The effects of equine somatotropin (eST) on follicular development and circulating plasma hormone profiles in cyclic mares treated during different stages of the estrous cycle.

The effects of exogenous equine somatotropin (eST) administration on ovarian activity and plasma hormone levels were evaluated on horse and pony mares. The objectives of this study were to determine the effects of eST on follicular development and circulating concentrations of leutinizing hormone (LH), estradiol, progesterone, and insulin-like growth factor I (IGF-I) in cyclic horse and pony mares. Sixteen mares received daily injections (i.m.) of eST at a concentration of 25 micrograms/kg body weight on either Days 6 through 12 (Treatment A) or 13 through 19 (Treatment B) postovulation. In addition, contemporary mares were similarly given the carrier vehicle and served as controls (Treatments C and D). Blood samples were collected at 24-hr intervals and ultrasonographic evaluations were performed on the ovaries of each mare at 48-hr intervals beginning on the first day of treatment and ending either on the day of ovulation or 5 d postovulation. Circulating levels of insulin-like growth factor-I (IGF-I) were increased in treated mares by Day 3 post-treatment (P < 0.05). Also, mares in Treatment B exhibited a decrease in plasma estradiol concentrations (P < 0.05) when compared with control mares on Days 1 through 5 postovulation of the post-treated estrous cycle. In addition, circulating leutinizing hormone levels were different for mares in Treatment A compared with controls on Days--8 through--1 pre-ovulation (P < 0.05). All follicles present on the ovaries of each mare were measured and placed into one of five categories based on their diameter. Neither the mean number of follicles per size category > or = 8 mm in diameter nor the mean follicular diameter within each size category differed among treatment and control mares. However, eST treatment significantly increased the number of follicles < or = 7 mm on the ovaries of mares treated early in the estrous cycle when compared with control mares on Days 3 and 7 post-treatment and at the onset of standing estrus.

Effect of feeding and feed deprivation on plasma concentrations of prolactin, insulin, growth hormone, and metabolites in horses.

Two experiments were conducted to determine 1) the prolactin response to different kinds of feedstuffs in stallions and 2) the effects of total feed deprivation on prolactin secretion in mares and its interaction with the prolactin response to feeding. Experiment 1 was performed with stallions as a 6 x 6 Latin square: A) no feed; B) pelleted feed fed to meet 82.5% of the horses' CP requirements; C) pelleted feed at 25% of the amount in B; D) pelleted feed as in B plus water ad libitum; E) cracked corn at the weight in B; and F) chopped alfalfa at the weight in B. The positive prolactin responses (P < .05) to feeding were similar for treatments B through F. The insulin response to feeding was highest (P < .05) in stallions fed water with the pelleted feed. In Exp. 2, 72 h of feed deprivation did not affect (P > .1) daily prolactin secretion. Feeding of a meal on the 3rd d of deprivation increased (P < .05) plasma prolactin, insulin, and glucose concentrations similarly in all mares. There was a positive growth hormone response (P < .1) after feeding in feed-deprived mares but not in fed mares. The prolactin response (P < .001) to thyrotropin-releasing hormone was greater (P = .083) for feed-deprived mares than for controls, whereas the response to sulpiride (P < .001) only tended to differ (P = .16) between groups. We conclude that prolactin secretion may be stimulated by aspects of eating other than the feedstuff itself. Total feed deprivation had little effect on the subsequent prolactin response to a meal or to other known secretagogues.

Prolactin involvement with the increase in seminal volume after sexual stimulation in stallions.

To test the hypothesis that prolactin mediates the increase in seminal volumes induced by sexual stimulation in stallions, semen was collected from six stallions every other day for 26 d. The last eight collection days were treatment days. For each stallion, four treatments were randomly assigned to the first four of the eight treatment collection days, and then repeated in reverse order on the last four collection days; 1) CONTROL: semen collected per normal procedures; 2) Sexually stimulated: stallions were presented to mares in a chute for 10 min before collection; 3) Bromocriptine (dopamine agonist) plus sexual stimulation: stallions were administered bromocriptine 10 min before 10 min of sexual stimulation prior to collection; and 4) Sulpiride: stallions were administered sulpiride (a dopamine antagonist) 25 min before collection. Prolactin concentrations in plasma were increased by sexual stimulation (P < .01) and sulpiride (P < .001) administration and were decreased (P < .01) when bromocriptine was administered before sexual stimulation. Sexual stimulation alone increased (P < .01) volume of gel-free semen relative to control values (102 vs 81 mL), and bromocriptine prevented this response (89 mL; P < .075 relative to sexual stimulation). Sulpiride had no effect (P > .1) on gel-free volume. Volume of gelatinous material, number of mounts, sperm concentration, motility, pH of gel-free semen, number of spermatozoa per ejaculate, and prolactin concentration in gel-free semen were not affected (P > .1) by treatment. Although the effect of bromocriptine indicated that prolactin may mediate the rise in seminal volume due to sexual stimulation, if it does, it must be in conjunction with other hormone(s) or factor(s), because increasing prolactin concentrations alone did not increase seminal volume in these stallions.

Comparison of various preparations of nuclear antigens by hemagglutination inhibition (HAI).

A clinical laboratory carrying out tests for antinuclear antibodies requires an efficient, reliable preparation method to produce a high yield of nuclear antigens at low cost and a very sensitive, specific assay method for antigen activity. Various tissues were employed for preparation of small nuclear ribonucleoprotein (snRNP) and Sm antigens for these purposes. Fresh calf thymus cells and nuclei, commercially available calf and rabbit thymus acetone powders, fresh rat kidney and liver cells were used as sources of antigens prepared similarly by methods published previously. Preparations of antigens from whole calf thymus cell extracts were prepared with and without inhibitors to protease and RNase. snRNP and Sm antigens were assayed at each preparation step by hemagglutination inhibition (HAI). Using HAI it was possible to routinely assay snRNP and Sm at nanogram/ml quantities which was 10(6) fold more sensitive than Ouchterlony immunodiffusion. Results were expressed as relative specific activity as compared with calf thymus nuclear extract prepared by conventional methods. Protease and RNase inhibitors did not significantly increase yields. Thymus was the best source of snRNP and Sm. Fresh calf thymus extract produced a good, stable, reliable quantity of antigens, whereas calf and rabbit thymus acetone powders provided antigen at higher specific activity with less labor but slightly lower yields. Thus, considering the total cost of preparations, commercial sources may be superior to fresh sources in the clinical laboratory setting. These studies also revealed the utility of the sensitive HAI test not only in the clinical laboratory but also for further research endeavors.