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A comprehensive microRNA expression profile of the backfat tissue from castrated and intact full-sib pair male pigs.

ABSTRACT: BACKGROUND: It is widely known that castration has a significant effect on the accumulation of adipose tissue. microRNAs (miRNAs) are known to be involved in fat deposition and to be regulated by the androgen-induced androgen receptor (AR). However, there is little understanding of the relationship between miRNAs and fat deposition after castration. In this study, the high-throughput SOLiD sequencing approach was used to identify and characterize miRNA expression in backfat from intact and castrated full-sib male 23-week-old pigs. The patterns of adipogenesis and fat deposition were compared between castrated and intact male pigs. RESULTS: A total of 366 unique miRNA genes were identified, comprising 174 known pre-miRNAs and 192 novel pre-miRNAs. One hundred and sixty-seven pre-miRNAs were common to both castrated (F3) and intact (F4) male pig small RNA libraries. The novel pre-miRNAs encoded 153 miRNAs/miRNA*s and 141 miRNAs/miRNA*s in the F3 and F4 libraries, respectively. One hundred and seventy-seven miRNAs, including 45 up- and 132 down-regulated, had more than 2-fold differential expression between the castrated and intact male pigs (p-value < 0.001). Thirty-five miRNAs were further selected, based on the expression abundance and differentiation between the two libraries, to predict their targets in KEGG pathways. KEGG pathway analyses suggested that miRNAs differentially expressed between the castrated and intact male pigs are involved in proliferation, apoptosis, differentiation, migration, adipose tissue development and other important biological processes. The expression patterns of eight arbitrarily selected miRNAs were validated by stem-loop reverse-transcription quantitative polymerase chain reaction. These data confirmed the expression tendency observed with SOLiD sequencing. miRNA isomiRs and mirtrons were also investigated in this study. Mirtrons are a recently described category of miRNA relying on splicing rather than processing by the microprocessor complex to generate the RNAi pathway. The functions of miRNAs important for regulating fat deposition were also investigated in this study. CONCLUSIONS: This study expands the number of fat-deposition-related miRNAs in pig. The results also indicate that castration can significantly affect the expression patterns of fat-related miRNAs. The differentially expressed miRNAs may play important roles in fat deposition after castration.

SUBMITTER: Bai Y

PROVIDER: S-EPMC3901342 | biostudies-other | 2014

REPOSITORIES: biostudies-other

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A comprehensive microRNA expression profile of the backfat tissue from castrated and intact full-sib pair male pigs.

Bai Ying Y Huang Jin-Ming JM Liu Gang G Zhang Ji-Bin JB Wang Jian-Ying JY Liu Cheng-Kun CK Fang Mei-Ying MY

BMC genomics 20140120

<h4>Background</h4>It is widely known that castration has a significant effect on the accumulation of adipose tissue. microRNAs (miRNAs) are known to be involved in fat deposition and to be regulated by the androgen-induced androgen receptor (AR). However, there is little understanding of the relationship between miRNAs and fat deposition after castration. In this study, the high-throughput SOLiD sequencing approach was used to identify and characterize miRNA expression in backfat from intact an ...[more]

PMID: 24443800

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Project description:BackgroundLong non-coding RNAs (lncRNAs) regulate adipose tissue metabolism, however, their function on testosterone deficiency related obesity in humans is less understood. For this research, intact and castrated male pigs are the best model animal because of their similar proportional organ sizes, cardiovascular systems and metabolic features.ResultsWe identified lncRNAs in subcutaneous adipose tissue by deep RNA-sequencing using the intact and castrated Huainan male pigs. The results showed that castration reduced serum testosterone but increased body fatness-related traits (serum triglyceride levels, backfat thickness, intramuscular fat content, and adipocyte size). Meanwhile, 343 lncRNAs from subcutaneous adipose tissue were identified, including 223 intergenic lncRNAs (lincRNAs), 68 anti-sense lncRNAs, and 52 intronic lncRNAs. It was predicted that there were 416 recognition sites for C/EBPα in the 303 lncRNA promoter region, and 13 adipogenesis-promoting miRNAs and five adipogenesis-depressing miRNAs target these lncRNAs. Eighteen lncRNAs, including nine up- and nine down-regulated had more than 2-fold differential expression between the castrated and intact male pigs (q-value < 0.05). Functional analysis indicated that these 18 lncRNAs and their target genes were involved in fatty acid, insulin, and the adipocytokine signaling pathway. We further analyzed the features of a conserved mouse lncRNA gene ENSMUST00000189966 and found it mainly expressed in the cell nucleus and target the Nuclear Receptor Subfamily 2 Group F Member 2 (NR2F2) gene. In 3 T3-L1 cells, differentiation down-regulated their expression, but dihydrotestosterone (DHT) significantly up-regulated their expression in a concentration-dependent manner (P < 0.05).ConclusionsThese results suggested that lncRNAs and their target genes might participated in the castration-induced fat deposition and provide a new therapeutic target for combatting testosterone deficiency-related obesity.

