Gene Expression In Piglets Research Articles

The Protective Effects of Astaxanthin (AST) in the Liver of Weaned Piglets.

During the weaning period, piglets are exposed to high levels of stress, which often causes problems with the digestive system. This stress also promotes the production of free radicals, resulting in oxidative stress. Astaxanthin (AST) stands out as one of the most potent antioxidants. Its resistance to light and heat makes it particularly valuable in compound feed production. This study was to determine the effect of AST impact on liver histology and gene expression in piglets. For our experiment, we used 16 weaned piglets of the PL breed, which we divided into two groups: Group I (control group with no AST supplementation) and Group II (supplemented with AST at 0.025 g/kg). Both feed mixtures were iso-proteins and iso-energetic, meeting the nutritional requirements of the piglets. The experiment lasted from day 35 to day 70 of the piglets' age, during which they had ad libitum access. The results indicate that the addition of AST prevents liver fibrosis due to reduced collagen deposition in the tissue. Analysis of gene expression supported these results. In the AST-supplemented group, we noted a decrease in NR1H3 expression, an increase in CYP7A1 expression, and reductions in the expression of NOTCH1 and CREB genes.

Dose-dependent effects of two inulin types differing in chain length on the small intestinal morphology, contractility and proinflammatory cytokine gene expression in piglets

ABSTRACTInulin is a linear fructose polymer which may affect small intestinal physiology. The effects of dietary level of two inulin types on morphology, contractility and proinflammatory cytokine gene expression in the small intestine of piglets were investigated. Fifty six piglets were divided into seven groups fed diets without inulin addition or with 1%, 2% or 3% of inulin with an average degree of polymerisation of 10 (IN10) or 23 (IN23). All diets were offered from day 10 of life for 40 d. Feeding IN10 diets did not affect villous height to crypt depth ratio in the duodenum, while in the jejunum the 2% IN10 diet increased it as compared to other groups. Jejunal muscle contractions induced by electrical field stimulation were impaired by the 2% and 3% IN10 diets. The ileal expression of interleukin-12p40 was decreased by the 2% IN10 diet. There was no effect of IN23 level on villous height to crypt depth ratio in any segment of the small intestine as well as on jejunal motility. The 2% and 3% IN23 diets decreased the jejunal expression of tumour necrosis factor-α. In conclusion, IN10 is more active in the small intestine than IN23. At the 2% dietary level, it increases absorptive area in the jejunum, but may slightly impair smooth muscle contractions.

L-carnitine and fat type in the maternal diet during gestation and lactation modify the fatty acid composition and expression of lipid metabolism-related genes in piglets.

This study examined the influence of adding different amounts of maternal dietary l-carnitine and two fat types on fatty acid (FA) composition and the expression of lipid metabolism-related genes in piglets. The experiment was designed as a 2×2 factorial with two fat types (3.5% soyabean oil, SO, and 3.5% fish oil, FO) and two levels of l-carnitine (0 and 100mg/kg) added to the sows' diets. A higher proportion of n-3 polyunsaturated fatty acids (PUFA) and a lower ratio of n-6/n-3 PUFA in sow milk and piglet tissues were observed in the FO groups than in the SO groups. Adding l-carnitine increased the proportion of C16:1 in sow milk and decreased n-3 PUFA in piglet subcutaneous fat. Hepatic peroxisome proliferator-activated receptor α (PPAR-α) was more abundantly expressed in piglets from the FO groups than from the SO groups (p<0.05), whereas stearoyl-CoA-desaturase (SCD), sterol regulatory element binding protein-1 (SREBP1) and ∆6-desaturase (D6D) genes were less expressed in the FO groups compared with piglets from the SO groups. The expression of fatty acid synthase (FAS) genes was decreased in the SO groups with l-carnitine compared to that of the other dietary treatments. No differences among dietary treatments were observed with regard to the expression of acetyl-CoA carboxylase (ACC). In conclusion, FO and l-carnitine supplementation in sows affect FA composition and hepatic gene expression in piglets.

