Impaired cued and contextual memory in NPAS2-deficient mice

Science

Volumn 288, Issue 5474, 2000, Pages 2226-2230

  Garcia, Joseph A ^a   Zhang, Di ^a   Estill, Sandi Jo ^a   Michnoff, Carolyn ^a   Rutter, Jared ^a   Reick, Martin ^a   Scott, Kristin ^a   Diaz Arrastia, Ramon ^a   McKnight, Steven L ^a  

^a UNIVERSITY OF TEXAS SOUTHWESTERN MEDICAL CENTER   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

BETA GALACTOSIDASE; HELIX LOOP HELIX PROTEIN; HYBRID PROTEIN; NEURONAL PAS DOMAIN PROTEIN 2; TRANSCRIPTION FACTOR; UNCLASSIFIED DRUG;

AMNESIA; ANIMAL BEHAVIOR; ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; ARTICLE; ASSOCIATIVE MEMORY; BRAIN MAPPING; BRAIN REGION; CONTROLLED STUDY; FEAR; GENE INSERTION; LONG TERM MEMORY; MOUSE; NONHUMAN; PRIORITY JOURNAL; PROTEIN DEFICIENCY; PROTEIN EXPRESSION; PROTEIN SYNTHESIS; REPORTER GENE;

ANIMALS; AVOIDANCE LEARNING; BASIC HELIX-LOOP-HELIX TRANSCRIPTION FACTORS; BEHAVIOR, ANIMAL; BETA-GALACTOSIDASE; BRAIN; CONDITIONING (PSYCHOLOGY); CUES; FEAR; GENE TARGETING; HELIX-LOOP-HELIX MOTIFS; LEARNING; LIMBIC SYSTEM; MALE; MEMORY; MICE; NERVE TISSUE PROTEINS; PROSENCEPHALON; RECOMBINANT FUSION PROTEINS; TOUCH; TRANS-ACTIVATION (GENETICS); TRANSCRIPTION FACTORS; TRANSFECTION;

ANIMALIA; VERTEBRATA;

EID: 0034705398     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.288.5474.2226     Document Type: Article

References (50)

