Pig Adipose Tissue Research Articles

Porcine transient receptor potential channel 1 promotes adipogenesis and lipid deposition

Adipose tissue, an important organ involved in energy metabolism and endocrine, is closely related to animal meat quality and human health. Transient receptor potential channel 1 (TRPC1), an ion transporter, is adipocytes’ major Ca2+ entry channel. However, its function in fat deposition is poorly understood, particularly in pigs, which are both an ideal model for human obesity research and a primary meat source for human diets. In the present investigation, our findings demonstrate a prominent expression of TRPC1 within the adipose tissue of pigs with a strong fat deposition ability. Functional analysis showed that TRPC1 promotes primary preadipocyte proliferation and adipogenic differentiation. In vivo, transgenic mice expressing porcine TRPC1 (Tg-pTRPC1) exhibited aggravated high-fat diet-induced obesity, hepatic steatosis, and insulin resistance. Moreover, TRPC1 may facilitate adipogenesis via activating PI3K/AKT and β-catenin signaling pathways. Our research underscores the pivotal role of porcine TRPC1 as a positive regulator in adipogenesis and lipid accumulation processes, providing a potential target for improving animal meat quality and treating obesity-related diseases in humans.

Transcriptome Analysis Reveals the Early Development in Subcutaneous Adipose Tissue of Laiwu Piglets.

Adipose tissue plays an important role in pig production efficiency. Studies have shown that postnatal development has a vital impact on adipose tissue; however, the mechanisms behind pig adipose tissue early-life programming remain unknown. In this study, we analyzed the transcriptomes of the subcutaneous adipose tissue (SAT) of 1-day and 21-day old Laiwu piglets. The results showed that the SAT of Laiwu piglets significantly increased from 1-day to 21-day, and transcriptome analysis showed that there were 2352 and 2596 differentially expressed genes (DEGs) between 1-day and 21-day SAT in male and female piglets, respectively. Expression of genes in glycolysis, gluconeogenesis, and glycogen metabolism such as pyruvate kinase M1/2 (PKM), phosphoenolpyruvate carboxy kinase 1 (PCK1) and amylo-alpha-1, 6-glucosidase, 4-alpha-glucanotransferase (AGL) were significantly different between 1-day and 21-day SAT. Genes in lipid uptake, synthesis and lipolysis such as lipase E (LIPE), acetyl-CoA carboxylase alpha (ACACA), Stearoyl-CoA desaturase (SCD), and 3-hydroxy-3-methylglutaryl-CoA synthase 1 (HMGCS1) were also differentially expressed. Functional analysis showed enrichment of DEGs in transcriptional regulation, protein metabolism and cellular signal transduction. The protein-protein interaction (PPI) networks of these DEGs were analyzed and potential hub genes in these pathways were identified, such as transcriptional factors forkhead box O4 (FOXO4), CCAAT enhancer binding protein beta (CEBPB) and CCAAT enhancer binding protein delta (CEBPD), signal kinases BUB1 mitotic checkpoint serine/threonine kinase (BUB1) and cyclin-dependent kinase 1 (CDK1), and proteostasis-related factors ubiquitin conjugating enzyme E2 C (UBE2C) and cathepsin D (CTSD). Moreover, we further analyzed the transcriptomes of SAT between genders and the results showed that there were 54 and 72 DEGs in 1-day and 21-day old SAT, respectively. Genes such as KDM5D and KDM6C showed gender-specific expression in 1-day and 21-day SAT. These results showed the significant changes in SAT between 1-day and 21-day in male and female Laiwu pigs, which would provide information to comprehensively understand the programming of adipose tissue early development and to regulate adipose tissue function.

