Abstract

Simple SummaryIn ruminants, the rumen epithelium plays an important role in nutrient absorption, metabolism and transport. MicroRNAs (miRNAs) have been reported to regulate the proliferation of diverse epithelial cells. In this study, we profiled the miRNA transcriptomes of goat rumens at four development stages and screened for candidate miRNAs related to rumen development. MiR-148a-3p was found to be highly expressed in the rumen tissues and induced the proliferation of GES-1 cells by targeting QKI. Our findings provide some insights into the functional roles of miRNAs in rumen growth and functional development in ruminants.The rumen is an important digestive organ of ruminants. From the fetal to adult stage, the morphology, structure and function of the rumen change significantly. However, the knowledge of the intrinsic genetic regulation of these changes is still limited. We previously reported a genome-wide expression profile of miRNAs in pre-natal goat rumens. In this study, we combined and analyzed the transcriptomes of rumen miRNAs during pre-natal (E60 and E135) and post-natal (D30 and D150) stages. A total of 66 differentially expressed miRNAs (DEMs) were identified in the rumen tissues from D30 and D150 goats. Of these, 17 DEMs were consistently highly expressed in the rumens at the pre-weaning stages (E60, E135 and D30), while down-regulated at D150. Noteworthy, annotation analysis revealed that the target genes regulated by the DEMs were mainly enriched in MAPK signaling pathway, Jak-STAT signaling pathway and Ras signaling pathway. Interestingly, the expression of miR-148a-3p was significantly high in the embryonic stage and down-regulated at D150. The potential binding sites of miR-148a-3p in the 3′-UTR of QKI were predicted by the TargetScan and verified by the dual luciferase report assay. The co-localization of miR-148a-3p and QKI through in situ hybridization was observed in the rumen tissues but not in the intestinal tracts. Moreover, the expression of miR-148a-3p in the epithelium was significantly higher than that in the other layers of the rumen, suggesting that miR-148a-3p is involved in the development of the rumen epithelial cells by targeting QKI. Subsequently, miR-148a-3p inhibitor was found to induce the proliferation of GES-1 cells. Taken together, our study identified DEMs involved in the development of the rumen and provides insights into the regulation mechanism of rumen development in goats.

Highlights

  • The rumen is the largest compartment of stomach in adult ruminants, which plays an important role in the digestion and absorption of nutrients

  • Inhibition of the expression of miR-148a-3p could significantly improve the proliferation capacity of GES-1 cells, suggesting that miR-148a-3p may be involved in the process of rumen development in goats

  • The expression characteristics of genome-wide microRNAs involved in rumen development and their potential influences on the functional transition of goat rumen epidermis were development and their potential influences on the functional transition of goat rumen epidermis were investigated

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Summary

Introduction

The rumen is the largest compartment of stomach in adult ruminants, which plays an important role in the digestion and absorption of nutrients. The development of the rumen in goats includes the development of tissue’s morphology and metabolic function. Some miRNAs have been shown to play crucial roles in skeletal myogenesis [9–11], fat deposition [12], adipogenic differentiation [13], lipid metabolism in mammary gland [14,15], testes development [16]. Previous studies have revealed that miR-148a-3p regulates the proliferation and apoptosis of skeletal muscle cells by post-transcriptional regulation of target gene in cattle [18] and chicken [19]. We investigated differentially expressed miRNAs (DEMs) identified between pre-weaning (D30) and post-weaning stage (D150), and jointly analyzed the RNA sequencing data from our previous study. Inhibition of the expression of miR-148a-3p could significantly improve the proliferation capacity of GES-1 cells, suggesting that miR-148a-3p may be involved in the process of rumen development in goats

Experimental Animals and Rumen Tissue Collection
Expression Vector Construction
Luciferase Reporter Assay
Cell Proliferation Assays
Dual-Color Fluorescence In Situ Hybridization
Statistical Analyses
Results
GO and KEGG Pathway Analyses of Putative Target Genes
Candidate miR-148a-3p Expression and Target Gene Analysis
Candidate miR-148a-3p
Discussion
Findings
Conclusions
Full Text
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