Reference: Swanson LW, Sawchenko PE, Rivier J, Vale WW, 1983: Experiment acronym: Swanson-1983(CRH)
Data types: quantitative
Mapping approach:
brain region is captured in a BAMS nomenclature other than that used in the original publication
Mapping details
Coordinates: none
Anatomical data presentation:
text
representative labeled images
drawings/template mappings of all sections
| Animals (subjects):
Number: 10 Sex: M
Age: not specified Weight: not specified
Housing conditions: normal housing
Annotation: normal housing | Method:
Neuron/glia identification method: not specified
Staining frequency: 1:4 or 1:8
Technique: immunohistochemistry
| Measurement 1:
Antigen: not specified
Antigen species: not specified
Source (producer): not specified
Primary antibody: not specified
Primary antibody species: rabbit
Antibody type (monoclonal, polyclonal): polyclonal
Secondary antibody: not specified
Secondary antibody species: not specified
Immunoglobulin class: not specified
Control: not specified
Visualization method: fluorescence
Visualization medium: slide
Annotation: Seven different antisera to synthetic ovine CRF (or to a fragment of CRF), which were raised in rabbits [7], were tested and all but one were found to stain cells and fibers in the rat. Since antiserum 24 ([Try22. Gly23]-CRF (1-23) conjugated to human alpha-globulin with bis-diazetized benzidine] has been found to interact with rat CRF particularly well in radioimmunoassays [Vale, unpublished observations], it was used extensively for the mapping studies reported here.
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| Reference: Sawchenko, P.E., Swanson, L.W. & Vale W.W., 1984: Experiment acronym: Sawchenko-1984
Data types: quantitative
qualitative
Mapping approach:
brain region is captured in a BAMS nomenclature other than that used in the original publication
Mapping details
Coordinates: none
Anatomical data presentation:
text
representative labeled images
representative drawings/mappings onto templates
| Animals (subjects):
Number: not specified Sex: M
Age: not specified Weight: not specified
Housing conditions: normal housing
Annotation: normal housing | Method:
Neuron/glia identification method: stain specific
Staining frequency: 1:5
Technique: immunohistochemistry
| Measurement 1:
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.
| Measurement 2:
Antigen: oxytocin
Antigen species: N/A
Source (producer): Dr. F. Vandesande and Dr.
K. Dierickx (University of Ghent, Belgium)
Primary antibody: oxytocin 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.
| Measurement 3:
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.
| Measurement 4:
Antigen: neurotensin
Antigen species: N/A
Source (producer): Dr. M. R. Brown (Peptide Biology Laboratory, The Salk
Institute)
Primary antibody: neurotensin 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: staining pattern
Visualization method: autoradiogram
Visualization medium: slide
Annotation: Dr. M. R. Brown (Peptide Biology Laboratory, The Salk Institute) and was prepared against neurotensin conjugated with glutaraldehyde to thyroglobulin. This serum (Nil-1) is directed against the C-terminal portion of the molecule (Brown et al., 1978) and was used at a dilution of 1:4000. All staining in the hypothalamus was blocked by the addition of synthetic neurotensin (15 mg/ml) to the serum.
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