1. H. P. Davis and L. R. Squire, Psychol. Bull. 96, 518 (1984).
2. R. Bourtchuladze et al., Cell 79, 59 (1994); J. H. Kogan et al., Curr. Biol. 7, 1 (1996).
3. R. Bourtchuladze et al., Cell 79, 59 (1994); J. H. Kogan et al., Curr. Biol. 7, 1 (1996).
4. T. Abel and E. Kandel, Brain Res. Rev. 26, 360 (1998).
5. Y. D. Zhou et al., Proc. Natl. Acad. Sci. U.S.A. 94, 713 (1997).
6. B. A. Campbell and N. E. Spear, Psychol. Rev. 79, 215 (1972); R. J. Green and M. E. Stanton, Behav. Neurosci. 103, 98 (1989); J. W. Rudy, Behav. Neurosci. 107, 887 (1993); J. W. Rudy and P. Morledge, Behav. Neurosci. 108, 227 (1994); N. E. Spear, J. S. Miller, J. A. Jagielo, Annu. Rev. Psychol. 41, 169 (1990); D. A. Wilson, Brain Res. 318, 61 (1984).
7. B. A. Campbell and N. E. Spear, Psychol. Rev. 79, 215 (1972); R. J. Green and M. E. Stanton, Behav. Neurosci. 103, 98 (1989); J. W. Rudy, Behav. Neurosci. 107, 887 (1993); J. W. Rudy and P. Morledge, Behav. Neurosci. 108, 227 (1994); N. E. Spear, J. S. Miller, J. A. Jagielo, Annu. Rev. Psychol. 41, 169 (1990); D. A. Wilson, Brain Res. 318, 61 (1984).
8. B. A. Campbell and N. E. Spear, Psychol. Rev. 79, 215 (1972); R. J. Green and M. E. Stanton, Behav. Neurosci. 103, 98 (1989); J. W. Rudy, Behav. Neurosci. 107, 887 (1993); J. W. Rudy and P. Morledge, Behav. Neurosci. 108, 227 (1994); N. E. Spear, J. S. Miller, J. A. Jagielo, Annu. Rev. Psychol. 41, 169 (1990); D. A. Wilson, Brain Res. 318, 61 (1984).
9. B. A. Campbell and N. E. Spear, Psychol. Rev. 79, 215 (1972); R. J. Green and M. E. Stanton, Behav. Neurosci. 103, 98 (1989); J. W. Rudy, Behav. Neurosci. 107, 887 (1993); J. W. Rudy and P. Morledge, Behav. Neurosci. 108, 227 (1994); N. E. Spear, J. S. Miller, J. A. Jagielo, Annu. Rev. Psychol. 41, 169 (1990); D. A. Wilson, Brain Res. 318, 61 (1984).
10. B. A. Campbell and N. E. Spear, Psychol. Rev. 79, 215 (1972); R. J. Green and M. E. Stanton, Behav. Neurosci. 103, 98 (1989); J. W. Rudy, Behav. Neurosci. 107, 887 (1993); J. W. Rudy and P. Morledge, Behav. Neurosci. 108, 227 (1994); N. E. Spear, J. S. Miller, J. A. Jagielo, Annu. Rev. Psychol. 41, 169 (1990); D. A. Wilson, Brain Res. 318, 61 (1984).
11. B. A. Campbell and N. E. Spear, Psychol. Rev. 79, 215 (1972); R. J. Green and M. E. Stanton, Behav. Neurosci. 103, 98 (1989); J. W. Rudy, Behav. Neurosci. 107, 887 (1993); J. W. Rudy and P. Morledge, Behav. Neurosci. 108, 227 (1994); N. E. Spear, J. S. Miller, J. A. Jagielo, Annu. Rev. Psychol. 41, 169 (1990); D. A. Wilson, Brain Res. 318, 61 (1984).
12. J. E. LeDoux, Annu. Rev. Psychol. 46, 209 (1995).
13. S. M. Dymecki, Gene 171, 197 (1996).
14. Cultured 293 cells were transfected with the indicated amount of expression plasmids (wild-type NPAS2, NPAS2-ΔbHLH, and/or BMAL) plus an NPAS2 reporter plasmid. Cells were incubated for 16 hours and then were harvested in luciferase lysis buffer [30 mM Tricine (pH 7.8), 8 mM magnesium acetate, 0.2 mM EDTA, 100 mM β-mercaptoethanol, and 1% Triton-X 100]. Luciferase activity (reported as relative light units) was measured with a Torcon AML-34 luminometer (Torcon Instruments, Torrance, CA).
15. J. B. Hogenesch, Y. Z. Gu, S. Jain, C. A. Bradfield, Proc. Natl. Acad. Sci. U.S.A. 95, 5474 (1998).
16. M. Ikeda and M. Nomura, Biochem. Biophys. Res. Commun. 233, 258 (1997).
17. 2, 1X phosphate-buffered saline (PBS), and X-Gal (1 mg/ml).
18. T. A. Woolsey and H. Van der Loos, Brain Res. 17, 205 (1970).
19. 2 saturated solution containing 2 g of sucrose, 20 mg of diaminobenzidine, and 20 mg of cytochrome c in 50 ml of 0.1 M PBS (pH 7.4). Sections were stained in the dark at 37°C for 4 to 6 hours or until a brown precipitate appeared.
20. M. Wong-Riley, Brain Res. 171, 11 (1979).
21. For analysis of NPAS2-lacZ expression in the SCN or pineal gland, vibratome (100-μm-thick) or frozen (30-μm-thick) brain sections from 1-month-old male NPAS2-lacZ (-/-) were examined. Positive staining was observed outside the SCN or pineal gland in NPAS2-lacZ-expressing areas on all sections.
