Effects of carbohydrates on the pharmacokinetics and biological activity of equine chorionic gonadotropin in vivo.

The sialylation of eCG was examined to determine its influence on the in vivo metabolism and biological activity of the molecule. Sialic acid was decrementally removed from eCG by incubation with agarose-linked neuraminidase for varying time periods. Pharmacokinetic parameters for the disappearance of 4,000 IU (267 micrograms) of three desialylated eCG preparations (20%, 53%, and 80% sialic acid removed) and control eCG were determined in sheep. The clearance rate of eCG increased (p less than 0.05) with each decrement of sialic acid. The removal of 53% sialic acid enhanced the distribution of eCG into the tissues, compared to control and 20% desialylated eCG (p less than 0.05), presumably because of increased lipid solubility and decreased molecular size. Desialylation to 53% did not alter the elimination half-life of eCG. The removal of 80% sialic acid resulted in the disappearance of eCG from the serum within 1 h, whereas control eCG was still present at 120 h. In vivo trials in rats disclosed that the control eCG preparation increased ovulatory rate at doses of 10-100 IU and ovarian weight at doses of 10-300 IU relative to saline-treated rats (p less than 0.01). The 20% desialylated eCG induced superovulatory and ovarian weight responses, but 100-500 IU were required to achieve the same result as that produced by control eCG. The 53% and 80% desialylated eCG preparations induced a mild superovulatory response (p less than 0.01) but no ovarian weight response. It was concluded that sialic acid was significant to the distribution and disappearance of eCG. The effects of carbohydrate removal on biological activity (e.g., superovulation) are primarily a function of clearance rate rather than tissue-specific phenomena.

Equine chorionic gonadotropin.

Cells from the chorionic girdle of the equine trophoblast invade the maternal endometrium at day 36 of gestation and become established as secretory elements known as the endometrial cups. These structures, which persist for 40-60 days, produce a gonadotropin which can be found in circulation until about day 130 of gestation. This glycoprotein has been identified in the horse and the donkey, with the former having received much better characterization. It consists of 2 noncovalently linked peptide chains; an alpha-subunit of 96 amino acids, which is common to that found in other horse glycoprotein hormones. The beta-subunit of 149 amino acids is identical to horse LH beta. Horse CG is the most heavily glycosylated of the known pituitary and placental glycoprotein hormones. The alpha-subunit has two and the beta-subunit one N-linked glycosylation site, and the beta-chain has in excess of four O-linked glycosylation sites. The N-linked glycans have some oligosaccharides that are not found on other glycoprotein hormones. The sialic component of glycosylation confers an exceptionally long half-life on CG compared to other glycoprotein hormones. Horse CG has LH-like activity in horse receptor and in vitro bioassays. In spite of the amino acid homology, it has lower LH activity than does horse LH. Its most intriguing, and as yet unexplained, characteristic is its pronounced FSH and LH activity in species other than the horse. Horse CG binds to FSH receptors of virtually all mammalian species, other than the horse, in which it has been tested and will produce biological effects peculiar to FSH. It has similar and potent interaction with LH receptors. The structural basis of this duality is not known but may be related to the region 90-110 of the beta-chain. Horse CG is believed to be constitutively expressed by the trophoblastic cells until the endometrial cups degenerate. The role of CG in equine gestation is not completely understood. It is believed to act as an LH-like hormone to induce supplementary ovulation and/or luteinization of follicles in the mare. It has not been established whether CG or the accessory corpora lutea are necessary for successful horse pregnancy. They may serve as a redundant system to assure that there is sufficient secretion of the primary corpus luteum to maintain pregnancy until the placenta assumes its role as the principal steroidogenic organ of gestation.

Effects of the dam on equine chorionic gonadotropin concentrations during pregnancy.

Breeding trials were designed to determine the influence of the mare on serum concentrations of equine chorionic gonadotropin (eCG) from Day 39 to Day 104 of gestation. Sires were ranked according to mean eCG concentrations found in the groups of randomly selected mares to which they were mated in 1983. Mares were ranked according to their mean eCG concentrations on Days 55, 71 and 85 of gestation (Day 0 = mating), in 1983 and 1985. In the 1986 breeding season, mares that had pregnancies characterized by high eCG levels were mated to sires previously associated with low eCG concentration pregnancies and low producing mares were mated to sires associated with high eCG concentration pregnancies. The highest eCG concentrations (12.8 micrograms/ml serum) were detected on Day 55 of gestation in mares mated to the low-ranked sire (P less than 0.01), indicating an influence of the mare on serum eCG concentration. A comparison of eCG production by individual mares in 1985, 1986 and 1987 showed that mares retained the same rank (P less than 0.001), regardless of the rank of the sire to which they were mated. It was concluded that the influence of the mare predominates in determining eCG concentration. In 1987, the highest ranked sire was mated to the highest eCG producing mares (ranked 1-20) and the second highest ranked sire was mated to mares ranked 21-40. Two distinct subgroups resulted, suggesting that the selection of sires and mares could be used to maximize the production of eCG.

In vitro binding and utilization of lipoproteins by luteal cells from ferrets treated with dopaminergic drugs during pseudopregnancy.

The effects of administration of dopaminergic drugs in vivo on the binding and utilization of lipoproteins for progesterone synthesis in vitro by ferret luteal cells were investigated. Pimozide, a dopamine antagonist, and bromoergocriptine (CB-154), a dopamine agonist, were administered to pseudopregnant ferrets to alter prolactin (PRL) concentrations daily beginning the day after ovulation. The control group received the vehicle solution only. Corpora lutea taken on Day 13 after ovulation were dissociated and the cells were incubated with canine lipoproteins, cyclic adenosine monophosphate (cAMP), and 5-cholesten-3 beta-25-diol (25-OH-cholesterol). Canine high-density lipoprotein (HDL) and low-density lipoprotein (LDL) stimulated progesterone accumulation by luteal cells from pimozide-treated animals but not from CB-154-treated ferrets. However, when 25-OH-cholesterol, which bypasses the LDL receptor, was provided as the substrate, steroidogenesis was stimulated in all groups. Together these observations suggest that dopaminergic alteration of PRL levels preferentially affects the utilization of lipoproteins. The uptake of canine HDL and LDL by luteal cells was saturable, and a high degree of cross-reactivity was observed. Heparin released surface-bound HDL and LDL, suggesting that HDL was binding to the LDL receptor. The quantity of LDL which could be released from luteal cells by heparin treatment was greater in animals treated with pimozide and decreased by treatment with CB-154, relative to luteal cells from control animals. It was concluded that the chronic administration of pimozide or CB-154 alters serum PRL levels in vivo, and influences the subsequent binding and utilization of lipoproteins by luteal cells in vitro. PRL may increase the number of LDL binding sites in luteal cells, thereby enhancing lipoprotein uptake for progesterone synthesis.