Endocrine profile and OPU-ICSI outcomes in mares: a comparative study.
Abstract: Despite recent improvements in equine ovum pick-up (OPU) combined with intracytoplasmic sperm injection (ICSI), there is still significant inter-individual variability. In this study, serum concentrations of the oocyte-secreted factors growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), as well as the GDF9/BMP15 complex, anti-Müllerian hormone (AMH), progesterone (P4), and free and conjugated estradiol-17β (E2), were determined in 59 mares (3-24 years) at the time of OPU. Mares were classified retrospectively, based on the number of embryos obtained, into low or high outcome groups, using two or three embryos as the threshold values. Results of OPU-ICSI were not affected by the mare's age, body condition score, cycle stage, number of aspirated follicles, number of recovered oocytes, or the recovery rate (P > 0.05 in all cases). Mares with <2 or ≥2 embryos had similar hormone levels (P > 0.05 in all cases), but there was a >90% likelihood of obtaining ≥2 embryos per session when P4 and E2 concentrations were considered simultaneously. Mares with ≥3 embryos/session had lower P4 and higher AMH (P < 0.05 in both cases). GDF9 and BMP15 were detectable in 14 and 12% of mares, respectively, while the GDF9/BMP15 complex was detected in 93% of mares, with no correlation to other factors. In conclusion, these preliminary findings suggest a negative correlation between high P4 levels and OPU-ICSI outcome in horses. Unassigned: We analyzed the relationship between the hormone profiles of mares at the time of egg collection and the outcome of intracytoplasmic sperm injection (ICSI), as ICSI success often varies significantly between individuals. There were no differences in the hormone profiles of mares producing fewer than two or two or more embryos. However, mares with three or more embryos/session showed significant differences in their hormone profiles compared to those producing fewer than three embryos/session. These changes were mainly related to steroid hormones produced by the ovary, such as progesterone and estradiol, and to the concentration of AMH. Our results suggest a possible influence of the hormone profile of the mare on the number of embryos obtained.
Publication Date: 2025-07-04 PubMed ID: 40539920PubMed Central: PMC12231760DOI: 10.1530/RAF-25-0027Google Scholar: Lookup
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- Journal Article
- Comparative Study
Summary
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This research explores the link between certain hormone levels in horses and the effectiveness of intracytoplasmic sperm injection, a method to treat infertility. The study finds that high levels of the hormones GDF9, BMP15, AMH, progesterone and estradiol-17β may hamper successful pregnancy outcomes.
Introduction to the Study
- The study was conducted to explore the inter-individual variability in the success rates of equine ovum pick-up (OPU), a fertility procedure, combined with intracytoplasmic sperm injection (ICSI), a technique where individual sperm are injected directly into an egg.
- The researchers took blood serum measurements from 59 female horses of a variety of ages (3 to 24 years) at the time of the OPU procedure. These measurements included concentrations of several hormones and proteins: the oocyte-secreted factors growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), the complex of GDF9 and BMP15, anti-Müllerian hormone (AMH), progesterone (P4), and free and conjugated estradiol-17β (E2).
Criteria for Categorizing Study Animals
- The horses in the study were divided into low and high outcome groups retrospectively. This grouping was based on the number of embryos obtained in each group after performing OPU-ICSI, with the threshold between the groups being two or three embryos.
Research Findings
- The success rates of the OPU-ICSI procedure wasn’t influenced by the age or body condition score of the horses. Similarly, the number of follicles aspirated, number of oocytes recovered, or the recovery rate did not correlate to the success rates.
- There was no significant difference across the hormone concentrations of horses that had less than two or equal to or more than two embryos. There was, however, a 90% probability of getting two or more embryos in each session when horses had particular concentrations of progesterone (P4) and estradiol-17β (E2) simultaneously.
- Horses yielding three or more embryos per session had lower P4 and higher AMH concentrations. Both these observations were statistically significant.
- GDF9 and BMP15 could be detected only in 14% and 12% of the horses, respectively. Almost all horses (93%) registered a presence of the GDF9/BMP15 complex, but these data appeared unrelated to the other measured factors in the study.
Conclusion of the Study
- The preliminary results of this study suggested that there’s an inverse relationship between higher levels of progesterone and the success rate of the OPU-ICSI procedure for treating horse infertility. Additional research is required to confirm and further explore this correlation.
Cite This Article
APA
Scarlet D, Schuler G, Malama E, Bollwein H, Bocci C, Colleoni S, Lazzari G, Galli C, Kowalewski MP.
