caa a22 4500 475792653 CHVBK 20180406123714.0 cr unu---uuuuu 170329e20000301xx s 000 0 eng 10.1023/A:1011113802642 doi (NATIONALLICENCE)springer-10.1023/A:1011113802642 Targeted Neutralization of Calcineurin, by Expression of an Inhibitor Peptide Under the Control of a Cholinergic Specific Promoter in PC12 Cells, Promotes Neurite Outgrowth in the Presence of NGF [Elektronische Daten] [Jorge Naciff, Karen King, John Dedman] We have characterized a region of the mouse vesicular acetylcholine transporter(VAChT)/choline acetyltransferase (ChAT) gene locus that serves as a cholinergic-specific promoter for the expression of both VAChT and ChAT genes, as well as a reporter gene (LacZ) in vivo. We have used this promoter to direct the expression of an inhibitor peptide, derived from the calcineurin (CalN) autoregulatory domain, to directly neutralize the function of CalN to define the role of this Ca2+/Calmodulin regulated phosphatase in neurite outgrowth. Targeted inhibition of CalN promotes neurite outgrowth in PC12 cells in the presence of NGF, as early as 24 h after transfection. Inhibition of CalN-mediated enhancement of neurite outgrowth in PC12 cells reaches a maximum effect within the first 4 to 6 days after transfection, and does not cause adverse effects when highly expressed for up to 12 days. Cyclosporin A, a nontargeted CalN inhibitor, increases the number of neurites in mock transfected cells by 1.5 fold, while in transfected PC12 cells, the expression of the CalN inhibitor peptide increases the neurite number by 1.8 fold. These data demonstrate that CalN is an important regulator of the neurotrophic response in cholinergic cells and may prove valuable in developing treatment strategies to promote recovery from neurological Injury. Plenum Publishing Corporation, 2000 Cholinergic-specific promoter nationallicence PC12 cells nationallicence calcineurin nationallicence neurite outgrowth nationallicence Naciff Jorge Department of Molecular and Cellular Physiology, University of Cincinnati, 231 Bethesda Ave., 45267-0576, Cincinnati, OH, U.S.A aut King Karen Department of Molecular and Cellular Physiology, University of Cincinnati, 231 Bethesda Ave., 45267-0576, Cincinnati, OH, U.S.A aut Dedman John Department of Molecular and Cellular Physiology, University of Cincinnati, 231 Bethesda Ave., 45267-0576, Cincinnati, OH, U.S.A aut Metabolic Brain Disease Kluwer Academic Publishers-Plenum Publishers 15/1(2000-03-01), 65-81 0885-7490 15:1<65 2000 15 11011 https://doi.org/10.1023/A:1011113802642 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1011113802642 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Naciff Jorge Department of Molecular and Cellular Physiology, University of Cincinnati, 231 Bethesda Ave., 45267-0576, Cincinnati, OH, U.S.A aut NATIONALLICENCE 700 1- King Karen Department of Molecular and Cellular Physiology, University of Cincinnati, 231 Bethesda Ave., 45267-0576, Cincinnati, OH, U.S.A aut NATIONALLICENCE 700 1- Dedman John Department of Molecular and Cellular Physiology, University of Cincinnati, 231 Bethesda Ave., 45267-0576, Cincinnati, OH, U.S.A aut NATIONALLICENCE 773 0- Metabolic Brain Disease Kluwer Academic Publishers-Plenum Publishers 15/1(2000-03-01), 65-81 0885-7490 15:1<65 2000 15 11011 Metadata rights reserved Springer special CC-BY-NC licence nationallicence SWISSBIB 475792610 BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer