Gel shift assays were used to study the formation of transcription factors of the AP-1 family in the CNS of Helix during the acquisition of a conditioned defensive reflex based on food aversion. Increases in the DNA-binding activity of AP-1 factors were seen 1-3 h after training. Modeling of "learning" in an in situ system (incubation of the CNS in the presence of serotonin and the Ca2+ ionophore A23187 or the protein kinase C activator phorbol ester (TPA)) also increased the DNA-binding activity of this transcription complex. The DNA-binding activity induced by serotonin acting alone was significantly less than that in controls, while that in the presence of A23187 and TPA was the same as or less than that in controls. The calcium/calmodulin-dependent protein kinase inhibitor KN62 produced significant suppression of the effects of simultaneous exposure to serotonin and calcium on the activation of transcription factors of the AP-1, while greater suppression was obtained with the mitogen-activated protein kinase (ERK) inhibitor PD98059. Cooperative induction of the activation of AP-1 transcription factors in the CNS of the common snail by the serotonin-induced and calcium-dependent regulatory systems may be a mechanism underlying the formation of conditioned defensive reflexes in these animals. Lesions of the formation of transcription factors of the AP-1 family in animals unable to learn defensive forms of behavioral plasticity may be explained in terms of the presence of inhibitory forms of transcription factors interacting with the SRE element.
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http://dx.doi.org/10.1023/a:1021175230674 | DOI Listing |
J Vis Exp
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Institute of Biochemistry and Molecular Biology, Hengyang Medical School, University of South China; National Health Commission Key Laboratory of Birth Defect Research and Preventio, Hunan Provincial Maternal and Child Health Care Hospital;
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