Functionally Distinct Pomc-Expressing Neuron Subpopulations in Hypothalamus Revealed by Intersectional Targeting
We report differences in translational profiles between POMCLepr+ and POMCGlp1r+ neurons. Overall design: 6 hypothalami from male and female (3m, 3f) POMCDre LeprCre ROSA26lSlrSrEGFPL10a+/- and 12 hypothalami from male and female (6m, 6f) POMCDre Glp1rCre ROSA26lSlrSrEGFPL10a+/- mice were pooled for RNA extraction. Mice were rapidly decapitated. The arcuate nucleus (ARC) was dissected manually, snap frozen in liquid Nitrogen and stored at -80°C. RNA was extracted from the samples using the RNA extraction protocol (see details below). The pooled hypothalami, served as the input and was subjected to immunoprecipitation of the EGFP-tagged ribosomes using an anti-EGFP antibody. RNA concentrations were measured by a Qbit machine, finally, mRNA sequencing was performed on input and pulldown RNA samples.