Validation of the sperm mobility assay in boars and stallions.

The sperm mobility assay used in the present study measures the rate of sperm penetration in a biologically inert cell-separation solution (Accudenz). When a sample of sperm is overlaid in a cuvette containing Accudenz, sperm penetrate the solution and absorbance of the sample can be measured with a spectrophotometer. This assay has been successfully used to select chicken and turkey semen donors. We validated this assay for semen from boars and stallions. Absorbance was measured after overlaying fresh semen from each species in prefilled cuvettes for 1, 5, 10, 15, 20, and 40 min. There were no significant differences when sperm were incubated in prewarmed cuvettes at 37, 39, or 41 degrees C. However, a minimum concentration of 5x10(7) viable sperm/mL was needed to evaluate the rate of sperm penetration in boars. Absorbance was half-maximal at 5.4 and 14.1 min for boar and stallion sperm, respectively. Frequency analysis suggested a normal distribution of mobility values for boar sperm. There were positive correlations between mobility values and several computer-aided sperm analysis (CASA) parameters. In addition, there was medium repeatability for multiple ejaculates from single males. We concluded that the mobility assay can be used for mammalian sperm and there seemed to be phenotypic variation among boars in mobility estimates.

Episodic gonadotropin secretion in the mature fowl: serial blood sampling from unrestrained male broiler breeders (Gallus domesticus).

Forty-week-old male broiler breeders were used in two experiments. Males were reared as recommended by the breeder, housed in individual cages, and cannulated to facilitate blood sampling. In experiment 1, blood samples were collected at 10- min intervals for 4 h commencing the day of cannulation (Day 0) and for 12 h on each of Days 1 and 2. In experiment 2, blood samples were collected at 10-min intervals for 8 h on Day 1. After centrifugation, plasma was stored at -20 degrees C until LH, FSH (experiment 1 and 2), testosterone, and corticosterone (experiment 1) concentrations were determined by RIA. Different statistical methods used to identify hormone secretion profiles revealed a characteristic pulsatile pattern of LH and FSH in plasma. However, LH pulses were more frequent and had greater amplitude than FSH pulses. Less than 32% of the FSH pulses were associated with LH episodes. Conversely, the association between LH and testosterone pulses averaged 83% in birds with testis weight greater than 10 g. Concentrations of corticosterone tended to increase after cannulation and remained elevated for only 3-4 h. Our data indicate that LH, FSH, and testosterone secretion is pulsatile in male broiler breeders. Additionally, LH pulses are associated with testosterone episodes but not with FSH pulses. The pulsatile pattern of FSH secretion, which is unique from those of LH, in adult males suggests that FSH secretion is independently regulated in the adult male fowl.

Molecular cloning and functional characterization of a vasotocin receptor subtype expressed in the pituitary gland of the domestic chicken (Gallus domesticus): avian homolog of the mammalian V1b-vasopressin receptor.

The neurohypophysial hormone arginine vasotocin (AVT) stimulates adrenocorticotropin hormone (ACTH) secretion from the avian anterior pituitary gland resulting in increased adrenal secretion of corticosterone in response to stress. Here, we report molecular cloning and functional characterization of a gene encoding an AVT receptor subtype, designated the VT2 receptor, that may mediate the stimulatory effect of AVT on ACTH secretion in birds. The open reading frame predicts a 425 amino acid polypeptide that includes seven segments of 19 to 24 hydrophobic amino acids, typical of guanine nucleotide-protein coupled receptors. Phylogenetic analysis revealed that the VT2 receptor shares highest identity with the mammalian V1b-vasopressin receptor subtype. Expressed VT2 receptors in COS7 cells mediate AVT-induced phosphatidylinositol turnover and Ca(2+) mobilization. In the domestic chicken, expression of VT2 receptor gene transcripts is limited to the pituitary gland. Based on similarities in sequence, site of expression and coupled signal transduction pathways, we conclude that the VT2 receptor is the avian homolog of the mammalian V1b-vasopressin receptor, and therefore may play an important role in the avian stress response.

Changes in plasma concentrations of luteinizing hormone, progesterone, and estradiol-17beta in peripubertal turkey hens under constant or diurnal lighting.

Possible circadian fluctuations and long-term changes in concentrations of reproductive hormones in peripubertal female birds is poorly documented in comparison with mammalian species. Our objective was to document changes in concentrations of several reproductive hormones the several days before and after initial pubertal preovulatory surges of LH in turkey hens photostimulated with either constant (24L:0D) or diurnal (14L:10D) lighting. The hens were cannulated for hourly blood sampling, starting 10 days after photostimulation and continuing until all hens had laid at least two eggs. First eggs were oviposited between 16 and 24 days after photostimulation, and egg production ranged from two to nine eggs/hen during the experimental period. With both lighting treatments, concentrations of LH declined slightly, concentrations of progesterone (P(4)) increased, and concentrations of estradiol-17beta (E(2)) were constant the 3-4 days prior to initial LH surges with no circadian fluctuations in hormone concentrations. Most (10 of 13) initial preovulatory surges of LH were coupled with ovulations, and all LH surges were coupled with P(4) surges. Those LH and P(4) surges not coupled with ovulations (blind surges) occurred with both lighting treatments, but the incidence of blind surges was higher with diurnal lighting. The interval between LH and P(4) surges was longer between the first and second surges than between subsequent surges, when the interval was approximately 26 h. The duration of LH surges (7.4 +/- 3.0 h) was shorter than that of P(4) surges (10.0 +/- 2.0 h). We conclude that, in the peripubertal female turkey, 1) prior to puberty (first LH-P(4) surges), there are no circadian fluctuations in concentrations of LH, P(4), and E(2), 2) 3 days prior to initial LH surges, E(2) concentrations are stable, LH concentrations decline slightly, and P(4) concentrations increase, and 3) surges of LH are coupled to surges of P(4) but LH-P(4) surges are not always coupled to ovipositions (blind surges), possibly because of internal ovulations.