메뉴 건너뛰기




Volumn 274, Issue 5293, 1996, Pages 1678-1683

Control of memory formation through regulated expression of a CaMKII transgene

Author keywords

[No Author keywords available]

Indexed keywords

DOXYCYCLINE; PROTEIN KINASE (CALCIUM,CALMODULIN) II;

EID: 0029807527     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.274.5293.1678     Document Type: Article
Times cited : (1140)

References (48)
  • 2
    • 0026847039 scopus 로고
    • W. B. Scoville and B. Milner, J. Neurol. Neurosurg. Psychiatry 20, 11 (1957); L. R. Squire, Psychol. Rev. 99, 195 (1992).
    • (1992) Psychol. Rev. , vol.99 , pp. 195
    • Squire, L.R.1
  • 4
    • 0015433570 scopus 로고    scopus 로고
    • D. C. Blanchard and R. J. Blanchard, J. Comp. Physiol. Psychol. 81, 281 (1972); M. Davis, in The Amygdala: Neurobiologial Aspects of Emotion, Memory, and Mental Dysfunction, J. P. Aggleton, Ed. (Wiley-Liss, New York, 1992), p. 255; J. E. LeDoux, ibid., p 339.
    • (1972) J. Comp. Physiol. Psychol. , vol.81 , pp. 281
    • Blanchard, D.C.1    Blanchard, R.J.2
  • 7
    • 0027092647 scopus 로고
    • S. G. N. Grant et al., Science 258, 1903 (1992).
    • (1992) Science , vol.258 , pp. 1903
    • Grant, S.G.N.1
  • 23
    • 10544220308 scopus 로고    scopus 로고
    • note
    • The CaMKIIα promoter consisted of 8.5 kb of genomic DNA upstream of the transcription initiation site of the mouse CaMKIIα gene, as well as 84 base pairs of the 5′ noncoding exon. Genomic DNA was isolated from a C57 B16/J mouse spleen cosmid library with a rat genomic probe consisting of a 0.4-kb Ava l fragment comprising the transcription-initiation region of rat CaMKIIα (40). The tTA gene from plasmid pUHD 15-1 (70) was flanked by an artificial intron and splice sites at the 5′ end (41) and by a polyadenylation signal from SV40 at the 3′ end. The cDNA with intron and polyadenylation signal was placed downstream of the 8.5-kb CaMKII promoter fragment. The cDNAs for Escherichia coll lacZ and mouse CaMKIIα were similarly flanked by the hybrid intron and polyadenylation signal and placed down-stream of the tet-O promoter element of plasmid pUHD 10-3(10). The CaMKIIα gene was a full-length cDNA (4.8 kb) isolated from a C57B16/J mouse brain cDNA library. The lacZ gene carried an SV40 large T antigen nuclear localization signal as well as the 3′ untranslated region (UTR) of CaMKIIα, which targets the mRNA to dendrites (42).
  • 24
    • 10544242478 scopus 로고    scopus 로고
    • note
    • 286 transcript did not localize to dendrites, presumably as a result of the loss of a sequence element in the 3′ UTR that is necessary for mRNA targeting to dendrites (42).
  • 25
    • 10544230085 scopus 로고    scopus 로고
    • note
    • Transverse slices (400 μm thick) of mouse hippocampus were prepared and placed in an interface slice chamber perfused with artificial cerebrospinal fluid as described (7). Field excitatory postsynaptic potentials (EPSPs) were elicited once per minute with fine tungsten bipolar stimulation electrodes (0.05-ms pulse duration). Stainless steel recording electrodes were placed in striatum radiatum. The stimulation strength was set to produce 50% of the maximum obtainable EPSP in each slice. Baseline synaptic response was collected for 20 min before the tetanus. The 10-Hz tetanus was delivered for 1.5 min at the same intensity as used in the baseline recording. For doxycycline treatment, animals were administered doxycycline (1 mg/ml) plus 5% sucrose in the drinking water for 2 to 3 weeks, and the slices were the exposed to doxycycline (1 ng/ml) in the perfusate. All animals were 2.5 to 6 months of age at the time of recording.
  • 26
    • 10544251719 scopus 로고    scopus 로고
    • note
    • It would also be useful to suppress transgene expression during development and then activate the gene only in the adult animal. However, we found that treatment of wild-type mice with doxycycline (1 mg/ml) during development impaired adult spatial memory and memory for fear conditioning. This result suggests that doxycycline itself produces a defect in neuronal development. We therefore used transgene suppression only in the adult animal in which the doxycycline treatment did not affect memory. Given the activation of the transgene throughout development, it is possible that the LTP and memory phenotypes observed with the transgene active in the adult animal result from a synergistic interaction between developmental and adult expression rather than a direct acute effect of transgene expression in the adult animal.
  • 27
    • 10544227636 scopus 로고    scopus 로고
    • note
