scholarly journals ISOLATION AND BIOCHEMICAL STUDY OF SECRETORY GRANULES FROM RAT PITUITARY GLANDS

1962 ◽  
Vol 15 (2) ◽  
pp. 159-172 ◽  
Author(s):  
J. F. Perdue ◽  
W. H. McShan
Keyword(s):  
Alkaline Protease ◽  
Density Gradient Centrifugation ◽  
Biochemical Study ◽  
Secretory Granules ◽  
Gradient Centrifugation ◽  
Succinic Dehydrogenase ◽  
Acid Protease ◽  
Alkaline Phosphatases ◽  
Rat Pituitary ◽  
Pituitary Glands

Secretory granules from anterior pituitary glands of young adult male castrate rats were isolated by differential centrifugation, microfiltration, and discontinuous density gradient centrifugation. The granules were obtained as pellets, sectioned, and studied with the electron microscope. A major part of the gonadotropin and a substantial amount of the TSH were associated with the isolated granules. Negligible amounts of growth hormone and prolactin were present as contaminants. Succinic dehydrogenase, glucose-6-phosphatase, acid protease, and acid and alkaline phosphatases were not found in the granules. Alkaline protease was the only enzyme found to be associated with the granules, and it is suggested, in the light of these results, that the alkaline protease may be involved in the release of the hormones.

Pituitary Follicular Cells: Their Origin, Possible Function and Fate

1974 ◽  
Vol 32 ◽  
pp. 34-35
Author(s):  
E. Horvath ◽  
K. Kovacs ◽  
G. Penz ◽  
C. Ezrin
Keyword(s):  
Fine Structure ◽  
Secretory Granules ◽  
Follicular Cells ◽  
Human Pituitary ◽  
Rat Pituitary ◽  
Pituitary Glands ◽  
Junctional Complexes

Follicular structures, in the rat pituitary, composed of cells joined by junctional complexes and possessing few organelles and few, if any, secretory granules, were first described by Farquhar in 1957. Cells of the same description have since been observed in several species including man. The importance of these cells, however, remains obscure. While studying human pituitary glands, we have observed wide variations in the fine structure of follicular cells which may lead to a better understanding of their morphogenesis and significance.

Subcellular localization of oxytocin in the ovine corpus luteum

1985 ◽  
Vol 63 (4) ◽  
pp. 309-314 ◽  
Author(s):  
G. E. Rice ◽  
G. D. Thorburn
Keyword(s):  
Subcellular Localization ◽  
Physicochemical Properties ◽  
Corpus Luteum ◽  
Density Gradient Centrifugation ◽  
Secretory Granules ◽  
Gradient Centrifugation ◽  
Particulate Fraction ◽  
Dense Granules ◽  
Oxytocin Release ◽  
Midluteal Phase

The subcellular localization of oxytocin within the corpus luteum of sheep was investigated using differential and density gradient centrifugation. Oxytocin was associated with a particulate fraction which sedimented to a density of 1.054 – 1.061 g/mL. The exclusion of [3H]oxytocin from this particulate fraction is indicative that particulate oxytocin represents endogenous compartmentalization. Particulate oxytocin, incubated in buffered medium at 37 °C, was stable for up to 1 h and the release of oxytocin was not affected by the pH of the incubation medium, over the range 5.5 – 8.5. Oxytocin release, however, was stimulated by incubating particle-bound oxytocin in buffered medium of low osmolality (<200 mosmol). These data are similar to the physicochemical properties reported for peptide-containing neurohypophysial secretory granules. Ultrastructural analysis of oxytocin-containing fractions revealed the presence of electron-dense granules (diameter, 200–250 nm). These data are suggestive that oxytocin, in the corpus luteum of sheep, is contained within a population of secretory granules which occur in high numbers during the midluteal phase of the oestrous cycle.

scholarly journals Cell Fractionation of Anterior Pituitary Glands from Beef and Pig

1959 ◽  
Vol 5 (1) ◽  
pp. 17-23 ◽  
Author(s):  
Frank S. Labella ◽  
J. H. U. Brown
Keyword(s):  
Alkaline Phosphatase ◽  
Anterior Pituitary ◽  
Secretory Granules ◽  
Succinic Dehydrogenase ◽  
Subcellular Fractions ◽  
Acid Proteinase ◽  
Cell Fractionation ◽  
Ph Optimum ◽  
Pituitary Glands ◽  
The Individual

