Activation of corticotropin-releasing factor in the limbic system during cannabinoid withdrawal

Science

Volumn 276, Issue 5321, 1997, Pages 2050-2054

  Rodríguez De Fonseca, Fernando ^a   Carrera, M Rocío A ^b   Navarro, Miguel ^a   Koob, George F ^b   Weiss, Friedbert ^b  

^a UNIVERSIDAD COMPLUTENSE DE MADRID   (Spain)

^b SCRIPPS RESEARCH INSTITUTE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CANNABIS; CORTICOTROPIN RELEASING FACTOR; DEXANABINOL; RIMONABANT;

ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; ARTICLE; CANNABIS ADDICTION; CONTROLLED STUDY; DRUG BLOOD LEVEL; DRUG DEPENDENCE; DRUG EFFECT; DRUG WITHDRAWAL; LIMBIC SYSTEM; NONHUMAN; PRIORITY JOURNAL; RAT;

AMYGDALA; ANIMALS; ANXIETY; BEHAVIOR, ANIMAL; BRAIN; CORTICOSTERONE; CORTICOTROPIN-RELEASING HORMONE; MALE; MICRODIALYSIS; PIPERIDINES; PROTO-ONCOGENE PROTEINS C-FOS; PYRAZOLES; RATS; RATS, WISTAR; RECEPTORS, CANNABINOID; RECEPTORS, DRUG; SUBSTANCE WITHDRAWAL SYNDROME; TETRAHYDROCANNABINOL;

EID: 0039534405     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.276.5321.2050     Document Type: Article

References (56)

1. J. A. Halikas, R. A. Weller, C. L. Morse, R. A. Hoffman, Int. J. Addict. 20, 701 (1985); L. Hollister, Pharmacol. Rev. 38, 1 (1986) and references therein; M. W. Fishman, J. F. Rosenbaum, D. I. Jabusaka, D. B. Carr, Res. Commun. Subst. Abuse 9, 219 (1988); J. D. Anthony, L. A. Warner, R. C. Kessler, Exp. Clin. Psychopharmacol. 2, 244 (1994); H. G. Pope and D. Yurgelun-Todd, JAMA 275, 521 (1996).
2. J. A. Halikas, R. A. Weller, C. L. Morse, R. A. Hoffman, Int. J. Addict. 20, 701 (1985); L. Hollister, Pharmacol. Rev. 38, 1 (1986) and references therein; M. W. Fishman, J. F. Rosenbaum, D. I. Jabusaka, D. B. Carr, Res. Commun. Subst. Abuse 9, 219 (1988); J. D. Anthony, L. A. Warner, R. C. Kessler, Exp. Clin. Psychopharmacol. 2, 244 (1994); H. G. Pope and D. Yurgelun-Todd, JAMA 275, 521 (1996).
3. J. A. Halikas, R. A. Weller, C. L. Morse, R. A. Hoffman, Int. J. Addict. 20, 701 (1985); L. Hollister, Pharmacol. Rev. 38, 1 (1986) and references therein; M. W. Fishman, J. F. Rosenbaum, D. I. Jabusaka, D. B. Carr, Res. Commun. Subst. Abuse 9, 219 (1988); J. D. Anthony, L. A. Warner, R. C. Kessler, Exp. Clin. Psychopharmacol. 2, 244 (1994); H. G. Pope and D. Yurgelun-Todd, JAMA 275, 521 (1996).
4. J. A. Halikas, R. A. Weller, C. L. Morse, R. A. Hoffman, Int. J. Addict. 20, 701 (1985); L. Hollister, Pharmacol. Rev. 38, 1 (1986) and references therein; M. W. Fishman, J. F. Rosenbaum, D. I. Jabusaka, D. B. Carr, Res. Commun. Subst. Abuse 9, 219 (1988); J. D. Anthony, L. A. Warner, R. C. Kessler, Exp. Clin. Psychopharmacol. 2, 244 (1994); H. G. Pope and D. Yurgelun-Todd, JAMA 275, 521 (1996).
