence for Nuclear and Cytoplasmic in Vitro Maturation: Effect of Follicle Size and Hormonal Environment1

The equine oocyte: Factors affecting meiotic and developmental competence

Chorionic Gonadotropin Secretion Is Associated with an Inhibition of Follicular Growth and an Improvement in Oocyte Competence for In Vitro Maturation in the Mare1

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