Project description:Intramuscular fat (IMF) is known to enhance beef palatability and can be markedly increased by castration. However, there is little understanding of the molecular mechanism underlying the IMF deposition after castration of beef cattle. We hypothesize that genetic regulators function differently in IMF from bulls and steers. Therefore, after detecting serum testosterone and lipid parameter, as well as the contents of IMF at 6, 12, 18 and 24 months, we have investigated differentially expressed (DE) microRNAs (miRNAs) and mRNAs in IMF of bulls and steers at 24 months of age in Qinchuan cattle using next-generation sequencing, and then explored the possible biopathways regulating IMF deposition. Serum testosterone levels were significantly decreased in steers, whereas IMF content, serum total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and triglycerides (TGs) were markedly increased in steers. Comparing the results of steers and bulls, 580 upregulated genes and 1,120 downregulated genes in IMF tissues were identified as DE genes correlated with IMF deposition. The upregulated genes were mainly associated with lipid metabolism, lipogenesis and fatty acid transportation signalling pathways, and the downregulated genes were correlated with immune response and intracellular signal transduction. Concurrently, the DE miRNAs-important players in adipose tissue accumulation induced by castration-were also examined in IMF tissues; 52 DE miRNAs were identified. The expression profiles of selected genes and miRNAs were also confirmed by quantitative real-time PCR (qRT-PCR) assays. Using integrated analysis, we constructed the microRNA-target regulatory network which was supported by target validation using the dual luciferase reporter system. Moreover, Ingenuity Pathway Analysis (IPA) software was used to construct a molecular interaction network that could be involved in regulating IMF after castration. The detected molecular network is closely associated with lipid metabolism and adipocyte differentiation, which is supported by functional identification results of bta-let-7i on bovine preadipocytes. These results provided valuable insights into the molecular mechanisms of the IMF phenotype differences between steers and bulls.

Project description:The optimized use of dietary nutrients and the accurate knowledge of the growth dynamics of body components is important for efficient pig production. This study aimed at evaluating the growth of carcass components and organs of Swiss Large White pigs from birth to 140 kg BW depending on the CP and amino acid (AA) supply. A total of 66 entire males (EM), 58 castrates (CA) and 66 female (FE) pigs were used. From 20 kg BW onwards, they had either ad libitum access to a control (C) or a diet (LP) with 20% lower CP, lysine, methionine + cystine, threonine and tryptophan content compared to C. The weight of organs, primal cuts and external fat were recorded in eight EM and eight FE; at 10 kg BW, on two EM, CA and FE each, and at 20 kg BW, on eight pigs from each sex. From 40 to 140 kg BW at 20 kg intervals, measurements were recorded on four pigs per sex and dietary treatment. The weight of each component was related to empty body (EB) using allometric regressions. Kidneys were heavier (P<0.05) in C- than LP-pigs and in EM than CA and FE. Above 21 kg EB weight, growth rate of LP-FE overpassed (P<0.05) the one of C-pigs. Consequently, LP-FE had heavier (P<0.05) lean cuts than C-pigs in the finisher period. However, LP-CA and LP-EM displayed lower (P<0.05) weights and growth rates of the lean cuts than C-CA and C-EM. Shoulder and loin weights and growth rates were reduced (P<0.05) in LP-pigs when compared to C-pigs. Growth rates of the ham were greater (P<0.05) in LP-FE than C-FE, whereas in LP-EM and LP-CA they were lower (P<0.05) than their C-counterparts. Total amounts of subcutaneous fat, backfat, ham fat and shoulder fat were lower (P<0.05) in C- than LP-pigs. The total amount of subcutaneous fat, backfat and shoulder fat of C-CA was, regardless of diet, greater (P<0.05) than of C-FE. In the LP group, this difference was even more pronounced. The amount of deposited ham fat was greater (P<0.05) in LP-CA than LP-FE, but not in C-CA v. C-FE. Differences in kidney weights suggested a greater nitrogen clearance required in C-pigs. Overall, dietary restriction and sex did not affect all body parts to the same extent. This study further suggests the possibility to reduce the CP and AA supply in FE without compromising the yield of primal lean cuts or increasing the amount of subcutaneous fat.

Dataset Information

A comprehensive microRNA expression profile of the backfat tissue from castrated and intact full-sib pair male pigs.

Publications

A comprehensive microRNA expression profile of the backfat tissue from castrated and intact full-sib pair male pigs.

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