Piglet weight gain during the first two weeks of lactation influences the immune system development

The aim of this study was to investigate the effect of the piglet growth during the first week of life on ileal expression of genes and on development of the immune system. Eight litters adjusted to 12 piglets were used. Within each litter, the piglet that showed the lowest weight gain (LWG; n = 8) and the one that showed the highest weight gain (HWG; n = 8) in their first week of life were enrolled. Peripheral blood mononuclear cells (PBMC) were isolated on days 8 and 16 to characterize cellular population profiles and to assess ex-vivo secretion of interleukin-10 (IL-10), IL-6 and tumor necrosis factor-α (TNF-α). On day 16, piglets were euthanized and ileum samples were collected to extract RNA for microarray analysis and gene expression by qPCR. As expected, growth performance of LWG piglet was impaired compared to HWG piglets (P < 0.05). From day 8 to 16, the percentage of CD21+ B cells significantly increased in blood of heavier HWG piglets while the percentage remained constant in smaller LWG piglets (P weight x day = 0.01). For the CD4+CD8α− Th cells, a marked increase was observed in LWG piglets from 8 to 16 days of age (P = 0.002) whereas no significant change occurred in HWG piglets. Percentages of CD14+ monocytes and other MHC-II+ cells were respectively higher and lower on day 8 compared to day 16 for both groups of piglets (P < 0.01). On day 8, LPS-activated PBMC from LWG piglets produced less IL-6 compared to HWG piglets (P < 0.05). Microarray analysis of gene expression in piglets’ ileum tissue indicated that several genes involed in defense response and response to oxidative stress were modulated differently in LWG compared to HWG. Gene analysis by Q-PCR confirmed microarray results and revealed that IL-10, SOD1, NOS2, NOD2, TLR4, TLR9, CD40 and CD74 expressions were significantly decreased (P < 0.05) in LWG in comparison to HWG piglets, while MYD88 and NFkBiA showed a tendency to decrease (0.05 ≤ P < 0.07). These results suggest that birth weight and milk intake affect the growth performances and the development of immunity by modulating the expression of genes associated with immunity and oxidative stress in piglets’ intestinal tissue, and by affecting the leukocyte populations involved in innate and cell-mediated immunity in nursing piglets. Therefore, impaired development of immune system in LWG piglets might have an impact on their resistance to infections later in life.

Erratum

Physiological ReportsVolume 4, Issue 16 e12954 ErratumOpen Access Erratum This article corrects the following: Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets Anders Bergström, Sanne S. Kaalund, Kerstin Skovgaard, Anders D. Andersen, Bente Pakkenberg, Ann Rosenørn, Ruurd M. van Elburg, Thomas Thymann, Gorm O. Greisen, Per T. Sangild, Volume 4Issue 14Physiological Reports First Published online: July 26, 2016 First published: 25 August 2016 https://doi.org/10.14814/phy2.12954AboutSections ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat In 1, the surname of author Per T. Sangild was misspelled as “Sanglid.” The online version of the article has been corrected. Reference 1Bergstorm, A., S. S. Kaalund, K. Skovgaard, A. D. Andersen, B. Pakkenberg, A. Rosenorn, R. M. van Elburg, T. Thymann, G. O. Greisen, and P. T. Sangild. 2016. Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets. Physiol Rep 4: e12871. Volume4, Issue16August 2016e12954 ReferencesRelatedInformation

Limited effects of preterm birth and the first enteral nutrition on cerebellum morphology and gene expression in piglets.

Preterm pigs show many signs of immaturity that are characteristic of preterm infants. In preterm infants, the cerebellum grows particularly rapid and hypoplasia and cellular lesions are associated with motor dysfunction and cognitive deficits. We hypothesized that functional brain delays observed in preterm pigs would be paralleled by both structural and molecular differences in the cerebellum relative to term born piglets. Cerebella were collected from term (n = 56) and preterm (90% gestation, n = 112) pigs at 0, 5, and 26 days after birth for stereological volume estimations, large‐scale qPCR gene expression analyses (selected neurodevelopmental genes) and western blot protein expression analysis (Sonic Hedgehog pathway). Memory and learning was tested using a T‐maze, documenting that preterm pigs showed delayed learning. Preterm pigs also showed reduced volume of both white and gray matter at all three ages but the proportion of white matter increased postnatally, relative to term pigs. Early initiation of enteral nutrition had limited structural or molecular effects. The Sonic Hedgehog pathway was unaffected by preterm birth. Few differences in expression of the selected genes were found, except consistently higher mRNA levels of Midkine, p75, and Neurotrophic factor 3 in the preterm cerebellum postnatally, probably reflecting an adaptive response to preterm birth. Pig cerebellar development appears more affected by postconceptional age than by environmental factors at birth or postnatally. Compensatory mechanisms following preterm birth may include faster white matter growth and increased expression of selected genes for neurotrophic factors and regulation of angiogenesis. While the pig cerebellum is immature in 90% gestation preterm pigs, it appears relatively mature and resilient toward environmental factors.

