Biomedical subjects

G Duchamp

Publications and source records attributed to G Duchamp.

26 records · Page 2Linked to original sources

PubMed1997

Equine oocyte competence for nuclear and cytoplasmic in vitro maturation: effect of follicle size and hormonal environment.

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.

Animals↗

PubMed1997

A comparison of the biochemical composition of equine follicular fluid and serum at four different stages of the follicular cycle.

Samples of blood and follicular fluid were recovered from 27 Welsh Pony mares at 4 distinct stages of follicular development. Eighteen biochemical parameters were measured in each sample, including sodium, potassium, chloride, glucose, urea, creatinine, calcium, inorganic phosphate, total bilirubin, total protein, albumin, magnesium, triglyceride, total cholesterol, nonesterified fatty acids, alkaline phosphatase, gamma-glutamyltransferase and aspartate aminotransferase. The concentrations of progesterone, 17beta oestradiol and testosterone, pH and osmolarity, were also measured in all the follicular fluid samples. The concentrations of all proteins measured were lower in follicular fluid than serum whereas the reverse was true in the case of the lipids. Analysis of variance indicated that serum and follicular fluid concentrations of most of the parameters measured varied in parallel.

Animals↗

PubMed1997

Follicular fluid is not a compulsory carrier of the oocyte at ovulation in the mare.

The aim of this study was to test the possibility that ovulation can occur from a preovulatory follicle emptied of its follicular fluid. Transport of the oocyte into the oviduct and fertilisation in 29% of cases demonstrated that ovulation can occur in the absence of follicular fluid but the higher fertility achieved in control mares (62.5%) suggested that follicular fluid does serve a role during ovulation, fertilisation and oviductal transport. Injection of horse oocytes into preovulatory follicles in mules after removal of the follicular fluid, followed by insemination of the mules with horse semen, resulted in the production of one horse x horse embryo.

Animals↗

PubMed1997

Transfer of immature oocytes to a preovulatory follicle: an alternative to in vitro maturation in the mare?

In the mare, success rates for the in vitro maturation of oocytes are low. Accordingly, we attempted to determine if immature oocytes could be matured in vivo by injecting them into a preovulatory follicle. Groups of 3-9 oocytes collected from donor mares were transferred under ultrasound control into the preovulatory follicle of a recipient mare that was treated with crude equine pituitary gonadotrophin (CEG) to induce ovulation. Just before ovulation (34 h post treatment) the preovulatory follicle of the recipient mare was punctured to collect both the transferred and the indigenous oocytes to analyse the stages of nuclear maturation. The transfer technique did not impair significantly the final maturation of the recipient preovulatory follicle. The indigenous oocytes within the recipient follicles were recognisable by their larger expanded cumulus of yellow colouration due to high hyaluronic acid content; 7/12 of these oocytes were mature (metaphase II). Around half (42/86; 49%) of the oocytes transferred to preovulatory follicles were recovered subsequently. Most of them showed cumulus expansion (41/42, 6 of which were rich in hyaluronic acid), 13 (32%) were mature, 15 (36%) were immature and 13 (32%) were degenerate. When the indigenous oocyte of the recipient mare was mature, 38% of the transferred oocytes were mature, this rate being no different from the in vitro maturation rate of 46%. This study showed that in vivo maturation of immature oocytes by transfer into a preovulatory follicle in a recipient mare is possible. The maturation rate is not different from the in vitro maturation rate. The technique allows the generation of mature oocytes that have an expanded cumulus rich in hyaluronic acid, similar to the situation in preovulatory oocytes. This result has not been obtained in vitro previously.

Animals↗

PubMed1997

Timing of in vivo maturation of equine preovulatory oocytes and competence for in vitro maturation of immature oocytes collected simultaneously.

The objects of this study were to monitor the development of the cumulus complex and nuclear maturation in oocytes recovered from preovulatory follicles following treatment to induce ovulation and to investigate the in vitro maturation competence of oocytes recovered from smaller nonpreovulatory follicles of varying size. All follicles > or =5 mm in pony mares were individually punctured at 0, 6, 12, 24 and 35 h after an injection of LH to induce ovulation. The recovery rates of oocytes were 64% from 55 preovulatory follicles, 22% from 32 subordinate follicles and 52% from 227 small follicles. Cumulus expansion of the preovulatory oocytes occurred at 12 h post LH treatment while the metaphase I and II components of nuclear maturation were not completed until 24 and 35 h post LH respectively. For nonpreovulatory follicles, the frequency of atresia and oocyte competence for in vitro nuclear maturation both increased with increasing follicular size.