5. J. A. Halikas, R. A. Weller, C. L. Morse, R. A. Hoffman, Int. J. Addict. 20, 701 (1985); L. Hollister, Pharmacol. Rev. 38, 1 (1986) and references therein; M. W. Fishman, J. F. Rosenbaum, D. I. Jabusaka, D. B. Carr, Res. Commun. Subst. Abuse 9, 219 (1988); J. D. Anthony, L. A. Warner, R. C. Kessler, Exp. Clin. Psychopharmacol. 2, 244 (1994); H. G. Pope and D. Yurgelun-Todd, JAMA 275, 521 (1996).
6. A. W. Zuardi, W. A. Shirakawa, E. Finkelfarb, I. G. Karniol, Psychopharmacology 76, 245 (1982).
7. American Psychiatric Association, Diagnostic and Statistical Manual of Mental Disorders (American Psychiatric Association, Washington, DC, ed. 3, 1994).
8. D. E. McMillan, W. L. Dewey, L. S. Harris, Ann. N.Y. Acad. Sci. 191, 83 (1971); S. Kaymakcalan, Bull. Narc. 25, 39 (1973); G. A. Wiesbeck et al., Addiction 91, 1469 (1996); J. H. Mendelson, N. K. Mello, B. W. Lex, S. Bavli, Am. J. Psychiatry 141, 1289 (1984).
9. D. E. McMillan, W. L. Dewey, L. S. Harris, Ann. N.Y. Acad. Sci. 191, 83 (1971); S. Kaymakcalan, Bull. Narc. 25, 39 (1973); G. A. Wiesbeck et al., Addiction 91, 1469 (1996); J. H. Mendelson, N. K. Mello, B. W. Lex, S. Bavli, Am. J. Psychiatry 141, 1289 (1984).
10. D. E. McMillan, W. L. Dewey, L. S. Harris, Ann. N.Y. Acad. Sci. 191, 83 (1971); S. Kaymakcalan, Bull. Narc. 25, 39 (1973); G. A. Wiesbeck et al., Addiction 91, 1469 (1996); J. H. Mendelson, N. K. Mello, B. W. Lex, S. Bavli, Am. J. Psychiatry 141, 1289 (1984).
11. D. E. McMillan, W. L. Dewey, L. S. Harris, Ann. N.Y. Acad. Sci. 191, 83 (1971); S. Kaymakcalan, Bull. Narc. 25, 39 (1973); G. A. Wiesbeck et al., Addiction 91, 1469 (1996); J. H. Mendelson, N. K. Mello, B. W. Lex, S. Bavli, Am. J. Psychiatry 141, 1289 (1984).
12. K. Tsou, S. L. Patrick, J. M. Walker, Eur. J. Pharmacol. 280, R-13 (1995).
13. M. Rinaldi-Carmona et al., FEBS Lett. 350, 240 (1994); V. Santucci, J. J. Storme, P. Soubrié, G. Le Fur, Life Sci. 58, 103 (1996).
14. M. Rinaldi-Carmona et al., FEBS Lett. 350, 240 (1994); V. Santucci, J. J. Storme, P. Soubrié, G. Le Fur, Life Sci. 58, 103 (1996).
15. W. A. Devane, F. A. Dysarz III, M. R. Johnson, L. S. Melvin, A. Hewlett, Mol. Pharmacol. 34, 605 (1988); M, Herkenham et al., Proc. Natl. Acad. Sci. U.S.A. 87, 1932 (1990); L. Matsuda, S. J. Lolait, M. J. Brownstein, A. C. Young, T. Bonner, Nature 346, 561 (1990).
16. W. A. Devane, F. A. Dysarz III, M. R. Johnson, L. S. Melvin, A. Hewlett, Mol. Pharmacol. 34, 605 (1988); M, Herkenham et al., Proc. Natl. Acad. Sci. U.S.A. 87, 1932 (1990); L. Matsuda, S. J. Lolait, M. J. Brownstein, A. C. Young, T. Bonner, Nature 346, 561 (1990).