Fresh anterior pituitary glands from beef and pig were separated by differential centrifugation into subcellular fractions. Nuclei and debris were obtained at 700 g for 15 minutes, secretory granules at 7000 g for 20 minutes, mitochondria at 34,000 g for 15 minutes, and microsomes at 78,000 g for 3 hours. Electron micrographs were taken of the individual fractions. Each fraction was analyzed for nitrogen, pentosenucleic acid (PNA), and phospholipide. Beef and pig anterior lobes were quite similar in their intracellular composition as seen in the subcellular fractions. Succinic dehydrogenase was localized in mitochondria, while alkaline phosphatase was concentrated in the microsomes. A proteinase with pH optimum at 8.2 was exclusively localized. in microsomal and supernatant fractions. Acid phosphatase, acid ribonuclease, and acid proteinase were distributed among the subcellular fractions in another pattern, indicating the presence of a particle type distinct from mitochondria and microsomes. The distribution of cytoplasmic PNA paralleled that of alkaline phosphatase.

scholarly journals ISOLATION AND BIOLOGICAL PROPERTIES OF SECRETORY GRANULES FROM RAT ANTERIOR PITUITARY GLANDS

1969 ◽  
Vol 43 (3) ◽  
pp. 564-574 ◽  
Author(s):  
Allen Costoff ◽  
W. H. McShan
Keyword(s):  
Electron Microscopy ◽  
Anterior Pituitary ◽  
Close Agreement ◽  
Secretory Granules ◽  
Cell Types ◽  
Pituitary Hormones ◽  
Biological Properties ◽  
Single Fraction ◽  
Rat Pituitary ◽  
Pituitary Glands

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.

Comparison of particle-associated progesterone and oxytocin in the ovine corpus luteum

1986 ◽  
Vol 108 (1) ◽  
pp. 109-116 ◽  
Author(s):  
G. E. Rice ◽  
G. Jenkin ◽  
G. D. Thorburn
Keyword(s):  
Corpus Luteum ◽  
Biogenic Amines ◽  
Density Gradient ◽  
Subcellular Distribution ◽  
Incubation Medium ◽  
Density Gradient Centrifugation ◽  
Secretory Granules ◽  
Gradient Centrifugation ◽  
Biochemical Characteristics ◽  
Oxytocin Release

ABSTRACT The subcellular distribution of progesterone and oxytocin within the ovine corpus luteum was investigated using differential and density gradient centrifugation. Progesterone and oxytocin were associated with particles which sedimented to a density of 1·049–1·054 g/ml and 1·054–1·061 g/ml respectively. Particle-associated progesterone did not, however, display physical or biochemical characteristics consistent with its storage within secretory granules. When particle-associated progesterone was incubated in HEPES buffer at 37 °C, 70% of the total progesterone was recovered in the incubation medium. The remaining stable particle-associated progesterone was not affected by treatments which stimulated oxytocin release and which have been shown to cause the release of peptides and biogenic amines from secretory granules. These results suggest that particle-associated progesterone represents the intercalation of progesterone into cell membranes and they do not support the hypothesis that progesterone is stored, in a protein-bound form, in luteal secretory granules. J. Endocr. (1986) 108, 109–116

FATE OF NEUROHYPOPHYSIAL GRANULE MEMBRANES AFTER SECRETION IN THE RAT: GENERATION OF A NEW APPARENT ORGANELLE BY FREEZING AND THAWING SECRETORY GRANULES

1981 ◽  
Vol 88 (1) ◽  
pp. 115-123
Author(s):  
JEAN H. LACEY ◽  
B. T. PICKERING
Keyword(s):  
Density Gradient Centrifugation ◽  
Secretory Granules ◽  
Gradient Centrifugation ◽  
Hormone Release ◽  
Freezing And Thawing ◽  
Granule Fraction ◽  
Two Peaks ◽  
Stimulation Of

The contents and membranes of the secretory granules in the rat neurohypophysis were labelled in vivo with [35S]cysteine and [3H]choline respectively. Density-gradient centrifugation of the labelled granules showed the membrane label to be distributed largely between two peaks: one associated with intact granules and one with the characteristics of empty granule envelopes. Stimulation of hormone release in vitro led to the movement of membrane label from the intact granule fraction to the other one, consistent with the recapture of membrane as large vesicles. Freezing and thawing the crude granule fraction, ostensibly to aid osmotic fracture, produced a single membrane component with a density intermediate between the two original fractions.

scholarly journals Distribution of bicarbonate-stimulated ATPase in rat intestinal epithelium.

