ABSTRACT
Rat anterior pituitary glands were dissociated with Pronase and the cells were separated by velocity sedimentation at unit gravity. After 30 min of incubation of the enriched gonadotropic cells with LH-RH, there was a significant increase in LH and FSH in the incubation medium. LH-RH (100 ng/ml) and 10(-3) M cAMP both caused significant increases in LH in the incubation medium after 24 hr of incubation.
Subject(s)
Bucladesine/pharmacology , Gonadotropin-Releasing Hormone/pharmacology , Pituitary Gland, Anterior/drug effects , Pituitary Gland/drug effects , Animals , Cell Separation , Follicle Stimulating Hormone/metabolism , In Vitro Techniques , Luteinizing Hormone/metabolism , Male , Pituitary Gland, Anterior/cytology , Pituitary Gland, Anterior/metabolism , Rats , Time FactorsABSTRACT
Highly purified ovine FSH and LH were treated with neuraminidase to remove sialic acid and the desialylated derivatives were examined for biological activity in hypophysectomized immature male and female rats. The male rats were hypophysectomized at 22 days of age and beginning on day 25 were injected sc twice daily for 4 days with native or neuraminidase-treated FSH (total dose, 15 or 60 mug) or LH (12 mug). The ventral prostates, seminal vesicles, and testes were then removed and weighed, and serum testosterone levels were measured by radioimmunoassay. The female rats were hypophysectomized on day 28 and beginning on day 35 were injected sc twice daily for 4 days with native or neuraminidase-treated FSH (8 mug) or saline. On the morning of day 39, the rats were given an ovulating dose of gonadotropin (8 mug native or neuraminidase-treated FSH, or 1.28 mug native or neuraminidase-treated LH) or 1.0 ml saline iv via tail vein. Twenty-four hours later ova were counted in the oviducts the ovaries were weighed, and serum levels of progesterone and 20alpha-dihydroprogesterone were determined by radioimmunoassay. Treatment of ovine LH with neuraminidase did not diminish the ability of this hormone to increase prostate and testes weights and serum testosterone levels. Desialylation also did not decrease the ability of LH to induce ovulation. Although native ovine FSH significantly increased the weights of the ventral prostate, seminal vesicles, and testes, and elevated plasma testosterone levels, the desialylated derivative was essentially inactive. Neuraminidase treatment also eliminated the ability of ovine FSH to increase ovarian weight, to induce ovulation, and to elevate serum progesterone and 20alpha-dihydroprogesterone. These results indicate that the LH-like activity of ovine FSH is an intrinsic property of the FSH molecule.
Subject(s)
Follicle Stimulating Hormone/metabolism , Hypophysectomy , Luteinizing Hormone/metabolism , Neuraminidase/pharmacology , Animals , Female , Follicle Stimulating Hormone/isolation & purification , Luteinizing Hormone/isolation & purification , Male , Organ Size , Ovary/anatomy & histology , Ovulation , Progesterone/blood , Rats , Sheep , Testosterone/bloodSubject(s)
Follicle Stimulating Hormone/isolation & purification , Sheep/metabolism , Amino Acids/analysis , Animals , Biological Assay , Carbohydrates/analysis , Chromatography, Gel , Chromatography, Ion Exchange , Electric Conductivity , Electrophoresis, Disc , Female , Follicle Stimulating Hormone/analysis , Follicle Stimulating Hormone/pharmacology , Glucosamine/analysis , Hypophysectomy , Macromolecular Substances , Male , Mannose/analysis , Molecular Weight , Ovarian Follicle/drug effects , Pituitary Gland/physiology , Prostate/drug effects , Rats , Seminal Vesicles/drug effects , Sex Factors , Testis/drug effects , Ultracentrifugation , UreaSubject(s)
Follicle Stimulating Hormone/isolation & purification , Amino Acids/analysis , Animals , Biological Assay , Carbohydrates/analysis , Chromatography, Gel , Chromatography, Ion Exchange , Electrophoresis, Disc , Female , Follicle Stimulating Hormone/analysis , Follicle Stimulating Hormone/pharmacology , Fucose/analysis , Galactose/analysis , Glucosamine/analysis , Hexosamines/analysis , Horses , Humans , Macromolecular Substances , Mannose/analysis , Molecular Weight , Neuraminic Acids/analysis , Pituitary Gland/analysis , Rats , Species Specificity , UreaSubject(s)
Luteinizing Hormone/isolation & purification , Amino Acids/analysis , Animals , Ascorbic Acid/metabolism , Biological Assay , Carbohydrates/analysis , Chorionic Gonadotropin/analysis , Chromatography, Gel , Chromatography, Ion Exchange , Electrophoresis, Disc , Female , Hexosamines/analysis , Hexoses/analysis , Horses , Humans , Luteinizing Hormone/analysis , Luteinizing Hormone/pharmacology , Macromolecular Substances , Molecular Weight , Ovary/metabolism , Species Specificity , UreaSubject(s)
Cytoplasmic Granules , Pituitary Gland, Anterior/cytology , Pituitary Gland/cytology , Animals , Cell Fractionation/methods , Centrifugation, Density Gradient , Chromatography , Cytoplasmic Granules/ultrastructure , Male , Microscopy, Electron , Rats , Silicon Dioxide , UltracentrifugationSubject(s)
Pituitary Gland/cytology , Animals , Biological Assay , Cell Separation , Follicle Stimulating Hormone/analysis , Glucosamine/metabolism , Luteinizing Hormone/analysis , Male , Methods , Microscopy , Microscopy, Electron , Pituitary Gland/analysis , Pituitary Gland/metabolism , Pronase , Radioimmunoassay , Rats , Tritium , TrypsinSubject(s)
