Identification of Differentially Expressed MicroRNAs and Their Potential Target Genes in Adipose Tissue from Pigs with Highly Divergent Backfat Thickness

Fatty traits are very important in pig production. However, the role of microRNAs (miRNAs) in fat deposition is not clearly understood. In this study, we compared adipose miRNAs from three full-sibling pairs of female Landrace pigs, with high and low backfat thickness, to investigate the associated regulatory network. We obtained an average of 17.29 million raw reads from six libraries, 62.27% of which mapped to the pig reference genome. A total of 318 pig miRNAs were detected among the samples. Among them, 18 miRNAs were differentially expressed (p-value < 0.05, |log2fold change| ≥ 1) between the high and low backfat groups; 6 were up-regulated and 12 were down-regulated. Functional enrichment of the predicted target genes of the differentially expressed miRNAs, indicated that these miRNAs were involved mainly in lipid and carbohydrate metabolism, and glycan biosynthesis and metabolism. Comprehensive analysis of the mRNA and miRNA transcriptomes revealed possible regulatory relationships for fat deposition. Negatively correlated mRNA–miRNA pairs included miR-137–PPARGC1A, miR-141–FASN, and miR-122-5p–PKM, indicating these interactions may be key regulators of fat deposition. Our findings provide important insights into miRNA expression patterns in the backfat tissue of pig and new insights into the regulatory mechanisms of fat deposition in pig.

Download Full-text

Expression of microRNAs in the hypothalamus of pregnant and non-pregnant goats

Czech Journal of Animal Science ◽

10.17221/113/2020-cjas ◽

2021 ◽

Vol 66 (No. 5) ◽

pp. 156-167

Author(s):

Lu Zhu ◽

Jingtong Huang ◽

Jing Jing ◽

Qi Zheng ◽

Qianyun Ji ◽

...

Keyword(s):

Target Genes ◽

Functional Enrichment ◽

Differentially Expressed ◽

Protein Coding ◽

Reproductive Process ◽

Animal Reproduction ◽

Differentially Expressed Mirnas ◽

Hypothalamic Regulation ◽

Basic Reference

MicroRNAs (miRNAs) play a significant role in animal reproduction by regulating the expression of protein-coding genes. The hypothalamus regulates the pregnancy cycle changes in goats; however, the action mechanism of miRNAs in this regulation remains to be investigated. In this study, we performed RNA sequencing of hypothalamus samples to establish a comprehensive miRNA profiling of pregnant and non-pregnant goats. A total of 384 miRNAs were identified in the hypothalamus of pregnant goats, of which 239 were newly discovered, and 390 miRNAs were detected in the hypothalamus of non-pregnant goats of which 192 were novel miRNAs. In addition, a total of 280 differentially expressed miRNAs are characterized, of which 171 were known miRNAs and 109 were novel miRNAs. Functional enrichment suggests that the predicted target genes of differentially expressed miRNAs may be involved in the reproductive process. This preliminary study revealed that let-7f-5p, miR-99a-5p and miR-100-5p may be involved in the hypothalamic regulation of pregnancy cycle changes in goats. These data provide a basic reference for subsequent studies on the regulatory role of miRNAs in mammalian pregnancy.

Download Full-text

Transcriptome analysis of miRNA and mRNA in the livers of pigs with highly diverged backfat thickness

Scientific Reports ◽

10.1038/s41598-019-53377-x ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 2

Author(s):

Kai Xing ◽

Xitong Zhao ◽

Hong Ao ◽

Shaokang Chen ◽

Ting Yang ◽

...

Keyword(s):

