Insulin receptors in tanycytes gate hypothalamic insulin uptake and regulate AgRP neuronal activity. Insulin receptors in tanycytes gate hypothalamic insulin uptake and regulate AgRP neuronal activity

We reveal a critical role of tanycyte insulin receptors in gating feeding state-dependent regulation of AgRP neurons and systemic insulin sensitivity, and show that insulin resistance in tanycytes contributes to the pleiotropic manifestations of obesity-associated insulin resistance Overall design: Bac-TRAP based RNA sequencing. 12 week old mice injected with AAV-Dio2-CreTW in lateral ventricle to express fluorescent L10-EGFP fusion protein in NCD-fed insulin receptor (IR) KO, WT or HFD-fed (for 8w) WT mice (NCD:IR fl/fl Rosa26-L10-EGFP tg/wt , IR wt/wt Rosa26-L10-EGFP tg/wt; HFD: IR wt/wt Rosa26-L10-EGFP tg/wt) in Cre dependent manner in tanycytes. 4 weeks later mediobasal hypothalamus was dissected. 5-7 mice (M&F) were pooled for replicate. Translating mRNA was pulled down via immunoprecipitation. Isolated RNA was subjected for RNA sequencing using Illumina platform. See full protocol below.