Dynamics of glycolipid domains in the plasma membrane of living cultured neurons, following protein kinase C activation: a study performed by excimer-formation imaging

被引:13
作者
Pitto, M
Palestini, P
Ferraretto, A
Flati, S
Pavan, A
Ravasi, D
Masserini, M
Bottiroli, G
机构
[1] Dipartimento Chim & Biochim Med, I-20133 Milan, Italy
[2] Univ Aquila, Dept Expt Med, I-67100 Laquila, Italy
[3] CNR, Histochem Ctr, I-27100 Pavia, Italy
关键词
fluorescence imaging; G(MI) ganglioside; glutamate; pyrene; signal transduction;
D O I
10.1042/0264-6021:3440177
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
Dynamic changes of glycolipid domains within the plasma membranes of cultured rat cerebellar granule cells have been investigated. For this purpose, a pyrene-labelled derivative of G(M1), ganglioside has been incorporated in the cell plasma membrane, and the rate of excimer formation, directly related to the formation of domains, has been studied by a fluorescence imaging technique (excimer-formation imaging). Fluorescence imaging showed that upon addition of 100 mu M glutamate, indirectly inducing the activation of protein kinase C (PKC), glycolipid concentration within domains increases in cell bodies. Comparable effects were exerted by the addition of PMA, directly inducing the activation of PKC. On the contrary, the phorbol ester was not effective in the presence of the specific PKC inhibitor, bisindolylmaleimide. These results suggest that glycolipid-enriched domains are dynamic supramolecular structures affected by membrane-associated events, such as PKC activation. Dynamic changes of domains could be important in modulating their postulated participation in a series of functions, including signal transduction and lipid/protein sorting.
引用
收藏
页码:177 / 184
页数:8
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