Effects of intrauterine growth retardation and maternal folic acid supplementation on hepatic mitochondrial function and gene expression in piglets

Piglets with intrauterine growth retardation (IUGR) or with normal birth weight (NBW) were selected to evaluate the effects of maternal folic acid supplementation on hepatic mitochondrial function and expression levels of genes involved in mitochondrial DNA (mtDNA) biogenesis and mitochondrial function. During gestation, primiparous Yorkshire sows were fed a Control diet (folic acid 1.3 mg/kg) or a folic acid-supplemented diet (folic acid 30 mg/kg) with 16 replicates per diet. During the 28-d lactation period, sows were fed a common diet. Compared with NBW piglets, hepatic ATP concentrations and mtDNA contents were decreased in IUGR piglets. Furthermore, IUGR piglets exhibited lower membrane potential and decreased oxygen consumption in liver mitochondria, but these parameters were not affected by maternal folic acid supplementation. Intrauterine growth retardation decreased mRNA expression abundance of peroxisomal proliferator-activated receptor-γ coactivator-1α, mitochondrial transcription factor A, uncoupling protein 3, and cytochrome c oxidase subunit I and IV. Impaired antioxidant capacity characterised by increased malondialdehyde content and decreased manganese-superoxide dismutase activity was also observed in IUGR pigs. In IUGR piglets, however, nearly all of these parameters were normalised to the level of NBW piglets when the maternal diet was supplemented with folic acid during pregnancy. Hence, maternal folic acid supplementation was proved to be an effective way to reverse the changes in gene expressions in IUGR pigs, which provided a possible nutritional strategy to improve growth development of IUGR individuals.

Differentiation of Porcine TLR4 Gene Expression in Piglets of Different Ages

Differentiation of Porcine TLR4 Gene Expression in Piglets of Different Ages

Histomorphology and small intestinal sodium-dependent glucose transporter 1 gene expression in piglets fed phytic acid and phytase-supplemented diets1

An experiment was conducted to determine the effect of dietary phytic acid (PA) and phytase supplementation on small intestinal histomorphology and Na-dependent glucose transporter 1 (SGLT1) gene expression in piglets. Twenty-four piglets with an average initial BW of 7.60 ± 0.73 kg were randomly assigned to 3 experimental diets, to give 8 piglets per diet. The diets were a casein-cornstarch-based diet that was supplemented with 0 or 2% PA, or 2% PA (as Na phytate) plus an Escherichia coli-derived phytase at 500 phytase units/kg. The basal diet was formulated to meet the 1998 NRC energy, digestible AA, mineral, and vitamin requirements for piglets. After 10 d of feeding, the piglets were killed to determine small intestinal histomorphology and small intestinal SGLT1 gene expression. Phytic acid supplementation did not affect (P > 0.1) villus height (VH) and the VH-to-crypt depth (CD) ratio, but did decrease (P < 0.05) CD in the jejunum. Phytase supplementation did not affect (P > 0.1) VH, CD, and the VH-to-CD ratio. Phytic acid supplementation reduced SGLT1 gene expression in the duodenum, jejunum, and ileum by 1.1-, 5.4-, and 2.4-fold, respectively. Phytase supplementation increased SGLT1 gene expression in the jejunum by 2.6-fold, but reduced SGLT1 gene expression in the duodenum and ileum by 2.0- and 4.0-fold, respectively. In conclusion, PA reduced CD in the jejunum and SGLT1 gene expression in the duodenum, jejunum, and ileum, whereas phytase supplementation increased the expression of SGLT1 in the jejunum. The reduced SGLT1 gene expression by PA implies that PA reduces nutrient utilization in pigs partly through reduced expression of SGLT1, which is involved in glucose and Na absorption. The increased expression of SGLT1 in the jejunum by phytase supplementation implies that phytase alleviated the negative effects of PA partly through increased expression of SGLT1.