17. W. A. Devane, F. A. Dysarz III, M. R. Johnson, L. S. Melvin, A. Hewlett, Mol. Pharmacol. 34, 605 (1988); M, Herkenham et al., Proc. Natl. Acad. Sci. U.S.A. 87, 1932 (1990); L. Matsuda, S. J. Lolait, M. J. Brownstein, A. C. Young, T. Bonner, Nature 346, 561 (1990).
18. R. K. Kubena, J. C. Peehack, H. Barry, Eur. J. Pharmacol. 14, 89 (1971); F. Rodríguez de Fonseca, M. A. Villanua, R. M. Muñoz, O. San-Martin-Clarke, M. Navarro, Neuroendocrinology 61, 714 (1995).
19. R. K. Kubena, J. C. Peehack, H. Barry, Eur. J. Pharmacol. 14, 89 (1971); F. Rodríguez de Fonseca, M. A. Villanua, R. M. Muñoz, O. San-Martin-Clarke, M. Navarro, Neuroendocrinology 61, 714 (1995).
20. G. F. Koob, Neuron 16, 893 (1996).
21. E. Merlo Pich et al., J. Neurosci. 15, 5439 (1995); R. M. Richter, G. F. Koob, F. Weiss, in preparation.
22. E. Merlo Pich et al., J. Neurosci. 15, 5439 (1995); R. M. Richter, G. F. Koob, F. Weiss, in preparation.
23. S. Rassnick, S. C. Heinrichs, K. T. Britton, G. F. Koob, Brain Res. 605, 25 (1992); M. A. Vargas, G. Bissette, M. J. Owens, C. L. Ehlers, C. B. Nemeroff, Ann. N. Y. Acad. Sci. 654, 145 (1992); G. F. Koob, A. Markou, F. Weiss, G. Schulteis, Semin. Neurosci. 5, 351 (1993).
24. S. Rassnick, S. C. Heinrichs, K. T. Britton, G. F. Koob, Brain Res. 605, 25 (1992); M. A. Vargas, G. Bissette, M. J. Owens, C. L. Ehlers, C. B. Nemeroff, Ann. N. Y. Acad. Sci. 654, 145 (1992); G. F. Koob, A. Markou, F. Weiss, G. Schulteis, Semin. Neurosci. 5, 351 (1993).
25. S. Rassnick, S. C. Heinrichs, K. T. Britton, G. F. Koob, Brain Res. 605, 25 (1992); M. A. Vargas, G. Bissette, M. J. Owens, C. L. Ehlers, C. B. Nemeroff, Ann. N. Y. Acad. Sci. 654, 145 (1992); G. F. Koob, A. Markou, F. Weiss, G. Schulteis, Semin. Neurosci. 5, 351 (1993).
26. F. R. de Fonseca et al., J. Pharmacol. Exp. Ther. 276, 56 (1996).
27. W. Vale, J. Spiess, C. Rivier, J. Rivier, Science 213, 1394 (1981).
28. G. F. Koob and F. E. Bloom, Fed. Proc. 44, 259 (1985); G. F. Koob, S. C. Heinrichs, F. Menzaghi, E. Merlo Pich, K. T. Britton, Semin. Neurosci. 6, 221 (1994); F. Menzaghi, S. C. Heinrichs, E. Merlo Pich, F. J. H. Tilders, G. F. Koob, Neurosci. Lett. 168, 139 (1994);
29. G. F. Koob and F. E. Bloom, Fed. Proc. 44, 259 (1985); G. F. Koob, S. C. Heinrichs, F. Menzaghi, E. Merlo Pich, K. T. Britton, Semin. Neurosci. 6, 221 (1994); F. Menzaghi, S. C. Heinrichs, E. Merlo Pich, F. J. H. Tilders, G. F. Koob, Neurosci. Lett. 168, 139 (1994);
30. G. F. Koob and F. E. Bloom, Fed. Proc. 44, 259 (1985); G. F. Koob, S. C. Heinrichs, F. Menzaghi, E. Merlo Pich, K. T. Britton, Semin. Neurosci. 6, 221 (1994); F. Menzaghi, S. C. Heinrichs, E. Merlo Pich, F. J. H. Tilders, G. F. Koob, Neurosci. Lett. 168, 139 (1994);
