Skip to main content
PMC home page
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1988 Sep;95(1):109–120. doi: 10.1111/j.1476-5381.1988.tb16554.x

Effects of non-sedative anxiolytic drugs on responses to GABA and on diazepam-induced enhancement of these responses on mouse neurones in cell culture.

P P De Deyn 1, R L Macdonald 1
PMCID: PMC1854132  PMID: 2905900

Abstract

1. Intracellular microelectrode recording techniques were performed on mouse spinal cord and cerebral hemisphere neurones grown in primary dissociated cell culture. The effects of several anxiolytics applied by local pressure ejection on responses to gamma-aminobutyric acid (GABA) evoked by iontophoresis were investigated. Responses to GABA were depolarizing since intracellular chloride ion concentration was increased by injection from potassium chloride (3M)-filled recording micropipettes and neurones were held at large negative membrane potentials (-70 to -90 mV). The agents studied were six 'non-sedative anxiolytics', CL 218,872 (3-methyl-6-(3-trifluoromethyl-phenyl)1,2,4-triazolo(4,3-b) pyridazine), PK 8165 (2-phenyl-4-(2-(4-piperidinyl)ethyl)-quinoline), PK 9084 (2-phenyl-4-(2-(3-piperidinyl)ethyl)-quinoline), CGS 9896 (2-(4-chlorophenyl)-2,5-dihydropyrazolo(4,3-c)quinoline-3(3H)-one) , ZK 91296 (ethyl 5-benzyloxy-4-methoxymethyl-beta-carboline-3-beta-carboxylate), buspirone (8-4-[4-(2-pyrimidinyl)-1-piperazinyl]butyl-8-azaspiro[4.5]decane- 7,9- dione), and two sedative anxiolytics, diazepam and zopiclone [( 6-(5-chloro-2-pyridyl)-6,7-dihydro-7-oxo-5H-pyrrolo[3,4-b]pyrazin- 5- yl]4-methyl-1-piperazinecarboxylate). 2. Direct effects on responses to GABA were studied for all drugs applied in varying concentrations. For the drugs which significantly altered responses to GABA, the effects of the benzodiazepine receptor antagonists Ro 15-1788 (ethyl-8-fluoro-5,6-dihydro-5-methyl-6-oxo-4H-imidazo(1,5a)-(1,4)benzodi azepine - 3-carboxylate) and CGS 8216 (2-phenylpyrazolo(4,3-c)-quinolin-3(5H)-one) were evaluated. For the drugs devoid of significant direct effect on responses to GABA, the influence on diazepam-induced enhancement of responses to GABA was evaluated. 3. Diazepam, zopiclone and CL 218,872 concentration-dependently and reversibly enhanced responses to GABA. Maximal enhancement was 82% for diazepam (500 nM), 64% for zopiclone (10 microM) and 20% for CL 218,872 (10 microM). PK 8165 effects varied with concentration, enhancing responses to GABA (up to 18%) at nM concentrations and reducing responses to GABA (up to 90%) at microM concentrations. CGS 9896, ZK 9126, PK 9084 and buspirone, in concentrations ranging from 1 nM to 10 microM, lacked significant direct effects on responses to GABA.(ABSTRACT TRUNCATED AT 400 WORDS)

Full text

PDF
113

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allgulander C. Dependence on sedative and hypnotic drugs. A comparative clinical and social study. Acta Psychiatr Scand Suppl. 1978;(270):1–102. [PubMed] [Google Scholar]
  2. Benavides J., Malgouris C., Flamier A., Tur C., Quarteronet D., Begassat F., Camelin J. C., Uzan A., Gueremy C., Le Fur G. Biochemical evidence that 2-phenyl-4[(4-piperidinyl) ethyl]quinoline, a quinoline derivative with pure anticonflict properties, is a partial agonist of benzodiazepine receptors. Neuropharmacology. 1984 Oct;23(10):1129–1136. doi: 10.1016/0028-3908(84)90229-6. [DOI] [PubMed] [Google Scholar]
  3. Bernard P. S., Bennett D. A., Pastor G., Yokoyama N., Liebman J. M. CGS 9896: agonist-antagonist benzodiazepine receptor activity revealed by anxiolytic, anticonvulsant and muscle relaxation assessment in rodents. J Pharmacol Exp Ther. 1985 Oct;235(1):98–105. [PubMed] [Google Scholar]