Fatty Acid ◽

Energy Metabolism ◽

Differentially Expressed Genes ◽

Regulatory Network ◽

Acid Synthesis ◽

Fat Deposition ◽

Differentially Expressed ◽

Backfat Thickness ◽

Lipid Deposition ◽

Differentially Expressed Mirnas

AbstractFat deposition is very important in pig production, and its mechanism is not clearly understood. MicroRNAs (miRNAs) play critical roles in fat deposition and energy metabolism. In the current study, we investigated the mRNA and miRNA transcriptome in the livers of Landrace pigs with extreme backfat thickness to explore miRNA-mRNA regulatory networks related to lipid deposition and metabolism. A comparative analysis of liver mRNA and miRNA transcriptomes from pigs (four pigs per group) with extreme backfat thickness was performed. We identified differentially expressed genes from RNA-seq data using a Cufflinks pipeline. Seventy-one differentially expressed genes (DEGs), including twenty-eight well annotated on the porcine reference genome genes, were found. The upregulation genes in pigs with higher backfat thickness were mainly involved in fatty acid synthesis, and included fatty acid synthase (FASN), glucokinase (GCK), phosphoglycerate dehydrogenase (PHGDH), and apolipoprotein A4 (APOA4). Cytochrome P450, family 2, subfamily J, polypeptide 34 (CYP2J34) was lower expressed in pigs with high backfat thickness, and is involved in the oxidation of arachidonic acid. Moreover, 13 differentially expressed miRNAs were identified. Seven miRNAs were associated with fatty acid synthesis, lipid metabolism, and adipogenic differentiation. Based on comprehensive analysis of the transcriptome of both mRNAs and miRNAs, an important regulatory network, in which six DEGs could be regulated by differentially expressed miRNAs, was established for fat deposition. The negative correlate in the regulatory network including, miR-545-5p and GRAMD3, miR-338 and FASN, and miR-127, miR-146b, miR-34c, miR-144 and THBS1 indicate that direct suppressive regulation may be involved in lipid deposition and energy metabolism. Based on liver mRNA and miRNA transcriptomes from pigs with extreme backfat thickness, we identified 28 differentially expressed genes and 13 differentially expressed miRNAs, and established an important miRNA-mRNA regulatory network. This study provides new insights into the molecular mechanisms that determine fat deposition in pigs.

Download Full-text

System Analysis of MIRNAs in Maize Internode Elongation

Biomolecules ◽

10.3390/biom9090417 ◽

2019 ◽

Vol 9 (9) ◽

pp. 417

Author(s):

Chuanxi Peng ◽

Xing Wang ◽

Tianyu Feng ◽

Rui He ◽

Mingcai Zhang ◽

...

Keyword(s):

Target Genes ◽

System Analysis ◽

Interaction Network ◽

Enrichment Analysis ◽

Functional Enrichment Analysis ◽

Functional Enrichment ◽

Differentially Expressed ◽

Internode Elongation ◽

Differentially Expressed Mirnas ◽

Post Transcriptional Regulation

MicroRNAs (miRNAs), the post-transcriptional gene regulators, are known to play an important role in plant development. The identification of differentially expressed miRNAs could better help us understand the post-transcriptional regulation that occurs during maize internode elongation. Accordingly, we compared the expression of MIRNAs between fixed internode and elongation internode samples and classified six differentially expressed MIRNAs as internode elongation-responsive miRNAs including zma-MIR160c, zma-MIR164b, zma-MIR164c, zma-MIR168a, zma-MIR396f, and zma-MIR398b, which target mRNAs supported by transcriptome sequencing. Functional enrichment analysis for predictive target genes showed that these miRNAs were involved in the development of internode elongation by regulating the genes respond to hormone signaling. To further reveal how miRNA affects internode elongation by affecting target genes, the miRNA–mRNA–PPI (protein and protein interaction) network was constructed to summarize the interaction of miRNAs and these target genes. Our results indicate that miRNAs regulate internode elongation in maize by targeting genes related to cell expansion, cell wall synthesis, transcription, and regulatory factors.

Download Full-text

Next-generation sequencing analysis reveals segmental patterns of microRNA expression in yak epididymis

Reproduction Fertility and Development ◽

10.1071/rd20113 ◽

2020 ◽

Vol 32 (12) ◽

pp. 1067

Author(s):

Wangsheng Zhao ◽

Eugene Quansah ◽

Meng Yuan ◽

Pengcheng Li ◽

Chuanping Yi ◽

...

Keyword(s):

Gene Expression ◽

Mirna Expression ◽

Expression Patterns ◽

Differentially Expressed ◽

Sequencing Analysis ◽

Differentially Expressed Mirnas ◽

Next Generation Sequencing Analysis ◽

Cauda Epididymidis ◽

Caput Epididymidis

MicroRNAs (miRNAs) have emerged as potent regulators of gene expression and are widely expressed in biological systems. In reproduction, they have been shown to have a significant role in the acquisition and maintenance of male fertility, whereby deletion of Dicer in mouse germ cells leads to infertility. Evidence indicates that this role of miRNAs extends from the testis into the epididymis, controlling gene expression and contributing to regional variations in gene expression. In this study, RNA sequencing technology was used to investigate miRNA expression patterns in the yak epididymis. Region-specific miRNA expression was found in the yak epididymis. In all, 683 differentially expressed known miRNAs were obtained; 190, 186 and 307 differentially expressed miRNAs were identified for caput versus corpus, corpus versus cauda and caput versus cauda region pairs respectively. Kyoto Encyclopedia of Genes and Genomes results showed endocytosis as the most enriched pathway across region pairs, followed by protein processing in the endoplasmic reticulum, phagosome, spliceosome and biosynthesis of amino acids in region pair-specific hierarchical order. Gene ontology results showed varied enrichment in terms including cell, biogenesis, localisation, binding and locomotion across region pairs. In addition, significantly higher miR-34c expression was seen in the yak caput epididymidis relative to the corpus and cauda epididymidis.

