TRAP experiment to determine gene expression changes in absence of AMPK activity in the VMN

The ventromedial hypothalamus (VMN) is implicated in the regulation of many aspects of energy homeostasis, including the counterregulatory response to hypoglycemia. Neurons of the VMN that are activated by hypoglycemia (glucose inhibited neurons (GI)) have been assumed to play a critical, although untested role in this response. Here, we used that expression of a dominant negative form of AMP-activated protein kinase (AMPK) in Sf1 neurons of the VMN which selectively suppressed GI neurons activity. We then performed Translating Ribosome Affinity Purification (TRAP Seq) to evaluate the transcriptional changes induced specifically in that cell population. Pools of six animals dissected hypothalamus were used for each sample, and an input control was generated. Immunoprecipitated material constitutes the outputs samples. In total, six samples were generated in control conditions (dnw_1_i, dnw_1_o, dnw_2_i, dnw_2_o, dnw_3_i, dnw_3_o) and six in dominant negative conditions (dn1_i, dn1_o, dn2_i, dn2_o, dn3_i, dn3_o).