Lysine 2-hydroxyisobutyrylation levels determined adipogenesis and fat accumulation in adipose tissue in pigs

BackgroundExcessive backfat deposition lowering carcass grade is a major concern in the pig industry, especially in most breeds of obese type pigs. The mechanisms involved in adipogenesis and fat accumulation in pigs remain unclear. Lysine 2-hydroxyisobutyrylation (Khib), is a novel protein post-translational modification (PTM), which play an important role in transcription, energy metabolism and metastasis of cancer cells, but its role in adipogenesis and fat accumulation has not been shown.ResultsIn this study, we first analyzed the modification levels of acetylation (Kac), Khib, crotonylation (Kcr) and succinylation (Ksu) of fibro-adipogenic progenitors (FAPs), myogenic precursors (Myo) and mesenchymal stem cells (MSCs) with varied differentiation potential, and found that only Khib modification in FAPs was significantly higher than that in MSCs. Consistently, in parallel with its regulatory enzymes lysine acetyltransferase 5 (KAT5) and histone deacetylase 2 (HDAC2) protein levels, the Khib levels increased quadratically (P < 0.01) during adipogenic differentiation of FAPs. KAT5 knockdown in FAPs inhibited adipogenic differentiation, while HDAC2 knockdown enhanced adipogenic differentiation. We also demonstrated that Khib modification favored to adipogenic differentiation and fat accumulation by comparing Khib levels in FAPs and backfat tissues both derived from obese-type pigs (Laiwu pigs) and lean-type pigs (Duroc pigs), respectively. Accordingly, the expression patterns of KAT5 and HDAC2 matched well to the degree of backfat accumulation in obese- and lean-type pigs.ConclusionsFrom the perspective of protein translational modification, we are the first to reveal the role of Khib in adipogenesis and fat deposition in pigs, and provided new clues for the improvement of fat accumulation and distribution as expected via genetic selection and nutritional strategy in obese-type pigs.

Differential expression of ADRB1 causes different responses to norepinephrine in adipocytes of Duroc-Landrace-Yorkshire pigs and min pigs

Research has shown that pigs from different regions exhibit varying responses to cold stimuli. Typically, cold stimuli induce browning of white adipose tissue mediated by adrenaline, promoting non-shivering thermogenesis. However, the molecular mechanisms underlying differential response of pig breeds to norepinephrine are unclear.The aim of this study was to investigate the differences and molecular mechanisms of the effects of norepinephrine (NE) treatment on adipocytes of Min pigs (a cold-resistant pig breed) and Duroc-Landrace-Yorkshire (DLY) pigs. Real time-qPCR, western blot, and immunofluorescence were performed following NE treatment on cell cultures of adipocytes originating from Min pigs (n = 3) and DLY pigs (n = 3) to assess the expressions of adipogenesis markers, beige fat markers, and mitochondrial biogenesis markers. The results showed that NE did not affect browning of adipocytes in DLY pigs, whereas promoted browning of adipocytes in Min pigs. Further, the expression of ADRB1 (Adrenoceptor Beta 1, ADRB1) was higher in subcutaneous adipose tissue and adipocytes of Min pigs than those of DLY pigs. Overexpression of ADRB1 in DLY pig adipocytes enhanced sensitivity to NE, exhibiting decreased adipogenesis markers, upregulated beige fat markers, and increased mitochondrial biogenesis. Conversely, adipocytes treated with ADRB1 antagonist in Min pigs resulted in decreased cellular sensitivity to NE. Further studies revealed differential CpG island methylation in ADRB1 promoter region, with lower methylation levels in Min pigs compared to DLY pigs. In conclusion, differential methylation of the ADRB1 promoter region leads to different ADRB1 expression, resulting in varying responsiveness to NE in adipocytes of two pig breeds. Our results provide new insights for further analysis of the differential cold responsiveness in pig breeds from different regions.

Network Meta-Analysis: Effect of Cold Stress on the Gene Expression of Swine Adipocytes ATGL, CIDEA, UCP2, and UCP3.