Download Full-text

Differentially Expressed miRNA-Gene Targets Related to Intramuscular Fat in Musculus Longissimus Dorsi of Charolais × Holstein F2-Crossbred Bulls

Genes ◽

10.3390/genes11060700 ◽

2020 ◽

Vol 11 (6) ◽

pp. 700

Author(s):

Bilal Ahmad Mir ◽

Henry Reyer ◽

Katrin Komolka ◽

Siriluck Ponsuksili ◽

Christa Kühn ◽

...

Keyword(s):

Target Genes ◽

Mirna Gene ◽

Target Prediction ◽

Microarray Experiment ◽

Intramuscular Fat ◽

Differentially Expressed ◽

Longissimus Dorsi ◽

P Value ◽

Mirna Expression Pattern ◽

Differentially Expressed Mirnas

Intramuscular fat (IMF) is a meat quality indicator associated with taste and juiciness. IMF deposition, influenced by genetic and non-genetic factors, occurs through a transcriptionally coordinated process of adipogenesis. MicroRNAs (miRNAs) are transcriptional regulators of vital biological processes, including lipid metabolism and adipogenesis. However, in bovines, limited data on miRNA profiling and association with divergent intramuscular fat content, regulated exclusively by genetic parameters, have been reported. Here, a microarray experiment was performed to identify and characterize the miRNA expression pattern in the Musculus longissimus dorsi of F2-cross (Charolais × German Holstein) bulls with high and low IMF. A total of 38 differentially expressed miRNAs (DE miRNAs), including 33 upregulated and 5 downregulated (corrected p-value ≤ 0.05, FC ≥ ±1.2), were reported. Among DE miRNAs, the upregulated miRNAs miR-105a/b, miR-695, miR-1193, miR-1284, miR-1287-5p, miR-3128, miR-3178, miR-3910, miR-4443, miR-4445 and miR-4745, and the downregulated miRNAs miR-877-5p, miR-4487 and miR-4706 were identified as novel fat deposition regulators. DE miRNAs were further analyzed, along with previously identified differentially expressed genes (DEGs) from the same samples and predicted target genes, using multiple bioinformatic approaches, including target prediction tools and co-expression networks, as well as Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment. We identified DE miRNAs and their gene targets associated with bovine intramuscular adipogenesis, and we provide a basis for further functional investigations.

Download Full-text

Genome-wide profiling of miRNA expression patterns in tubal endometriosis

Reproduction ◽

10.1530/rep-18-0631 ◽

2019 ◽

Vol 157 (6) ◽

pp. 525-534 ◽

Cited By ~ 4

Author(s):

Hang Qi ◽

Guiling Liang ◽

Jin Yu ◽

Xiaofeng Wang ◽

Yan Liang ◽

...

Keyword(s):

Pathway Analysis ◽

Mirna Expression ◽

Gene Networks ◽

Target Genes ◽

Expression Profiles ◽

Expression Patterns ◽

Mirna Gene ◽

Differentially Expressed ◽

Signal Pathways ◽

Differentially Expressed Mirnas

MicroRNA (miRNA) expression proﬁles in tubal endometriosis (EM) are still poorly understood. In this study, we analyzed the differential expression of miRNAs and the related gene networks and signaling pathways in tubal EM. Four tubal epithelium samples from tubal EM patients and five normal tubal epithelium samples from uterine leiomyoma patients were collected for miRNA microarray. Bioinformatics analyses, including Ingenuity Pathway Analysis (IPA), Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, were performed. Quantitative real-time polymerase chain reaction (qRT-PCR) validation of five miRNAs was performed in six tubal epithelium samples from tubal EM and six from control. A total of 17 significantly differentially expressed miRNAs and 4343 potential miRNA-target genes involved in tubal EM were identified (fold change >1.5 and FDR-adjustedPvalue <0.05). IPA indicated connections between miRNAs, target genes and other gynecological diseases like endometrial carcinoma. GO and KEGG analysis revealed that most of the identified genes were involved in the mTOR signaling pathway, SNARE interactions in vesicular transport and endocytosis. We constructed an miRNA-gene-disease network using target gene prediction. Functional analysis showed that the mTOR pathway was connected closely to tubal EM. Our results demonstrate for the first time the differentially expressed miRNAs and the related signal pathways involved in the pathogenesis of tubal EM which contribute to elucidating the pathogenic mechanism of tubal EM-related infertility.

