title: Mitochondrial impairment, apoptosis and autophagy in a rat brain as immediate and long-term effects of perinatal phencyclidine treatment - influence of restraint stress.
abstract: Phencyclidine (PCP) acts as a non-competitive antagonist of glutamatergic N-methyl-d-aspartate receptor. Its perinatal administration to rats causes pathophysiological changes that mimick some pathological features of schizophrenia (SCH). Numerous data indicate that abnormalities in mitochondrial structure and function could be associated with the development of SCH. Mitochondrial dysfunction could result in the activation of apoptosis and/or autophagy. The aim of this study was to assess immediate and long-term effects of perinatal PCP administration and acute restraint stress on the activity of respiratory chain enzymes, expression of apoptosis and autophagy markers and ultrastructural changes in the cortex and hippocampus of the rat brain. Six groups of rats were subcutaneously treated on 2nd, 6th, 9th and 12th postnatal days (P), with either PCP (10mg/kg) or saline (0.9% NaCl). One NaCl and one PCP group were sacrificed on P13, while other two NaCl and PCP groups were sacrificed on P70. The remaining two NaCl and PCP groups were subjected to 1h restraint stress prior sacrifice on P70. Activities of respiratory chain enzymes were assessed spectrophotometrically. Expression of caspase 3 and AIF as markers of apoptosis and Beclin 1, p62 and LC3, as autophagy markers, was assessed by Western blot. Morphological changes of cortical and hippocampal ultrastructure were determined by transmission electron microscopy. Immediate effects of perinatal PCP administration at P13 were increased activities of complex I in the hippocampus and cytochrome c oxidase (COX) in the cortex and hippocampus implying mitochondrial dysfunction. These changes were followed by increased expression of apoptotic markers. However the measurement of autophagy markers at this time point has revealed decrease of this process in cortex and the absence of changes in hippocampus. At P70 the activity of complex I was unchanged while COX activity was significantly decreased in cortex and increased in the hippocampus. Expressions of apoptotic markers were still significantly higher in PCP perinatally treated rats in all investigated structures, but the changes of autophagy markers have indicated increased level of autophagy also in both structures. Restraint stress on P70 has caused increase of COX activity both in NaCl and PCP perinatally treated rats, but this increase was lower in PCP group. Also, restraint stress resulted in decrease of apoptotic and increase of autophagy processes especially in the hippocampus of PCP perinatally treated group. The presence of apoptosis and autophagy in the brain was confirmed by transmission electron microscopy. In this study we have demonstrated for the first time the presence of autophagy in PCP model of SCH. Also, we have shown increased sensitivity of PCP perinatally treated rats to restraint stress, manifested in alterations of apoptotic and autophagy markers. The future studies are necessary to elucidate the role of mitochondria in the pathophysiology of SCH and putative significance for development of novel therapeutic strategies.
authors.affiliation: Institute of Clinical and Medical Biochemistry, School of Medicine, University of Belgrade, Pasterova 2, 11000 Belgrade, Serbia.
authors.forename: Gordana
authors.initials: G
authors.lastname: Jevtić
authors.affiliation: Institute of Clinical and Medical Biochemistry, School of Medicine, University of Belgrade, Pasterova 2, 11000 Belgrade, Serbia.
authors.forename: Tatjana
authors.initials: T
authors.lastname: Nikolić
authors.affiliation: Institute of Histology and Embryology, School of Medicine, University of Belgrade, Višegradska 26, 11000 Belgrade, Serbia.
authors.forename: Aleksandar
authors.initials: A
authors.lastname: Mirčić
authors.affiliation: Institute of Clinical and Medical Biochemistry, School of Medicine, University of Belgrade, Pasterova 2, 11000 Belgrade, Serbia.
authors.forename: Tihomir
authors.initials: T
authors.lastname: Stojković
authors.affiliation: Institute of Clinical and Medical Biochemistry, School of Medicine, University of Belgrade, Pasterova 2, 11000 Belgrade, Serbia.
authors.forename: Milica
authors.initials: M
authors.lastname: Velimirović
authors.affiliation: Institute of Microbiology and Immunology, School of Medicine, University of Belgrade, Dr Subotića 1, 11000 Belgrade, Serbia.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Trajković
authors.affiliation: Institute of Clinical and Medical Biochemistry, School of Medicine, University of Belgrade, Pasterova 2, 11000 Belgrade, Serbia.
authors.forename: Ivanka
authors.initials: I
authors.lastname: Marković
authors.affiliation: Department of Pathological Physiology, School of Medicine, University of Belgrade, Dr Subotića 9, 11000 Belgrade, Serbia.
authors.forename: Alexander M
authors.initials: AM
authors.lastname: Trbovich
authors.affiliation: Institute of Clinical and Medical Biochemistry, School of Medicine, University of Belgrade, Pasterova 2, 11000 Belgrade, Serbia.
authors.forename: Nevena V
authors.initials: NV
authors.lastname: Radonjić
authors.affiliation: Institute of Clinical and Medical Biochemistry, School of Medicine, University of Belgrade, Pasterova 2, 11000 Belgrade, Serbia. Electronic address: natasapetronijevic@yahoo.com.
authors.forename: Nataša D
authors.initials: ND
authors.lastname: Petronijević
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051033:Apoptosis Inducing Factor / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D002540:Cerebral Cortex / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology*; D006624:Hippocampus / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D010622:Phencyclidine / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D012149:Restraint, Physical
chemical_list: D051033:Apoptosis Inducing Factor; D018691:Excitatory Amino Acid Antagonists; D053148:Caspase 3; D010622:Phencyclidine
keywords: Apoptosis; Autophagy; Phencyclidine; Schizophrenia; Stress