Resources.
Single-cell RNA-sequencing of mouse periaqueductal gray (PAG) neurons using Smart-seq2.
We optimized Voltage-Seq which combines, all-optical physiology, spatial mapping, on-site classification, and RNA-transcriptomics to robustly increase the throughput of synaptic connectivity testing and targeted molecular classification of postsynaptic neurons. Single-cell RNA-sequencing was performed on spatial- and voltage-recorded neurons in the mouse PAG. Uniform Manifold Approximation and Projection (UMAP) clustered cells as excitatory or inhibitory, and further differential expression analyses highlighted putative marker genes of GABAergic neurons. These sequencing results were in agreement with in-situ hybridization (ISH) and neuronal activity recordings.
VoltView 1.0 to analyze all-optical Voltron imaging with Example Data.
VoltVew 1.0 is an analysis package written in MATLAB by the Fuzik Laboratory at Karolinska Institute."VoltView_README.pdf" describes the installation of VoltView 1.0 and its dependencies (MATLAB 2021, bfmatlab, Anaconda3, Cellpose 1.0 in Python 3.8).
Patch-Seq RNA-transcriptome of hypothalamus-habenula projection neurons using Smart-seq2.
Excitatory projections from the lateral hypothalamic area (LHA) to the lateral habenula (LHb) drive aversive responses. We used Patch-seq guided multimodal classification to define the structural and functional heterogeneity of the LHA-LHb pathway. Our classification identified six glutamatergic neuron types with unique electrophysiological properties, molecular profiles, and projection patterns. We found that genetically-defined LHA-LHb neurons signal distinct aspects of emotional or naturalistic behaviors: Esr1+ LHA-LHb neurons induce aversion, whereas Npy+ LHA-LHb neurons control rearing behavior.