A caged Ab reveals an immediate/instructive effect of BDNF during hippocampal synaptic potentiation
A caged Ab reveals an immediate/instructive effect of BDNF during hippocampal synaptic potentiation
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DOI：
10.1073/pnas.251326998
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发表时间：
2001-12-04
影响因子：
11.1
通讯作者：
Bonhoeffer, T
中科院分区：
文献类型：
--
作者：
Kossel, AH;Cambridge, SB;Bonhoeffer, T
神经营养因子已被证明参与中枢神经系统突触的功能强化。虽然它们在这一过程中的重要性是无可争议的，但神经营养因子是否是突触强化的真正介质或仅仅是重要的辅助因子仍然没有解决。为了解决这个问题，我们已经设计了一种方法，通过用光敏保护化合物“笼住”抗BDNF的功能阻断mAb，以高时间分辨率来检测内源性脑源性神经营养因子（BDNF）。使用不同的测定来显示Ab的这种失活通过UV光是可逆的。当应用未修饰的Ab时，在急性海马切片的CA 1区域中tau爆发刺激后的突触增强显著低于笼化Ab。重要的是，在诱导突触增强的过程中，笼状抗体的光活化导致增强作用显著降低。因此，我们的实验加强了内源性BDNF在诱导突触可塑性过程中具有快速作用的观点。结果还表明，笼状抗体可以提供一种工具，用于精确的时空控制内源性蛋白质水平。
Neurotrophins have been shown to be involved in functional strengthening of central nervous system synapses. Although their general importance in this process is undisputed, it remains unresolved whether neurotrophins are truly mediators of synaptic strengthening or merely important cofactors. To address this question, we have devised a method to inactivate endogenous brain-derived neurotrophic factor (BDNF) with high time resolution by "caging" a function-blocking mAb against BDNF with a photosensitive protecting compound. Different assays were used to show that this inactivation of the Ab is reversible by UV light. Synaptic potentiation after tau -burst stimulation in the CA1 region of acute hippocampal slices was significantly less when applying the unmodified Ab compared with the caged Ab. Importantly, photoactivation of the caged Ab during the time of induction of synaptic enhancement led to a marked decrease in potentiation. Our experiments therefore strengthen the view that endogenous BDNF has fast effects during induction of synaptic plasticity. The results additionally show that caged Abs can provide a tool for precise spatiotemporal control over endogenous protein levels.