  4. Blanchard J. C., Boireau A., Garret C., Julou L. In vitro and in vivo inhibition by zopiclone of benzodiazepine binding to rodent brain receptors. Life Sci. 1979 Jun 25;24(26):2417–2420. doi: 10.1016/0024-3205(79)90449-1. [DOI] [PubMed] [Google Scholar]
  5. Blanchard J. C., Boireau A., Julou L. Brain receptors and zopiclone. Pharmacology. 1983;27 (Suppl 2):59–69. doi: 10.1159/000137912. [DOI] [PubMed] [Google Scholar]
  6. Boast C. A., Snowhill E. W., Simke J. P. CGS 8216 and CGS 9896, novel pyrazoloquinoline benzodiazepine ligands with benzodiazepine agonist and antagonist properties. Pharmacol Biochem Behav. 1985 Oct;23(4):639–644. doi: 10.1016/0091-3057(85)90431-9. [DOI] [PubMed] [Google Scholar]
  7. Braestrup C., Nielsen M. Anxiety. Lancet. 1982 Nov 6;2(8306):1030–1034. doi: 10.1016/s0140-6736(82)90059-9. [DOI] [PubMed] [Google Scholar]
  8. Brown C., Martin I., Jones B., Oakley N. In vivo determination of efficacy of pyrazoloquinolinones at the benzodiazepine receptor. Eur J Pharmacol. 1984 Aug 3;103(1-2):139–143. doi: 10.1016/0014-2999(84)90200-0. [DOI] [PubMed] [Google Scholar]
  9. Choi D. W., Farb D. H., Fischbach G. D. Chlordiazepoxide selectively augments GABA action in spinal cord cell cultures. Nature. 1977 Sep 22;269(5626):342–344. doi: 10.1038/269342a0. [DOI] [PubMed] [Google Scholar]
  10. Costa E., Guidotti A. Endogenous ligands for benzodiazepine recognition sites. Biochem Pharmacol. 1985 Oct 1;34(19):3399–3403. doi: 10.1016/0006-2952(85)90709-9. [DOI] [PubMed] [Google Scholar]
  11. Costa E., Guidotti A. Molecular mechanisms in the receptor action of benzodiazepines. Annu Rev Pharmacol Toxicol. 1979;19:531–545. doi: 10.1146/annurev.pa.19.040179.002531. [DOI] [PubMed] [Google Scholar]
  12. De Deyn P. P., Macdonald R. L. CGS 9896 and ZK 91296, but not CGS 8216 and RO 15-1788, are pure benzodiazepine receptor antagonists on mouse neurons in culture. J Pharmacol Exp Ther. 1987 Jul;242(1):48–55. [PubMed] [Google Scholar]
  13. Duriez R., Barthelemy C., Rives H., Courjaret J., Gregoire J. Traitement des troubles du sommeil par la zopiclone. Essais cliniques en double insu contre placebo. Therapie. 1979 May-Jun;34(3):317–325. [PubMed] [Google Scholar]
  14. Ehlert F. J., Ragan P., Chen A., Roeske W. R., Yamamura H. I. Modulation of benzodiazepine receptor binding: insight into pharmacological efficacy. Eur J Pharmacol. 1982 Feb 26;78(2):249–253. doi: 10.1016/0014-2999(82)90246-1. [DOI] [PubMed] [Google Scholar]
  15. FINK G. B., SWINYARD E. A. Comparison of anticonvulsant and psychopharmacologic drugs. J Pharm Sci. 1962 Jun;51:548–551. doi: 10.1002/jps.2600510612. [DOI] [PubMed] [Google Scholar]
  16. File S. E., Lister R. G. Quinolines and anxiety: anxiogenic effects of CGS 8216 and partial anxiolytic profile of PK 9084. Pharmacol Biochem Behav. 1983 Feb;18(2):185–188. doi: 10.1016/0091-3057(83)90361-1. [DOI] [PubMed] [Google Scholar]
  17. File S. E., Pellow S., Wilks L. The sedative effects of CL 218,872, like those of chlordiazepoxide, are reversed by benzodiazepine antagonists. Psychopharmacology (Berl) 1985;85(3):295–300. doi: 10.1007/BF00428190. [DOI] [PubMed] [Google Scholar]
  18. File S. E. Sedative effects of PK 9084 and PK 8165, alone and in combination with chlordiazepoxide. Br J Pharmacol. 1983 May;79(1):219–223. doi: 10.1111/j.1476-5381.1983.tb10515.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. File S. E., Simmonds M. A. Interactions of two phenylquinolines with picrotoxin and benzodiazepines in vivo and in vitro. Eur J Pharmacol. 1984 Jan 27;97(3-4):295–300. doi: 10.1016/0014-2999(84)90463-1. [DOI] [PubMed] [Google Scholar]