(2025).
Endocrine profile and OPU-ICSI outcomes in mares: a comparative study.
Reprod Fertil, 6(3).
https://doi.org/10.1530/RAF-25-0027 Publication
Researcher Affiliations
- Institute of Veterinary Anatomy, Vetsuisse Faculty Zurich, Zurich, Switzerland.
- Clinic of Reproductive Medicine, Vetsuisse Faculty Zurich, Zurich, Switzerland.
- AgroVet-Strickhof, Vetsuisse Faculty, Lindau, Switzerland.
- Veterinary Clinic for Reproductive Medicine and Neonatology, Justus-Liebig-University, Giessen, Germany.
- Clinic of Reproductive Medicine, Vetsuisse Faculty Zurich, Zurich, Switzerland.
- AgroVet-Strickhof, Vetsuisse Faculty, Lindau, Switzerland.
- Clinic of Reproductive Medicine, Vetsuisse Faculty Zurich, Zurich, Switzerland.
- AgroVet-Strickhof, Vetsuisse Faculty, Lindau, Switzerland.
- Avantea, Cremona, Italy.
- Avantea, Cremona, Italy.
- Avantea, Cremona, Italy.
- Avantea, Cremona, Italy.
- Institute of Veterinary Anatomy, Vetsuisse Faculty Zurich, Zurich, Switzerland.
MeSH Terms
- Animals
- Horses / blood
- Horses / physiology
- Sperm Injections, Intracytoplasmic / veterinary
- Female
- Estradiol / blood
- Pregnancy
- Progesterone / blood
- Growth Differentiation Factor 9 / blood
- Anti-Mullerian Hormone / blood
- Oocyte Retrieval / veterinary
- Bone Morphogenetic Protein 15 / blood
- Retrospective Studies
- Treatment Outcome
Conflict of Interest Statement
The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the work reported.
References
This article includes 57 references
- Arıkan FB, Sagsoz N. Effects of obesity on the serum BMP15, GDF9, and kisspeptin concentrations in women of reproductive age.. J Med Biochem 42 392–400.
- Armstrong DG, Mcevoy TG, Baxter G. Effect of dietary energy and protein on bovine follicular dynamics and embryo production in vitro: associations with the ovarian insulin-like growth factor system.. Biol Reprod 64 1624–1632.
- Ball BA, Conley AJ, Maclaughlin DT. Expression of anti-Müllerian hormone (AMH) in equine granulosa-cell tumors and in normal equine ovaries.. Theriogenology 70 968–977.
- Brown KA, Boerboom D, Bouchard N. Human chorionic gonadotropin-dependent regulation of 17beta-hydroxysteroid dehydrogenase type 4 in preovulatory follicles and its potential role in follicular luteinization.. Endocrinology 145 1906–1915.
- Brown KA, Doré M, Lussier JG. Human chorionic gonadotropin-dependent up-regulation of genes responsible for estrogen sulfoconjugation and export in granulosa cells of luteinizing preovulatory follicles.. Endocrinology 147 4222–4233.
- Buratini J, Dellaqua TT, Dal Canto M. The putative roles of FSH and AMH in the regulation of oocyte developmental competence: from fertility prognosis to mechanisms underlying age-related subfertility.. Hum Reprod Update 28 232–254.
- Campos-Chillon F, Farmerie TA, Bouma GJ. Effects of aging on gene expression and mitochondrial DNA in the equine oocyte and follicle cells.. Reprod Fertil Dev 27 925–933.
- Catandi GD, Fresa KJ, Cheng MH. Follicular metabolic alterations are associated with obesity in mares and can be mitigated by dietary supplementation.. Sci Rep 14 7571.
- Choi Y-H, Velez IC, Macías-García B. Effect of clinically-related factors on in vitro blastocyst development after equine ICSI.. Theriogenology 85 1289–1296.
- Claes A, Stout TAE. Success rate in a clinical equine in vitro embryo production program.. Theriogenology 187 215–218.
- Claes A, Ball BA, Scoggin KE. The interrelationship between anti-Müllerian hormone, ovarian follicular populations and age in mares.. Equine Vet J 47 537–541.
- Conley AJ, Scholtz EL, Legacki EL. 5α-dihydroprogesterone concentrations and synthesis in non-pregnant mares.. J Endocrinol 238 25–32.
- Cuervo-Arango J, Claes AN, Stout TA. A retrospective comparison of the efficiency of different assisted reproductive techniques in the horse, emphasizing the impact of maternal age.. Theriogenology 132 36–44.
