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Titolo:
A SYSTEM FOR CHARACTERIZING CELLULAR AND MOLECULAR EVENTS IN PROGRAMMED NEURONAL CELL-DEATH
Autore:
PITTMAN RN; WANG SL; DIBENEDETTO AJ; MILLS JC;
Indirizzi:
UNIV PENN,SCH MED,DEPT PHARMACOL 6084 PHILADELPHIA PA 19104
Titolo Testata:
The Journal of neuroscience
fascicolo: 9, volume: 13, anno: 1993,
pagine: 3669 - 3680
SICI:
0270-6474(1993)13:9<3669:ASFCCA>2.0.ZU;2-S
Fonte:
ISI
Lingua:
ENG
Soggetto:
NERVE GROWTH-FACTOR; TROPHIC-FACTOR DEPRIVATION; KILLS POSTMITOTIC NEURONS; CAENORHABDITIS-ELEGANS; SYMPATHETIC NEURONS; IMMATURE THYMOCYTES; PROTEIN-SYNTHESIS; DNA METHYLATION; RNA-SYNTHESIS; PC12 CELLS;
Keywords:
APOPTOSIS; PROGRAMMED CELL DEATH; PC12-CELLS; DIFFERENTIATION; NGF; RNA SYNTHESIS; PROTEIN SYNTHESIS; TRANSCRIPTION DEPENDENCE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
52
Recensione:
Indirizzi per estratti:
Citazione:
R.N. Pittman et al., "A SYSTEM FOR CHARACTERIZING CELLULAR AND MOLECULAR EVENTS IN PROGRAMMED NEURONAL CELL-DEATH", The Journal of neuroscience, 13(9), 1993, pp. 3669-3680

Abstract

A model system has been established in which PC12 cells are convertedto neuronal-like cells that undergo transcription-dependent cell death following removal of NGF. Nineteen sublines of PC12 cells were tested to establish parameters for making cells dependent on NGF for survival. In most sublines, a relatively small percentage of cells become dependent on NGF for survival, and following removal of NGF, most of thecells begin proliferating in serum-containing medium. In several sublines, however, a significant percentage of cells die following removalof NGF. One of these sublines, PC6-3, can be grown under conditions in which 90% of the cells undergo transcription-dependent cell death following removal of NGF in either serum-free or serum-containing medium. Fourteen hours after removing NGF, 50% of the cells are committed todie, while initial morphological signs of cell death as determined bytime-lapse videomicroscopy occur 2-6 hr later and include loss of neurites followed by a 1-3 hr period of active membrane ''blebbing'' and protrusions. Cell death can be blocked by the RNA synthesis inhibitor actinomycin D, the protein synthesis inhibitor cycloheximide, KCl, basic fibroblast growth factor, or dibutryl-cAMP, but not by epidermal growth factor, leupeptin, or the endonuclease inhibitor aurintricarboxylic acid (ATA). Removal of NGF activates an endonuclease that causes nucleosomal laddering of the DNA; however, endonuclease activity does not appear to be required for cell death. In agreement with previous studies (Batistatou and Greene, 1991; Rukenstein et al., 1991) demonstrating that naive PC12 cells undergo transcription-independent cell deathwhen shifted into serum-free medium in the absence of growth factors,all cell lines tested except for one die when cultured in RPMI mediumlacking growth factors. DNA fragmentation is a prominent feature of transcription-independent cell death, and death can be blocked with NGF, ATA, and dibutryl-cAMP but not with actinomycin D or KCl. The PC12 model system described here should be useful for identifying cell deathgenes and for characterizing cellular and molecular events in programmed neuronal cell death.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/11/20 alle ore 11:37:35