Download Full-text

Identification of microRNAs in Silver Carp (Hypophthalmichthys molitrix) Response to Hypoxia Stress

Animals ◽

10.3390/ani11102917 ◽

2021 ◽

Vol 11 (10) ◽

pp. 2917

Author(s):

Qiaoxin Wang ◽

Xiaohui Li ◽

Hang Sha ◽

Xiangzhong Luo ◽

Guiwei Zou ◽

...

Keyword(s):

Rna Sequencing ◽

Small Rna ◽

Target Genes ◽

Silver Carp ◽

Functional Enrichment ◽

Differentially Expressed ◽

Small Rna Sequencing ◽

Hypophthalmichthys Molitrix ◽

Hypoxia Stress ◽

Differentially Expressed Mirnas

Hypoxia is one of the serious stresses in fish culture, which can lead to physical and morphological changes, and cause injury and even death to fish. Silver carp (Hypophthalmichthys molitrix) is an important economic fish and widely distributed in China. MicroRNA is a kind of endogenous non-coding single-stranded small RNA, which is involved in cell development, and immune response and gene expression regulation. In this study, silver carp were kept in the closed containers for hypoxia treatment by spontaneous oxygen consumption. The samples of heart, brain, liver and gill were collected, and the total RNAs extracted separately from the four tissues were mixed in equal amounts according to the concentration. Afterwards, the RNA pool was constructed for high-throughput sequencing, and based on the small RNA sequencing, the differentially expressed microRNAs were identified. Furthermore, their target gene prediction and enrichment analyses were carried out. The results showed that a total of 229 known miRNAs and 391 putative novel miRNAs were identified, which provided valuable resources for further study on the regulatory mechanism of miRNAs in silver carp under hypoxia stress. The authors verified 16 differentially expressed miRNAs by qRT-PCR, and the results were consistent with small RNA sequencing (sRNA-seq). The predicted target genes number of differentially expressed miRNAs was 25,146. GO and KEGG functional enrichment analysis showed that these target genes were mainly involved in the adaption of hypoxia stress in silver carp through biological regulation, catalytic activity and apoptosis. This study provides references for further study of interaction between miRNAs and target genes, and the basic data for the response mechanism under hypoxia stress in silver carp.

Download Full-text

Identification of miRNAs and Their Target Genes Involved in Cucumber Fruit Expansion Using Small RNA and Degradome Sequencing

Biomolecules ◽

10.3390/biom9090483 ◽

2019 ◽

Vol 9 (9) ◽

pp. 483 ◽

Cited By ~ 1

Author(s):

Sun ◽

Luo ◽

Chang ◽

Li ◽

Zhou ◽

...

Keyword(s):

Small Rna ◽

High Throughput Sequencing ◽

Target Genes ◽

Expression Patterns ◽

Differentially Expressed ◽

Sequencing Data ◽

Degradome Sequencing ◽

Complex Biological Process ◽

Cucumber Fruit ◽

Differentially Expressed Mirnas

Fruit expansion is an essential and very complex biological process. Regulatory roles of microRNAs (miRNAs) and miRNA–mRNA modules in the cucumber fruit expansion are not yet to be investigated. In this work, 1253 known and 1269 novel miRNAs were identified from nine cucumber fruit small RNA (sRNA) libraries through high-throughput sequencing. A total of 105 highly differentially expressed miRNAs were recognized in the fruit on five days post anthesis with pollination (EXP_5d) sRNA library. Further, expression patterns of 11 differentially expressed miRNAs were validated by quantitative real-time PCR (qRT-PCR). The expression patterns were similar to sRNAs sequencing data. Transcripts of 1155 sequences were predicted as target genes of differentially expressed miRNAs by degradome sequencing. Gene Ontology (GO) enrichment showed that these target genes were involved in 24 biological processes, 15 cell components and nine molecular functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis demonstrated that these target genes were significantly enriched in 19 pathways and the enriched KEGG pathways were associated with environmental adaptation, signal transduction and translation. Based on the functional prediction of miRNAs and target genes, our findings suggest that miRNAs have a potential regulatory role in cucumber fruit expansion by targeting their target genes, which provide important data for understanding the miRNA-mediated regulatory networks controlling fruit expansion in cucumber. Specific miRNAs could be selected for further functional research and molecular breeding in cucumber.

