Influence of culture conditions on monoamine oxidase A and B activity in rat astrocytes

被引:6
作者
Carlo, P [1 ]
DelRio, M [1 ]
Violani, E [1 ]
Sciaba, L [1 ]
Picotti, GB [1 ]
机构
[1] UNIV MILAN,DEPT PHARMACOL CHEMOTHERAPY & TOXICOL,I-20122 MILAN,ITALY
关键词
astrocyte cultures; monoamine oxidase A and B; serum; stripped serum; chemically defined medium;
D O I
10.1002/cbf.645
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Astroglial cells dispersed from newborn rat hemispheres were established in medium supplemented with 20 per cent fetal calf serum (FBS) and then grown to a confluent monolayer in the presence of 10 per cent FBS or charcoal-stripped FBS (CS). Type 1 astrocytes were subcultured and either maintained under the same conditions of the primary cultures or converted to serum-free chemically defined medium (CDM). No differences were found in either MAO A or MAO B activity of astrocytes grown in the presence of FBS or CS after 15 and 21 days in vitro (day 1 and 6 of subculture). In contrast, on day 21 both MAO A and MAO B activities were markedly higher in astrocytes subcultured in CDM compared with cells maintained in serum-supplemented medium. This difference appeared to be due to increased number of enzyme molecules, since kinetic analysis showed an increase in V-max of both MAO isoenzymes in serum-free medium, but no change in K-m. Consistently, the recovery of MAO A and MAO B activity after irreversible enzyme inhibition by clorgyline and deprenyl was faster in CDM than in FBS-supplemented medium, indicating enhanced enzyme synthesis under serum-free condition. Estimates of half-lives for the recovery of MAO A and MAO B activity indicated that, under both culture conditions, type A activity had a higher turnover rate than type B. The effect of CDM on astrocyte MAO does not appear to be due to selection of a subpopulation of cells, but rather linked to a morphological change (differentiation) with increased synthesis of both MAO isoenzymes.
引用
收藏
页码:19 / 25
页数:7
相关论文
共 40 条
[1]   SERUM-FREE CELL-CULTURE - A UNIFYING APPROACH [J].
BARNES, D ;
SATO, G .
CELL, 1980, 22 (03) :649-655
[2]   PHENOL RED IN TISSUE-CULTURE MEDIA IS A WEAK ESTROGEN - IMPLICATIONS CONCERNING THE STUDY OF ESTROGEN-RESPONSIVE CELLS IN CULTURE [J].
BERTHOIS, Y ;
KATZENELLENBOGEN, JA ;
KATZENELLENBOGEN, BS .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (08) :2496-2500
[3]   SELECTIVE CULTURE OF NEURONS FROM RAT CEREBRAL-CORTEX - MORPHOLOGICAL CHARACTERIZATION, GLUTAMATE UPTAKE AND RELATED ENZYMES DURING MATURATION IN VARIOUS CULTURE MEDIA [J].
BORG, J ;
SPITZ, B ;
HAMEL, G ;
MARK, J .
DEVELOPMENTAL BRAIN RESEARCH, 1985, 18 (1-2) :37-49
[4]   GROWTH OF A RAT NEUROBLASTOMA CELL LINE IN SERUM-FREE SUPPLEMENTED MEDIUM [J].
BOTTENSTEIN, JE ;
SATO, GH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1979, 76 (01) :514-517
[5]   PKC ACTIVITY AND PKC-ALPHA MESSENGER-RNA CONTENT ARE REDUCED IN SERUM-DEPRIVED HUMAN NEUROBLASTOMA-CELLS WITHOUT CONCOMITANT INDUCTION OF DIFFERENTIATION [J].
CARLSON, RO ;
MARTIN, DM ;
FELDMAN, EL ;
AGRANOFF, BW .
EXPERIMENTAL CELL RESEARCH, 1993, 207 (02) :340-347
[6]   THYROID-HORMONE DEIODINASES IN PURIFIED PRIMARY GLIAL-CELL CULTURES [J].
CAVALIERI, RR ;
GAVIN, LA ;
COLE, R ;
DEVELLIS, J .
BRAIN RESEARCH, 1986, 364 (02) :382-385
[7]   [H-3]RO 16-6491, A SELECTIVE PROBE FOR AFFINITY LABELING OF MONOAMINE-OXIDASE TYPE-B IN HUMAN-BRAIN AND PLATELET MEMBRANES [J].
CESURA, AM ;
IMHOF, R ;
TAKACS, B ;
GALVA, MD ;
PICOTTI, GB ;
DAPRADA, M .
JOURNAL OF NEUROCHEMISTRY, 1988, 50 (04) :1037-1043
[8]  
DARBRE P, 1983, CANCER RES, V43, P349
[9]  
EDELSTEIN SB, 1981, BIOCHEM BIOPH RES CO, V98, P836, DOI 10.1016/0006-291X(81)91187-6
[10]   MONOAMINE OXIDASE-A AND OXIDASE-B ARE DIFFERENTIALLY REGULATED BY GLUCOCORTICOIDS AND AGING IN HUMAN-SKIN FIBROBLASTS [J].
EDELSTEIN, SB ;
BREAKEFIELD, XO .
CELLULAR AND MOLECULAR NEUROBIOLOGY, 1986, 6 (02) :121-150