cGMP-dependent protein kinase type I promotes CREB/CRE-mediated gene expression in neurons of the lateral amygdala.

The process transforming newly learned information into stable long-term memory is called memory consolidation and, like the underlying long-term synaptic plasticity, critically depends on de novo RNA and protein synthesis. We have shown recently that the cGMP-dependent protein kinase Type I (cGKI) plays an important role for the consolidation of amygdala-dependent fear memory and long-term potentiation (LTP) in the lateral amygdala. Signalling downstream of cGKI at the level of transcriptional regulation remained unclear. A transcription factor of major importance for learning and memory is the cAMP-response element binding protein (CREB). The representation of fear memory in the lateral amygdala strikingly depends on the activity of CREB in individual neurons. Moreover, findings from in vitro experiments demonstrate CREB phosphorylation by cGK. In the hippocampus, CREB phosphorylation increases following activation of NO/cGMP signalling contributing to the late phase of LTP. To demonstrate a link from cGKI to activation of CREB and CREB-dependent transcription in neurons of the lateral amygdala as a possible mechanism for cGKI-mediated fear memory consolidation, we examined the effect of cGMP on activation of CREB/CRE using immunohistochemical staining specific for phospho-CREB and a reporter gene in control and cGKI-deficient mice, respectively. Supporting our hypothesis, marked CREB phosphorylation and CRE-mediated transcription was induced by cGMP in the lateral amygdala of control mice, but not in cGKI-deficient mice. It has been proposed that activation of cGKI is followed by its nuclear translocation that would allow direct phosphorylation of CREB. Therefore, we examined the cellular localisation of cGKI in neurons of the lateral amygdala in the presence of cGMP by double staining for cGKI and a nuclear marker in sections from areas showing prominent CREB phosphorylation, and did not observe prominent nuclear translocation of the enzyme. In summary, we provide evidence that cytosolic cGKI can support fear memory consolidation and LTP in neurons of the lateral amygdala via activation of CREB and CRE-dependent transcription.

Signaling through cGMP-dependent protein kinase I in the amygdala is critical for auditory-cued fear memory and long-term potentiation.

Long-term potentiation (LTP) of inputs relaying sensory information from cortical and thalamic neurons to principal neurons in the lateral amygdala (LA) is thought to serve as a cellular mechanism for associative fear learning. Nitric oxide (NO), a messenger molecule widely implicated in synaptic plasticity and behavior, has been shown to enhance LTP in the LA as well as consolidation of associative fear memory. Additional evidence suggests that NO-induced enhancement of LTP and amygdala-dependent learning requires signaling through soluble guanylyl cyclase (sGC) and cGMP-dependent protein kinase (cGK). Mammals possess two genes for cGK: the prkg1 gene gives rise to the cGK type I isoforms, cGKIalpha and cGKIbeta, and the prkg2 gene encodes the cGK type II. Reportedly, both cGKI and cGKII are expressed in the amygdala, and cGKII is involved in controlling anxiety-like behavior. Because selective pharmacological tools for individual cGK isoforms are lacking, we used different knock-out mouse models to examine the function of cGKI and cGKII for LTP in the LA and pavlovian fear conditioning. We found robust expression of the cGKI specifically in the LA with cGKIbeta as the prevailing isoform. We further show a marked reduction of LTP at both thalamic and cortical inputs to the LA and a selective impairment of auditory-cued fear memory in cGKI-deficient mutants. In contrast, cGKII null mutants lack these phenotypes. Our data suggest a function of cGKI, likely the beta isoform, in the LA, supporting synaptic plasticity and consolidation of fear memory.