Division of Biological Sciences, Center for Structural and Functional Neuroscience, The University of Montana, Missoula, MT, USA.
Brain Mind Institute, Swiss Federal Institute of Technology (EPFL), Lausanne, Switzerland.
J Neurosci Methods. 2022 Apr 15;372:109540. doi: 10.1016/j.jneumeth.2022.109540. Epub 2022 Feb 24.
Throughout the animal kingdom, GABA is the principal inhibitory neurotransmitter of the nervous system. It is essential for maintaining the homeostatic balance between excitation and inhibition required for the brain to operate normally. Identification of GABAergic neurons and their GABA release sites are thus essential for understanding how the brain regulates the excitability of neurons and the activity of neural circuits responsible for numerous aspects of brain function including information processing, locomotion, learning, memory, and synaptic plasticity, among others.
Since the structure and features of GABA synapses are critical to understanding their function within specific neural circuits of interest, here we developed and characterized a conditional marker of GABAergic synaptic vesicles for Drosophila, 9XV5-vGAT.
9XV5-vGAT is validated for conditionality of expression, specificity for localization to synaptic vesicles, specificity for expression in GABAergic neurons, and functionality. Its utility for GABAergic neurotransmitter phenotyping and identification of GABA release sites was verified for ellipsoid body neurons of the central complex. In combination with previously reported conditional SV markers for acetylcholine and glutamate, 9XV5-vGAT was used to demonstrate fast neurotransmitter phenotyping of subesophageal ganglion neurons.
This method is an alternative to single cell transcriptomics for neurotransmitter phenotyping and can be applied to any neurons of interest represented by a binary transcription system driver.
A conditional GABAergic synaptic vesicle marker has been developed and validated for GABA neurotransmitter phenotyping and subcellular localization of GABAergic synaptic vesicles.
在整个动物王国中,GABA 是神经系统的主要抑制性神经递质。它对于维持大脑正常运作所需的兴奋与抑制之间的动态平衡至关重要。因此,鉴定 GABA 能神经元及其 GABA 释放部位对于理解大脑如何调节神经元的兴奋性以及负责众多大脑功能(包括信息处理、运动、学习、记忆和突触可塑性等)的神经回路的活动至关重要。
由于 GABA 突触的结构和特征对于理解它们在特定神经回路中的功能至关重要,因此我们在这里开发并鉴定了一种用于果蝇的 GABA 能突触小泡的条件性标记物 9XV5-vGAT。
9XV5-vGAT 的表达条件性、定位于突触小泡的特异性、在 GABA 能神经元中的表达特异性和功能性得到了验证。它对于 GABA 能神经递质表型鉴定和 GABA 释放部位的鉴定在中央复合体的椭圆体神经元中得到了验证。与先前报道的用于乙酰胆碱和谷氨酸的条件性 SV 标记物结合使用,9XV5-vGAT 被用于证明食管下神经节神经元的快速神经递质表型鉴定。
与单细胞转录组学相比,该方法是一种用于神经递质表型鉴定的替代方法,可应用于任何具有二元转录系统驱动的感兴趣神经元。
已经开发并验证了一种条件性 GABA 能突触小泡标记物,用于 GABA 神经递质表型鉴定和 GABA 能突触小泡的亚细胞定位。