31. R. M. Richter, E. Merlo Pich, G. F. Koob, F. Weiss, ibid. 187, 169 (1995).
32. Male Wistar rats (250 to 300 g) were used. HU-210 was provided by R. Mechoulam (Hebrew University of Jerusalem). SR 141716A was obtained through SANOFI Recherche (Montpellier, France). Both drugs were prepared in a vehicle solution of saline, propylene glycol, and Tween 80 (90:5:5). Doses were selected on the basis of full dose-response studies (6, 12). Drugs were administered intraperitoneally in a volume of 1 ml per kilogram of body weight. Animals assigned to the cannabinoid withdrawal condition received daily injections of HU-210 (100 μg/kg) for 14 days.
33. Intracranial microdialysis for CRF was performed as described (10). Fractions of the perfusate were collected at 20-min intervals in polyethylene tubes on ice. Five fractions were collected for determination of basal CRF efflux. Animals were then injected with either HU-210 (100 μg/kg), SR 141716A (3 mg/kg), or vehicle. Rats that had been pretreated with HU-210 (100 μg/kg) for 14 days (the "cannabinoid withdrawal" group) received either SR 141716A (3 mg/ kg) or vehicle. Sampling continued for six to eight fractions after drug treatments were terminated, at which time the microdialysis probes were perfused for 60 min with artificial cerebrospinal fluid containing the depolarizing agent 4-aminopyridine (5 mM) (4-AP, Sigma) to confirm the neurogenic origin of CRF. At the end of each experiment, rats were injected with a lethal dose of pentobarbital; their brains were perfused and fixed in 4% paraformaldehyde and then frozen, sectioned, and stained with cresyl violet.
34. 1,29 = 11.75, P < 0.002 compared with basal concentrations) except after single treatments with HU-210 or SR 141716A. Means ± SEM of basal CRF concentrations versus CRF concentrations after treatment with 4-AP (fmol of CRF-IR per 50 μl) were as follows: 0.99 ± 0.22 versus 2.32 ± 0.9 (vehicle); 1.2 ± 0.29 versus 1.57 ± 0.45 (HU-210); 1.07 ± 0.19 versus 1.75 ± 0.56 (SR 141716A); 0.85 ± 0.2 versus 1.51 ± 0.21 (long-term HU-210); and 0.69 ± 0.12 versus 1.30 ± 0.19 (SR 141716A after long-term HU-210).
35. 1,29 = 11.75, P < 0.002 compared with basal concentrations) except after single treatments with HU-210 or SR 141716A. Means ± SEM of basal CRF concentrations versus CRF concentrations after treatment with 4-AP (fmol of CRF-IR per 50 μl) were as follows: 0.99 ± 0.22 versus 2.32 ± 0.9 (vehicle); 1.2 ± 0.29 versus 1.57 ± 0.45 (HU-210); 1.07 ± 0.19 versus 1.75 ± 0.56 (SR 141716A); 0.85 ± 0.2 versus 1.51 ± 0.21 (long-term HU-210); and 0.69 ± 0.12 versus 1.30 ± 0.19 (SR 141716A after long-term HU-210).
36. Cannabinoid withdrawal signs were measured with counted signs (such as wet-dog shakes, compulsive grooming, and scratching sequences) and observed signs (such as ptosis, piloerection, teeth chattering, salivation, and diarrhea). Counted signs (total number of events) were summed with observed signs (events observed over a specified observation time) and subjected to parametric statistical analysis, because sums are normally distributed.
