J. P. Cassady

Conjugated linoleic acid evokes de‐lipidation through the regulation of genes controlling lipid metabolism in adipose and liver tissue

Conjugated linoleic acid (CLA) is a unique lipid that elicits dramatic reductions in adiposity in several animal models when included at ≤ 1% of the diet. Despite a flurry of investigations, the precise mechanisms by which conjugated linoleic acid elicits its dramatic effects in adipose tissue and liver are still largely unknown. In vivo and in vitro analyses of physiological modifications imparted by conjugated linoleic acid on protein and gene expression suggest that conjugated linoleic acid exerts its de‐lipidating effects by modulating energy expenditure, apoptosis, fatty acid oxidation, lipolysis, stromal vascular cell differentiation and lipogenesis. The purpose of this review shall be to examine the recent advances and insights into conjugated linoleic acids effects on obesity and lipid metabolism, specifically focused on changes in gene expression and physiology of liver and adipose tissue.

Detection of transcriptional difference of porcine imprinted genes using different microarray platforms

BackgroundPresently, multiple options exist for conducting gene expression profiling studies in swine. In order to determine the performance of some of the existing microarrays, Affymetrix Porcine, Affymetrix Human U133+2.0, and the U.S. Pig Genome Coordination Program spotted glass oligonucleotide microarrays were compared for their reproducibility, coverage, platform independent and dependent sensitivity using fibroblast cell lines derived from control and parthenogenic porcine embryos.ResultsArray group correlations between technical replicates demonstrated comparable reproducibility in both Affymetrix arrays. Glass oligonucleotide arrays showed greater variability and, in addition, approximately 10% of probes had to be discarded due to slide printing defects. Probe level analysis of Affymetrix Human arrays revealed significant variability within probe sets due to the effects of cross-species hybridization. Affymetrix Porcine arrays identified the greatest number of differentially expressed genes amongst probes common to all arrays, a measure of platform sensitivity. Affymetrix Porcine arrays also identified the greatest number of differentially expressed known imprinted genes using all probes on each array, an ad hoc measure of realistic performance for this particular experiment.ConclusionWe conclude that of the platforms currently available and tested, the Affymetrix Porcine array is the most sensitive and reproducible microarray for swine genomic studies.

Phenotypic and genetic correlations between gilt estrus, puberty, growth, composition, and structural conformation traits with first-litter reproductive measures

The objective was to estimate correlations of gilt estrus, puberty, growth, composition, and structural conformation traits with first-litter reproductive measures. Four groups of gilts (n = 1,225; Genetic Improvement Services of NC, Newton Grove, NC) entered the NC Swine Evaluation Station (Clayton, NC) averaging 162 d of age and were observed daily for symptoms of estrus. Once symptoms of first estrus were observed in 70% of gilts, recording of symptoms of estrus in all gilts occurred every 12 h for 30 d, utilizing fence-line boar contact. Subjective estrous traits were maximum and total strength of standing reflex, as observed with and without the presence of a boar, and strength of vulva reddening and swelling. Objective estrous traits consisted of vulva redness, vulva width, length of estrus, and age at puberty. Growth and composition traits included BW at puberty, days to 114 kg, and 10th rib backfat and LM area at 114 kg and at puberty. Subjective structural conformation traits were muscle mass, rib width, front leg side view, rear leg side view, front legs front view, rear legs rear view, and locomotion. First-litter sow traits included if gilt farrowed (Stay), age at first farrowing (AFF), total number of piglets born (TNB), and weaning to conception interval (WCI). Variance components were estimated using an animal model with AIREMLF90 for linear traits and THRGIBBS1F90 for categorical traits. Heritability estimates for Stay, AFF, and TNB were 0.14, 0.22, and 0.02, respectively. Genetic correlations between length of estrus, the standing reflex traits, and age at puberty with Stay were 0.34, 0.34 to 0.74, and -0.27, respectively, and with AFF were -0.11, -0.04 to -0.41, and 0.76, respectively. Days to 114 kg had genetic associations with Stay, AFF, and TNB of 0.52, -0.25, and -0.08, respectively. Backfat at 114 kg had genetic correlations with Stay, AFF, and TNB of -0.29, 0.14, and 0.47, respectively. Vulva redness and TNB were negatively correlated phenotypically (r = -0.14) and genetically (r = -0.53). Associations between structural conformation traits with Stay, AFF, TNB, and WCI were generally low to moderate and favorable. Selection for longer length of estrus, stronger standing reflex, or younger age at puberty would increase the proportion of gilts that farrow and reduce age at first farrowing.

