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. 2010 Jul 23:1345:156-63.
doi: 10.1016/j.brainres.2010.05.048. Epub 2010 May 24.

Differences in hippocampal CREB phosphorylation in trace fear conditioning of two inbred mouse strains

Yoo Kyeong Hwang  1 , Jae-Chun Song, Seol-Heui Han, Jeiwon Cho, Dani R Smith, Michela Gallagher, Jung-Soo Han
Affiliations

Differences in hippocampal CREB phosphorylation in trace fear conditioning of two inbred mouse strains

Yoo Kyeong Hwang et al. Brain Res. .

Abstract

The effects of genetic background on fear trace conditioning were evaluated in relation to phosphorylated levels of cAMP response element-binding protein (CREB) in the hippocampus using two different inbred strains of mice, C57BL/6 and DBA/2. The male mice received a trace fear conditioning protocol and unpaired control groups were included to assess nonassociative effects on test performance. Both C57BL/6 and DBA/2 mice with paired training displayed higher freezing responses during testing than those with unpaired training, respectively. The C57BL/6 mice with paired training also displayed higher freezing responses to the tone-CS during testing than the DBA/2 mice with paired training. Because much evidence implicates the hippocampus as an important neural substrate for trace fear conditioning, the engagement of the hippocampus was examined after testing by measuring levels of CREB and phosphorylated CREB (pCREB). The results revealed that hippocampal CREB levels in both strains of mice were not significantly altered according to the type of training (unpaired vs. paired). However, the hippocampal pCREB levels were significantly higher in the paired training group than the unpaired control group in C57BL/6 mice, but not in DBA/2 mice. These findings indicate that hippocampal pCREB is closely tied to this form of associative conditioning only in C57BL/6 mice and that different neural substrates may support trace conditioning in C57BL/6 and DBA/2 strains.

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Figures

Fig. 1
Fig. 1
Schematic representation of the protocol for trace fear conditioning. The third inset in day 2 (Training) and day 3 (Testing) shows the time interval used for the analysis of freezing levels. The testing in day 3 was conducted in a novel test chamber. Note that the times represented by the dashed lines are not repeated.
Fig. 2
Fig. 2
Representative immunoblots of hippocampal CREB and pCREB from C57BL/6 (A) and DBA/2 (B) mice 30 min after the last tone-CS presentation on day 3.
Fig. 3
Fig. 3
Quantification of hippocampal CREB and pCREB levels (mean±S.E.M) from C57BL/6 and DBA/2 mice 30 min after the last tone-CS presentation on day 3. Data are expressed as the ratio of CREB/actin (A) and pCREB/actin (B) in the hippocampus. The overall hippocampal CREB levels of the C57BL/6 mice were statistically higher than those of the DBA/2 mice (#, A). (*) indicates significantly greater hippocampal pCREB following tone-CS presentationin comparison with the unpaired training in C57BL/6 mouse strains (B).
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