Immunohistochemical Localization of ACTH and Prolactin in the Pituitary Gland of Adult Migratory Sockeye Salmon (Oncorhynchus nerka)

1969 ◽  
Vol 26 (7) ◽  
pp. 1837-1846 ◽  
Author(s):  
B. A. McKeown ◽  
A. P. van Overbeeke
Keyword(s):  
Sockeye Salmon ◽  
Fluorescein Isothiocyanate ◽  
Cell Types ◽  
Pituitary Hormones ◽  
Immunohistochemical Localization ◽  
Granule Density ◽  
Pituitary Glands ◽  
Antibody Complex ◽  
Ovine Prolactin ◽  
Antigen Antibody

Antibodies to porcine adrenocorticotrophic hormone (ACTH), Synacthen (synthetic corticotrophin, Ciba), and ovine prolactin were prepared in rabbits and the antisera were tested for specificity against several pituitary hormones. The gamma-globulin fractions of the antisera were conjugated with fluorescein isothiocyanate and the labelled antibodies were "purified" by column chromatography.Fresh-frozen sections of pituitary glands of adult migratory sockeye salmon were incubated with the antibody solutions and examined with a fluorescence microscope. The resulting antigen–antibody complex could be localized by re-photographing the same or alternate sections after fixation and staining. Anti-ACTH and anti-Synacthen appeared to be bound specifically by the epsilon cells, whereas anti-prolactin reacted with the eta cells of the rostral pars distalis. Pituitary glands collected at various stations along the migratory route, including one seawater sample, showed the same reactivity. Other glands were prepared for histological examination. Microspectrophotometric analysis of cell types showed that the granule density of the ACTH cells increased gradually during the later part of migration. In the prolactin cells, no change in granulation could be detected during entrance into the river or subsequent spawning migration.

Immunohistochemical Localization of Prolactin in the Pituitary Gland of the Pigeon, Columba livia

1972 ◽  
Vol 50 (11) ◽  
pp. 1021-1025 ◽  
Author(s):  
B. A. McKeown
Keyword(s):  
Pituitary Gland ◽  
Fluorescein Isothiocyanate ◽  
Columba Livia ◽  
Immunohistochemical Localization ◽  
Frozen Sections ◽  
Specific Antibodies ◽  
Cephalic Lobe ◽  
Fresh Frozen ◽  
Pituitary Glands ◽  
Ovine Prolactin

Specific antibodies to ovine prolactin were produced in rabbits. The antisera were conjugated to fluorescein isothiocyanate and "purified" by column chromatography. Antibodies, thus labelled, were applied to fresh-frozen sections of pigeon pituitary glands. Fluorescence was observed in specific cells of the cephalic lobe. After staining, these cells were identified as the erythrosinophilic eta cells.

scholarly journals Ultrastructural immunohistochemical localization of anti-invasion factor (AIF) in bovine cartilage matrix.

1982 ◽  
Vol 30 (6) ◽  
pp. 524-531 ◽  
Author(s):  
F H Wezeman ◽  
G V Childs
Keyword(s):  
Staining Technique ◽  
Ultrastructural Localization ◽  
Immunohistochemical Localization ◽  
Cartilage Matrix ◽  
Fixed Tissue ◽  
Antibody Complex ◽  
Protein Components ◽  
Antigen Antibody ◽  
Unlabeled Antibody ◽  
Bovine Cartilage

Rabbit antibodies prepared against bovine cartilage anti-invasion factor (AIF) were tested for their affinity toward antigenic sites in glutaraldehyde-fixed bovine hyaline cartilage matrix. Ultrastructural localization of the antigen-antibody complex was accomplished by the unlabeled antibody peroxidase-antiperoxidase staining technique. Unextracted and salt-extracted (1 M NaCl or 3 M GuHCl) cartilage slices were incubated with anti-AIF antibodies at a working dilution of 1:20,000. Staining occurred in unextracted matrix distributed throughout the tissue, but with regional variation in the lacunar matrix. Significantly less stain was noted in extracted tissues. The results suggest that at least certain protein components in AIF are morphologically associated with matrix complexes in aldehyde-fixed tissue.

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.

IMMUNOHISTOCHEMISTRY OF INDIVIDUAL ADENOHYPOPHYSIAL CELLS

1971 ◽  
Vol 68 (1_Suppl) ◽  
pp. S168-S189 ◽  
Author(s):  
P. Leleux ◽  
C. Robyn
Keyword(s):  
Present Report ◽  
Mammalian Species ◽  
Fluorescein Isothiocyanate ◽  
Cell Types ◽  
Pituitary Hormones ◽  
Thyroid Stimulating Hormone ◽  
Human Pituitary ◽  
Cross Reactions ◽  
Immunohistochemical Studies ◽  
Stimulating Hormone