22. J. N. Crawley and R. Paylor, Horm. Behav. 31, 197 (1997).
23. 2 NPAS2-lacZ (+/-) mice. Male progeny were separated at 21 days of age and randomly housed with respect to genotype in groups of four or five. Mice were maintained on a cycle of 12 hours of light and 12 hours of dark (lights on at 0600 hours) and were provided with unrestricted food and water. Testing was in blinded sessions composed of equal numbers (18 to 24 per group) of age-matched (21 to 23 weeks old at onset) NPAS2-lacZ (-/-) and (+/+) littermates. One hour before behavioral testing (0800 hours), mice were transferred to the behavioral testing room for acclimatization. Part 1 of the CCF test involved a 9-min exploration of the apparatus. In part 2 (training), the mice were placed in the same apparatus 6 hours after part 1 and were allowed to explore for 3 min. The conditioned stimulus, 30 s of 80 dB white noise, followed and ended with the unconditioned stimulus, a 1-s 0.5-mA shock. This was followed by a 30-s observation interval. The conditioned stimulus-unconditioned stimulus pairing was repeated for a total of three times, making the entire test duration 6 min. In part 3, the mice were returned to the identical testing apparatus 0.5 hour [CCF 0.5 hour/contextual (Fig. 4B)] or 24 hours [CCF 24 hours/ contextual (Fig. 4A)] after the training period for a 3-min assessment of contextual learning. For 24-hour cued memory, the animals were assessed for freezing behavior 45 min after part 3, first in the absence then in the presence of the acoustic cued stimulus. The subjects from part 3 were split into two groups so that testing for cued memory was either in a novel environment [novel versus novel + cued (Fig. 4C)] or in a tactile environment [tactile versus tactile + cued (Fig. 4D)]. The novel environment was an identical apparatus as used in part 1, except it was painted black (versus unpainted aluminum), scented with 0.1% acetic acid (versus ethanol), and contained a black Plexiglas (acrylic plastic) floor insert. The tactile environment was identical, except that the black Plexiglas floor insert was omitted, thereby retaining the wire grid flooring used in the training session. The mice were allowed to explore the novel or tactile chamber for 3 min followed by another 3-min period in the presence of the acoustic stimulus (cued). During each test phase, the animals were scored at 10-s intervals by two independent observers for freezing defined as total immobilization except for respiratory movement. The total number of freezes for each individual according to session was generated, and then, a mean was calculated according to genotype for a summary measure. Comparisons were made by planned comparisons (StatView, SAS Institute, Cary, NC). For analysis of 24-hour contextual learning (Fig. 4A) or 0.5-hour contextual learning (Fig. 4B), the number of freezes for the 3-min contextual interval (contextual) was compared to the 3-min exploration period during training (train). For analysis of the cued portion, the number of freezes during the 3-min interval preceding the acoustic stimuli [novel (Fig. 4C) or tactile (Fig. 4D)] was compared to the number of freezes during the 3-min acoustic stimulus interval [novel + cued (Fig. 4C) or tactile + cued (Fig. 4D)]. The indicated CCF findings were significant for rejection of the null hypothesis at P ≤ 0.05 with either parametric or nonparametric stochastic comparisons. Error bars indicate 95% confidence intervals. Similar results were obtained from both observers. The data from one observer are shown in Fig. 4.
24. R. J. Blanchard and D. C. Blanchard, J. Comp. Physiol. Psychol. 67, 370 (1969); M. S. Fanselow, Pavlov J. Biol. Sci. 15, 177 (1980).
25. R. J. Blanchard and D. C. Blanchard, J. Comp. Physiol. Psychol. 67, 370 (1969); M. S. Fanselow, Pavlov J. Biol. Sci. 15, 177 (1980).