37. The defensive withdrawal test was conducted as described [L. K. Takahashi, N. H. Kalin, J. A. van de Burgt, J. E. Sherman, Behav. Neurosci. 103, 648 (1989)]. The apparatus consisted of an illuminated (350 lux) opaque open field (100 by 100 by 40 cm) marked with squares (20 by 20 cm). The field contained a cylindrical chamber (17 by 10 cm) open at one end and positioned 20 cm from one corner of the field. Rats were placed inside the chamber for 15 min and scored for (i) latency to leave the chamber (emergence latency), (ii) total time spent in the chamber, (iii) mean time spent in the chamber per entry, and (iv) motor activity (rearing and square crossings outside the chamber). Drugs were injected 30 min before the test. All animals were habituated to the apparatus before testing.
38. Plasma corticosterone concentration was monitored in separate groups of rats that were exposed to the same treatment as the animals in the microdialysis experiments. Rats were decapitated 3 hours after treatment. Corticosterone was measured by radioimmunoassay (8) with a specific antibody from Bio Clin (Cardiff, England). The detection limit was 62 pg/ml.
39. Three hours after cannabinoid treatments, rats were quickly perfused with 0.9% saline followed by 2% paraformaldehyde in isotonic sodium phosphate buffer (PBS, pH 7.4). Brains were removed, fixed in the perfusion buffer for 24 hours, stored for 3 to 7 days in a 30% solution of sucrose in PBS, sliced in 40-μm sections (Cryocut 1800; Leica, Foster City, CA), and collected in PBS. The Fos protein was quantified by immunohistochemistry analysis with affinity-purified rabbit antibodies to a peptide corresponding to human Fos amino acid residues 3 to 16 (Santa Cruz Biotechnology, Santa Cruz, CA) that was not reactive to Fos-B and Fra-1 proteins. Sections were incubated with goat antiserum to rabbit antibody in 0.3% Triton X-100 in PBS solution for 2 hours at room temperature, followed by Fos antiserum (diluted 1:1000) in 0.3% Triton X-100 containing 0.1% bovine serum albumin in PBS for 20 hours at 4°C [A. E. Ryabinin, K. R. Melia, M. Cole, E. E. Bloom, M. C. Wilson, J. Neurosci. 15, 721 (1995)].
40. J. I. Morgan and T. Curran, Annu. Rev. Neurosci. 14, 421 (1991); M. Pfahl, Endocr. Rev. 14, 651 (1993).
41. J. I. Morgan and T. Curran, Annu. Rev. Neurosci. 14, 421 (1991); M. Pfahl, Endocr. Rev. 14, 651 (1993).
42. S. Ceccatelli, M. J. Villar, M. Goldstein, T. Hökfelt, Proc. Natl. Acad. Sci. U.S.A. 86, 9569 (1989); E. Bullit, J. Comp. Neurol. 296, 517 (1990); J. Honkaniemi, Brain Res. 598, 107 (1992).
43. S. Ceccatelli, M. J. Villar, M. Goldstein, T. Hökfelt, Proc. Natl. Acad. Sci. U.S.A. 86, 9569 (1989); E. Bullit, J. Comp. Neurol. 296, 517 (1990); J. Honkaniemi, Brain Res. 598, 107 (1992).
44. S. Ceccatelli, M. J. Villar, M. Goldstein, T. Hökfelt, Proc. Natl. Acad. Sci. U.S.A. 86, 9569 (1989); E. Bullit, J. Comp. Neurol. 296, 517 (1990); J. Honkaniemi, Brain Res. 598, 107 (1992).
45. F. J. L. Arnold et al., Neuroscience 51, 377 (1992); T. Imaki, T. Shibasaki, X. Q. Wang, H. Demura, Neuroendocrinology 61, 445 (1995).
46. F. J. L. Arnold et al., Neuroscience 51, 377 (1992); T. Imaki, T. Shibasaki, X. Q. Wang, H. Demura, Neuroendocrinology 61, 445 (1995).
47. M. D. Hayward, R. S. Duman, E. J. Nestler, Brain Res. 525, 256 (1990); R. L. Stometta, F. E. Norton, P. G. Guyenet, ibid. 624, 19 (1993); A. M. Beckmann, I. Matsumoto, P. A. Wilce, Neuropharmacology 34, 1183 (1995).
