Reference: Sawchenko, P.E., Swanson, L.W. & Vale W.W., 1984: Experiment acronym: Sawchenko-1984
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
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.
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| Reference: Watts, A.G. & Sanchez-Watts, G., 1995: Experiment acronym: Watts-CRH/AVP(PEG)
Data types: quantitative
Mapping approach:
brain region is captured in original published nomenclature, in BAMS
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: 6 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): Stratageme Corp
Probe sequence: 700 bp RsaI-RsaI CRH
Sequence species: rat
Probe sequence orientation: antisense
Control: sense
Labelling method: radiolabelling
Visualization method: DIG
Visualization medium: slide
Annotation: ....a 700 bp RsaI-RsaI fragment of the rat CRH gene (rCRH1) corresponding to part of the 3 end of exon 1 and the entire translated region of exon 2 (Frim et al., 1990) was subcloned into Bluescript (Stratagene Corp., La Jolla, CA) and linearized with EcoRl to generate an antisense digoxigenin-UTP labelled ppCRH cRNA probe...Additional experiments (data not given) have shown that neither RNase pretreatment followed by hybridization with antisense strand probes nor hybridization with sense strand probes gives any hybridization..Digoxigenin-UTP was visualized with the reagents from the digoxigenin nucleic acid detection kit (The Genius System, Boehringer-Mannheim Corp.).
| Measurement 2:
Measured nucleic acid: mRNA
Source (producer): Promega Gemini
Probe sequence: cDNA AVP 1004
Sequence species:
Probe sequence orientation: antisense
Control: sense
Labelling method: not specified
Visualization method: autoradiogram
Visualization medium: slide
Annotation: The pp- AVP sequence was generated from the cDNA AW 1004 described by Carrazana et al. (1988)....Additional experiments (data not given) have shown that neither RNase pretreatment followed by hybridization with antisense strand probes nor hybridization with sense strand probes gives any hybridization..
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| Experiment acronym: Watts-CRH/pENK(PEG)
Data types: quantitative
Mapping approach:
brain region is captured in original published nomenclature, in BAMS
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: 6 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): Stratageme Corp
Probe sequence: 700 bp RsaI-RsaI CRH
Sequence species: rat
Probe sequence orientation: antisense
Control: sense
Labelling method: radiolabelling
Visualization method: autoradiogram
Visualization medium: slide
Annotation: ....a 700 bp RsaI-RsaI fragment of the rat CRH gene (rCRH1) corresponding to part of the 3 end of exon 1 and the entire translated region of exon 2 (Frim et al., 1990) was subcloned into Bluescript (Stratagene Corp., La Jolla, CA) and linearized with EcoRl to generate an antisense digoxigenin-UTP labelled ppCRH cRNA probe...Additional experiments (data not given) have shown that neither RNase pretreatment followed by hybridization with antisense strand probes nor hybridization with sense strand probes gives any hybridization..Digoxigenin-UTP was visualized with the reagents from the digoxigenin nucleic acid detection kit (The Genius System, Boehringer-Mannheim Corp.)
| Measurement 2:
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)...
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| Experiment acronym: Watts-NT/pENK (PEG)
Data types: quantitative
Mapping approach:
brain region is captured in original published nomenclature, in BAMS
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: 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)...
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