Although it is now known that certain neurons can produce, store, and release multiple neurotransmitters, their locations, abundance, and functions remain elusive. We developed intersectional genetic strategies to identify multi-transmitter neurons based on the expression of neurotransmitter-specific genes. Here we present our procedures for whole-brain mapping of GABA/glutamate co-releasing neurons. We also detail our technique for labeling GABA/glutamate neurons in specific brain regions with adeno-associated virus (AAV). Our protocol can be readily extended to other types of multi-transmitter neurons. For complete details on the use and execution of this protocol, please refer to Xu et al. (2022).1.
Keywords: Genetics; Neuroscience.
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