Reduction of morphine abstinence in mice with a mutation in the gene encoding CREB

Science

Volumn 273, Issue 5275, 1996, Pages 657-659

  Maldonado, Rafael ^a   Blendy, Julie A ^b   Tzavara, Eleni ^c   Gass, Peter ^b   Roques, Bernard P ^a   Hanoune, Jacques ^c   Schütz, Günter ^b  

^a Laboratoire de Pharmacochimie   (France)

^b GERMAN CANCER RESEARCH CENTER DKFZ   (Germany)

^c HENRI MONDOR HOSPITAL   (France)

Author keywords

[No Author keywords available]

Indexed keywords

ADENYLATE CYCLASE; CYCLIC AMP; CYCLIC AMP RESPONSIVE ELEMENT BINDING PROTEIN;

ANALGESIA; ANIMAL CELL; ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; BINDING SITE; CONTROLLED STUDY; DRUG DEPENDENCE; DRUG WITHDRAWAL; GENE EXPRESSION; GENE MUTATION; GENETIC TRANSCRIPTION; IMMEDIATE EARLY GENE; MORPHINE ADDICTION; MOUSE; NONHUMAN; OPIATE ADDICTION; PRIORITY JOURNAL; REVIEW; SIGNAL TRANSDUCTION; WITHDRAWAL SYNDROME;

ANIMALIA;

EID: 0029764770     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5275.657     Document Type: Review

References (26)

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22. We have recently generated a mouse mutant in which the entire DNA binding domain of the CREB gene has been deleted, resulting in a null allele. These mice die at birth and are currently being investigated in our laboratory (D. Rudolph and G. Schütz, unpublished observations).
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25. Animals were killed by transcardiac perfusion with 4% (w/v) paraformaldehyde 2 hours after naloxone-precipitated withdrawal. Their brains were then fixed overnight in the same fixative before vibratome sectioning. Expression of the c-FOS and c-JUN proteins was examined in coronal 50-μM vibratome sections at the level of the locus coeruleus (Fig. 3, A through D) and amygdala (Fig. 3, E through H). All antibodies were generated in rabbits immunized with bacterially expressed fusion proteins. All cDNAs used to construct the fusion proteins were of mouse origin. Specificity and characterization of the antibodies had been described previously [K. Kovary and R. Bravo, Mol. Cell Biol. 11, 4466 (1991)]. Sections were incubated in normal goat serum [2% in phosphate-buffered saline (PBS) and 0.2% Triton ×-100] for 1 hour, and then in the primary antisera for 36 hours at 4°C. The primary antisera were diluted as follows: anti-c-FOS, 1:40,000; and anti-c-JUN, 1:40,000 (16). Immunoreactivity was visualized by the avidin biotin complex method (Vectastin, Vector Labs, Burlingame, CA). Sections were developed in 0.02% diaminobenzidine with 0.02% hydrogen peroxide, and a subset of sections were counterstained with hemalum.
26. We thank C. Durieux for assistance in binding studies and C. Dupuis and M. Bock for manuscript drafting. Supported by the Deutsche Forschungsgemeinschaft through SFB 229, the B102-CT93-0319 Programme of the Commission of the European Communities, the Caisse Nationale d'Assurance et Maladie des Travailleurs Salaries, and the Biomed and Health Research Programme (931721) of the Commission of the European Communities. E.T. was supported by the I. S. Latsis Foundation.