TET3 epigenetically controls feeding and stress response behaviors via AGRP neurons

Abstract

The TET family of dioxygenases promote DNA demethylation by oxidizing 5-methylcytosine to 5-hydroxymethylcytosine (5hmC). Hypothalamic agouti-related peptide–expressing (AGRP-expressing) neurons play an essential role in driving feeding, while also modulating nonfeeding behaviors. Besides AGRP, these neurons produce neuropeptide Y (NPY) and the neurotransmitter GABA, which act in concert to stimulate food intake and decrease energy expenditure. Notably, AGRP, NPY, and GABA can also elicit anxiolytic effects. Here, we report that in adult mouse AGRP neurons, CRISPR-mediated genetic ablation of Tet3 , not previously known to be involved in central control of appetite and metabolism, induced hyperphagia, obesity, and diabetes, in addition to a reduction of stress-like behaviors. TET3 deficiency activated AGRP neurons, simultaneously upregulated the expression of Agrp , Npy , and the vesicular GABA transporter Slc32a1 , and impeded leptin signaling. In particular, we uncovered a dynamic association of TET3 with the Agrp promoter in response to leptin signaling, which induced 5hmC modification that was associated with a chromatin-modifying complex leading to transcription inhibition, and this regulation occurred in both the mouse models and human cells. Our results unmasked TET3 as a critical central regulator of appetite and energy metabolism and revealed its unexpected dual role in the control of feeding and other complex behaviors through AGRP neurons.

DNA demethylation in the hypothalamus promotes transcription of Agtr1a and Slc12a2 and hypertension development

“…The membrane was then incubated overnight at 4 °C with the following primary antibodies diluted in in TBST: rabbit anti-DNMT1 (1:500, #24206-1-AP, Proteintech Group, Inc), rabbit anti-DNMT3A (1:1000, #3598S, Cell Signaling Technology), rabbit anti-DNMT3B (1:1000, #72335, Cell Signaling Technology), rabbit anti-MeCP2 (1:1000, #C15310088, Diagenode), rabbit anti-TET1 (1:500, #GTX124207, GeneTex, Inc), rabbit anti-TET2 (1:500, #C15410255, Diagenode Inc), and mouse anti-TET3 (1:1000, #61395, Active Motif). The specificity of these primary antibodies has been previously validated ( 66 , 67 , 68 , 69 , 70 , 71 , 72 ). For the protein loading control, a rabbit anti-GAPDH antibody (1:1000, #5174S, Cell Signaling Technology) was used.…”

Neuropeptide Y Peptide Family and Cancer: Antitumor Therapeutic Strategies

“…Y2R and Y4R stimulate Gq/11 alpha subunit dissociation to increase intracellular calcium levels and protein kinase C (PKC) activation [ 60 ]. YR signaling routes may coexist and connect with other receptor-triggered pathways turning on downstream biochemical signaling changes and leading to adaptive changes controlling cell behavior [ 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 ] ( Figure 7 ).…”

DNA Methylation in the Hypothalamic Feeding Center and Obesity

“…Tet3 expression in mouse AgRP neurons is elevated under fed conditions compared with fasted conditions; furthermore, TET3 plays an indispensable role in leptin-induced repression of Agrp expression by inducing 5hmC modifications in the Agrp promoter, which stabilizes the association of signal transducer and activator of transcription 3 (STAT3) and the corepressor complex with the Agrp promoter. 53 Tet3 deletion in AgRP neurons increases body weight through increased food intake and reduction in energy expenditure. 53 Bobcat339, a synthetic small molecule that decreases TET3 in AgRP neurons, prevents weight loss in food-restricted conditions by increasing food intake, and attenuating compulsive wheel running.…”