Propargylamines Induce Antiapoptotic New Protein Synthesis in Serum- and Nerve Growth Factor (NGF)-Withdrawn, NGF-Differentiated PC-12 Cells

Abstract

ABSTRACT(Ϫ)-Deprenyl and structurally related propargylamines increase neuronal survival independently of monoamine oxidase B (MAO-B) inhibition, in part by decreasing apoptosis. We found that deprenyl and two other propargylamines, one of which does not inhibit monoamine oxidase B, increased survival in trophically withdrawn 6-day nerve growth factor (NGF)-and 9-day NGF-differentiated PC-12 cells but not in NGF naive or 3-day NGF-differentiated PC-12 cells. Four days of prior NGF exposure were required for the propargylamine-mediated antiapoptosis. Studies using actinomycin D, cycloheximide, and camptothecin revealed that the maintenance of both transcription and translation, particularly between 2 and 6 h after trophic withdrawal, was required for propargylamine-mediated antiapoptosis. Metabolic labeling of newly synthesized proteins for two-dimensional protein gel autoradiography and scintillation counting showed that the propargylamines either increased or reduced the levels of new synthesis or induced de novo synthesis of a number of different proteins, most notably proteins in the mitochondrial and nuclear subfractions. Western blotting for whole cell or subcellular fraction lysates showed that the timing of new protein synthesis changes or subcellular redistribution of apoptosis-related proteins induced by the propargylamines were appropriate to antiapoptosis. The apoptosisrelated proteins included superoxide dismutases (SOD1 and SOD2), glutathione peroxidase, c-JUN, and glyceraldehyde-3-phosphate dehydrogenase. Most notable were the prevention of apoptotic decreases in BCL-2 levels and increases in mitochondrial BAX levels. In general, (Ϫ)-deprenyl-related propargylamines appear to reduce apoptosis by altering the levels or subcellular localization of proteins that affect mitochondrial membrane permeability, scavenge oxidative radicals, or participate in specific apoptosis signaling pathways. The propargylamine (Ϫ)-deprenyl (DEP) inhibits monoamine oxidase B (MAO-B). DEP was first shown to reduce the death of primate nigrostriatal dopaminergic neurons exposed to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) (Cohen et al., 1984) and to slow the clinical progress of human Parkinson's disease (Parkinson, 1993). Both actions appeared to depend on MAO-B inhibition. Subsequently, DEP and deprenylrelated propargylamines (DRPs) were demonstrated to reduce neuronal loss independently of MAO-B inhibition in a variety of experimental models including cortical catecholaminergic neurons exposed to N-(-2-chloroethyl)-Nethyl-2-bromobenzylamine, murine or primate substantia nigra dopaminergic neurons exposed to MPTP, rat facial motoneurons after axotomy, dopaminergic cells treated with the 1-methyl-4-phenylpyridinium ion (MPP ϩ ) or nitric oxide, and hippocampal neurons exposed to kainate (see for details and references). The MAO-B-independent increases in neuronal survival by DRPs were shown to involve decreased apoptosis in a number of the above models (e.g., kainate-exposed hippocampal neurons, nitric oxide, o...

Paraquat leads to dopaminergic neural vulnerability in organotypic midbrain culture

“…Recently, deprenyl and dopamine D2 receptor agonist have been reported that they increase the levels of neurotrophins (Guo et al, 2002;Mizuta et al, 2000;Tatton et al, 2002).…”

Protective Effect of Glycyrrhizin on 1-Methyl-4-phenylpyridinium-Induced Mitochondrial Damage and Cell Death in Differentiated PC12 Cells

“…PC12 cells were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated horse serum, 5% heat-inactivated fetal bovine serum (FBS), 100 U/ml penicillin, and 100 g/ml streptomycin according to the manual of the cell line bank. Cells were differentiated by treating with 100 ng/ml 7S nerve growth factor for 9 days (Tatton et al, 2002). Cells were washed with RPMI 1640 medium containing 1% FBS 24 h before experiments and replated onto the 96-and 24-well plates.…”

Anti‐apoptotic effect of Mao‐B inhibitor PF9601N [N‐(2‐propynyl)‐2‐(5‐benzyloxy‐indolyl) methylamine] is mediated by p53 pathway inhibition in MPP+‐treated SH‐SY5Y human dopaminergic cells

“…Some of their reported effects include up-regulation of prosurvival genes (Maruyama et al 2002;Tatton et al 2002;Weinreb et al 2004), abrogation of mitochondrial swelling and prevention of loss of mitochondrial membrane potential (Wadia et al 1998;Zhang et al 1999;Battaglia et al 2006) as well as prevention of GAPDH up-regulation and nuclear accumulation (Tatton et al 2003). In order to elucidate the molecular mechanisms underlying the neuroprotective actions of PF9601N, we have assessed its possible antiapoptotic effects using the human dopaminergic SH-SY5Y cell line exposed to MPP + , a well-documented in vitro model of apoptosis and PD (Fall and Bennett 1999).…”