Regulation of avidin accumulation by prostaglandins in chick oviduct culture

Regulation of avidin accumulation by prostaglandins (PGs) and their inhibitors was studied in chick oviduct organ culture. Avidin was induced neither by progesterone nor PGF2 alpha in the oviduct of immature chicks. By progesterone and PGs, a high avidin synthesis was induced when the chicks received diethylstilbestrol (DES) for 7 days. Enhanced avidin production was observed by PGF2 alpha, PGE1 and PGE2, whereas PGA2 and PGB2 had a slight inhibitory effect and PGA1 and PGB1 had no effect on avidin production. PGF2 alpha was most effective at a concentration of 10-20 micrograms/ml. The effects of progesterone and PGF2 alpha were not additive. Mefenamic acid, at concentrations of 40 and 60 micrograms/ml, inhibited 50 and 85%, respectively, of the avidin synthesis induced by progesterone, whereas the inhibition of the total protein synthesis was only 20%, and this only by the higher concentration of the drug. Tolfenamic and meclofenamic acid were also inhibitory in the case of progestin-induced avidin synthesis. These studies indicate that the PGs (F2 alpha, E1 and E2) might be involved in the avidin induction in the chick differentiated oviduct. The specific inhibition of the progesterone-dependent avidin synthesis by the PG inhibitors suggests that PGs may be connected with the progesterone action in the oviduct. We propose that the avidin synthesis by the chick oviduct might be considered as a model system for studying PG effects on the synthesis of a specific protein.     

Progesterone-dependent and -independent avidin production in chick oviduct organ culture

Progesterone-dependent and -independent avidin inductions were found in chick oviduct culture. The effects of oestrogen priming and actinomycin D on these inductions were studied. Priming comprised daily diethylstilboestrol (DES) injections in vivo and one day's incubation in vitro with DES before experimental treatment. After 0 or 1 day of DES pretreatment no measurable amount of avidin was found. Avidin was present in progesterone-untreated incubated tissues after 4 days' DES pretreatment. Incubation with progesterone for 24 h increased avidin levels only after 4 or more days of oestrogen priming. No induction of avidin by actinomycin D similar to that found in vivo was observed in vitro. Actinomycin D inhibited both progesterone-dependent and -independent avidin production, but this effect diminished as oestrogen pretreatment was prolonged. Actinomycin D also significantly reduced total oviductal RNA synthesis. It is concluded that oestrogen priming enhances both progesterone-dependent and -independent avidin production in vitro and that both inductions are partially dependent on new RNA synthesis.     

The chick oviduct in tissue culture. I. Initial characterization of growing primary oviduct tissue cultures

A simple three-enzyme treatment of collagenase, dispase and hyaluronidase on finely minced chick oviduct yields clumps of 50-150 cells. These cells attach to collagen-treated dishes and survive in culture for at least 2 weeks without subculturing. Oviduct cell cultures can also be induced to grow. Estradiol or epidermal growth factor (EGF) induce a 40% increase in cells in 4 days when cultures are grown in serum levels that do not support growth. Serum from estrogen-stimulated chicks promotes rapid cellular proliferation (doubling times of 1-2 days). Sera from estrogen withdrawn chicks, laying hen or horse do not support as rapid proliferation. The oviduct growth-promoting factors in serum from estrogen-stimulated chicks are not steroids or fibroblast growth factors (FGF). Removal of steroids from these sera by charcoal treatment or delipidization does not decrease the rate of growth. The addition of 1-100 nM estradiol does not increase a serum's ability to promote growth. Purified FGF or platelet-derived growth factor (PDGF) do not induce oviduct proliferation. These results were reproduced in oviduct cell cultures started from estrogen-stimulated and withdrawn chicks as well as laying hens. Thus the factors in serum from estrogen-stimulated chicks that promote rapid oviduct growth are induced by estrogen treatments in vivo, but do not seem to be only steroids.     

Testosterone binding in the chick oviduct

A novel androgen receptor was observed in estrogen-stimulated chick oviducts but not in unstimulated oviducts. This binding component showed a preference for androgens and could therefore be distinguished from oviduct receptors for estadiol and progesterone. Testosterone was tightly bound having a dissociation constant (Kd) of 2.7 × 10^?10 M. Sucrose gradient centrifugation, under low ionic strength conditions, showed testosterone to be bound as an 85 complex. These binding properties, plus the estrogen dependency of this component, suggest its role as a biological receptor for androgens.     

Estrogen dependent ciliogenesis in the chick oviduct

Both the hormone dependency and the morphological details of estrogen-dependent ciliogenesis in the shell gland of the chick oviduct were investigated. Ciliogenesis was initiated on day 3 of estrogen treatment, and progressively more cells became differentiated until, on day 10, 55% ciliation occurred with 17-estradiol (1 mg/day) and 75% ciliation occurred with diethylstilbestrol (1 mg/day). Simultaneous administration of progesterone with diethylstilbestrol (1 mg each/day for 10 days) caused a 50% depression in the number of ciliated cells on day 10. The rate of ciliogenesis was found to be affected by progesterone and the type of estrogen administered. The minimum stimulatory dose of estradiol was found to be between 0.01 mg/day and 0.05 mg/day. Ciliogenic cells were first recognized by the appearance of pro-basal bodies in the apical portion of the cell. Pro-basal body maturation and cilium formation were the same as those described for the chick trachea. Ciliogenesis in the chick was found to be homologous to estrogen-dependent ciliogenesis in various mammalian oviducts.     

Augmentation of progesterone receptor concentration by progesterone and estrogen treatments in the chick oviduct

Cytosol receptors for progesterone in the chick oviduct were measured by charcoal-adsorbtion assay by using ORG 2058 as a ligand after long-term administration of progesterone and diethylstilbestrol (DES). Steroid administration was carried out by using daily injections or silastic capsules. DES treatment increased the progesterone receptor concentration (from 11500 to 21500 sites per cell, day 14). Progesterone also augmented the concentration of its own receptors (from 11500 to 29000 sites per cell, day 14). In the experiments with capsule administration the same trend was seen. This indicates that both diethylstilbestrol and progesterone are able to increase the concentration of progesterone specific cytosol receptors in the non-differentiated chick oviduct.     

Differential binding of antiestrogens by rat uterine and chick oviduct cytosol

Analysis of the interactions of two synthetic estrogen antagonists, tamoxifen and CI 628, with rat uterine and chick oviduct cytosol revealed significant differences in the antiestrogen binding properties of these tissues. In the rat uterus CI 628, tamoxifen and estradiol were bound to a similar number of saturable binding sites and estradiol could completely inhibit the binding of tritiated antiestrogens to these sites. In contrast, high affinity, saturable antiestrogen binding sites in chick oviduct were present at three times the concentration of estradiol binding sites and estradiol could only partially inhibit the binding of tritiated antiestrogens to these sites. It is concluded that antiestrogens bind to the estrogen receptor in both tissues and that chick oviduct has an additional saturable antiestrogen binding site distinct from the classical estrogen receptor site.     

Estrogen-like effects of combined dexamethasone and tamoxifen in the chick oviduct

The effects of dexamethasone alone on withdrawn chick oviduct weight, DNA, protein content and progesterone receptor concentration were barely detectable, whereas ovalbumin and conalbumin synthesis were increased. When dexamethasone and tamoxifen were combined, a marked increase of total proteins, including egg white proteins, DNA and wet weight occurred. Progesterone receptor also was increased. The most striking result was the stimulation of DNA polymerase-α activity by combined dexamethasone and tamoxifen, whereas either compound was completely ineffective.