Abstract
We have identified a novel member of the Ndr subfamily of serine/threonine protein kinases, Ndr2, as a gene product that is induced in the mouse amygdala during fear memory consolidation and examined a possible function of this kinase in neural differentiation. Expression of Ndr2 mRNA was detected in various cortical and subcortical brain regions, as well as non-neuronal tissues. Its expression in the amygdala was increased 6 h after Pavlovian fear conditioning training and returned to control levels within 24 h. To study intracellular localization and functions of Ndr2, EGFP::Ndr2 fusion proteins were expressed in rat pheochromocytoma (PC12) cells and acutely isolated cortical neurons, thereby revealing an association of Ndr2 with the actin cytoskeleton in somata, neurites and filopodia, in spines and at sites of cell contact. Co-precipitation and pull-down experiments support this finding. Evidence for an involvement of Ndr2 in actin-mediated cellular functions further comes from the observation of decreased cell spreading and changes in neurite outgrowth that were associated with protein serine phosphorylation in transfected PC12 cells. Together, our data suggest that Ndr2 is an interesting candidate gene for the regulation of structural processes in differentiating and mature neuronal cells.
Highlights
Stimulus-dependent modifications of cell structure and connectivity in the central nervous system are basic elements of long term memory formation
We have identified a novel member of the nuclear Dbf2-related (Ndr) subfamily of serine/threonine protein kinases, Ndr2, as a gene product that is induced in the mouse amygdala during fear memory consolidation and examined a possible function of this kinase in neural differentiation
To address the cellular and molecular processes that underlie memory consolidation and neural plasticity, we study gene products that are expressed in the mouse amygdala after classic fear conditioning [2]
Summary
A fragment of the 3Ј-untranslated region of the Ndr mRNA (clone #VIIE6) was identified through suppression subtractive hybridization of cDNA from the amygdala of fear-conditioned and control mice (Fig. 1A) und subsequent screening with cDNA blots (Fig. 1B). Experiments employed for the identification and expression analysis of clone #VIIE6 have been described previously [2]. All animal experiments were performed in accordance with regulations through Japanese and German
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