Intersectional, anterograde transsynaptic targeting of the neurons receiving monosynaptic inputs from two upstream regions

Abstract

A brain region typically receives inputs from multiple upstream areas. However, currently, no method is available to selectively access neurons that receive monosynaptic inputs from two upstream regions. Here, we devised a method to genetically label such neurons in mice by combining the anterograde transsynaptic spread of adeno-associated virus serotype 1 (AAV1) with intersectional gene expression. Injections of AAV1s expressing either Cre or Flpo recombinases and the Cre/Flpo double-dependent AAV into two upstream regions and the downstream region, respectively, were used to label the postsynaptic neurons receiving inputs from the two upstream regions. We demonstrated this labelling in two distinct circuits: the retina/primary visual cortex to the superior colliculus and the bilateral motor cortex to the dorsal striatum. Systemic delivery of the intersectional AAV allowed for unbiased detection of the labelled neurons throughout the brain. This strategy may help analyse the interregional integration of information in the brain.