ABSTRACT
Mkrn3, the maternally imprinted gene encoding the makorin RING-finger protein-3, has recently emerged as putative pubertal repressor, as evidenced by central precocity caused by MKRN3 mutations in humans; yet, the molecular underpinnings of this key regulatory action remain largely unexplored. We report herein that the microRNA, miR-30, with three binding sites in a highly conserved region of its 3' UTR, operates as repressor of Mkrn3 to control pubertal onset. Hypothalamic miR-30b expression increased, while Mkrn3 mRNA and protein content decreased, during rat postnatal maturation. Neonatal estrogen exposure, causing pubertal alterations, enhanced hypothalamic Mkrn3 and suppressed miR-30b expression in female rats. Functional in vitro analyses demonstrated a strong repressive action of miR-30b on Mkrn3 3' UTR. Moreover, central infusion during the juvenile period of target site blockers, tailored to prevent miR-30 binding to Mkrn3 3' UTR, reversed the prepubertal down-regulation of hypothalamic Mkrn3 protein and delayed female puberty. Collectively, our data unveil a novel hypothalamic miRNA pathway, involving miR-30, with a prominent role in the control of puberty via Mkrn3 repression. These findings expand our current understanding of the molecular basis of puberty and its disease states.
Subject(s)
Hypothalamus/metabolism , MicroRNAs/physiology , Sexual Maturation/genetics , Ubiquitin-Protein Ligases/genetics , Animals , Binding Sites , Cell Line , Female , Gene Expression Regulation, Developmental , Male , MicroRNAs/metabolism , Rats , Sequence Analysis, DNAABSTRACT
The influence of pomegranate juice (PJ, replacing water as solvent) and citric acid (CA) on properties of pectin films was studied. PJ provided the films with a bright red color, and acted as a plasticizer. Increasing PJ/water ratio from 0/100 to 100/0 resulted in enhanced elongation (from 2% to 20%), decreased strength (from 10 to <2 MPa) and modulus (from 93 to <10 MPa), increased water vapor permeability (WVP, from 3 to 9 g.mm.kPa(-1).h(-1).m(-2)), and decreased insoluble matter (IM, from 35% to 24%). Although a crosslinking effect by CA was not confirmed, it has been suggested to occur from its effects on films. CA noticeably increased IM (from <10% to almost 40%); moreover, when measured on a dry film basis, the CA effects presented a noticeable tendency to increases strength and modulus, and to decrease WVP. The red color density was decreased by CA, suggesting a destabilization of anthocyanins.