    • On the Barnes circular maze (8), the mice (2.5 to 6 months of age) were tested once a day until they met the criterion (five cut of six sessions with three or fewer errors) or until 40 days had elapsed. The order of holes searched was recorded by an observer who was blind to genotype and doxycycline condition, and from these data the number of errors was determined. Errors were defined as searches of any hole that did not have the tunnel beneath it. Searches included nose pokes and head deflections over the hole. At the end of each session the search strategy used was recorded by the observer. The spatial search strategy was operationally defined as reaching the escape tunnel with both error and distance scores ≤3. Distance was calculated by counting the number of holes between the first hole searched within a session and the escape tunnel. A one-factor analysis of variance (ANOVA) (gender) revealed no significant effect of gender for either transgenic or wild-type mice, so the data were collapsed across this variable. For the error data, a three-factor ANOVA (genotype, doxycycline, and session block) with one repeated measure was used. For the spatial search strategy data, the two groups of B22 transgenic mice were compared with a two-way ANOVA (doxycycline and session block) with one repeated measure.
  • 28
    • 10544232183 scopus 로고    scopus 로고
    • note
    • In the conditioned fear task (8), freezing was defined as a total lack of movement with the exception of respiration and was measured by an experimenter who was blind to genotype and doxycycline condition. The percentage of time spent freezing to context and cue was calculated. No significant effect of gender was observed in the B22 or B21 transgenic mice or the wild-type mice, so the data were collapsed across this variable. Freezing to context and cue on testing day was analyzed by fwo three-factor ANOVAs (genotype, line, and doxycycline) that were used to compare the B22 and B21 transgenic and wild-type mice. Two one-way ANOVAs were used to compare the amount of freezing 6 weeks later to cue and context in B21 transgenics on doxycycline, B21 transgenics switched to water, and wild-type mice.
  • 31
    • 0025143633 scopus 로고
    • H. Kita and S. T. Kitai, J. Comp. Neurol. 298, 40 (1990); N. S. Canteras, R. B. Somerly, L. W. Swanson, ibid. 360, 213 (1995).
    • (1990) J. Comp. Neurol. , vol.298 , pp. 40
    • Kita, H.1    Kitai, S.T.2
  • 33
    • 10544256486 scopus 로고    scopus 로고
    • note
    • Pain sensitivity was measured in B21 transgenic (n = 4) and wild-type mice (n = 6). The mice were placed individually in a mouse operant chamber with a metal grid floor and given 1-s foot shocks of increasing intensity (for example, 1, 2, 3, 4 mA . . . ). An experimenter who was blind to the genotype of the mice recorded the intensity of foot shock required to elicit each of the following three responses: startles, vocalizations, and jumps. A t test for each response revealed no significant effect of genotype.
  • 34
    • 10544249239 scopus 로고    scopus 로고
    • note
    • Unconditioned freezing in the presence of an intruder was measured in B21 transgenic (n = 8) and wild-type mice (n = 10) in a Nalgene plastic metabolism cage. The mice and intruder were placed in the upper and lower chambers, respectively. The chambers were separated by a metal grid floor. A seven-week-old male Sprague-Dawley rat served as the intruder and was placed in the lower chamber 10 min before introduction of the mouse. The amount of unconditioned freezing occurring during the first 120 s after the mouse was introduced was measured by an experimenter who was blind to genotype. A (test revealed no significant effect of genotype.
  • 38
    • 0028579743 scopus 로고
    • D. L. Pettit, S. Perlman, R. Malinow, ibid. 266, 1881 (1994); P.-M. Lledo et al., Proc. Natl. Acad. Sci. U.S.A. 92, 11175 (1995).
    • (1994) Science , vol.266 , pp. 1881
    • Pettit, D.L.1    Perlman, S.2    Malinow, R.3
  • 39
  • 40
    • 10544226476 scopus 로고    scopus 로고
    • note
    • 286 transgene in the CA1 region of the B22 mice was patchy; that is, some neurons expressed the transgene well, whereas in other neurons expression was absent. This patchy expression precluded an assessment of LTP in this line of mice by means of field recordings, which sample many synapses from different neurons in a region. However, it is assumed that in those neurons where the transgene was strongly expressed in these mice, a shift in the LTP/LTD frequency threshold would occur.
  • 48
    • 10544228033 scopus 로고    scopus 로고
    • note
    • We thank R. Axel and T. Jessell for critically reading this manuscript; J. Finkelstein for maintaining and genotyping the mice; R. Shin, V. Winder, and L. Varshavsky for help with behavioral experiments; C. Lam for help with figures; H. Ayers and I. Trumpet for typing the manuscript; and M. Osman for animal care. This research was supported by the Howard Hughes Medical Institute and the National Institute of Mental Health.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.