Li Y X, Xu Y, Ju D, Lester H A, Davidson N, Schuman E M
Division of Biology 156-29, California Institute of Technology, Pasadena, CA 91125, USA.
Proc Natl Acad Sci U S A. 1998 Sep 1;95(18):10884-9. doi: 10.1073/pnas.95.18.10884.
We have developed a method to analyze the relative contributions of pre- and postsynaptic actions of a particular gene product in neurons in culture and potentially in slices using adenovirus-mediated gene transfer. A recombinant virus directed the expression of both a GFP reporter protein and TrkB.T1, a C-terminal truncated dominant negative TrkB neurotrophin receptor. When expressed in the presynaptic cell at synapses between embryonic hippocampal neurons in culture, the dominant negative TrkB.T1 inhibited two forms of synaptic potentiation induced by the neurotrophin brain-derived neurotrophic factor (BDNF): (i) greater evoked synaptic transmission and (ii) higher frequency of spontaneous miniature synaptic currents. These inhibition effects are not seen if the transgene is expressed only in the postsynaptic cell. We conclude that BDNF-TrkB signal transduction in the presynaptic terminal leads to both types of potentiation and is therefore the primary cause of synaptic enhancement by BDNF in these neurons.
我们开发了一种方法,可利用腺病毒介导的基因转移,分析特定基因产物在培养神经元以及可能在脑片中的突触前和突触后作用的相对贡献。一种重组病毒可指导绿色荧光蛋白(GFP)报告蛋白和TrkB.T1(一种C末端截短的显性负性TrkB神经营养因子受体)的表达。当在培养的胚胎海马神经元之间的突触前细胞中表达时,显性负性TrkB.T1抑制了神经营养因子脑源性神经营养因子(BDNF)诱导的两种突触增强形式:(i)更大的诱发突触传递和(ii)更高频率的自发微小突触电流。如果转基因仅在突触后细胞中表达,则不会出现这些抑制作用。我们得出结论,突触前终末中的BDNF-TrkB信号转导导致了这两种增强类型,因此是BDNF在这些神经元中增强突触的主要原因。