Metadata

Experiment acronym Annotation Collator Author checked
Sawchenko-1984CRH-oxytocin colocalization study in the rat PVH.Mihail Botano

Animals (subjects)
Sex: M
Number of animals: not specified
Age: not specified
Mass: not specified
Unit of mass: not specified
Housing conditions: normal housing
Annotation: Adult male albino rats of the Sprague-Dawley strain were used in all experiments.
27, 14
Experimental method
Experiment type: immunohistochemistry
Neuron/glia identification method: stain specific
Staining frequency: 1:5

Experimental details:
Antigen: CRH
Antigen species: not specified
Source (producer): specified elsewhere
Primary antibody: rabbit antiserum
Primary antibody species: rabbit
Antibody type (monoclonal, polyclonal): polyclonal
Secondary antibody: goat-antirabbit
Secondary antibody species: goat
Immunoglobulin class: IgG
Control: staining pattern
Visualization method: autoradiogram
Visualization medium: slide
Annotation: Up to five one-in-five series of ...sections through the PVH were saved and then prepared for indirect immunofluorescence staining of cells that cross-react with antisera against CRF, neurotensin, oxytocin, or vasopressin using a conventional method (Sawchenko and Swanson, 1981) based upon the localization of a primary antiserum raised in rabbits using an affinity-purified, fluorescein-conjugated goat anti-rabbit IgG. To ensure the validity of our localization of CRFimmunoreactive neurons, three sera against CRF human alpha-globulin conjugates were used. Two of these (C24 and C30) are directed against the C- and N-terminal portions, respectively, of the ovine CRF molecule (Swanson et al., 1983; Vale et al., 1983). The third was prepared against a synthetic rat CRF (Rivier et al., 1983) conjugate. Each of these sera was used at a final dilution of 1:3000, and pre-incubation of each with 22 mg ml-l of the respective immunogen blocked specific staining of cells and fibers in the hypothalamus. All sera used for both normal immunohistochemistry and blocking experiments contained 10 mg/ml of the “carrier” protein used for immunization.
Antigen: vasopression
Antigen species: N/A
Source (producer): K. Dierickx (University of Ghent, Belgium)
Primary antibody: vasopressin antiserum
Primary antibody species: not specified
Antibody type (monoclonal, polyclonal): monoclonal
Secondary antibody: not specified
Secondary antibody species: not specified
Immunoglobulin class: not specified
Control: adsorption
Visualization method: autoradiogram
Visualization medium: slide
Annotation: Antisera against oxytocin and vasopressin were provided by Dr. F. Vandesande and Dr. K. Dierickx (University of Ghent, Belgium) and were cross-adsorbed in the solid phase against the heterologous antigen according to a procedure outlined elsewhere (Swaab and Pool, 1975). Specific staining with each antiserum was blocked when immunoprocessing was carried out using sera that had been pre-incubated with an excess (15 mg/ml) of the homologous synthetic peptide and then diluted to the working concentration of 1:2000.

Anatomy and histology
Section plane: coronalAngle: not specifiedCutting method: not specified Preservation: freezing Thickness: 20 micrometer
Staining type: thionin Sampling: 1:5 Annotation: Up to five one-in-five series of 20 micrometers thick frozen sections through the PVH were saved and then prepared for indirect immunofluorescence staining of cells that cross-react with antisera against CRF, neurotensin, oxytocin, or vasopressin using a conventional method (Sawchenko and Swanson, 1981) based upon the localization of a primary antiserum raised in rabbits using an affinity-purified, fluorescein-conjugated goat anti-rabbit IgG. The distributions of labeled cells were mapped onto projection drawings made from adjacent series of sections counterstained with thionin, and the region around the PVH was photographed at x 100 magnification using Ilford XP-1 film.

Mapping details
Coordinates: noneData presentation:
text
representative labeled images
representative drawings or mappings onto templates
Mapping approach: