High-resolution and cell-type-specific photostimulation mapping shows weak excitatory versus strong inhibitory inputs in the bed nucleus of the stria terminalis

The bed nucleus of the stria terminalis (BNST) is a key component of the extended amygdala, and has been implicated in anxiety and addiction. As individual neurons function within neural circuits, it is important to understand local microcircuits and larger network connections of identified neuronal types, and understand how maladaptive changes in the BNST neural networks are induced by stress and drug use. However, due to limitations of classic anatomical and physiological methods, local circuit organization of synaptic input to specific BNST neuron types is not well understood. In this study, we report on our application of high-resolution and cell-type specific photostimulation methodology developed in our laboratory to local circuit mapping in the BNST. Under calibrated experimental conditions, laser photostimulation via glutamate uncaging or channelrhodopsin-2 photoactivation evokes spiking of BNST neurons perisomatically, without activating spikes from axons of passage or distal dendrites. Whole cell recordings combined with spatially restricted photostimulation of presynaptic neurons at many different locations over a large region allow high resolution mapping of presynaptic input sources to single recorded neurons in the BNST. We constructed maps of synaptic inputs impinging onto corticotrophin releasing hormone-expressing (CRH+) BNST neurons in the dorsolateral BNST, and found that the CRH+ neurons receive predominant local inhibitory synaptic connections with very weak excitatory connections. Through cell-type specific optogenetic stimulation mapping, we generated maps of somatostatin-expressing-neuron specific inhibitory inputs to BNST neurons. Taken together, the photostimulation-based techniques offer us powerful tools for determining the functional organization of local circuits of specific BNST neuron types.

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