The impact of the voltage sensing phosphatase (VSP) dimers on neurons

这项研究将调查被称为神经元的脑细胞如何使用电信号和化学信号进行交流。具体地说，神经元中的一种成分，称为电压感应磷酸酶或VSP，使用电荷直接改变神经元内部的化学物质。然而，VSP如何改变神经元之间的通信仍是未知的，对这一过程的更深入了解可能会澄清大脑如何通过神经元之间的通信来处理视觉、听觉和触觉等信息。这项研究的结果可能也适用于大脑以外的其他组织，这些组织已知使用电信号来调节细胞功能。参与该项目的研究生和本科生将接受如何批判性思考、解决问题和进行实验的培训。该项目的方方面面将通过儿童和成人的实践经验与公众分享。为了接触到蒙大拿州农村社区中更广泛的受众，还将使用视频会议来分享该项目及其背后的基本科学。膜电位和磷脂酰肌醇磷酸(PIP)是神经元中的关键信号，调节突触传递、离子通道、生长和迁移。VSP是已知的唯一一种直接连接这两种信号的蛋白质，它使用一个电压感应域来激活一个磷酸酶结构域，然后以一种电压依赖的方式使PIP去磷酸化。VSP可以显著影响神经元信号转导，因为它比其他磷酸酶在更快的时间尺度上被激活。然而，VSP对神经功能的影响尚不清楚。本研究旨在确定VSP二聚化是否作为VSP活性和特异性的调节机制。首先，二聚体将被破坏，以确定VSP二聚体与单体对功能的影响。其次，将确定VSP异二聚化对催化的影响。方法将包括电生理学、荧光、成像、生化技术和分子生物学。这项研究对于理解VSP背后的分子机制，增加该领域对电压感应蛋白质的理解，并作为设计新的电压门控工具(如传感器和酶)的基础，是至关重要的。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This research will investigate how brain cells called neurons use both electrical and chemical signals to communicate. Specifically, one component in neurons, called the voltage sensing phosphatase or VSP, uses an electrical charge to directly change the chemistry inside the neuron. However, how VSP then changes neuron communication is still unknown and a greater understanding of this process may clarify how brains process information such as vision, hearing and touch, by communication between neurons. The results from this research may also be applicable to other tissues apart from the brain that are known to use electrical signals to regulate cellular function. Graduate and undergraduate students involved in this project will be trained on how to think critically, solve problems and conduct experiments. Aspects of the project will be shared with the public through hands-on experiences for children and adults. To reach a broader audience within the rural communities of Montana, video conferencing will also be used to share the project and the underlying science behind it. Membrane potential and phosphatidylinositol phosphates (PIPs) are critical signals in neurons, regulating synaptic transmission, ion channels, growth and migration. VSP is the only known protein to directly link both signals, using a voltage sensing domain to activate a phosphatase domain that then dephosphorylates PIPs in a voltage dependent manner. VSP could significantly influence neuronal signaling because it is activated on a faster time scale than other phosphatases. However, the impact of VSP on neuronal function is still unknown. This research aims to determine whether VSP dimerization serves as a regulatory mechanism for VSP activity and specificity. First, the dimer will be disrupted to determine the impact of VSP dimers versus monomers on function. Second, the consequences of VSP heterodimerization on catalysis will be determined. Methods will include electrophysiology, fluorescence, imaging, biochemical techniques, and molecular biology. This research is critical for understanding the molecular mechanism behind VSP, increasing the field’s understanding of voltage sensing proteins and serving as a basis for engineering new voltage-gated tools such as sensors and enzymes.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.