Fyn-kinase as a determinant of ethanol sensitivity: Relation to NMDA- Receptor function

Science

Volumn 278, Issue 5338, 1997, Pages 698-701

  Miyakawa, Tsuyoshi ^a   Yagi, Takeshi ^a   Kitazawa, Hiromasa ^a   Yasuda, Masahiro ^a   Kawai, Nobufumi ^a   Tsuboi, Kenta ^a   Niki, Hiroaki ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

4 AMINOBUTYRIC ACID A RECEPTOR; ALCOHOL; N METHYL DEXTRO ASPARTIC ACID RECEPTOR; PROTEIN TYROSINE KINASE;

ALCOHOL ABUSE; ANIMAL BEHAVIOR; ANIMAL EXPERIMENT; ARTICLE; CENTRAL NERVOUS SYSTEM; CONTROLLED STUDY; HIPPOCAMPUS; MOUSE; NEUROTRANSMISSION; NONHUMAN; PRIORITY JOURNAL; PROTEIN PHOSPHORYLATION; SUBSTANCE ABUSE;

ANIMALS; CENTRAL NERVOUS SYSTEM DEPRESSANTS; ETHANOL; EXCITATORY POSTSYNAPTIC POTENTIALS; FLURAZEPAM; HIPPOCAMPUS; HYPNOTICS AND SEDATIVES; MALE; MICE; MICE, KNOCKOUT; MOTOR ACTIVITY; N-METHYLASPARTATE; PHOSPHORYLATION; PHOSPHOTYROSINE; PROTEIN-TYROSINE KINASES; PROTO-ONCOGENE PROTEINS; PROTO-ONCOGENE PROTEINS C-FYN; RECEPTORS, N-METHYL-D-ASPARTATE;

ANIMALIA;

EID: 0030680941     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.278.5338.698     Document Type: Article

References (31)

