Regulation of presynaptic phosphatidylinositol 4,5-biphosphate by neuronal activity.
Regulation of presynaptic phosphatidylinositol 4,5-biphosphate by neuronal activity.
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DOI：
10.1083/jcb.200102098
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发表时间：
2001-07-23
期刊：
影响因子：
--
通讯作者：
Smith SJ
中科院分区：
文献类型：
--
作者：
Micheva KD;Holz RW;Smith SJ
磷脂酰肌醇4，5-二磷酸（PIP 2）参与多种细胞过程，包括突触囊泡再循环。然而，人们对这种磷脂在神经元中的空间分布及其动力学知之甚少。在本研究中，我们通过在培养的海马神经元中瞬时表达与绿色荧光蛋白（GFP）融合的磷脂酶C（PLC）-δ1普列克底物蛋白同源（PH）结构域，来关注这些问题。该PH结构域特异性地且以高亲和力结合PIP 2。实时共聚焦成像显示，在静息细胞中，PH-GFP主要定位在质膜上。有趣的是，在静止神经元中没有观察到PH-GFP与突触囊泡的关联，表明在成熟的突触囊泡上不存在可检测的PIP 2。海马神经元的电刺激导致质膜处的PH-GFP信号的减少，最可能是由于PLC介导的PIP 2水解。这是伴随着在大多数突触前终端的细胞质PH-GFP信号的显着增加，本地化最有可能在新鲜的内吞膜。进一步的研究表明，PH-GFP信号的增加依赖于N-甲基-D-天冬氨酸受体的激活和随后的一氧化氮（NO）的产生。因此，突触前末梢的PIP 2似乎通过NO的逆行作用受到突触后活动的调节。
Phosphatidylinositol 4,5-biphosphate (PIP2) has been implicated in a variety of cellular processes, including synaptic vesicle recycling. However, little is known about the spatial distribution of this phospholipid in neurons and its dynamics. In this study, we have focused on these questions by transiently expressing the phospholipase C (PLC)-δ1 pleckstrin homology (PH) domain fused to green fluorescent protein (GFP) in cultured hippocampal neurons. This PH domain binds specifically and with high affinity to PIP2. Live confocal imaging revealed that in resting cells, PH-GFP is localized predominantly on the plasma membrane. Interestingly, no association of PH-GFP with synaptic vesicles in quiescent neurons was observed, indicating the absence of detectable PIP2 on mature synaptic vesicles. Electrical stimulation of hippocampal neurons resulted in a decrease of the PH-GFP signal at the plasma membrane, most probably due to a PLC-mediated hydrolysis of PIP2. This was accompanied in the majority of presynaptic terminals by a marked increase in the cytoplasmic PH-GFP signal, localized most probably on freshly endocytosed membranes. Further investigation revealed that the increase in PH-GFP signal was dependent on the activation of N-methyl-D-aspartate receptors and the consequent production of nitric oxide (NO). Thus, PIP2 in the presynaptic terminal appears to be regulated by postsynaptic activity via a retrograde action of NO.
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DOI：
10.1073/pnas.92.23.10472
发表时间：
1995-11-07
影响因子：
11.1
作者：
LEMMON, MA;FERGUSON, KM;SCHLESSINGER, J
通讯作者：
SCHLESSINGER, J
影响因子：
5.3
作者：
BETZ, WJ;MAO, F;BEWICK, GS
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BEWICK, GS
影响因子：
9.2
作者：
Jost, M;Simpson, F;Schmid, SL
通讯作者：
Schmid, SL
影响因子：
7.5
作者：
Corvera, S;D'Arrigo, A;Stenmark, H
通讯作者：
Stenmark, H
影响因子：
4.1
作者：
EICHBERG, J;DAWSON, RMC
通讯作者：
DAWSON, RMC