Category Archives: Plasminogen

Prolactin Regulation of Tissue: MATERIALS AND METHODS(4)

Fibrin Overlay and Reverse Fibrin Autography (RFA)

Assay

The fibrin-agar indicator gels were prepared as previously described. The fibrin-agar gel contained 50 mg/ ml plasminogen as zymogen for PA. Fibrinogen (2.4 mg/ ml) and thrombin (0.5 U/ml) were also added to allow the formation of fibrin as the substrate for plasmin. After electrophoresis, gels were laid carefully onto the fibrin-agar gel and were incubated at 37°C in a humid chamber until the lysis zones became visible, indicating the presence of PAs. For detection of PAI-I activity, the samples were analyzed by RFA. In addition to containing plasminogen, fibrinogen, and thrombin, RFA indicator gels contained 0.01 U/ ml urokinase to allow autolysis. The development of an opaque, lysis-resistant zone indicated the presence of PAI-I activity. asthma inhalers

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Prolactin Regulation of Tissue: MATERIALS AND METHODS(3)

MATERIALS AND METHODS(3)

At the end of the incubation, the cells were washed once with 0.5 ml fresh McCoy’s 5a medium and further incubated in fresh serum-free medium for 48 h in the presence or absence of LH (100 ng/ml) and PRL (10-1000 ng/ml) alone or in combination. For extraction of tPA and PAI-I mRNA, 2-3 X 106 viable granulosa cells were precultured for 6 h in poly-D-lysine-coated glass slides (22 X 22 mm) in 1 ml of McCoy’s 5a medium containing 5% calf serum supplemented with the same concentrations of L-glutamine penicillin and streptomycin. After being washed, the cells were further incubated for the designated times in 1 ml fresh serum-free McCoy’s 5a medium in the presence or absence of the hormones. All the cell cultures were performed in a humidified 95% air: 5% CO2 incubator at 37°C. Media were collected at the end of incubation, adjusted to 0.01% Tween 80, and kept at -20°C until assayed for PAI-I activity. buy prednisone

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Prolactin Regulation of Tissue: MATERIALS AND METHODS(2)

T7 RNA polymerase was obtained from Promega-Biotech, and [a-32P]uridine triphosphate (UTP) was from New England Nuclear (Boston, MA). Restriction enzymes were purchased from Boehringer Mannheim (Bromma, Sweden), and collagenase was obtained from Worthington Biochemical Corporation (Freehold, NJ). Ovine FSH (NIH-FSH-S15, FSH activity 20 X NIH FSH-S1 U/mg, LH activity 0.04 X NIH-LH-SI U/mg) was obtained from the National Hormone and Pituitary Distribution Program, NIADDK; ovine PRL (NIAMDD-OPRL-15, 30.5 IU/mg) was obtained from the National Institute of Arthritis, Metabolism and Digestive Diseases); and hCG CR-121 (13 550 IU/mg) was obtained from the National Institute of Child Health and Human Development (NICHHD). buy asthma inhaler

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Prolactin Regulation of Tissue: MATERIALS AND METHODS(1)

MATERIALS AND METHODS(1)

Materials

McCoy’s 5a medium (modified, without serum), peni-cillin-streptomycin solution, L-glutamine, and fetal calf serum were purchased from Gibco (Paisley, Scotland); eCG, LH, poly-D-lysine (M 70 000-150 000), Triton X-100, ni-troblue tetrazolium (NBT), 5-bromo-4-chloro-3-indolyl phosphate (BCIP), trypan blue dye, and thrombin were obtained from Sigma Chemical Company (St. Louis, MO); acrylamide Л^-methylene-bis-acrylamide, SDS, tetra-methylene diamine (TEMED), ammonium persulfate, and Coomassie brilliant blue dye were from Bio-Rad Laboratories (Richmond, CA); nitrocellulose filters were obtained from Schleicher and Schuell GmBH (Dassal, Germany). flovent inhaler

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Prolactin Regulation of Tissue Type Plasminogen Activator and Plasminogen Activator Inhibitor Type-I Gene Expression(2)

Many studies have indicated that gonadotropin-induced ovulation is associated with an increase in ovarian plasminogen activator (PA) levels. Using an in vitro perfused rabbit ovarian system, Hamada et al. showed that the presence of PRL in the perfusion medium inhibited hCG-induced ovulation. Yoshimura et al. further demonstrated that PA activity induced by hCG in the perfused rabbit ovary was significantly inhibited by high doses of PRL in the perfusate.

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Prolactin Regulation of Tissue Type Plasminogen Activator and Plasminogen Activator Inhibitor Type-I Gene Expression(1)

Type-I Gene Expression(1)

Prolactin (PRL), a hormone with partial structural homology to growth hormone, is secreted by the lactotroph cells of the anterior pituitary and is primarily involved in the stimulation of milk production. However, PRL is also necessary for ovarian function. Specific PRL receptors have been found in the ovaries of several mammalian species. An antigonadal or antigonadotropic action of this hormone has been demonstrated in several species. Buy Asthma Inhalers Online

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