RAPID PHOSPHORYLATION OF PHOSPHOLIPASE C-GAMMA-1 BY BRAIN-DERIVED NEUROTROPHIC FACTOR AND NEUROTROPHIN-3 IN CULTURES OF EMBRYONIC RAT CORTICAL-NEURONS

被引:75
作者
WIDMER, HR [1 ]
KAPLAN, DR [1 ]
RABIN, SJ [1 ]
BECK, KD [1 ]
HEFTI, F [1 ]
KNUSEL, B [1 ]
机构
[1] NCI,FREDERICK CANC RES & DEV CTR,ABL BASIC RES PROGRAM,FREDERICK,MD 21701
关键词
PHOSPHOLIPASE-C; TYROSINE PHOSPHORYLATION; PHOSPHOINOSITIDE METABOLISM; NEURON CULTURES; NERVE GROWTH FACTOR; BRAIN-DERIVED NEUROTROPHIC FACTOR; NEUROTROPHIN-3; K-252B; TRK;
D O I
10.1111/j.1471-4159.1993.tb03496.x
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
Phospholipase C-gamma1 (PLC-gamma1) is involved at an early step in signal transduction of many hormones and growth factors and catalyzes the hydrolysis of phosphatidylinositol (PI) 4,5-bisphosphate to diacylglycerol and inositol trisphosphate, two potent intracellular second messenger molecules. The transformation of PC12 cells into neuron-like cells induced by nerve growth factor is preceded by a rapid stimulation of PLC-gamma1 phosphorylation and PI hydrolysis. The present study analyzed the effects of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) on phosphorylation of PLC-gamma1 in primary cultures of embryonic rat brain cells. BDNF and NT-3 stimulated the phosphorylation of PLC-gamma1, followed by hydrolysis of PI. The stimulation of PLC-gamma1 phosphorylation occurred within 20 s after addition of BDNF or NT-3 and lasted up to 30 min, with a peak after 4 min. ED50 values were similar for BDNF and NT-3, with approximately 25 ng/ml. Phosphorylation of PLC-gamma1 by BDNF and NT-3 was found in cultures from all major brain areas. K-252b, a compound known to inhibit selectively neurotrophin actions by interfering with the phosphorylation of trk-type neurotrophin receptors, prevented the BDNF- and NT-3-stimulated phosphorylation of PLC-gamma1. Receptors of the trk type were coprecipitated with anti-PLC-gamma1 antibodies. The presence of irk-B mRNA in the cultures was substantiated by northern blot analysis. The action of BDNF and NT-3 seems to be neuron specific because no phosphorylation of PLC-gamma1 was observed in cultures of nonneuronal brain cells. The results provide evidence that developing neurons of the cerebral cortex and other brain areas are responsive to BDNF and NT-3, and they indicate that the transduction mechanism of BDNF and NT-3 in the brain involves rapid phosphorylation of PLC-gamma1 followed by PI hydrolysis.
引用
收藏
页码:2111 / 2123
页数:13
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