ABSTRACT Immunohistochemistry is the intracellular detection of antigens by the use of specific antibodies labelled with a tracer. The choice of the tracer is such that the sites of the antigen-antibody reactions can be visualized by microscopic examination. The present report refers to the human pituitary where most of the immunohistochemical identifications of adenohypophysial cells were conducted with antibodies specific of their hormonal content and labelled with fluorescein isothiocyanate as tracer. Such immunohistochemical identifications had to be correlated to the morphological nomenclatures of the glandular cells based on histochemical stainings. Confusion has been introduced in these nomenclatures by the definition of three to eight cell types using different criteria and different terminologies. In the present report, owing to this absence of standardization, the comparative evaluation of immunohistochemical data have been based on Romeis (1940) and Pearse & van Noorden (1963) nomenclatures. There is strong experimental evidence supporting the localization of adrenocorticotrophic hormone (ACTH) in the basophils of Romeis β type (R-mucoids of Pearse). Somatotrophic hormone (STH) has been consistently found in the acidophils of Romeis a type. In the human, there is no direct evidence to support the localization of prolactin (LTH) in the acidophils of Romeis ε type. Luteinizing hormone (LH) and follicle stimulating hormone (FSH) have both been located in the basophils of Romeis δ type (S-mucoids of Pearse). Further investigations into the human and also into other mammalian species are required to determine if the gonadotrophic hormones have different localizations on the cellular or on the subcellular level. The immunohistochemical localizations of thyroid stimulating hormone (TSH) and melanocyte stimulating hormone (MSH) have not been convincingly achieved. The conclusions drawn from immunohistochemical studies of the adenohypophysis are essentially limited by the cross reactions existing between STH and prolactin, between ACTH and MSH and between LH, FSH and TSH. More experimental data on these immunological cross reactions are still required before more accurate morphological discriminations can be achieved between the cell types secreting STH and prolactin, between those secreting ACTH and MSH and between those secreting LH, FSH and TSH. In addition, when hormones of the adenohypophysis are chemically and/or antigenically closely related, the cells responsible for their secretion are morphologically very similar too. Finally, immunohistochemical studies revealed the lack of species specificity of the pituitary hormones. Extensive cross reactions have been shown between human STH and STH of all mammalian species studied so far. Consistent cross reactions were also found between human gonadotrophins and those of several mammalian species.

PURIFICATION AND CHARACTERIZATION OF CAPRINE PROLACTIN

1974 ◽  
Vol 60 (2) ◽  
pp. 359-367 ◽  
Author(s):  
A. S. McNEILLY ◽  
P. ANDREWS
Keyword(s):  
Simple Procedure ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Product Yield ◽  
Pituitary Hormones ◽  
Major Product ◽  
Purification And Characterization ◽  
Pituitary Glands ◽  
Ovine Prolactin

SUMMARY Prolactin was isolated from frozen goat pituitary glands by a simple procedure involving gel filtration and chromatography on DEAE-cellulose. The major product (yield, 2·5 mg/g pituitary tissue) had high pigeon crop sac-stimulating activity (27 i.u./mg) and was free of growth hormone and other pituitary hormones. The molecular weight was similar to that of ovine prolactin. Caprine prolactin was immunologically indistinguishable from ovine prolactin in radioimmunoassays, in which ovine prolactin antiserum and either ovine or caprine prolactin labelled with 125I were used. The results indicate that caprine and ovine prolactin are closely related and that radioimmunoassay for ovine prolactin may be used to estimate caprine prolactin in serum.

Fluorometric estimation of antigens (antibodies)

1965 ◽  
Vol 20 (10) ◽  
pp. 974-976 ◽  
Author(s):  
A. Robert Neurath
Keyword(s):  
Rabies Virus ◽  
Fluorescein Isothiocyanate ◽  
Virus Antigen ◽  
Paper Chromatogram ◽  
Quenching Of Fluorescence ◽  
Γ Globulins ◽  
Antibody Complex ◽  
Antigen Protein ◽  
Antigen Antibody ◽  
Millipore Filters

A method for the titration of antigens based on separation by filtration through millipore filters of the complex of antigen with fluorescein isothiocyanate-labeled antibodies from free labeled antibodies and nonspecific γ-globulins is described. The fluorescence of the antigen-antibody complex is measured directly on the millipore filters using the Turner fluorometer equipped with a paper chromatogram door. The limit of sensitivity of the method is about 0.05 μg of antigen protein. Quenching of fluorescence by nonlabeled antibodies can be used for the titration of antisera. An example of the application of the method for titration of rabies virus antigen is also presented.

