Intracellular localization and functional effects of mutant Protein kinase D expression in hippocampal neurons

Abstract

Event Abstract Back to Event Intracellular localization and functional effects of mutant Protein kinase D expression in hippocampal neurons Flóra Vajda1*, Angelika Hausser2 and Katalin Schlett1 1 Eötvös Loránd University, Department of Physiology and Neurobiology, Hungary 2 University of Stuttgart, Inst. Cell Biology and Immunology, Germany Protein Kinase D (PKD) is a serine/threonine kinase known to regulate various cellular processes in non neuronal cells. Despite the fact that PKD isoforms are expressed in the central nervous system already from early embryonic stages, so far little is known about PKD-directed cellular processes in neurons. In the present study, several fluorescently tagged PKD mutants were introduced into mouse embryonal hippocampal neurons. Upon transfection by an EGFP tagged dominant-negative kinase-inactive form of PKD1 (kdPKD1-EGFP) or a construct lacking the kinase domain, overexpressed proteins were found to be excluded from the axons and to be restricted to the somatodendritic compartment of pyramidal neurons. Similar uneven intracellular localization was observed when PKD1 mutants lacking the PH (Pleckstrin homology) domain or amino acids 80-146 were used. These findings suggested that beside kinase activity, the N terminal region also regulates PKD’s axonal entry. Functional effects of PKD mutant overexpression on axonal growth cone motility were also analyzed by fluorescent fast live cell imaging. Growth cones of neurons overexpressing wild type PKD1 seemed to be more rigid with less filopodial movement, which is concordant with the role of PKD in cytoskeletal remodeling. In vivo experiments were also carried out by using a transgenic mouse line expressing kdPKD1-EGFP in a forebrain-specific manner upon doxycyclin treatment to describe kdPKD1-EGFP expression pattern after 2, 4, 6 and 8 weeks of tetracycline induction. In accordance with our in vitro results, kdPKD1-EGFP expression was restricted to the somatodendritic compartment of pyramidal neurons in the hippocampus and in the entorhinal cortex. Surprisingly, kdPKD1-EGFP expression was detected in the axons of the primary sensory neurons in the olfactory bulb. Further experiments are planned to elucidate PKD1 features causing distinct distribution of the mutant enzyme in different cell types in the brain.. Conference: IBRO International Workshop 2010, Pécs, Hungary, 21 Jan - 23 Jan, 2010. Presentation Type: Poster Presentation Topic: Cellular neuroscience Citation: Vajda F, Hausser A and Schlett K (2010). Intracellular localization and functional effects of mutant Protein kinase D expression in hippocampal neurons. Front. Neurosci. Conference Abstract: IBRO International Workshop 2010. doi: 10.3389/conf.fnins.2010.10.00173 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 30 Apr 2010; Published Online: 30 Apr 2010. * Correspondence: Flóra Vajda, Eötvös Loránd University, Department of Physiology and Neurobiology, Budapest, Hungary, floravajda@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Flóra Vajda Angelika Hausser Katalin Schlett Google Flóra Vajda Angelika Hausser Katalin Schlett Google Scholar Flóra Vajda Angelika Hausser Katalin Schlett PubMed Flóra Vajda Angelika Hausser Katalin Schlett Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.