| Animals (subjects) | | Sex: M | | Number of animals: 6 | | Age: not specified | | Mass: 280-320 | | Unit of mass: | | Housing conditions: normal housing | | Annotation: Adult male Sprague-Dawley rats (280-320 g BW at injection) were maintained on a 12 hour light/ 12 hour dark photoperiod (lights on 0700 hours) with water and rat chow available ad libitum. They were allowed 7 days' acclimatization to the animal quarters before we proceeded with the experiment. |
| | Experimental method | | Experiment type: in situ hybridization | | Neuron/glia identification method: stain specific | | Staining frequency: 1:8 |
Experimental details:
| 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.)
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