Reference: Watts, A.G. & Sanchez-Watts, G., 1995: Experiment acronym: Watts-NT/pENK (PEG)
Data types: quantitative
Mapping approach: Mapping details
Coordinates: none
Anatomical data presentation:
text
representative labeled images
representative drawings/mappings onto templates
| Animals (subjects):
Number: 3 Sex: not specified
Age: not specified Weight: 280-320g
Housing conditions: not specified
Annotation: normal housing | Method:
Neuron/glia identification method: stain specific
Staining frequency: 1:8
Technique: in situ hybridization
| Measurement 1:
Measured nucleic acid: mRNA
Source (producer): Promega Gemini
Probe sequence: proenkephalin
Sequence species:
Probe sequence orientation: antisense
Control: sense
Labelling method: radiolabelling
Visualization method: autoradiogram
Visualization medium: slide
Annotation: Sections were hybridized with 35SUTP- labelled complementary RNA (cRNA) probes transcribed from cDNA sequences for part of the mRNAs encoding proenkephalin (pENK)....All 35S-UTP probes were synthesized using the Riboprobe Gemini System I1 kit (Promega Inc., Madison, WI) and the appropriate RNA polymerase using a modification of the method of Melton et al. (1984)...
| Measurement 2:
Measured nucleic acid: mRNA
Source (producer): not specified
Probe sequence: pp-neurotensin/neuromedin
Sequence species:
Probe sequence orientation: antisense
Control: sense
Labelling method: radiolabelling
Visualization method: autoradiogram
Visualization medium: slide
Annotation: Sections were hybridized with 35SUTP- labelled complementary RNA (cRNA) probes transcribed from cDNA sequences for part of the mRNAs encoding pp-neurotensin/neuromedin (pENK)....All 35S-UTP probes were synthesized using the Riboprobe Gemini System I1 kit (Promega Inc., Madison, WI) and the appropriate RNA polymerase using a modification of the method of Melton et al. (1984)...
|
|
| 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: not specified
Age: not specified Weight: not specified
Housing conditions: not specified
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.
|
|
|