Follicle Stimulating Hormone/analysis , Amino Acids/analysis , Animals , Aspartic Acid/analysis , Cystine/analysis , Fucose/analysis , Glutamates/analysis , Glycoproteins , Hexosamines/analysis , Hexoses/analysis , Horses , Lysine/analysis , Methods , Neuraminic Acids/analysis , Phenylalanine/analysis , Proteins/analysis , Threonine/analysisSubject(s)
Corpus Luteum/physiology , Horses/physiology , Immune Sera , Pituitary Gland/immunology , Adrenal Glands/anatomy & histology , Animals , Antigens , Corpus Luteum/anatomy & histology , Estrus , Female , Follicle Stimulating Hormone , Immunization, Passive , Immunodiffusion , Luteinizing Hormone , Male , Neutralization Tests , Organ Size , Ovarian Follicle/anatomy & histology , Ovary/anatomy & histology , Pituitary Gland/anatomy & histology , Pregnancy , Rats , Sheep/immunology , Thyroid Gland/anatomy & histology , Uterus/anatomy & histologySubject(s)
Amino Acids/analysis , Carbohydrates/analysis , Luteinizing Hormone/analysis , Animals , Carboxypeptidases , Cattle , Chromatography, Gel , Dansyl Compounds , Fucose/analysis , Galactosamine/analysis , Glucosamine/analysis , Hexoses/analysis , Horses , Humans , Hydrazines , Isoleucine/analysis , Leucyl Aminopeptidase , Luteinizing Hormone/isolation & purification , Neuraminic Acids/analysis , Phenylalanine/analysis , Serine/analysis , Sheep , Species SpecificitySubject(s)
Amino Acid Sequence , Carbohydrates/analysis , Follicle Stimulating Hormone/analysis , Animals , Dansyl Compounds , Fucose/analysis , Glutamates/analysis , Glycine/analysis , Glycoproteins/analysis , Hexosamines/analysis , Hexoses/analysis , Hydrazines , Neuraminic Acids/analysis , Serine/analysis , SheepSubject(s)
Luteinizing Hormone/isolation & purification , Pituitary Gland/analysis , Amino Acids/analysis , Animals , Ascorbic Acid , Biological Assay , Carbohydrates/analysis , Chromatography, Gel , Chromatography, Ion Exchange , Electrophoresis, Disc , Female , Follicle Stimulating Hormone/analysis , Hypophysectomy , Immunodiffusion , Isoelectric Focusing , Luteinizing Hormone/pharmacology , Male , Ovary/drug effects , Pituitary Gland/physiology , Prostate/drug effects , Rabbits , Rats , Sheep , UltracentrifugationSubject(s)
Follicle Stimulating Hormone/isolation & purification , Luteinizing Hormone/isolation & purification , Pituitary Gland/analysis , Acetone , Animals , Ascorbic Acid , Biological Assay , Chemical Precipitation , Chromatography, Gel , Chromatography, Ion Exchange , Dialysis , Electrophoresis, Disc , Ethanol , Female , Horses , Hydrogen-Ion Concentration , Immunodiffusion , Immunoelectrophoresis , Isoelectric Focusing , Male , Methods , Molecular Weight , Organ Size , Ovary/drug effects , Phosphorous Acids , Prostate/anatomy & histology , Prostate/drug effects , Rabbits , Rats , Sodium Chloride , UltracentrifugationSubject(s)
Follicle Stimulating Hormone , Pituitary Gland/analysis , Amino Acids/analysis , Animals , Antibodies , Antigens , Biological Assay , Chemical Precipitation , Chromatography , Chromatography, Gel , Chromatography, Ion Exchange , Electrophoresis, Disc , Ethanol , Female , Follicle Stimulating Hormone/analysis , Follicle Stimulating Hormone/isolation & purification , Follicle Stimulating Hormone/pharmacology , Hydrogen-Ion Concentration , Hydroxyapatites , Hypophysectomy , Immunodiffusion , Immunoelectrophoresis , Isoelectric Focusing , Male , Methods , Molecular Weight , Organ Size , Ovary/drug effects , Phosphoric Acids , Pituitary Gland/physiology , Prostate/drug effects , Quaternary Ammonium Compounds , Rats , Seminal Vesicles/drug effects , Sheep , Stimulation, Chemical , Sulfates , Testis/drug effectsSubject(s)
Pituitary Hormones, Anterior/isolation & purification , Adrenocorticotropic Hormone/isolation & purification , Animals , Electrophoresis, Disc , Follicle Stimulating Hormone/isolation & purification , Growth Hormone/isolation & purification , Luteinizing Hormone/isolation & purification , Male , Prolactin/isolation & purification , Rats , Thyrotropin/isolation & purificationABSTRACT
A method is described for the isolation of secretory granules from rat anterior pituitary glands. The method consists of differential and isopycnic gradient centrifugations, followed by filtration of the zones containing granules on Nuclepore filters to remove mitochondria. Highly purified granules were obtained as indicated by electron microscopy. Major parts of the thyrotropin (TSH) and adrenocorticotropin (ACTH) were recovered in a single fraction of granules as were follicle-stimulating (FSH) and luteinizing (LH) hormones. The somatotropin (STH) and prolactin (LTH) were recovered in separate granule fractions. The major parts of the six different hormones were associated with their respective granule fractions as shown by bioassays specific for each of the hormones. The diameters of granules in sections of intact rat pituitary glands and in isolated pellets were measured, and the means and ranges were in close agreement. These results contribute to the identification of the cell types which produce the different pituitary hormones.