1. R. A. Harris, Ann. N.Y. Acad. Sci. 625, 508 (1991); J. C. Crabbe, J. K. Belknap, K. J. Buck, Science 264, 1715 (1994).
2. R. A. Harris, Ann. N.Y. Acad. Sci. 625, 508 (1991); J. C. Crabbe, J. K. Belknap, K. J. Buck, Science 264, 1715 (1994).
3. J. C. Crabbe et al., Nature Genet. 14, 98 (1996).
4. R. A. Harris et al., Proc. Natl. Acad. Sci. U.S.A. 92, 3658 (1995).
5. Y. T. Wang and M. W. Salter, Nature 369, 233 (1994); X.-M. Yu, R. Askalan, G. J. Keil II, M. W. Salter, Science 275, 674 (1997).
6. Y. T. Wang and M. W. Salter, Nature 369, 233 (1994); X.-M. Yu, R. Askalan, G. J. Keil II, M. W. Salter, Science 275, 674 (1997).
7. T. Suzuki and K. Okumura-Noji, Biochem. Biophys. Res. Commun. 216, 582 (1995).
8. S. J. Moss, G. H. Gorrie, A. Amato, T. G. Smart, Nature 377, 344 (1995); C. F. Valenzuela et al., Mol. Brain Res. 31, 165 (1995).
9. S. J. Moss, G. H. Gorrie, A. Amato, T. G. Smart, Nature 377, 344 (1995); C. F. Valenzuela et al., Mol. Brain Res. 31, 165 (1995).
10. D. M. Lovinger, G. White, F. F. Weight, Science 243, 1721 (1989); J. Neurosci. 10, 1372 (1990); K. A. Wafford, D. M. Burnett, T. V. Dunwiddie, R. A. Harris, Science 249, 291 (1990).
11. D. M. Lovinger, G. White, F. F. Weight, Science 243, 1721 (1989); J. Neurosci. 10, 1372 (1990); K. A. Wafford, D. M. Burnett, T. V. Dunwiddie, R. A. Harris, Science 249, 291 (1990).
12. D. M. Lovinger, G. White, F. F. Weight, Science 243, 1721 (1989); J. Neurosci. 10, 1372 (1990); K. A. Wafford, D. M. Burnett, T. V. Dunwiddie, R. A. Harris, Science 249, 291 (1990).
13. z mice, 34.8 ± 3.2 min; P > 0.05]. Testing was double-blind [T. Miyakawa, T. Yagi, S. Watanabe, H. Niki, Mol. Brain Res. 27, 179 (1994)]. Averaged values in the text and figures are mean ± SEM, unless otherwise noted. The EtOH (Wako Pure Chemicals), Osaka, Japan was dissolved in physiological saline [20% (v/v)] and administered intraperitoneally in all behavioral and biochemical experiments. Flurazepam was dissolved in 0.2 N HCl, diluted with saline, and administered intraperitoneally. The duration of LORR was measured as described (3).
14. z mice, 34.8 ± 3.2 min; P > 0.05]. Testing was double-blind [T. Miyakawa, T. Yagi, S. Watanabe, H. Niki, Mol. Brain Res. 27, 179 (1994)]. Averaged values in the text and figures are mean ± SEM, unless otherwise noted. The EtOH (Wako Pure Chemicals), Osaka, Japan was dissolved in physiological saline [20% (v/v)] and administered intraperitoneally in all behavioral and biochemical experiments. Flurazepam was dissolved in 0.2 N HCl, diluted with saline, and administered intraperitoneally. The duration of LORR was measured as described (3).
15. z mice, 34.8 ± 3.2 min; P > 0.05]. Testing was double-blind [T. Miyakawa, T. Yagi, S. Watanabe, H. Niki, Mol. Brain Res. 27, 179 (1994)]. Averaged values in the text and figures are mean ± SEM, unless otherwise noted. The EtOH (Wako Pure Chemicals), Osaka, Japan was dissolved in physiological saline [20% (v/v)] and administered intraperitoneally in all behavioral and biochemical experiments. Flurazepam was dissolved in 0.2 N HCl, diluted with saline, and administered intraperitoneally. The duration of LORR was measured as described (3).
16. Blood samples of about 10 μl were obtained from the tail 5, 20, 60, 120, and 240 min after administration of 3.5 g/kg EtOH. Blood EtOH concentration was quantified with the use of a Sigma Alcohol reagents kit with some minor modification of instruction.
17. A. W. Thurston Jr. and S. D. Shukla, Biochem. Biophys. Res. Commun. 185, 1062 (1992).
18. R. Roivainen, B. Hundle, R. O. Messing, Proc. Natl. Acad. Sci. U.S.A. 92, 1891 (1995).
19. z 5 min after saline administration). Five animals per condition were used.
20. I. S. Moon, M. L. Apperson, M. B. Kennedy, Proc. Natl. Acad. Sci. U.S.A. 91, 3954 (1994).
21. M. Menegoz, L-F. Lau, D. Herve, R. L. Huganir, J.-A. Girault, Neuroreport 7, 125 (1995).
22. K. Rosenblum, D. E. Berman, S. Hazvi, R. Lamprecht, Y. Dudai, J. Neurosci. 17, 5129 (1997).
23. J. A. P. Rostas et al., Proc. Natl. Acad. Sci. U.S.A. 93, 10452 (1996); K. Rosenblum, Y. Dudai, G. Richter-Levin, ibid., p. 10457.
24. J. A. P. Rostas et al., Proc. Natl. Acad. Sci. U.S.A. 93, 10452 (1996); K. Rosenblum, Y. Dudai, G. Richter-Levin, ibid., p. 10457.
25. Cell lysates from hippocampus obtained 5 min after saline or EtOH (3.5 g/kg) administration were homogenized in lysis buffer [10 mM tris-HCl (pH 7.4), 1% Triton X-100, 0.1% sodium deoxycholate, 0.1% SDS, 0.15 M NaCl, 1 mM EDTA, 1 mM sodium orthovanadate, and protease inhibitors as listed in (12)], sonicated, centrifuged to remove insoluble material, and precleared with protein G-Sepharose. Samples were incubated for 1 hour at 4°C with antibody to NMDARε1 or antibody to NMDARε2 and then incubated for 1 hour at 4°C with added protein G-Sepharose. After five washes, pellets were boiled for 5 min in 2x SDS sample buffer, subjected to SDS-PAGE, immunoblotted, and quantified as described above with antibody to NMDARε1, to NMDARε2, or to P-Tyr. Six animals per condition were used.
26. C. A. Grover, G. D. Frye, W. H. Griffith, Brain Res. 642, 70 (1994).
27. 4 (0.1 mM). Schaffer collateral/commissural afferents were stimulated with 0.1-Hz pulses. The extracellular field EPSPs were recorded in the stratum radiatum of the CA1 subfield, with glass micropipettes filled with Ringer solution. The maximal rate of change in field EPSP within a time window selected around the rising phase was calculated. To obtain NMDA-mediated EPSPs, we perfused slices with 10 μM 6-cyano-7-nitroquinoxaline-2.3-dione (CNQX) (Tocris, Bristol, UK). The EtOH (60 to 70 mM), DL-2-aminophosphonovalerate (APV) (Sigma, 10 μM), and ifenprodil (Wako, 10 μM) were applied in the perfusing medium. Little or no inhibitory effect of EtOH on EPSPs mediated by non-NMDA receptors could be noted in the presence of APV, as reported (7).
28. K. Williams, Mol. Pharmacol. 44, 851 (1993).
29. The tolerance to EtOH inhibition seemed longer lasting in physiological data (Fig. 3) than the up-regulation of tyrosine phosphorylation (Fig. 2). This discrepancy may be due to the difference in the preparation used (whole tissues versus slices) or in EtOH administration procedures. The other possibility is that the up-regulation of tyrosine phosphorylation is a trigger for acute tolerance to EtOH. Concerning tyrosine phosphorylation of the NMDARζ1 subunit, 115-kd bands immunoprecipitated with antibody to NMDARζ1 (Santa Cruz Biotechnology, Santa Cruz, CA) were scarcely stained by anti P-Tyr in our supplementary experiments, as others reported [L.-F. Lau and R. L. Huganir, J. Biol. Chem. 270, 20036 (1995)]. No changes in this hardly detectable band nor in the amount of NMDARζ1 protein were found by EtOH administration.
30. B. Tabakoff and P. L. Hoffman, Neuron 16, 909 (1996).
31. We thank M. Mishina and K. Sakimura for providing antibodies against NMDARε1 and ε2. Flurazepam was kindly provided by Kyorin Pharmaceutical company. This work was supported by Grants-in-Aid for Scientific Research to H.N., N. K., and T.Y. from the Japanese Ministry of Education, Science, Sports, and Culture and by a fellowship grant to T.M. from the Japan Society for the Promotion of Science.