Dataset Information

Adipose Tissue Gene Expression of Entire Male, Immunocastrated and Surgically Castrated Pigs.

ABSTRACT: Differences in adipose tissue deposition and properties between pig male sex categories, i.e., entire males (EM), immunocastrates (IC) and surgical castrates (SC) are relatively well-characterized, whereas the underlying molecular mechanisms are still not fully understood. To gain knowledge about the genetic regulation of the differences in adipose tissue deposition, two different approaches were used: RNA-sequencing and candidate gene expression by quantitative PCR. A total of 83 differentially expressed genes were identified between EM and IC, 15 between IC and SC and 48 between EM and SC by RNA-sequencing of the subcutaneous adipose tissue. Comparing EM with IC or SC, upregulated genes related to extracellular matrix dynamics and adipogenesis, and downregulated genes involved in the control of lipid and carbohydrate metabolism were detected. Differential gene expression generally indicated high similarity between IC and SC as opposed to EM, except for several heat shock protein genes that were upregulated in EM and IC compared with SC. The candidate gene expression approach showed that genes involved in lipogenesis were downregulated in EM compared with IC pigs, further confirming RNA-sequencing results.

SUBMITTER: Poklukar K

PROVIDER: S-EPMC7916650 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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Adipose Tissue Gene Expression of Entire Male, Immunocastrated and Surgically Castrated Pigs.

Poklukar Klavdija K Čandek-Potokar Marjeta M Vrecl Milka M Batorek-Lukač Nina N Fazarinc Gregor G Kress Kevin K Stefanski Volker V Škrlep Martin M

International journal of molecular sciences 20210210 4

Differences in adipose tissue deposition and properties between pig male sex categories, i.e., entire males (EM), immunocastrates (IC) and surgical castrates (SC) are relatively well-characterized, whereas the underlying molecular mechanisms are still not fully understood. To gain knowledge about the genetic regulation of the differences in adipose tissue deposition, two different approaches were used: RNA-sequencing and candidate gene expression by quantitative PCR. A total of 83 differentially ...[more]

PMID: 33578947

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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: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.

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.

Project description:Breeding efforts over the last decades altered markedly empty body (EB) composition of pigs. This study aimed to re-evaluate the dynamics of changes in the composition and deposition rate of fat, protein and amino acids (AA) in the EB from birth to 140 kg BW depending on the dietary CP and AA supply in a current pig genotype. In the experiment 66 entire male, 58 castrated and 66 female Swiss Large White pigs were used. From 20 kg BW onwards, they had either ad libitum access to a control (C) diet or a diet (LP) compared to diet C only 80% of CP, lysine, methione+cystine, threonine and tryptophan. The EB composition was determined at birth on eight boars and eight females, at 10 and 20 kg BW on two boars, two castrates and two females, and at 20 kg intervals from 40 to 140 kg BW, on four pigs per gender and dietary treatment. Each EB fraction was weighed and analysed for protein, fat and AA profile. The AA-to-lysine ratio was calculated and the different chemical component contents were fitted to allometric regressions. Overall, C-boars had the greatest EB protein and AA content and deposition rates, and lowest fat content and deposition rates. At the beginning of the grower period, LP-castrates and females displayed the lowest protein and AA and the highest fat deposition rates. However, compared with their counterparts in the C-group, in LP-castrates and females protein and AA deposition rates were greater above 64 and 40 kg EB weight, respectively, whereas fat deposition rates was lower above 80 kg EB weight. Thus, there seems a great potential to optimise protein and AA efficiency especially in the finisher period in castrates and females. Important individual variations were found in the essential AA-to-lysine ratio of the EB. Phenylalanine and threonine-to-lysine ratios decreased with increasing EB weight. Valine- and threonine-to-lysine ratios in C-castrates and C-females were 5% and 4% greater than recently reported by the National Research Council (NRC) whereas cysteine-, methionine- and tyrosine-to-lysine ratios were lower by 34%, 25% and 10%, respectively. The clear differences found between the EB AA-to-lysine ratios in the present study and the NRC might partly be explained by the genotype and the temporal changes in the relative weight of each EB fraction or changes in the AA profile. Nevertheless, these findings on changes in the essential AA profile of tissue protein warrant further studies.

Dataset Information

Adipose Tissue Gene Expression of Entire Male, Immunocastrated and Surgically Castrated Pigs.

Publications

Adipose Tissue Gene Expression of Entire Male, Immunocastrated and Surgically Castrated Pigs.

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