Cold stress significantly affects gene expression in adipocytes; studying this phenomenon can help reveal the pathogeneses of conditions such as obesity and insulin resistance. Adipocyte triglyceride lipase (ATGL); cell death-inducing deoxyribonucleic acid (DNA) fragmentation factor subunit alpha (DFFA)-like effector (CIDEA); and uncoupling protein genes UCP1, UCP2, and UCP3 are the most studied genes in pig adipose tissues under cold stress. However, contradictory results have been observed in gene expression changes to UCP3 and UCP2 when adipose tissues under cold stress were examined. Therefore, we conducted a meta-analysis of 32 publications in total on the effect of cold stress on the expression of ATGL, CIDEA, UCP2, and UCP3. Our results showed that cold stress affected the expression of swine adipocyte genes; specifically, it was positively correlated with the expression of UCP3 in swine adipocytes. Conversely, expression of ATGL was negatively affected under cold stress conditions. In addition, the loss of functional UCP1 in pigs likely triggered a compensatory increase in UCP3 activity. We also simulated the docking results of UCP2 and UCP3. Our results showed that UCP2 could strongly bind to adenosine triphosphate (ATP), meaning that UCP3 played a more significant role in pig adipocytes.

Heavy Pigs Reared for Italian Dry-Cured Products: Does Immunocastration Influence the Fatty Acid Profile of Loins and Backfat?

The Italian pig sector requires heavy pigs (raised for at least nine months and slaughtered at >160 kg). In order to avoid boar taint and lower the impact on welfare, immunocastration provides an alternative to surgical castration. This study investigated the effects of immunocastration compared to surgical castration on the chemical composition and fatty acid profile of loins (longissimus dorsi muscle) and adipose tissue in Italian heavy pigs raised for dry-cured ham. Twenty-four male pigs were subjected to surgical castration (n = 12) or immunocastration (n = 12). Carcass parameters were monitored at slaughter, and samples of longissimus dorsi muscle and subcutaneous fat were analysed. This study showed no significant differences in carcass characteristics and proximate composition of fresh meat. However, variations were observed in the fatty acid profiles of meat and adipose tissue between groups. Notably, saturated fatty acids, particularly stearic acid (18:0), were higher in the intramuscular fat (IMF) of the immunocastrated pigs compared to the surgically castrated pigs. Conversely, monounsaturated fatty acids, predominantly oleic acid (18:1n-9), were higher in the IMF from the surgically castrated pigs compared to the immunocastrated pigs. While immunocastration may offer benefits in terms of animal growth and carcass composition, it could lead to unfavourable lipid changes in fresh loin meat for Italian heavy pigs.

The Role of β3-Adrenergic Receptors in Cold-Induced Beige Adipocyte Production in Pigs.

After exposure to cold stress, animals enhance the production of beige adipocytes and expedite thermogenesis, leading to improved metabolic health. Although brown adipose tissue in rodents is primarily induced by β3-adrenergic receptor (ADRB3) stimulation, the activation of major β-adrenergic receptors (ADRBs) in pigs has been a topic of debate. To address this, we developed overexpression vectors for ADRB1, ADRB2, and ADRB3 and silenced the expression of these receptors to observe their effects on the adipogenic differentiation stages of porcine preadipocytes. Our investigation revealed that cold stress triggers the transformation of subcutaneous white adipose tissue to beige adipose tissue in pigs by modulating adrenergic receptor levels. Meanwhile, we found that ADRB3 promotes the transformation of white adipocytes into beige adipocytes. Notably, ADRB3 enhances the expression of beige adipose tissue marker genes, consequently influencing cellular respiration and metabolism by regulating lipolysis and mitochondrial expression. Therefore, ADRB3 may serve as a pivotal gene in animal husbandry and contribute to the improvement of cold intolerance in piglets.

Transcriptome analysis of adipose tissue and muscle of Laiwu and Duroc pigs

Fat content is an important trait in pig production. Adipose tissue and muscle are important sites for fat deposition and affect production efficiency and quality. To regulate the fat content in these tissues, we need to understand the mechanisms behind fat deposition. Laiwu pigs, a Chinese indigenous breed, have significantly higher fat content in both adipose tissue and muscle than commercial breeds such as Duroc. In this study, we analyzed the transcriptomes in adipose tissue and muscle of 21-d-old Laiwu and Duroc piglets. Results showed that there were 828 and 671 differentially expressed genes (DEG) in subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT), respectively. Functional enrichment analysis showed that these DEG were enriched in metabolic pathways, especially carbohydrate and lipid metabolism. Additionally, in the longissimus muscle (LM) and psoas muscle (PM), 312 and 335 DEG were identified, demonstrating enrichment in the cell cycle and metabolic pathways. The protein–protein interaction (PPI) networks of these DEG were analyzed and potential hub genes were identified, such as FBP1 and SCD in adipose tissues and RRM2 and GADL1 in muscles. Meanwhile, results showed that there were common DEG between adipose tissue and muscle, such as LDHB, THRSP, and DGAT2. These findings showed that there are significant differences in the transcriptomes of the adipose tissue and muscle between Laiwu and Duroc piglets (P < 0.05), especially in metabolic patterns. This insight serves to advance our comprehensive understanding of metabolic regulation in these tissues and provide targets for fat content regulation.