  20. Gee K. W., Brinton R. E., Yamamura H. I. PK 8165 and PK 9084, two quinoline derivatives with anxiolytic properties, antagonize the anticonvulsant effects of diazepam. Brain Res. 1983 Mar 28;264(1):168–172. doi: 10.1016/0006-8993(83)91138-1. [DOI] [PubMed] [Google Scholar]
  21. Gee K. W., Morelli M., Yamamura H. I. The effect of temperature on CL 218872 and propyl beta-carboline-3-carboxylate inhibition of [3H]-flunitrazepam binding in rat brain. Biochem Biophys Res Commun. 1982 Apr 29;105(4):1532–1537. doi: 10.1016/0006-291x(82)90962-7. [DOI] [PubMed] [Google Scholar]
  22. Gee K. W., Yamamura H. I. A novel pyrazoloquinoline that interacts with brain benzodiazepine receptors: characterization of some in vitro and in vivo properties of CGS 9896. Life Sci. 1982 Jun 28;30(26):2245–2252. doi: 10.1016/0024-3205(82)90251-x. [DOI] [PubMed] [Google Scholar]
  23. Goldberg H. L., Finnerty R. J. The comparative efficacy of buspirone and diazepam in the treatment of anxiety. Am J Psychiatry. 1979 Sep;136(9):1184–1187. doi: 10.1176/ajp.136.9.1184. [DOI] [PubMed] [Google Scholar]
  24. Goldberg H. L., Finnerty R. Comparison of buspirone in two separate studies. J Clin Psychiatry. 1982 Dec;43(12 Pt 2):87–91. [PubMed] [Google Scholar]
  25. Heyer E. J., Nowak L. M., Macdonald R. L. Membrane depolarization and prolongation of calcium-dependent action potentials of mouse neurons in cell culture by two convulsants: bicuculline and penicillin. Brain Res. 1982 Jan 28;232(1):41–56. doi: 10.1016/0006-8993(82)90609-6. [DOI] [PubMed] [Google Scholar]
  26. Hoehn-Saric R. Neurotransmitters in anxiety. Arch Gen Psychiatry. 1982 Jun;39(6):735–742. doi: 10.1001/archpsyc.1982.04290060075015. [DOI] [PubMed] [Google Scholar]
  27. Hunkeler W., Möhler H., Pieri L., Polc P., Bonetti E. P., Cumin R., Schaffner R., Haefely W. Selective antagonists of benzodiazepines. Nature. 1981 Apr 9;290(5806):514–516. doi: 10.1038/290514a0. [DOI] [PubMed] [Google Scholar]
  28. Julou L., Bardone M. C., Blanchard J. C., Garret C., Stutzmann J. M. Pharmacological studies on zopiclone. Pharmacology. 1983;27 (Suppl 2):46–58. doi: 10.1159/000137911. [DOI] [PubMed] [Google Scholar]
  29. Karobath M., Supavilai P., Borea P. A. Distinction of benzodiazepine receptor agonists and inverse agonists by binding studies in vitro. Adv Biochem Psychopharmacol. 1983;38:37–45. [PubMed] [Google Scholar]
  30. Keane P. E., Simiand J., Morre M. The quinolines PK 8165 and PK 9084 possess benzodiazepine-like activity in vitro but not in vivo. Neurosci Lett. 1984 Mar 9;45(1):89–93. doi: 10.1016/0304-3940(84)90334-3. [DOI] [PubMed] [Google Scholar]
  31. Klepner C. A., Lippa A. S., Benson D. I., Sano M. C., Beer B. Resolution of two biochemically and pharmacologically distinct benzodiazepine receptors. Pharmacol Biochem Behav. 1979 Oct;11(4):457–462. doi: 10.1016/0091-3057(79)90125-4. [DOI] [PubMed] [Google Scholar]
  32. Klockgether T., Schwarz M., Turski L., Sontag K. H. ZK 91296, an anticonvulsant beta-carboline which lacks muscle relaxant properties. Eur J Pharmacol. 1985 Apr 16;110(3):309–315. doi: 10.1016/0014-2999(85)90557-6. [DOI] [PubMed] [Google Scholar]
  33. Lamb R. J., Griffiths R. R. Precipitated and spontaneous withdrawal in baboons after chronic dosing with lorazepam and CGS 9896. Drug Alcohol Depend. 1984 Sep;14(1):11–17. doi: 10.1016/0376-8716(84)90013-9. [DOI] [PubMed] [Google Scholar]