1977 ◽  
Vol 73 (1) ◽  
pp. 257-260 ◽  
Author(s):  
C H van Os ◽  
A K Mircheff ◽  
E M Wright
Keyword(s):  
Atpase Activity ◽  
Brush Border ◽  
Plasma Membranes ◽  
Intestinal Epithelial Cells ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Succinic Dehydrogenase ◽  
Intestinal Epithelial ◽  
Mitochondrial Marker ◽  
Direct Role

This study reports on the distribution of bicarbonate-stimulated ATPase in rat intestinal epithelial cells. Brush-border membranes and basolateral membranes were separated from each other and from mitochondrial and other intracellular membranes by differential and density gradient centrifugation. Bicarbonate-sensitive ATPase activity followed the mitochondrial marker succinic dehydrogenase closely throughout all the centrifugation steps. The low HCO3--ATPase activity in purified brush-border and basolateral plasma membranes could be accounted for quantitatively by the small mitochondrial contamination. Consequently, there are no grounds for postulating that this enzyme has a direct role in H+ or HCO3- transport across the rat small intestine.

scholarly journals Localization of a 16,000-dalton fragment of the common precursor of adrenocorticotropin and beta-lipotropin in the rat and human pituitary gland.

1980 ◽  
Vol 86 (3) ◽  
pp. 825-830 ◽  
Author(s):  
J Guy ◽  
R Leclerc ◽  
G Pelletier
Keyword(s):  
Secretory Granules ◽  
Immunohistochemical Localization ◽  
Pars Intermedia ◽  
Electron Microscope Level ◽  
Human Pituitary ◽  
Human Pituitary Gland ◽  
Common Precursor ◽  
Rat Pituitary ◽  
Pituitary Glands ◽  
The Common

To clearly identify cells and organelles containing the common precursor (31,000 dalton) for both adrenocorticotropin (ACTH) and beta-lipotropin (beta-LPH), an immunohistochemical localization of a fragment (16,000 dalton) of the precursor that is not common to beta-LPH and ACTH was conducted in rat and human pituitary glands. With the help of specific antibodies that do not cross-react with beta-LPH and ACTH, the 16,000-dalton fragment was localized in the cells that also produce ACTH and beta-LPH in both the pars distalis and pars intermedia of the rat pituitary. At the electron microscope level, the secretory granules that contain ACTH were also stained for 16,000-dalton fragment. In the human pituitary, the 16,000-dalton fragment was also observed in all the secretory granules of lipocorticotrophs. These results suggest that, after enzymatic cleavage, fragment(s) of the common precursor and/or the whole common precursor remain packaged within the secretory granules with peptides of known activity.

Arginine esterase from isolated dog prostate secretory granules is fully active enzymatically

1985 ◽  
Vol 63 (12) ◽  
pp. 1603-1607 ◽  
Author(s):  
Gilles Frenette ◽  
Jean Y. Dubé ◽  
Jean R. Marcotte ◽  
Roland R. Tremblay
Keyword(s):  
Enzymatic Activity ◽  
Seminal Plasma ◽  
Density Gradient Centrifugation ◽  
Secretory Granules ◽  
Gradient Centrifugation ◽  
Protein Substrates ◽  
Inactive Form ◽  
Specific Enzymatic Activity ◽  
The One

We have isolated secretory granules from dog prostate homogenates and have determined whether a major portion of arginine esterase was localized in this fraction and if it was enzymatically active. Secretory granules were purified by density gradient centrifugation on sucrose, metrizamide, or Percoll. A major proportion of whole prostate homogenate arginine esterase was found in the granule fractions. Furthermore, the specific enzymatic activity in the granules was similar to the one observed in seminal plasma. No evidence could be found for the existence of significant amount of a zymogen inactive form of arginine esterase. These results suggest that arginine esterase could be active within the secretory granules in vivo and that it could hydrolyze protein substrates contained in this organelle.

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