The landscape of super-enhancer regulates remote target gene transcription through loop domains in adipose tissue of pig

BackgroundA super-enhancer (SE) is a huge cluster of multiple enhancers that control the key genes for cell identity and function. The rise of advanced chromatin immunoprecipitation sequencing (ChIP-seq) technology such as Cleavage Under Targets and Tagmentation (CUT&Tag) allows more SEs to be discovered. However, SE studies in Luchuan and Duroc pigs are very rare in animal husbandry. ResultsWe used the CUT&Tag technique to identify 145 and 378 SEs from the adipose tissues of Luchuan and Duroc pigs, respectively. There were significant differences in the peak coverage ratio of SE peaks in the gene promoter region between the two breeds. Not only that, peak signals at the start and end point of the SE peak profile showed obvious spikes. The proximal target genes of SE were highly expressed compared with the background genes and the typical enhancer target genes. Subsequently, in conjoint analysis with high-throughput chromosome conformation capture sequencing (Hi-C seq) data, we predicted the remote regulatory genes of SE and found that their expression level was related to the distance of SE extended to the loop's anchor, but not the length of loops. According to our prediction model, SEs can maintain promoter accessibility of partial remote target genes through loop domains. Finally, a batch of SEs closely related to fat metabolism traits were obtained by performing a coalition analysis of quantitative trait loci and SE data. ConclusionsThis work enabled us to obtain hundreds of SEs from Luchuan and Duroc pigs. Our model provides a new method for predicting the SE remote target genes based on loop domains, and to further explore the potential role of super-enhancer in the regulation of fat metabolism.

H3K27ac modification and transcription characteristics of adipose and muscle tissues in Chuxiang Black pig.

The establishment of high-quality pork breeds for improving meat quality in the pig industry is needed. The Chuxiang Black (CX) pig is a new breed developed from Chinese local pigs and Western lean pigs that has a high proportion of lean meat and excellent meat quality. However, the characteristics of cis-regulatory elements in CX pigs are still unknown. In this study, cis-regulatory elements of muscle and adipose tissues in CX pigs were investigated using ChIP-seq and RNA sequencing. Compared with the reported cis-regulatory elements of muscle and adipose tissues, 1768 and 1012 highly activated enhancers and 433 and 275 highly activated promoters in CX muscle and adipose tissues were identified, respectively. Motif analysis showed that transcription factors, such as MEF2A and MEF2C, were core regulators of highly activated enhancers and promoters in muscle. Similarly, the transcription factors JUNB and CUX1 were identified as essential for highly activated enhancers and promoters in CX adipose tissue. These results enrich the resources for the analysis of cis-regulatory elements in the pig genome and provide new basic data for further meat quality improvement through breeding in pigs.

Leucine improves the growth performance, carcass traits, and lipid nutritional quality of pork in Shaziling pigs