  34. Le Fur G., Mizoule J., Burgevin M. C., Ferris O., Heaulme M., Gauthier A., Guérémy C., Uzan A. Multiple benzodiazepine receptors: evidence of dissociation between anticonflict and anticonvulsant properties by PK 8165 and PK 9084 (two quinoline derivatives). Life Sci. 1981 Mar 30;28(13):1439–1448. doi: 10.1016/0024-3205(81)90375-1. [DOI] [PubMed] [Google Scholar]
  35. Lippa A. S., Coupet J., Greenblatt E. N., Klepner C. A., Beer B. A synthetic non-benzodiazepine ligand for benzodiazepine receptors: a probe for investigating neuronal substrates of anxiety. Pharmacol Biochem Behav. 1979 Jul;11(1):99–106. doi: 10.1016/0091-3057(79)90304-6. [DOI] [PubMed] [Google Scholar]
  36. Lippa A. S., Critchett D., Sano M. C., Klepner C. A., Greenblatt E. N., Coupet J., Beer B. Benzodiazepine receptors: cellular and behavioral characteristics. Pharmacol Biochem Behav. 1979 May;10(5):831–843. doi: 10.1016/0091-3057(79)90342-3. [DOI] [PubMed] [Google Scholar]
  37. MacDonald R. L., Barker J. L. Enhancement of GABA-mediated postsynaptic inhibition in cultured mammalian spinal cord neurons: a common mode of anticonvulsant action. Brain Res. 1979 May 11;167(2):323–336. doi: 10.1016/0006-8993(79)90826-6. [DOI] [PubMed] [Google Scholar]
  38. Macdonald R., Barker J. L. Benzodiazepines specifically modulate GABA-mediated postsynaptic inhibition in cultured mammalian neurones. Nature. 1978 Feb 9;271(5645):563–564. doi: 10.1038/271563a0. [DOI] [PubMed] [Google Scholar]
  39. McElroy J. F., Fleming R. L., Feldman R. S. A comparison between chlordiazepoxide and CL 218,872--a synthetic nonbenzodiazepine ligand for benzodiazepine receptors on spontaneous locomotor activity in rats. Psychopharmacology (Berl) 1985;85(2):224–226. doi: 10.1007/BF00428419. [DOI] [PubMed] [Google Scholar]
  40. Morelli M., Gee K. W., Yamamura H. I. The effect of GABA on in vitro binding of two novel non-benzodiazepines, PK 8165 and CGS 8216, to benzodiazepine receptors in the rat brain. Life Sci. 1982 Jul 5;31(1):77–81. doi: 10.1016/0024-3205(82)90403-9. [DOI] [PubMed] [Google Scholar]
  41. Möhler H., Okada T. Benzodiazepine receptor: demonstration in the central nervous system. Science. 1977 Nov 25;198(4319):849–851. doi: 10.1126/science.918669. [DOI] [PubMed] [Google Scholar]
  42. Newton R. E., Casten G. P., Alms D. R., Benes C. O., Marunycz J. D. The side effect profile of buspirone in comparison to active controls and placebo. J Clin Psychiatry. 1982 Dec;43(12 Pt 2):100–102. [PubMed] [Google Scholar]
  43. Nicoll R. A., Eccles J. C., Oshima T., Rubia F. Prolongation of hippocampal inhibitory postsynaptic potentials by barbiturates. Nature. 1975 Dec 18;258(5536):625–627. doi: 10.1038/258625a0. [DOI] [PubMed] [Google Scholar]
  44. Nishi S., Minota S., Karczmar A. G. Primary afferent neurones: the ionic mechanism of GABA-mediated depolarization. Neuropharmacology. 1974 Mar;13(3):215–219. doi: 10.1016/0028-3908(74)90110-5. [DOI] [PubMed] [Google Scholar]
  45. Nowak L. M., Young A. B., Macdonald R. L. GABA and bicuculline actions on mouse spinal cord and cortical neurons in cell culture. Brain Res. 1982 Jul 22;244(1):155–164. doi: 10.1016/0006-8993(82)90913-1. [DOI] [PubMed] [Google Scholar]
  46. Oakley N. R., Jones B. J., Straughan D. W. The benzodiazepine receptor ligand CL218,872 has both anxiolytic and sedative properties in rodents. Neuropharmacology. 1984 Jul;23(7A):797–802. doi: 10.1016/0028-3908(84)90114-x. [DOI] [PubMed] [Google Scholar]
  47. Pellow S., File S. E. Evidence that the beta-carboline, ZK 91296, can reduce anxiety in animals at doses well below those causing sedation. Brain Res. 1986 Jan 15;363(1):174–177. doi: 10.1016/0006-8993(86)90674-8. [DOI] [PubMed] [Google Scholar]