48. M. D. Hayward, R. S. Duman, E. J. Nestler, Brain Res. 525, 256 (1990); R. L. Stometta, F. E. Norton, P. G. Guyenet, ibid. 624, 19 (1993); A. M. Beckmann, I. Matsumoto, P. A. Wilce, Neuropharmacology 34, 1183 (1995).
49. M. D. Hayward, R. S. Duman, E. J. Nestler, Brain Res. 525, 256 (1990); R. L. Stometta, F. E. Norton, P. G. Guyenet, ibid. 624, 19 (1993); A. M. Beckmann, I. Matsumoto, P. A. Wilce, Neuropharmacology 34, 1183 (1995).
50. 1,32 = 3.08, 0.05 < P < 0.1].
51. In the defensive withdrawal test, a single dose of HU-210 in drug-naïve rats also produced an anxiety-like effect. An important factor in the subjective reaction to cannabinoids is dosage (1). Low doses of HU-210 abolish the behavioral response to novelty and inhibit the HPA stress response, whereas higher doses, particularly under conditions of novelty, have the opposite effect (12), as in the test described here. Comparative analysis of the patterns of Fos expression in the withdrawal and short-term HU-210 treatment conditions demonstrated an overlap as well as a dissociation of affected brain regions (Table 2), implicating the involvement of different neural substrates in the anxiety-like response induced by a single high dose of cannabinoid as opposed to withdrawal from long-term cannabinoid exposure. In the central amygdala, Fos expression appeared dispersed after a single injection of HU-210, whereas after antagonist-induced withdrawal Fos-positive nuclei were densely distributed (Fig. 2). In the BNST, immunopositive cells were found in a more medial-anterior gradient during cannabinoid withdrawal, whereas Fos activation was more prominent in the lateral dorsal region after short-term cannabinoid exposure (Table 2). In the hypothalamus, the PVN exhibited less Fos immunoreactivity during cannabinoid withdrawal compared with the effects of a single treatment with cannabinoid agonist. Thus, HPA activation after a single exposure to HU-210 in drug-naïve rats appears to be mediated directly by the PVN, whereas the increase in plasma corticosterone concentrations during withdrawal may involve activation of the central amygdala, transmitted to the PVN through its direct connections or by the BNST, which, in turn, may also activate the PVN. Because both the central amygdala and PVN are thought to be involved in anxiety-like behavioral responses to stress (13, 14), these observations suggest that the balance between the contributions of both structures after acute cannabinoid treatment or antagonist-induced withdrawal may result in the particular behavioral reactivity to the novelty condition in the defensive withdrawal test.
52. P. J. Larsen and D. Mikkelsen, J. Neurosci. 15, 2609 (1995).
53. U. Whanschaffe, U. Ebert, W. Löscher, Brain Res. 615, 295 (1993).
54. M. Navarro et al., Neuroreport 8, 491 (1997).
55. F. Rodríguez de Fonseca, M. R. A. Carrera, M. Navarro, G. F. Koob, F. Weiss, data not shown.
56. Supported by National Institute on Drug Abuse grant DA 08426 (F.W.); National Institute of Diabetes, Digestive and Kidney Diseases grant DK 26741 (G.F.K. and M.R.A.C.); and Comisión Interministerial de Ciencia y Tecnología grant SAF 94/0465, multidisciplinary grant PR218/94-5670, and Comunidad Autónoma de Madrid grant CAM-AE00340/95 (M.N. and F.R.d.F.). F.R.d.F. is a research fellow of the Fundación Jaime del Amo, Universidad Complutense de Madrid. We thank M. Wilson for providing facilities and help with Fos immunohistochemistry, R. Mechoulam for HU-210, M. A. Villanúa and R. M. Muñoz for measuring corticosterone, Y. Martin for assistance with behavioral procedures, and R. Schroeder for technical assistance with the CRF radioimmunoassay.