Leucine is involved in promoting fatty acid oxidation and lipolysis, mediating lipid metabolism and energy homeostasis, thus it has been widely used in livestock production. However, the effects of leucine on fat deposition and nutrition in Shaziling pigs remain unclear. A total of 72 Shaziling pigs (150 days old, weight 35.00 ± 1.00 kg) were randomly divided into 2 groups and fed with basal diet (control group) or basal diet containing 1% leucine (leucine group) for 60 days. The results showed that leucine significantly increased the average daily feed intake but decreased the ratio of feed to gain (P < 0.05), increased the loin muscle area and serum glucose content (P < 0.05) of Shaziling pigs. Besides, leucine regulated the re-distribution of fatty acids from adipose tissue to muscle as it significantly increased the contents of C18:1n-9 and C22:6n-3 (DHA) in the longissimus thoracis while decreased the contents of C22:5n-3 (DPA), C20:5n-3 (EPA), and DHA in the adipose tissue of Shaziling pigs (P < 0.05). Lipidomic analysis showed that the contents of phosphatidylethanolamines (PEs), cardiolipins (CLs), and phosphatidylglycerols (PGs) in the longissimus thoracis and the contents of lysophosphatidylethanolamines (LPEs), ceramides (Cers), phosphatidylinositols (PIs) in adipose tissue of Shaziling pigs were decreased in leucine group (P < 0.05). Collectively, this study clarified that dietary addition of 1% leucine have a better effect on growth performance and the deposition of beneficial fatty acids in the muscle of Shaziling pigs, which is conductive to the production of high quality and healthy pork. In addition, leucine altered the lipid composition of muscle and fat in Shaziling pigs. The related results provide a theoretical basis and application guidance for regulating fat deposition in Shaziling pigs, which is important for the healthy breeding of Shaziling pigs.

Trimethylamine oxide supplementation differentially regulates fat deposition in liver, longissimus dorsi muscle and adipose tissue of growing-finishing pigs

Trimethylamine oxide (TMAO) is a microbiota-derived metabolite, and numerous studies have shown that it could regulate fat metabolism in humans and mice. However, few studies have focused on the effects of TMAO on fat deposition in growing-finishing pigs. This study aimed to investigate the effect of TMAO on fat deposition and intestinal microbiota in growing-finishing pigs. Sixteen growing pigs were randomly divided into 2 groups and fed with a basal diet with 0 or 1 g/kg TMAO for 149 d. The intestinal microbial profiles, fat deposition indexes, and fatty acid profiles were measured. These results showed that TMAO supplementation had a tendency to decrease lean body mass (P < 0.10) and significantly increased backfat thickness (P < 0.05), but it did not affect growth performance. TMAO significantly increased total protein (TP) concentration, and reduced alkaline phosphatase (ALP) concentration in serum (P < 0.05). TMAO increased the α diversity of the ileal microbiota community (P < 0.05), and it did not affect the colonic microbial community. TMAO supplementation significantly increased acetate content in the ileum, and Proteobacteria and Escherichia–Shigella were significantly enriched in the TMAO group (P < 0.05). In addition, TMAO decreased fat content, as well as the ratio of linoleic acid, n-6 polyunsaturated fatty acids (PUFA), and PUFA in the liver (P < 0.05). On the contrary, TMAO increased intramuscular fat content of the longissimus dorsi muscle, whereas the C18:2n6c ratio was increased, and the n-6 PUFA:PUFA ratio was decreased (P < 0.05). In vitro, 1 mmol/L TMAO treatment significantly upregulated the expression of FASN and SREBP1 in C2C12 cells (P < 0.05). Nevertheless, TMAO also increased adipocyte area and decreased the CPT-1B expression in subcutaneous fat (P < 0.05). Taken together, TMAO supplementation regulated ileal microbial composition and acetate production, and regulated fat distribution and fatty acid composition in growing-finishing pigs. These results provide new insights for understanding the role of TMAO in humans and animals.

Evaluation of thermodynamic bioeffects of long-pulsed 1064 nm laser in the photothermal lipolysis.