Download Full-text

A combined microRNA and target protein-based panel for predicting the probability and severity of uraemic vascular calcification: a translational study

Cardiovascular Research ◽

10.1093/cvr/cvaa255 ◽

2020 ◽

Cited By ~ 2

Author(s):

Chia-Ter Chao ◽

Hsiang-Yuan Yeh ◽

You-Tien Tsai ◽

Chih-Kang Chiang ◽

Huei-Wen Chen

Keyword(s):

Vascular Calcification ◽

Target Genes ◽

Ex Vivo ◽

Target Protein ◽

Functional Enrichment ◽

Differentially Expressed ◽

Target Proteins ◽

Differentially Expressed Mirnas

Abstract Aims Vascular calcification (VC) increases the future risk of cardiovascular events in uraemic patients, but effective therapies are still unavailable. Accurate identification of those at risk of developing VC using pathogenesis-based biomarkers is of particular interest and may facilitate individualized risk stratification. We aimed to uncover microRNA (miRNA)-target protein-based biomarker panels for evaluating uraemic VC probability and severity. Methods and results We created a three-tiered in vitro VC model and an in vivo uraemic rat model receiving high phosphate diet to mimic uraemic VC. RNAs from the three-tiered in vitro and in vivo uraemic VC models underwent miRNA and mRNA microarray, with results screened for differentially expressed miRNAs and their target genes as biomarkers. Findings were validated in original models and additionally in an ex vivo VC model and human cells, followed by functional assays of identified miRNAs and target proteins, and tests of sera from end-stage renal disease (ESRD) and non-dialysis-dependent chronic kidney disease (CKD) patients without and with VC. Totally 122 down-regulated and 119 up-regulated miRNAs during calcification progression were identified initially; further list narrowing based on miRNA–mRNA pairing, anti-correlation, and functional enrichment left 16 and 14 differentially expressed miRNAs and mRNAs. Levels of four miRNAs (miR-10b-5p, miR-195, miR-125b-2-3p, and miR-378a-3p) were shown to decrease throughout all models tested, while one mRNA (SULF1, a potential target of miR-378a-3p) exhibited the opposite trend concurrently. Among 96 ESRD (70.8% with VC) and 59 CKD patients (61% with VC), serum miR-125b2-3p and miR-378a-3p decreased with greater VC severity, while serum SULF1 levels increased. Adding serum miR-125b-2-3p, miR-378a-3p, and SULF1 into regression models for VC substantially improved performance compared to using clinical variables alone. Conclusion Using a translational approach, we discovered a novel panel of biomarkers for gauging the probability/severity of uraemic VC based on miRNAs/target proteins, which improved the diagnostic accuracy.

Download Full-text

MicroRNA-Mediated Responses to Chromium Stress Provide Insight Into Tolerance Characteristics of Miscanthus sinensis

Frontiers in Plant Science ◽

10.3389/fpls.2021.666117 ◽

2021 ◽

Vol 12 ◽

Author(s):

Gang Nie ◽

Zongchao Liao ◽

Minyi Zhong ◽

Jie Zhou ◽

Jiabang Cai ◽

...

Keyword(s):

Heavy Metal ◽

Target Genes ◽

Gene Annotation ◽

Function Analysis ◽

Critical Role ◽

Differentially Expressed ◽

Miscanthus Sinensis ◽

Differentially Expressed Mirnas ◽

Mirna Sequencing

Chromium (Cr) is a heavy metal in nature, which poses a potential risk to toxicity to both animals and plants when releasing into the environment. However, the regulation of microRNA (miRNA)-mediated response to heavy metal Cr has not been studied in Miscanthus sinensis. In this study, based on high-throughput miRNA sequencing, a total of 104 conserved miRNAs and 158 nonconserved miRNAs were identified. Among them, there were 45 differentially expressed miRNAs in roots and 13 differentially expressed miRNAs in leaves. The hierarchical clustering analysis showed that these miRNAs were preferentially expressed in a certain tissue. There were 833 differentially expressed target genes of 45 miRNAs in roots and 280 differentially expressed target genes of 13 miRNA in leaves. After expression trend analysis, five significantly enriched modules were obtained in roots, and three significantly enriched trend blocks in leaves. Based on the candidate gene annotation and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) function analysis, miR167a, novel_miR15, and novel_miR22 and their targets were potentially involved in Cr transportation and chelation. Besides, miR156a, miR164, miR396d, and novel_miR155 were identified as participating in the physiological and biochemical metabolisms and the detoxification of Cr of plants. The results demonstrated the critical role of miRNA-mediated responses to Cr treatment in M. sinensis, which involves ion uptake, transport, accumulation, and tolerance characteristics.

Download Full-text