MiR-505-3p, a New Repressor of Puberty Onset in Female Mice

Abstract

Puberty onset is a complex trait regulated by multiple genetic and environmental factors. In this study, we narrowed a puberty-related QTL on chromosome X in female mice to a 1.7 Mb region and deduced that miR-505-3p was the functional gene.peer-reviewed) is the author/funder. All rights reserved. No reuse allowed without permission.The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/271718 doi: bioRxiv preprint first posted online Mar. 13, 2018; 2 In a GT1-7 cell line with stable overexpression of miR-505-3p (pGT1-7), both ribosomes and ribosome biogenesis pathways were affected, and the expression of some puberty-related genes was down-regulated. The amount of mRNA and protein products of the Srsf1 gene decreased by 50 percent, and the expression of puberty-related genes was rescued by the overexpression of Srsf1. With the down regulation of Srsf1 expression through shRNA, the mRNA accumulation of puberty-related genes decreased simultaneously in the GT1-7 cell line. The results of RIP-seq showed that SF2, the protein of the Srsf1 gene, primarily bound ribosome protein (RP) mRNAs in GT1-7 cells.miR-505-3p knockout female mice showed earlier vaginal opening, higher serum gonadotrophin levels and higher expression of puberty-related and Srsf1 genes in the hypothalamus than their wild-type littermates. B6 female mice with ectopic expression of miR-505-3p in the hypothalamus showed significant growth retardance and later VO than wild types.These results suggest that miR-505-3p may regulate puberty onset via the Srsf1 gene and RP expression, which reveals a new regulatory pathway in mammalian puberty onset involving microRNA, SF2 and ribosome proteins.Key words: puberty onset, positional cloning, miR-505-3p, GT1-7, CRISPR/Cas9 Author summaryIn this study, we identified miR-505-3p in a puberty-related QTL on the X chromosome as a female puberty onset repressor using a positional cloning strategy. GT1-7 cell lines stably overexpressing miR-505-3p (pGT1-7) showed Kiss1 and GnRH down-regulation. We also identified Srsf1 as the functional target gene of miR-505-3p in GT1-7 cells. Compared to wild-type mice, miR-505-3p knockout female mice showed puberty onset four days sooner, along with the overexpression of miR-505-3p in the hypothalamus 2 days later. Thus, miR-505-3p is a new repressor of puberty onset in female mice.

Proteomic Analysis of Hypothalamus and Pituitary Gland in Pre and Postpubertal Brahman Heifers

“…Further, ribosomal DA proteins in the interaction network, such as RPS5, RPS3 and RPLP1, were also listed in the networks constructed from endometrial gene expression of low and high fertility heifers ( Killeen et al, 2014 ). Recently, miR-503-3p was proposed as a new repressor of the initiation of puberty in female mice ( Tong et al, 2018 ). The stable overexpression of miR-503-3p in the GT1-7 cell line can influence ribosome biogenesis pathways and result in down-regulation of puberty-related genes ( Tong et al, 2018 ).…”

“…Recently, miR-503-3p was proposed as a new repressor of the initiation of puberty in female mice ( Tong et al, 2018 ). The stable overexpression of miR-503-3p in the GT1-7 cell line can influence ribosome biogenesis pathways and result in down-regulation of puberty-related genes ( Tong et al, 2018 ). The ribosomal protein RPL22 inhibited the expression of Lin28B, a puberty-related gene ( Ong et al, 2009 ; Rao et al, 2012 ).…”