Staurosporine-induced apoptosis in cultured chick embryonic neurons is reduced by polyethylenimine of low molecular weight used as a coating substrate

被引:13
作者
Ahlemeyer, B
Fischer, D
Kissel, T
Krieglstein, J
机构
[1] Univ Marburg, Inst Pharmakol & Toxikol, Fachbereich Pharm, D-35032 Marburg, Germany
[2] Univ Marburg, Fachbereich Pharm, Inst Pharmazeut Technol & Biopharm, D-35032 Marburg, Germany
关键词
extracellular matrix; neuron; apoptosis; staurosporine; polyethylenimine; polylysine; serum deprivation; cytoskeleton;
D O I
10.1016/S0168-0102(00)00128-0
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
The survival of neurons largely depends on adhesion to extracellular matrix proteins. This study investigated the influence of polycationic macromolecules of different molecular weights used as coating substrates on apoptosis in primary cultures of chick embryonic neurons. Coating of the culture flasks with positively charged polyethylenimine (PEI) of 12, 32 and 1616 kDa led to different susceptibilities of the neurons to apoptosis induced by staurosporine and serum deprivation. In cultures grown as usual on polylysine (PL)-coated flasks, we found, after 24 h of incubation in medium with serum (controls), serum-free medium and staurosporine (200 nM)-containing serum-free medium, 15, 35 and 63% apoptotic neurons, respectively as evaluated by nuclear staining with Hoechst 33258. Using 12-kDa PEI as a coating substrate, only 11, 15 and 47% apoptotic neurons could be determined in controls, serum-deprived and staurosporine-treated cultures, respectively. No change in the percentage of apoptotic neurons was found after 24 h of serum deprivation or treatment with staurosporine in cultures grown on 32-kDa PEI compared with cultures grown on PL. However, in staurosporine-treated cultures grown on 1616-kDa PEI, the percentage of apoptotic neurons was even higher than in cultures grown on PL. Immunostaining using neurofilament (NF) antibodies revealed that the reduction of staurosporine-induced apoptosis using 12-kDa PEI instead of PL as a coating substrate was accompanied by a reduced disruption and aggregation of the neurofilaments. Thus, the usage of the newly synthesized 12-kDa PEI as a coating substrate enhanced neuronal resistance to apoptosis. (C) 2000 Elsevier Science Ireland Ltd and the Japan Neuroscience Society. All rights reserved.
引用
收藏
页码:245 / 253
页数:9
相关论文
共 36 条
[1]  
[Anonymous], 1994, ZELL GEWEBEKULTUR
[2]  
Atabay C, 1996, J NEUROSCI RES, V43, P465
[3]  
Bahr BA, 1997, J NEUROSCI, V17, P1320
[4]   INDUCTION OF A COMMON PATHWAY OF APOPTOSIS BY STAUROSPORINE [J].
BERTRAND, R ;
SOLARY, E ;
OCONNOR, P ;
KOHN, KW ;
POMMIER, Y .
EXPERIMENTAL CELL RESEARCH, 1994, 211 (02) :314-321
[5]   Suppression of apoptosis by basement membrane requires three-dimensional tissue organization and withdrawal from the cell cycle [J].
Boudreau, N ;
Werb, Z ;
Bissell, MJ .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (08) :3509-3513
[6]  
BOZYCZKO D, 1986, J NEUROSCI, V6, P1241
[7]   Soluble integrin ligands and growth factors independently rescue neuroblastoma cells from apoptosis under nonadherent conditions [J].
Bozzo, C ;
Bellomo, G ;
Silengo, L ;
Tarone, G ;
Altruda, F .
EXPERIMENTAL CELL RESEARCH, 1997, 237 (02) :326-337
[8]   Neural apoptosis [J].
Bredesen, DE .
ANNALS OF NEUROLOGY, 1995, 38 (06) :839-851
[9]   IN-VITRO CYTOTOXICITY OF MACROMOLECULES IN DIFFERENT CELL-CULTURE SYSTEMS [J].
CHOKSAKULNIMITR, S ;
MASUDA, S ;
TOKUDA, H ;
TAKAKURA, Y ;
HASHIDA, M .
JOURNAL OF CONTROLLED RELEASE, 1995, 34 (03) :233-241
[10]  
DANIELS K, 1991, J CELL SCI, V100, P249