Influence of gonadotropins on ovarian follicle growth and development in vivo and in vitro.

Gonadotropins are the key regulators of ovarian follicles development. They are applied in therapeutic practice in assisted reproductive technology clinics. In the present review we discuss the basic gonadotropic hormones - recombinant human follicle-stimulating hormone, its derivatives, luteinizing hormone and gonadotropin serum of pregnant mares, their origin, and application in ovarian follicle systems in in vitro culture systems.

Luteoprotective role of equine chorionic gonadotropin (eCG) during pregnancy in the mare.

The effects of repeated cloprostenol administration were compared in mares impregnated by horses and mares impregnated by donkeys in order to assess the role of eCG on the development of pregnancy-associated resistance to the luteolytic and abortifacient effects of PGF2α. Eleven mares impregnated by donkey (mule pregnancy) and 9 mares impregnated by horse (horse pregnancy) were used. Six mares with mule pregnancy and four with horse pregnancy were injected with cloprostenol (0.25 mg) when they were between day 65 and day 75 of pregnancy, and the treatment was repeated 48, 72 and 96 h latter. The rest of the mares remained as controls. Concentrations of eCG were 10 times higher (p < 0.001) in mares impregnated by horses than in mares impregnated by donkeys, and they were not affected by cloprostenol treatment. Luteolysis was completed 30 h after the first cloprostenol injection in mule pregnancies, while mares with horse pregnancies required 96 h and three cloprostenol injections to complete luteolysis. Regression analysis revealed significant associations between eCG concentrations at time 0 and the time required for completion of luteolysis (p < 0.001), foetal death (p < 0.01) and foetal expulsion (p < 0.05). It is concluded that high eCG concentrations in mares impregnated by horses protect the corpora lutea of pregnancy against the luteolytic effects of PGF2α. Low eCG concentrations in mares carrying mule foetuses afford them less protection against the luteolytic effect of PGF2α, and this may be a cause of the increased foetal mortality that occurs between days 60 and 90 of pregnancy in these mares.

Follicle development in mares.

The mare provides a unique experimental model for studying follicle development in monovular species. Development of antral follicles in horses is characterized by the periodic growth of follicular waves which often involve the selection of a single dominant follicle. If properly stimulated, the dominant follicle will complete development and eventually ovulate a fertile oocyte. Regulation of follicular wave emergence and follicle selection involves an interplay between circulating gonadotropins and follicular factors that ensures that individual follicles are properly stimulated to grow (or to regress) at any given stage of follicular wave development. Periodic development of follicular waves continuously occurs during most of post-natal life in the mare and is influenced by factors such as stage of oestrous cycle, season, pregnancy, age, breed and individual so that different types of follicular waves (minor or major, ovulatory or anovulatory) and different levels of activity within waves may develop under different physiological conditions. Changes in gonadotropin levels and/or in the sensitivity of follicles to circulating gonadotropins seem to account largely for these physiological variations in follicle development.

Involvement of equine chorionic gonadotropin (eCG) carbohydrate side chains in its bioactivity; lessons from recombinant hormone expressed in insect cells.

Natural eCG consists of as much as 45% carbohydrate side chains. The present paper deals with the analysis of the roles of the N- and O-linked saccharides of this hormone in the different steps of its activity and its possible replacement by recombinant eCG expressed in baculovirus-insect cell systems.

Effects of follicle-stimulating hormone and ovarian steroids during in vitro meiotic maturation on fertilization of rat oocytes.

The present study examined the effects of gonadotropins and ovarian steroids during in vitro meiotic maturation of rat oocytes on their ability to undergo in vitro fertilization. Fully grown oocytes were isolated from antral follicles of immature rats and cultured as oocyte-cumulus cell complexes (OCC) under conditions in which completion of meiotic maturation occurs spontaneously. They were then exposed to spermatozoa under conditions in which oocytes matured in vivo exhibit high fertilization rates. Compared with oocytes from pregnant mare's serum gonadotropin (PMSG) or follicle-stimulating hormone (FSH)-treated rats, a similar proportion of the oocytes (greater than 80%) from untreated rats underwent germinal vesicle breakdown, but such oocytes had a lower rate of fertilization (70% vs. 20%). The presence of FSH during in vitro maturation restored the fertilization rate for oocytes from untreated rats, while a cytochrome P450 inhibitor, aminoglutethimide phosphate abolished this beneficial effect of FSH. The addition of progesterone during the in vitro maturation period duplicated the beneficial effect of FSH on fertilization rate of oocytes from untreated rats; oestradiol-17 beta was less effective in this regard, and 5 alpha-dihydrotestosterone was ineffective. These findings indicate that FSH and progesterone, although having no apparent effect on nuclear maturation of the oocyte, play an important role during oocyte maturation in enabling normal fertilization to occur.

[Endometrial cups in horses]. / De endometriumcups bij het paard.

Endometrial cups in horses are outgrowths appearing in the uterine wall of the pregnant horn between approximately 38 days and 150 of gestation. The cups are structures which vary in shape from oval to irregular and have distinct raised edges, showing an ulcer-like form. The maximum measurements are approximately 5 X 2.5 X 2.5 cm. Microscopic examination shows that they consist of large epithelioid decidual-like cells having large nucleoli. Pregnant Mare Endometrium Gonadotrophin (PMEG) is produced in the endometrial cups. PMEG may be regarded as a precursor of Pregnant Mare Serum Gonadotrophin (PMSG), which can be detected in the blood of mares. The exact function of PMSG is still obscure; some possible functions are suggested in the present paper. After approximately 70 days of gestation, regression starts. The regression of the cups is an immunological process in which the cellular branch of the immunological system plays an important role. Humoral immunity plays a more protective role in the process of regression, probably as a result of the production of 'blocking' antibodies.