Time course differential gene expression in response to porcine circovirus type 2 subclinical infection

This study was aimed at characterizing the potential differences in gene expression in piglets inoculated with Porcine circovirus type 2 (PCV2), the essential causative agent of postweaning multisystemic wasting syndrome. Seven-day-old caesarean-derived, colostrum-deprived piglets were distributed into two groups: control (n = 8) and pigs inoculated with 105.2 TCID50 of the Burgos PCV2 isolate (n = 16). One control and three inoculated pigs were necropsied on days 1, 2, 5, and 8 post-infection (p.i.). The remaining pigs (four of each group) were sequentially bled on days 0, 7, 14, 21, and 29 p.i. (necropsy). Total RNA from the mediastinal lymph node (MLN) and lysed whole blood (LWB) samples were hybridized to Affymetrix Porcine GeneChip®. Forty-three probes were differentially expressed (DE) in MLN samples (FDR < 0.1, fold change > 2) and were distributed into three clusters: globally down-regulated genes, and up-regulated genes at early (first week p.i.) and late (day 29 p.i.) stages of infection. In LWB samples, maximal differences were observed at day 7 p.i., with 54 probes DE between control and inoculated pigs. Main Gene Ontology biological processes assigned to up-regulated genes were related to the immune response. Six common genes were found in both types of samples, all of which belonged to the interferon signaling antiviral effector pathway. Down-regulated genes were mainly related to cell adhesion and migration in MLN, and cellular organization and biogenesis in LWB. Microarray results were validated by quantitative real-time PCR. This study provides, for the first time, the characterization of the early and late molecular events taking place in response to a subclinical PCV2 infection.

Transgenic overexpression of insulin‐like growth factor‐I (IGF‐I) in milk influences intestinal development and gene expression in piglets.

Piglets suckling transgenic sows (TG) that overexpress IGF-I in milk have greater (p<0.05) ileal mucosal weight, protein and DNA content, villus height, disaccharidase activities and type 1 IGF receptor abundance at d21 postpartum than piglets suckling non-TG (CON) sows. Herein, porcine-specific microarrays were used to examine gene expression in ileal crypt and villus cells isolated by laser capture microdissection. Following amplification, mRNA samples were labeled with Cy dyes and hybridized onto microarrays and data were analyzed using the limma package in R. In piglets suckling TG, 2 villus and 3 crypt genes were up-regulated and 1 crypt gene was down-regulated at a FDR-adjusted p<0.05. Five villus and 8 crypt genes tended (p<0.1) to be up-regulated and 4 crypt and 2 villus genes down-regulated. Differentially expressed genes displayed the following biological processes: cell signaling, proliferation, transcription, translation, apoptosis, immune function, metabolism, and structure. Six genes were selected for quantitative real-time PCR and the directionality and fold-change differences were in agreement with microarray results. Coordinate up-regulation of an anti-proliferative gene and down-regulation of a cell cycle regulator in the crypts of piglets suckling TG support reduced proliferation as an adaptation to elevated milk IGF-I. Funded by: AG-00-35206-9537 and IL C-FAR Sentinel Program.

Perinatal ontogeny of porcine growth hormone receptor gene expression is modulated by thyroid status.

The ontogeny of growth hormone receptors (GHR) represents a critical stage in growth and metabolism. We have investigated the perinatal ontogeny of hepatic and skeletal muscle GHR gene expression in piglets, and its modulation by GH and thyroid hormones. Test piglets were rendered hypothyroid in late gestation by feeding the sow a high-glucosinolate rapeseed meal. Plasma and tissue samples were obtained from test and control piglets at various ages between 80 days of fetal life (80F) and 2 days postnatally. Plasma hormone levels were determined by radioimmunoassay and GHR mRNA by RNase protection assays. In controls, plasma thyroxine (T4) and 3,5,3'-triiodothyronine (T3) levels increased between 80F and birth and the early postnatal period was characterized by a marked surge in plasma T3. Test piglets were hypothyroid at 110F with total T4, total T3 and free T3 levels being reduced by 28, 53 and 33% respectively. By contrast, the postnatal increase in T3 was more marked in test than in control animals. Plasma GH levels decreased over the perinatal period and there was no effect of treatment. Hepatic GHR mRNA was at the lower limit of detection at 80F but by 110F was expressed in both groups of animals. However, fetal hypothyroidism at 110F resulted in a marked 70% decrease in hepatic GHR mRNA (p < 0.01). The higher postnatal rise in T3 in test piglets was accompanied by a recovery of hepatic GHR mRNA levels. By contrast with liver, skeletal muscle (longissimus dorsi) expressed high levels of GHR mRNA at 80F and hypothyroidism induced a 68% increase in GHR mRNA (p < 0.001). The present results suggest that thyroid hormones may modulate the perinatal ontogeny of GHR gene expression, in addition to other hormonal factors, and that this modulation is tissue-specific.