- Cuervo-Arango J, Claes AN, Stout TAE. Mare and stallion effects on blastocyst production in a commercial equine ovum pick-up-intracytoplasmic sperm injection program.. Reprod Fertil Dev 31 1894–1903.
- Dong J, Albertini DF, Nishimori K. Growth differentiation factor-9 is required during early ovarian folliculogenesis.. Nature 383 531–535.
- El-Fouly MA, Cook B, Nekola M. Role of the ovum in follicular luteinization.. Endocrinology 87 286–293.
- Ferré LB, Alvarez-Gallardo H, Romo S. Transvaginal ultrasound-guided oocyte retrieval in cattle: state-of-the-art and its impact on the in vitro fertilization embryo production outcome.. Reprod Domest Anim 58 363–378.
- Fonte JS, Alonso MA, Junior MPM. Successful equine in vitro embryo production by ICSI – effect of season, mares’ age, breed, and phase of the estrous cycle on embryo production.. Theriogenology 223 47–52.
- Galli C, Colleoni S, Claes A. Overnight shipping of equine oocytes from remote locations to an ART laboratory enables access to the flexibility of ovum pick up-ICSI and embryo cryopreservation technologies.. J Equine Vet Sci 100 82.
- Galli C, Colleoni S, Duchi R. Male factors affecting the success of equine in vitro embryo production by ovum pickup-intracytoplasmic sperm injection in a clinical setting.. J Equine Vet Sci 43 S6–S10.
- Galloway SM, Mcnatty KP, Cambridge LM. Mutations in an oocyte-derived growth factor gene (BMP15) cause increased ovulation rate and infertility in a dosage-sensitive manner.. Nat Genet 25 279–283.
- Gastal EL, Gastal MO, Wiltbank MC. Follicle deviation and intrafollicular and systemic estradiol concentrations in mares.. Biol Reprod 61 31–39.
- Gilchrist RB, Lane M, Thompson JG. Oocyte-secreted factors: regulators of cumulus cell function and oocyte quality.. Hum Reprod Update 14 159–177.
- Ginther OJ, Utt MD, Bergfelt DR. Controlling interrelationships of progesterone/LH and estradiol/LH in mares.. Anim Reprod Sci 95 144–150.
- Henneke DR, Potter GD, Kreider JL. Relationship between condition score, physical measurements and body fat percentage in mares.. Equine Vet J 15 371–372.
- Henneke DR, Potter GD, Kreider JL. Body condition during pregnancy and lactation and reproductive efficiency of mares.. Theriogenology 21 897–909.
- Hoffmann B, Kyrein HJ, Ender ML. An efficient procedure for the determination of progesterone by radioimmunoassay applied to bovine peripheral plasma.. Horm Res 4 302–310.
- Hoffmann B, Höveler R, Hasan SH. Ovarian and pituitary function in dogs after hysterectomy.. J Reprod Fertil 96 837–845.
- Hoffmann B, Gentz F, Failing K. Investigations into the course of progesterone-oestrogen- and eCG-concentrations during normal and impaired pregnancy in the mare.. Reprod Domest Anim 31 717–723.
- Jacob JC, Gastal EL, Gastal MO. Temporal relationships and repeatability of follicle diameters and hormone concentrations within individuals in mares.. Reprod Domest Anim 44 92–99.
- Juengel JL, Hudson NL, Berg M. Effects of active immunization against growth differentiation factor 9 and/or bone morphogenetic protein 15 on ovarian function in cattle.. Reproduction 138 107–114.
- Klein R, Schams D, Failing K. Investigations on the re-establishment of the positive feedback of oestradiol during anoestrus in the bitch.. Reprod Domest Anim 38 13–20.
- Koskinen E, Kuntsi H, Lindeberg H. Predicting ovulation in the mare on the basis of follicular growth and serum oestrone sulphate and progesterone levels.. Zentralbl Veterinarmed A 36 299–304.
- Lazzari G, Wrenzycki C, Herrmann D. Cellular and molecular deviations in bovine in vitro-produced embryos are related to the large offspring syndrome.. Biol Reprod 67 767–775.
- Lazzari G, Colleoni S, Crotti G. Laboratory production of equine embryos.. J Equine Vet Sci 89 103097.
- Legacki EL, Scholtz EL, Ball BA. Concentrations of sulphated estrone, estradiol and dehydroepiandrosterone measured by mass spectrometry in pregnant mares.. Equine Vet J 51 802–808.