To evaluate the lipolysis effect of air cooling assisted long-pulsed 1064 laser for improving local adiposity. The second-level (pulse duration of 0.3-60 s) long-pulsed Nd:YAG 1064 nm laser (LP1064 nm) with or without forced-air cooling was used to irradiate ex-vivo subcutaneous adipose tissue (SAT)of pig or human and in-vivo inguinal fat tissue of Sprague Dawley rats. The temperature of skin surface as well as 5 mm deep SAT was monitored by a plug-in probe thermal couple, and the former was confined to 39°C or 42°C during the treatment. Histological analysis of SAT response was evaluated by SAT sections stained with hematoxylin-eosin and oil red O. Ultra-microstructure changes were examined by transmission electron microscopy. A pilot study on human subject utilizing LP1064 nm laser with air cooling was conducted. The changes in gross abdomen circumference and ultrasonic imaging were studied. Histological examination showed that LP1064 nm laser treatment induced adipocyte injury and hyperthermic lipolysis both in- and ex-vivo. It was also confirmed by clinical practice on patients. By real-time temperature monitoring, we found that in comparison with LP1064 nm laser alone, additional air cooling could increase the temperature difference between epidermis and SAT, promoting heat accumulation deep in fat tissue, as well as providing better protection for epidermis. LP1064 nm laser provided reliable adipose tissue thermolysis when the temperature of skin surface was sustained at 39°C or 42°C for 10 min. Application of air-cooling during the laser treatment achieved better effect and safety of photothermal lipolysis. LP1064 nm laser, as a noninvasive device, has comparable thermal lipolysis effect as other common heat-generating devices.

Dynamic changes in the transcriptome of tRNA-derived small RNAs related with fat metabolism

The prevalence of obesity and overweight is steadily rising, posing a significant global challenge for humanity. The fundamental cause of obesity and overweight lies in the abnormal accumulation of adipose tissue. While numerous regulatory factors related to fat deposition have been identified in previous studies, a considerable number of regulatory mechanisms remain unknown. tRNA-derived small RNAs (tsRNAs), a novel class of non-coding RNAs, have emerged as significant regulators in various biological processes. In this study, we obtained small RNA sequencing data from subcutaneous white adipose tissue and omental white adipose tissue of lean and obese pigs. In addition, we similarly obtained tsRNAs profiles from scapular brown adipose tissue (BAT), inguinal white adipose tissue (iWAT) and epigonadal white adipose tissue (eWAT) of normal mice. Finally, we successfully identified a large number of expressed tsRNAs in each tissue type and identified tsRNAs conserved in different adipose tissues of pigs and mice. These datasets will be a valuable resource for elucidating the epigenetic mechanisms of fat deposition.

THU051 GH Resistance And Adipose Tissue: Multispecies Measurement Of Morphology And Transcriptome

Abstract Disclosure: J.A. Young: None. A. Hinrichs: None. S.R. Bell: None. M. Soneson: None. D.K. Geitgey: None. E. Wolf: None. E.O. List: None. J.J. Kopchick: None. D.E. Berryman: None. Laron Syndrome (LS) is a condition caused by mutations in the growth hormone (GH) receptor gene that renders the individual GH resistant. LS was first described in humans in the 1960’s, and since then, animal models of LS have been developed, including pigs (GHR-KO pigs). One of the notable characteristics of LS, in both humans and animals, is obesity without some of its comorbidities, such as increased cancer and diabetes. The aim of this study was to examine the subcutaneous and visceral adipose tissue of GH resistant pigs and a human with LS using histology and transcriptomics and make cross-species and cross-depot comparisons where possible. Adipocyte sizing analysis showed that GH resistance resulted in a trend towards increased cell sizes in human subcutaneous (SubQ) and visceral (Visc) depots, and a significant increase in pig Visc. Interestingly, the human adipocyte size histogram was similar to that of the pig across genotypes and depots. When examining the adipose transcriptome, PCA analysis showed a strong species clustering, but not a strong depot or genotype clustering. Differentially expressed gene (DEG) analysis comparing GHRKO pigs to their respective wild type controls showed that IGF1 expression was decreased in both depots, as expected, and 38 other genes were also significantly altered in GHR-KO pigs compared to controls, in both depots. Enrichment analysis showed a decrease in an extracellular matrix term in the GHR-KO SubQ compared to controls, while terms associated with fatty acid and carbohydrate metabolism were increased in GHR-KO SubQ. When comparing GHR-KO visceral to WT, terms associated with IGF signaling and longevity regulation had decreased expression, while immune activation and cell motility were increased. Pathway analysis generally agreed with these results, but also showed a decrease in multiple signaling pathways including AMPK and PI3K-Akt in both depots. Paradoxically, the JAK-STAT signaling pathway was increased in GHR-KO Visc compared to controls. Depot differences were also explored using this dataset. Within each genotype, the gene expression of the SubQ and Visc depots resulted in no significant DEGs between depots in the WT pigs, while GHRKO pigs had 24 DEGs between SubQ and Visc. Enrichment analysis and pathway analysis showed no significant results in either depot, presumably due to the low number of DEGs. In conclusion, GHR-KO pigs had expected changes in adipocyte size and expression of metabolic and extracellular matrix genes, but other unexpected changes were also present, including changes to cell motility-related genes. There were few differences between depots within each genotype, indicating that depots are equally affected by GH. Further studies are required to understand the functional changes to adipose tissue caused by LS and the effectiveness of pigs as an LS model. Presentation: Thursday, June 15, 2023

