Placental miR-340 mediates vulnerability to activity based anorexia in mice

Abstract

Anorexia nervosa (AN) is a devastating eating disorder characterized by self-starvation that mainly affects women. Its etiology is unknown, which impedes successful treatment options leading to a limited chance of full recovery. Here, we show that gestation is a vulnerable window that can influence the predisposition to AN. By screening placental microRNA expression of naive and prenatally stressed (PNS) fetuses and assessing vulnerability to activity-based anorexia (ABA), we identify miR-340 as a sexually dimorphic regulator involved in prenatal programming of ABA. PNS caused gene-body hypermethylation of placental miR-340, which is associated with reduced miR-340 expression and increased protein levels of several target transcripts, GR, Cry2 and H3F3b. MiR-340 is linked to the expression of several nutrient transporters both in mice and human placentas. Using placenta-specific lentiviral transgenes and embryo transfer, we demonstrate the key role miR-340 plays in the mechanism involved in early life programming of ABA.

Sex dependent impact of gestational stress on predisposition to eating disorders and metabolic disease

“…To create these groups, we exposed pregnant ICR/CD1 dams to a chronic variable stress (prenatal stress - PNS) protocol for most of the pregnancy period, beginning a day after the detection of the copulation plug (gestation day (GD)1.5) and finishing on GD16.5 ( Figure 1 A). PNS dams gained less weight and had higher corticosterone (CORT) levels towards the end of the protocol ( Figure 1 B,C), which was already reported in a previous study [23] . PNS fetuses also displayed higher CORT levels but showed similar body weight at birth compared to controls (CTRLs; Figure 1 C,D).…”

“…PNS fetuses also displayed higher CORT levels but showed similar body weight at birth compared to controls (CTRLs; Figure 1 C,D). Maternal behavior did not differ between the groups, excluding this environmental factor as a potential confounding variable ( Figure 1 E, [23] ). Body weight follow-up of the offspring showed no differences between the groups in either sex ( Figure 1 F,K).…”

“…To assess the susceptibility and metabolic response of PNS offspring to ABA, we exposed the adolescent offspring (age 30 days) to a modified version of the ABA protocol [23] , [24] . The ABA model mimics the choice of exercise over eating, relying on limited food intake while allowing unlimited access to running wheels (RW) [25] , [26] .…”

“…The limited period of food access allows them to retain the majority of their BW as long as they engage in compensatory food intake when accessible, instead of excessive wheel running. The ABA protocol is characterized by adolescent vulnerability and sex specificity to females [23] , [27] , [28] , hyperactivity [23] , [25] , [29] , dramatic weight loss as a consequence of decreased food intake [23] , [24] , [30] , and circadian disruption in the running pattern [23] , [28] . Briefly, we habituated the animals to voluntary wheel running for one week with free access to food, followed by four days of free access to wheels combined with gradual FR in the form of 3–4 h of food access a day during the dark cycle ( Figure 3 A).…”

Nuclear receptor corepressor 1 represses cardiac hypertrophy

“…After primary and secondary antibody incubation, protein expression was visualized using ECL Western Blotting Substrate (Thermo Fisher Scientific) and quantified using ImageJ software. The primary antibodies were anti‐Actin (1:2,000, A7811, Santa Cruz), anti‐α‐Actinin (1:1,000, A7811, Sigma‐Aldrich), anti‐Flag (1:1,000, F3165, Sigma‐Aldrich), anti‐GFP (1:1,000, A10262, Thermo Fisher Scientific), anti‐HA (1:1,000, H3663, Sigma‐Aldrich or 3724, Cell Signaling), anti‐HDAC9 (1:200, sc‐398003, Santa Cruz; Schroeder et al , ), anti‐MEF2a (1:200, sc‐17785, Santa Cruz; Reineke et al , ), anti‐NCoR1 (1:500, 5948, Cell Signaling; Simcox et al , ), and anti‐HDAC4 (1:500, ab12172, Abcam or 2072, Cell Signaling; Wein et al , ).…”