Animals↗

PubMed1991

Ovarian response in mares to prolonged treatment with exogenous equine pituitary gonadotrophins.

Twelve anoestrous mares were treated with an intravaginal sponge containing 0.5 g allyl trembolone (Regumate; Roussel UCLAF, Paris) and 50 mg oestradiol benzoate for 7 days, followed by daily intramuscular (i.m.) injections of 25 mg crude equine pituitary extract (CEG), with (n = 6) or without (n = 6) 0.25 mg porcine growth hormone (pGH). No difference in ovarian response to this superovulation treatment was observed between the 2 groups (2.2 +/- 0.4 vs 2.3 +/- 0.4 ovulations per mare, respectively). CEG treatment was then combined with allyl trembolone (40 mg per os per day) and prolonged in the 12 mares, with (n = 6) or without (n = 6) a 7-day interruption of CEG treatment 7 days after the mares first ovulated. Ovarian responses showed that an interruption in gonadotrophin treatment was not necessary to allow the occurrence of a new follicular wave, but the interruption did seem to generate the growth of a more synchronous cohort of follicles. Ovulation rate in the second wave was higher than in the first although there was no effect of treatment interruption on ovulation rate in the second wave, which occurred during progestagen treatment and gave rise to persistent corpora lutea (36.5 +/- 5.9 days). The 6 mares that did not experience an interruption in treatment continued to receive daily CEG injections and oral progestagen. Two periods (Days 0-47 and Days 48-94) were compared for analysis of follicular growth.(ABSTRACT TRUNCATED AT 250 WORDS)

Anestrus↗

PubMed1991

In vitro fertilization in the horse. A retrospective study.

Since the first successful collection of oocytes by non-surgical puncture, there have been numerous attempts to fertilize them but few segmented embryos have resulted. The latest attempts at follicular puncture (Palmer et al., 1987) provided 159 oocytes. Oocytes found broken (18%) were probably already broken, or at least fragile, before puncture. The 41 oocytes were fertilized only with semen treated with Ionophore A23187. Following ionophore treatment of semen, 16 ova segmented (of 113 inseminated oocytes) indicating fertilization, and another 7 showed signs of fertilization but not segmentation. Our basic protocol, when applied to 60 oocytes, yielded 11 (18%) embryos and 5 (8%) that were fertilized without segmentation. Modifications to the protocol produced no improvement in the results. Eight embryos fertilized in vitro were transferred into the ampulla of 8 recipient mares. One pregnancy resulted and foaling occurred June 14, 1990. This is the first equine pregnancy to continue to full term following in vitro fertilization.

Animals↗

PubMed1987

Alternative solutions to hCG induction of ovulation in the mare.

Injection of hCG (2000-2500 i.u., i.v.) to mares when a follicle reaches 35 mm induces ovulation between 24 and 48 h. However, repeated injections induce antibodies against hCG. We report attempts to induce ovulation without this inconvenience. We called 'response' an ovulation between 24 and 48 h after treatment. The typical response to hCG was obtained in 73% (N = 145) of treated mares. After immunization against hCG, the response (0%, N = 10) was less than in nonimmunized controls (100%, N = 9). Simultaneous injection of dexamethasone and hCG resulted in induction of ovulation (71%, N = 14). However, simultaneous hCG + dexamethasone resulted in antibody formation similar to that induced by hCG alone, when injected repeatedly every 21 days. Neither GnRH (2 mg i.m.) nor partly purified pig LH (26 mg i.v. or s.c.) induced ovulation consistently (40%, N = 30 and 31%, N = 16). Crude horse gonadotrophin (60 mg i.v. or s.c.) induced ovulation (86%, N = 14). Fertility was not different from control (61%, N = 13 vs 40%, N = 10). Crude horse gonadotrophin also induced ovulation in mares previously immunized against hCG (78%, N = 9): 50 or 25 mg gave satisfactory response (86%, N = 29 and 57%, N = 40). We conclude that crude horse gonadotrophin is a good alternative to hCG for the induction of ovulation in mares.

Animals↗