Growth hormone insensitivity and adipose tissue: tissue morphology and transcriptome analyses in pigs and humans.

Growth hormone receptor knockout (GHR-KO) pigs have recently been developed, which serve as a large animal model of Laron syndrome (LS). GHR-KO pigs, like individuals with LS, are obese but lack some comorbidities of obesity. The purpose of this study was to examine the histological and transcriptomic phenotype of adipose tissue (AT) in GHR-KO pigs and humans with LS. Intraabdominal (IA) and subcutaneous (SubQ) AT was collected from GHR-KO pigs and examined histologically for adipocyte size and collagen content. RNA was isolated and cDNA sequenced, and the results were analyzed to determine differentially expressed genes that were used for enrichment and pathway analysis in pig samples. For comparison, we also performed limited analyses on human AT collected from a single individual with and without LS. GHR-KO pigs have increased adipocyte size, while the LS AT had a trend towards an increase. Transcriptome analysis revealed 55 differentially expressed genes present in both depots of pig GHR-KO AT. Many significant terms in the enrichment analysis of the SubQ depot were associated with metabolism, while in the IA depot, IGF and longevity pathways were negatively enriched. In pathway analysis, multiple expected and novel pathways were significantly affected by genotype, i.e. KO vs. controls. When GH related gene expression was analyzed, SOCS3 and CISH showed species-specific changes. AT of GHR-KO pigs has several similarities to that of humans with LS in terms of adipocyte size and gene expression profile that help describe the depot-specific adipose phenotype of both groups.

Transcriptome analysis of norepinephrine-induced lipolysis in differentiated adipocytes of Bama pig

Sympathetic innervation of white adipose tissue (WAT) plays a key role in the regulation of lipid metabolism. Sympathetic activation promotes release of norepinephrine (NE), which binds to adrenergic receptors on adipocytes, promoting adipocyte lipolysis and enhanced oxidative metabolism. However, the mechanism by which sympathetic nerves regulate lipid metabolism in pig adipose tissue remains unclear. We used NE to simulate the process of sympathetic driving in pig adipocytes. RNA sequencing (RNA-seq) was used to determine the gene expression profile of pig adipocytes responding to NE stimulation. Our data suggests that the lipolytic signaling pathway is activated in pig adipocytes upon acute stimulation of NE, resulting in enhanced lipid metabolism and lipolysis, consistent with the phenomena found in humans and mice. Specifically, differentially expressed protein coding genes (PCGs) (SIRT4, SLC27A1) are mainly associated with functions that inhibit fatty acid oxidation and promote lipid synthesis. Similarly, we investigated the changes in regulatory transcripts such as long non-coding RNAs (lncRNAs) and transcripts of uncertain coding potential (TUCP) in response to NE and found that differentially expressed lncRNAs (lncG47338, lncG30660, lncG29516, lncG3790) and TUCP (TUCP_G38001) were co-expressed with target genes related to the promotion of fatty acid β-oxidation, lipolysis and oxidative metabolism, thus acting as regulators. These results indicate a broad suite of gene expression alterations in response to NE stimulation and promote the understanding of the molecular mechanisms by which NE regulates lipid metabolism in pigs.

