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Production of live foals from sperm-injected oocytes harvested from pregnant mares.
Abstract: In vitro fertilization in horses has been less successful than anticipated owing to: (i) the inability to collect large numbers of good quality oocytes; (ii) alterations in the zona pellucida that occur during in vitro maturation of equine oocytes; and (iii) inadequate preparation of equine sperm cells. In addition, studies in humans, mice and cattle have indicated that high concentrations of glucose in culture media may inhibit embryonic development in vitro and this may also be a problem for development of equine embryos in vitro. The aims of the present study were: (i) to achieve fertilization of equine oocytes by sperm injection; and (ii) to determine whether culture media containing low concentrations of glucose are beneficial for the development of early stage equine IVF-derived embryos. In Expt 1, in vitro matured oocytes obtained from pregnant mares were subjected to intracytoplasmic sperm injection (ICSI), subzonal sperm injection (SUZI) or one of three control treatments. The cleavage rates were greater for oocytes subjected to ICSI (39%) than for oocytes subjected to SUZI (6%) (P < 0.05). The transfer of two embryos into one recipient mare resulted in the presence of an embryonic vesicle in the uterine body at day 14 after ICSI, but it was lost subsequently between days 16 and 18 after ICSI. In Expt 2, oocytes were subjected to ICSI and cultured for 48 h in either TCM-199 or P-1(TM) medium (glucose- and phosphate-free) supplemented with 15% fetal bovine serum. The cleavage rates for embryos cultured in the two culture media were different (47% and 63% in TCM-199 and P-1(TM), respectively; P < 0.10). In addition, four grade 1 embryos were transferred surgically into the oviducts of four recipient mares 48 h after ICSI. Three pregnancies were identified by ultrasonography by the presence of an embryonic vesicle in the uterine body by day 16 after ICSI. Two of these pregnancies proceeded to term, resulting in the birth of two healthy fillies, one at day 319 and the other at day 328 of gestation.
Publication Date: 2000-01-01 PubMed ID: 20681164 The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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This research article explores the use of sperm injection techniques in the fertilization of equine (horse) oocytes (egg cells). It investigates two key objectives: achieving fertilization through sperm injection, and determining the benefits of low-glucose culture media for developing early-stage equine embryos.
Context and Aims
- The study acknowledges challenges in successful in vitro fertilization in horses such as difficulty in gathering good quality oocytes, alterations in the zona pellucida (outer layer of the egg) during the maturation process in vitro, and inadequate preparation of equine sperm cells.
- Similar studies indicate that high glucose concentration in culture media can hinder embryonic development in vitro, an issue that might also affect equine embryos. Therefore, part of the study investigates the benefits of culture media containing low glucose concentrations for developing early stage equine embryos.
- The main objectives were to carry out fertilization through sperm injection and to assess the effect of media with low glucose concentration on embryo development.
Experiment 1
- In the first experiment, matured oocytes from pregnant mares were subjected to intracytoplasmic sperm injection (ICSI) or subzonal sperm injection (SUZI). Three control treatments were also carried out simultaneously.
- The results revealed better cleavage rates with ICSI (39%) compared to SUZI (6%) highlighting the effectiveness of ICSI method.
- However, an embryo produced through ICSI and transferred to a recipient mare was lost between days 16 and 18 post-ICSI, pointing to a potential issue with the early embryonic development in vitro.
Experiment 2
- In the second experiment, oocytes were subjected to ICSI and then cultured for 48 hours in two different media: TCM-199 and P-1(TM) (glucose- and phosphate-free), both supplemented with 15% fetal bovine serum.
- The experiment showed different cleavage rates in both media (47% in TCM-199 and 63% in P-1(TM)), hinting that media without glucose might be more beneficial for embryo cleavage.
- Four grade 1 embryos from this process were surgically transferred into the oviducts of four recipient mares after 48-hours from ICSI. Of these, three mares showed pregnancies by day 16 post-ICSI, verified through ultrasonography.
- Two of these pregnancies proceeded to term, resulting in the birth of two healthy fillies hence illustrating the potential of this method.
Cite This Article
APA
Cochran R, Meintjes M, Reggio B, Hylan D, Carter J, Pinto C, Paccamonti D, Graff KJ, Godke RA.
(2000).
Production of live foals from sperm-injected oocytes harvested from pregnant mares.
J Reprod Fertil Suppl(56), 503-512.
Publication
Researcher Affiliations
- Department of Animal Science, Louisiana State University Agriculture Center, Baton Rouge, LA 70803, USA.
MeSH Terms
- Animals
- Embryo Transfer / veterinary
- Female
- Fertilization in Vitro / veterinary
- Horses / physiology
- Male
- Oocytes / physiology
- Pregnancy
- Sperm Capacitation / physiology
- Sperm Injections, Intracytoplasmic / veterinary
- Spermatozoa / physiology
Citations
This article has been cited 1 times.- Zhou X, Yin M, Jiang W, Jiang M, Li S, Li H, Chen X. Electrical activation of rabbit oocytes increases fertilization and embryo development by intracytoplasmic sperm injection using sperm from deceased male. J Assist Reprod Genet 2013 Dec;30(12):1605-10.
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