26. J. LeDoux, Biol. Psychiatry 44, 1229 (1998); M. Davis, Biol. Psychiatry 44, 1239 (1998); L. Cahill and J. L. McGaugh, Trends Neurosci. 21, 294 (1998).
27. J. LeDoux, Biol. Psychiatry 44, 1229 (1998); M. Davis, Biol. Psychiatry 44, 1239 (1998); L. Cahill and J. L. McGaugh, Trends Neurosci. 21, 294 (1998).
28. J. LeDoux, Biol. Psychiatry 44, 1229 (1998); M. Davis, Biol. Psychiatry 44, 1239 (1998); L. Cahill and J. L. McGaugh, Trends Neurosci. 21, 294 (1998).
29. J. E. LeDoux, P. Cicchetti, A. Xagoraris, L. M. Romanski, J. Neurosci. 10, 1062 (1990); J. E. LeDoux, C. Farb, D. A. Ruggiero, J. Neurosci. 10, 1043 (1990).
30. J. E. LeDoux, P. Cicchetti, A. Xagoraris, L. M. Romanski, J. Neurosci. 10, 1062 (1990); J. E. LeDoux, C. Farb, D. A. Ruggiero, J. Neurosci. 10, 1043 (1990).
31. J. J. Kim, R. A. Rison, M. S. Fanselow, Behav. Neurosci. 107, 1093 (1993); R. G. Phillips and J. E. LeDoux, Behav. Neurosci. 106, 274 (1992); N. R. Selden, B. J. Everitt, L. E. Jarrard, T. W. Robbins, Neuroscience 42, 335 (1991).
32. J. J. Kim, R. A. Rison, M. S. Fanselow, Behav. Neurosci. 107, 1093 (1993); R. G. Phillips and J. E. LeDoux, Behav. Neurosci. 106, 274 (1992); N. R. Selden, B. J. Everitt, L. E. Jarrard, T. W. Robbins, Neuroscience 42, 335 (1991).
33. J. J. Kim, R. A. Rison, M. S. Fanselow, Behav. Neurosci. 107, 1093 (1993); R. G. Phillips and J. E. LeDoux, Behav. Neurosci. 106, 274 (1992); N. R. Selden, B. J. Everitt, L. E. Jarrard, T. W. Robbins, Neuroscience 42, 335 (1991).
34. J. J. Kim and M. S. Fanselow, Science 256, 675 (1992).
35. K. P. Corodimas and J. E. LeDoux, Behav. Neurosci. 109, 613 (1995).
36. L. M. Romanski and J. E. LeDoux, Cereb. Cortex 3, 499 (1993); Cereb. Cortex 3, 515 (1993).
37. L. M. Romanski and J. E. LeDoux, Cereb. Cortex 3, 499 (1993); Cereb. Cortex 3, 515 (1993).
38. B. Caldarone et al., Nature Genet 17, 335 (1997).
39. R. Trullas and P. Skolnick, Psychopharmacology 111, 323 (1993).
40. M. P. Antoch et al., Cell 89, 655 (1997); D. P. King et al., Cell 89, 641 (1997).
41. M. P. Antoch et al., Cell 89, 655 (1997); D. P. King et al., Cell 89, 641 (1997).
42. K. Oishi, K. Sakamoto, T. Okada, T. Nagase, N. Ishida, Biochem. Biophys. Res. Commun. 253, 199 (1998); S. Honma et al., Biochem. Biophys. Res. Commun. 250, 83 (1998); H. Abe et al., Neurosci. Lett. 258, 93 (1998).
43. K. Oishi, K. Sakamoto, T. Okada, T. Nagase, N. Ishida, Biochem. Biophys. Res. Commun. 253, 199 (1998); S. Honma et al., Biochem. Biophys. Res. Commun. 250, 83 (1998); H. Abe et al., Neurosci. Lett. 258, 93 (1998).
44. K. Oishi, K. Sakamoto, T. Okada, T. Nagase, N. Ishida, Biochem. Biophys. Res. Commun. 253, 199 (1998); S. Honma et al., Biochem. Biophys. Res. Commun. 250, 83 (1998); H. Abe et al., Neurosci. Lett. 258, 93 (1998).
45. S. Amir and J. Stewart, Neuroscience 86, 345 (1998).
46. M. R. Rosenzweig, Annu. Rev. Psychol. 47, 1 (1996); W. Singer, Science 270, 758 (1995).
47. M. R. Rosenzweig, Annu. Rev. Psychol. 47, 1 (1996); W. Singer, Science 270, 758 (1995).
48. Single-letter abbreviations for the amino acid residues are as follows: A, Ala; D, Asp; E, Glu; I, Ile; K, Lys; M, Met; Q, Gln; R, Arg; S, Ser; and V, Val.
49. K. B. J. Franklin and G. Paxinos, The Mouse Brain in Stereotaxic Coordinates (Academic Press, New York, 1997).
50. We thank D. Russell for critical input; K. Fay, L. Watkins, M. Barnard, J. Ritter, and C. Stricker for technical assistance; C. Sinton, H. Gershenfeld, and M. Garry for suggestions on behavioral tests; W. Frawley for assistance with statistical analyses; C. Sidle for graphics arts preparation; and L. Gerich for manuscript preparation. This work was supported by funds from the University of Texas Southwestern Medical Center Program of Excellence in Postgraduate Research and the National Alliance for Research on Schizophrenia and Affective Disorders (to J.A.G.); by NIH grants AG16450, AG12297, and NS01763 (to R.D.-A.) and DK52031 and MH59388 (to S.L.M.); and by unrestricted endowment funds from an anonymous donor (to S.L.M.).