  48. Petersen E. N., Jensen L. H., Honoré T., Braestrup C., Kehr W., Stephens D. N., Wachtel H., Seidelman D., Schmiechen R. ZK 91296, a partial agonist at benzodiazepine receptors. Psychopharmacology (Berl) 1984;83(3):240–248. doi: 10.1007/BF00464788. [DOI] [PubMed] [Google Scholar]
  49. Petrack B., Czernik A. J., Cassidy J. P., Bernard P., Yokoyama N. Benzodiazepine receptor ligands with opposing pharmacologic actions. Adv Biochem Psychopharmacol. 1983;38:129–137. [PubMed] [Google Scholar]
  50. Ransom B. R., Neale E., Henkart M., Bullock P. N., Nelson P. G. Mouse spinal cord in cell culture. I. Morphology and intrinsic neuronal electrophysiologic properties. J Neurophysiol. 1977 Sep;40(5):1132–1150. doi: 10.1152/jn.1977.40.5.1132. [DOI] [PubMed] [Google Scholar]
  51. Riblet L. A., Taylor D. P., Eison M. S., Stanton H. C. Pharmacology and neurochemistry of buspirone. J Clin Psychiatry. 1982 Dec;43(12 Pt 2):11–18. [PubMed] [Google Scholar]
  52. Rickels K., Weisman K., Norstad N., Singer M., Stoltz D., Brown A., Danton J. Buspirone and diazepam in anxiety: a controlled study. J Clin Psychiatry. 1982 Dec;43(12 Pt 2):81–86. [PubMed] [Google Scholar]
  53. Skerritt J. H., Macdonald R. L. Benzodiazepine Ro 15-1788: electrophysiological evidence for partial agonist activity. Neurosci Lett. 1983 Dec 30;43(2-3):321–326. doi: 10.1016/0304-3940(83)90208-2. [DOI] [PubMed] [Google Scholar]
  54. Skerritt J. H., Macdonald R. L. Benzodiazepine receptor ligand actions on GABA responses. Benzodiazepines, CL 218872, zopiclone. Eur J Pharmacol. 1984 May 18;101(1-2):127–134. doi: 10.1016/0014-2999(84)90038-4. [DOI] [PubMed] [Google Scholar]
  55. Skerritt J. H., Werz M. A., McLean M. J., Macdonald R. L. Diazepam and its anomalous p-chloro-derivative Ro 5-4864: comparative effects on mouse neurons in cell culture. Brain Res. 1984 Sep 17;310(1):99–105. doi: 10.1016/0006-8993(84)90013-1. [DOI] [PubMed] [Google Scholar]
  56. Squires R. F., Benson D. I., Braestrup C., Coupet J., Klepner C. A., Myers V., Beer B. Some properties of brain specific benzodiazepine receptors: new evidence for multiple receptors. Pharmacol Biochem Behav. 1979 May;10(5):825–830. doi: 10.1016/0091-3057(79)90341-1. [DOI] [PubMed] [Google Scholar]
  57. Squires R. F., Brastrup C. Benzodiazepine receptors in rat brain. Nature. 1977 Apr 21;266(5604):732–734. doi: 10.1038/266732a0. [DOI] [PubMed] [Google Scholar]
  58. Stephens D. N., Kehr W., Schneider H. H., Schmiechen R. Beta-carbolines with agonistic and inverse agonistic properties at benzodiazepine receptors of the rat. Neurosci Lett. 1984 Jun 29;47(3):333–338. doi: 10.1016/0304-3940(84)90535-4. [DOI] [PubMed] [Google Scholar]
  59. Trifiletti R. R., Snyder S. H. Anxiolytic cyclopyrrolones zopiclone and suriclone bind to a novel site linked allosterically to benzodiazepine receptors. Mol Pharmacol. 1984 Nov;26(3):458–469. [PubMed] [Google Scholar]
  60. Wood P. L., Loo P., Braunwalder A., Yokoyama N., Cheney D. L. In vitro characterization of benzodiazepine receptor agonists, antagonists, inverse agonists and agonist/antagonists. J Pharmacol Exp Ther. 1984 Dec;231(3):572–576. [PubMed] [Google Scholar]
  61. Yokoyama N., Ritter B., Neubert A. D. 2-Arylpyrazolo[4,3-c]quinolin-3-ones: novel agonist, partial agonist, and antagonist of benzodiazepines. J Med Chem. 1982 Apr;25(4):337–339. doi: 10.1021/jm00346a002. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Pharmacology are provided here courtesy of The British Pharmacological Society

RESOURCES