- Li Y, Li RQ, Ou SB. Increased GDF9 and BMP15 mRNA levels in cumulus granulosa cells correlate with oocyte maturation, fertilization, and embryo quality in humans.. Reprod Biol Endocrinol 12 81.
- Makawiti DW, Allen WE, Kilpatrick MJ. Changes in oestrone sulphate concentrations in peripheral plasma of Pony mares associated with follicular growth, ovulation and early pregnancy.. J Reprod Fertil 68 481–487.
- Metcalfa E, Masterson K, Battaglia D. Comparison of maturation, cleavage and blastocyst rates in equine oocytes recovered by transvaginal aspiration from estrogen-subordinate follicles versus non-estrogen-subordinate follicles.. Clin Theriogenology 9 449.
- Mottershead DG, Sugimura S, Al-Musawi SL. Cumulin, an oocyte-secreted heterodimer of the transforming growth factor-β family, is a potent activator of granulosa cells and improves oocyte quality.. J Biol Chem 290 24007–24020.
- Otsuka F, Mctavish KJ, Shimasaki S. Integral role of GDF-9 and BMP-15 in ovarian function.. Mol Reprod Dev 78 9–21.
- Palmer E, Terqui M. The measurement of total plasma oestrogens during the follicular phase of the mare’s oestrous cycle.. Theriogenology 7 331–338.
- Papas M, Govaere J, Peere S. Anti-müllerian hormone and OPU-ICSI outcome in the mare.. Animals 11 2004.
- Peng J, Li Q, Wigglesworth K. Growth differentiation factor 9: bone morphogenetic protein 15 heterodimers are potent regulators of ovarian functions.. Proc Natl Acad Sci U S A 110 E776–E785.
- Rico C, Fabre S, Médigue C. Anti-müllerian hormone is an endocrine marker of ovarian gonadotropin-responsive follicles and can help to predict superovulatory responses in the cow.. Biol Reprod 80 50–59.
- Riepsamen AH, Chan K, Lien S. Serum concentrations of oocyte-secreted factors BMP15 and GDF9 during IVF and in women with reproductive pathologies.. Endocrinology 160 2298–2313.
- Riepsamen AH, Donoghoe MW, Baerwald A. Exploratory analysis of serum concentrations of oocyte biomarkers growth differentiation factor 9 and bone morphogenetic protein 15 in ovulatory women across the menstrual cycle.. Fertil Steril 116 546–557.
- Riepsamen AH, Donoghoe MW, Indran IR. Serum GDF9 and BMP15 as potential markers of ovarian function in women with and without polycystic ovary syndrome.. Clin Endocrinol 98 567–577.
- Ruiz LL, Alvarez N, Nuñez I. Effect of body condition on the developmental competence of IVM/IVF bovine oocytes.. Theriogenology 45 292.
- Samie KA, Kowalewski MP, Schuler G. Roles of GDF9 and BMP15 in equine follicular development: in vivo content and in vitro effects of IGF1 and cortisol on granulosa cells.. BMC Vet Res 21 292.
- Sanfins A, Rodrigues P, Albertini DF. GDF-9 and BMP-15 direct the follicle symphony.. J Assist Reprod Genet 35 1741–1750.
- Scarlet D, Ille N, Ertl R. Glucocorticoid metabolism in equine follicles and oocytes.. Domest Anim Endocrinol 59 11–22.
- Scarlet D, Wulf M, Kuhl J. Anti-Müllerian hormone profiling in prepubertal horses and its relationship with gonadal function.. Theriogenology 117 72–77.
- Shamsa A, Gilchrist RB, Robertson DM. Oocyte-secreted serum biomarkers GDF9 and BMP15 in women with endometriosis.. Reprod Sci 30 1521–1527.
- Townson DH, Pierson RA, Ginther OJ. Characterization of plasma progesterone concentrations for two distinct luteal morphologies in mares.. Theriogenology 32 197–204.
- Vernunft A, Schwerhoff M, Viergutz T. Anti-Muellerian hormone levels in plasma of Holstein-Friesian heifers as a predictive parameter for ovum pick-up and embryo production outcomes.. J Reprod Dev 61 74–79.
- Walbornn SR, Felix M, Schnobrich MR. Effect of day of estrus cycle at time of transvaginal follicle aspiration for oocyte recovery on rates of in vitro maturation and blastocyst production after intracytoplasmic sperm injection.. J Am Vet Med Assoc 260 1683–1689.
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