Abstract

BackgroundMicroRNAs (miRNAs) have been implicated in the regulation of milk protein synthesis and development of the mammary gland (MG). However, the specific functions of miRNAs in these regulations are not clear. Therefore, the elucidation of miRNA expression profiles in the MG is an important step towards understanding the mechanisms of lactogenesis.ResultsTwo miRNA libraries were constructed from MG tissues taken from a lactating and a non-lactating Holstein dairy cow, respectively, and the short RNA sequences (18–30 nt) in these libraries were sequenced by Solexa sequencing method. The libraries included 885 pre-miRNAs encoding for 921 miRNAs, of which 884 miRNAs were unique sequences and 544 (61.5%) were expressed in both periods. A custom-designed microarray assay was then performed to compare miRNA expression patterns in the MG of lactating and non-lactating dairy cows. A total of 56 miRNAs in the lactating MG showed significant differences in expression compared to non-lactating MG (P<0.05). Integrative miRNA target prediction and network analysis approaches were employed to construct an interaction network of lactation-related miRNAs and their putative targets. Using a cell-based model, six miRNAs (miR-125b, miR-141, miR-181a, miR-199b, miR-484 and miR-500) were studied to reveal their possible biological significance.ConclusionOur study provides a broad view of the bovine MG miRNA expression profile characteristics. Eight hundred and eighty-four miRNAs were identified in bovine MG. Differences in types and expression levels of miRNAs were observed between lactating and non-lactating bovine MG. Systematic predictions aided in the identification of lactation-related miRNAs, providing insight into the types of miRNAs and their possible mechanisms in regulating lactation.

Highlights

  • MicroRNAs have been implicated in the regulation of milk protein synthesis and development of the mammary gland (MG)

  • Determination of bovine MG period Hematoxylin-eosin staining (HE) and immunofluorescence (IF) were employed to verify the microstructure differences of the lactating and non-lactating MG tissues used in constructing miRNA libraries

  • Alpha-casein was detected using immunofluorescence using an anti-α-casein antibody followed by HRP-conjugated goat anti-rabbit IgG and analysis using fluorescence microscopy. (E) Expression of αs1-casein mRNA in lactating and non-lactating bovine mammary glands using real-time PCR

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Summary

Introduction

MicroRNAs (miRNAs) have been implicated in the regulation of milk protein synthesis and development of the mammary gland (MG). The specific functions of miRNAs in these regulations are not clear. The elucidation of miRNA expression profiles in the MG is an important step towards understanding the mechanisms of lactogenesis. The complex initiation of MG lactation has been extensively studied over the years at the genetic, physiological and morphological levels because of its important functions [6]. Only a few studies have assessed the potential implication of miRNAs in MG lactogenesis. Studies on the regulation of miRNA expression profiles in bovine MG during lactation are still in their infancy. Identifying MG miRNA expression profiles is an important approach to explore the mechanism of lactation initiation and to identify biomarkers for lactogenesis

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