Single cell sequencing of Stellate Ganglion of mice retrolabeled from interscapular brown adipose tissue and forelimb

To identify the molecular signature of the sympathetic neurons innervating interscapular brown adipose tissue (iBAT) in mice, we isolated, sorted, and sequenced single cells from the stellate ganglia (SG) of mice three days after injections of CTB-555 in iBAT and CTB-488 in forelimb. Each cell was analyzed for their emission after excitation at 488 and 555nm, and was assigned to either 488-positive, 555-positive, or none. Overall design: Six C57BL/6J male mice between six and eight weeks of age were injected with CTB-488 in their forelimb and with CTB-555 in their iBAT. Three days later, SGs were harvested, dissociated, and single cells were sorted in 96-well plates. Two mice were used for each set, for a total of three sets. Ganglia were pooled within sets. Four plates per set were collected, except for the third set, where only three plates were collected, for a total of 1039 cells across 11 plates. Finally, plate-based single cell sequencing was performed.