Visualization of Antibody Transport in Rat Visceral Yolk-Sac Placenta

1982 ◽  
Vol 40 ◽  
pp. 246-247
Author(s):  
William P. Jollie
Keyword(s):  
Phosphate Buffer ◽  
Yolk Sac ◽  
Female Rats ◽  
Thin Sections ◽  
Visceral Yolk Sac ◽  
Antibody Complex ◽  
Cytochemical Reaction ◽  
Hind Foot ◽  
Antigen Antibody ◽  
Yolk Sac Placenta

A technique has been developed for visualizing antibody against horseradish peroxidase (HRP) in rat visceral yolk sac, the placental membrane across which passive immunity previously has been shown to be transferred from mother to young just prior to birth. Female rats were immunized by injecting both hind foot pads with 1 mg HRP emulsified in complete Freund's adjuvant. They were given a booster of 0.5mg HRP in 0.1 ml normal saline i.v. after one week, then bred and autopsied at selected stages of pregnancy, viz., 12, 1 7 and 22 days post coitum, receiving a second booster, injected as above, five days before autopsy. Yolk sacs were removed surgically and fixed immediately in 2% paraformaldehye, 1% glutaraldehye in 0.1 M phosphate buffer with 0.01% CaCl2 at pH 7.4, room temperature, for 3 hr, rinsed 3X in 0.1 M phosphate buffer plus 5% sucrose, then exposed to 1 mg HRP in 1 ml 0.1 M phosphate buffer at pH 7.4 for 1 hr. They were refixed in aldehydes, as above, for 1 5 min (to assure binding of antigen-antibody complex). Following buffer washes, the tissues were incubated in 3 mg diaminobenzidine tetrahydrochloride and 0.01% H2O2 in 0.05 M Tris-HCl buffer for 30 min. After brief buffer washes, they were postfixed in 2% OsO4. in phosphate buffer at pH 7.4, 4°C for 2 hr, dehydrated through a graded series of ethanols, and embedded in Durcupan. Thin sections were observed and photographed without contrast-enhancement with heavy metals. Cytochemical reaction product marked the site of HRP (i.e., antigen) which, in turn, was present only where it was bound with anti-HRP antibody.

INHIBITION OF LIPOLYTIC ACTIVITY OF SYNTHETIC β1–24 AND β1–39 CORTICOTROPHIN BY ANTI-ACTH ANTISERUM

1966 ◽  
Vol 51 (1) ◽  
pp. 88-94 ◽  
Author(s):  
A. Villanueva ◽  
S. J. H. Ashcroft ◽  
J. P. Felber
Keyword(s):  
Guinea Pig ◽  
Lipolytic Activity ◽  
Peptide Hormones ◽  
Fat Pad ◽  
Double Antibody ◽  
Epididymal Fat ◽  
Antibody Complex ◽  
Radio Immunoassay ◽  
Antigen Antibody

ABSTRACT The synthetic ACTH peptides β1–39 and β1–24 stimulated lipolysis as determined by the rat epididymal fat pad in vitro. The stimulating effect of these peptides was diminished by prior incubation of the peptides with antibodies produced by the guinea-pig against ACTH. The stimulating effect of these hormones was also diminished by the double antibody system used in the radio-immunoassay of ACTH and other peptide hormones, in which incubation with antiserum is followed by precipitation of the antigen-antibody complex by rabbit anti-guinea-pig-γ-globulin.

IDENTIFICATION OF LATS AS AN ANTIGEN-ANTIBODY COMPLEX

1973 ◽  
Vol 71 (4_Suppl) ◽  
pp. S11
Author(s):  
K. Schemmel ◽  
L. Weisbecker ◽  
H. Norden ◽  
V. Mokmol ◽  
V. Becker ◽  
...  
Keyword(s):  
Antibody Complex ◽  
Antigen Antibody

scholarly journals Immunohistochemical localization of phosphoenolpyruvate carboxykinase in adult and developing mouse tissues.

1990 ◽  
Vol 38 (2) ◽  
pp. 171-178 ◽  
Author(s):  
D B Zimmer ◽  
M A Magnuson
Keyword(s):  
Nervous System ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Submaxillary Gland ◽  
Cell Types ◽  
Immunohistochemical Localization ◽  
Mouse Tissues ◽  
Multiple Cell ◽  
Cell Type Specific ◽  
Immunohistochemical Techniques ◽  
Developing Nervous System

We used immunohistochemical techniques to analyze the cell distribution of phosphoenolpyruvate carboxykinase (PEPCK) in adult and developing mouse tissues. PEPCK immunoreactivity was detected in many tissues, including some that had not been previously reported to contain PEPCK enzyme activity (bladder, stomach, ovary, vagina, parotid gland, submaxillary gland, and eye). In some multicellular tissues, PEPCK immunoreactivity was observed in multiple cell types. Several tissues (spleen, thyroid, and submaxillary gland) contained no detectable PEPCK immunoreactivity. During development, PEPCK immunoreactivity was associated with the developing nervous system and somites in 15-day embryos. At prenatal day 18, PEPCK immunoreactivity was detected only in the nervous system. At prenatal day 20, PEPCK immunoreactivity was observed in many of the tissues that contain PEPCK in the adult, with the exception of liver, lung, and stomach. PEPCK immunoreactivity was detected in liver at postnatal day 1, lung at postnatal day 7, and stomach after postnatal day 21. The only tissue in which PEPCK immunoreactivity decreased during development was the pancreas, where PEPCK immunoreactivity was detected at prenatal day 20 and was present until postnatal day 21. These results suggest that PEPCK expression is cell-type specific, more widespread than previously thought, and differentially expressed during development.

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