Dietary supplementation with vegetable oils with low n-6:n-3 polyunsaturated fatty acid ratios improves the intramuscular fat and fatty acid composition of growingfinishing pigs

This study investigated the effects of different n-6:n-3 polyunsaturated fatty acid (PUFA) ratios in pig diets using vegetable oil supplementation on growth performance, carcass characteristics, meat quality, and fatty acid composition in the longissimus dorsi muscle (LM) and subcutaneous adipose tissue (SAT) of pigs. Fiftyfour cross-bred growing pigs (Large White × Landrace × Duroc; 45.03 ± 1.72 kg) were randomly assigned to one of three isoenergetic diets: the 1) basal diet (control), 2) RAPO diet (diet supplemented with 4.5% rapeseed oil), and 3) MIXO diet (diet supplemented with 2.25% rapeseed oil and 2.25% linseed oil), with n-6:n-3 PUFA ratios of approximately 13:1, 7:1, and 2:1, respectively. The experiment lasted 42 days. The RAPO and MIXO diets did not affect the growth performance and carcass characteristics of pigs, while the content of low-density lipoprotein (LDL), triglycerides (TGs), and glucose (GLU) in the serum decreased substantially. Intramuscular fat (IMF) content increased by 19.25% and 20.11% in the LM of pigs fed the RAPO and MIXO diets, respectively, and lower cooking loss and drip loss of meat were observed only in pigs fed the MIXO diet. The RAPO and MIXO diets decreased the stearic acid and palmitoleic acid levels, increased the total PUFA levels (including those of α-linolenic and linoleic acid), and decreased the n-6:n-3 PUFA ratios in LM and SAT. However, the MIXO diet was more effective in improving the fatty acid composition. A mixed oil diet with an n6:n-3 PUFA ratio of 2:1 is an effective measure for improving meat quality.

Methylation Genome-Wide Profiling in Lowly and Highly Efficient Somatic Cell Nuclear Transfer in Pigs

Swine is a common model organism for biomedical research. Epigenetic reprogramming in somatic cell nuclear transfer (SCNT) embryos does not fully recapitulate the natural DNA demethylation events at fertilisation. This study aimed to conduct genome-wide methylation profiling to detect differentially methylated regions (DMRs) responsible for epigenetic differences in stem cells that displayed high and low efficiency of SCNT and to elucidate the low efficiency of cloning rate in pigs. Adipose tissue mesenchymal stem cells (AMSC)s lines were isolated from adipose tissue of adult male pigs (n = 20; high-efficiency cells = 10; and low-efficiency cells = 10). Reduced representation bisulfite sequencing (RRBS) was performed on an Illumina HiSeq1500. Paired-end reads were filtered to remove the adapter contamination, and low-quality reads using TrimGalore! Filtered reads were mapped to the reference genome using Bismark. MethylKit was used to identify differentially methylated regions (DMRs) (bases and tiles), showing statistically significant differential methylation between high and low-efficiency AMSCs. Hierarchical cluster analysis according to methylation patterns clearly defined groups with low and high cloning efficiency. We report 3704 bases with statistically significant differences in methylation and 10062 tiles with statistically significant differences in methylation. Most differentially methylated sites are intergenic 62%, 31% are intronic, 4% are in exons, and 4% in promoters. Moreover, 37% of differentially methylated sites are located in known CpG islands (CGIs), and 4% in CpG island shores (CGSs).

Investigating livestock management in the early Neolithic archaeological site of Cabecicos Negros (Almería, Spain) from the organic residue analysis in pottery

This paper seeks to reconstruct the management of food resources in the early Neolithic site of Cabecicos Negros in southeastern Spain. For this purpose, we have studied 29 potsherds from Cabecicos Negros (Andalusia, Spain). Applying the methods of gas chromatography and mass spectrometry we were able to recompose the daily use of the sherds related to the consumption and storage of food products. Among the results obtained in this work, we were able to show new evidence of the exploitation of dairy products in the south of the Iberian Peninsula, as well as provide information on the exploitation and management of the early domestic animals herds. To improve the archaeological results obtained, isotopic results were compared with a modern reference of 53 fat samples from the adipose tissue of domestic pigs and wild boars.