Genetic relationships among pig behavior, growth, backfat, and loin muscle area.

The objective of this study was to estimate repeatabilities and heritabilities for measures of pig behavior and their relationship with performance. Measures of behavior and performance included the backtest, resident-intruder test, human approach test (HAT), novel object test (NOT), d 1 BW, backfat depth (BF), loin muscle area (LMA), ADG in the farrowing house, ADG, 21-d BW, and 140-d BW (W). Each behavioral trait was measured twice. The study consisted of 95 litters from 31 sires with an average of 3 litters per sire (n >or= 457). Between 7 and 14 d of age, the backtest was conducted by placing each pig in a supine position for 60 s. Total time spent struggling (TTS) and total number of attempts to struggle (TAS) were recorded. The resident intruder test involved 2 nursery pigs, a resident pig and an unfamiliar intruder pig. The resident pen was divided in half by a solid partition. A resident pig was placed in the test area, and an intruder pig was then introduced. Latency until an attack occurred (LAT) and total number of attacks over 2 tests (RIS) were recorded. Amount of time taken for each finishing pig to make snout contact with an unfamiliar human or object was recorded. Dam and sire effects influenced all traits (P < 0.01). Sex and pen affected LAT, RIS, HAT, and NOT (P < 0.10). Repeatabilities of TTS, TAS, RIS, LAT, HAT, and NOT were 0.38, 0.21, 0.07, 0.08, 0.17, and 0.11, respectively. The phenotypic correlations of TTS with TAS and HAT with NOT were 0.61 and 0.34, respectively. Phenotypic correlation between RIS and LAT was -0.85. Total time spent struggling and TAS tended to be phenotypically correlated with 21-d BW and ADG in the farrowing house. Total attempts to struggle were phenotypically correlated with BF (0.15). Latency until an attack occurred was phenotypically correlated with LMA (0.23). Resident intruder score was phenotypically correlated with ADG (-0.13), W (-0.13), and LMA (-0.21) and estimated to be lowly heritable (h(2) = 0.12). Heritabilities of TTS and TAS were 0.31 and 0.53, respectively. Genetic correlation of TAS with ADG and W was 0.38. Genetic correlations of TTS with BF, W, and TAS were 0.14, 0.18, and 0.81, respectively. The backtest is a heritable and repeatable measure of a behavioral characteristic in pigs that is phenotypically and genetically correlated with performance.

Trans-10, cis-12-conjugated linoleic acid alters hepatic gene expression in a polygenic obese line of mice displaying hepatic lipidosis.

The trans-10, cis-12 isomer of conjugated linoleic acid (CLA) causes a rapid reduction of body and adipose mass in mice. In addition to changes in adipose tissue, numerous studies have reported alterations in hepatic lipid metabolism. Livers of CLA-fed mice gain mass, partly due to lipid accumulation; however, the precise molecular mechanisms are unknown. To elucidate these mechanisms, we examined fatty acid composition and gene expression profiles of livers from a polygenic obese line of mice fed 1% trans-10, cis-12-CLA for 14 days. Analysis of gene expression data led to the identification of 1393 genes differentially expressed in the liver of CLA-fed male mice at a nominal P value of .01, and 775 were considered significant using a false discovery rate (FDR) threshold of .05. While surprisingly few genes in lipid metabolism were impacted, pathway analysis found that protein kinase A (PKA) and cyclic adenosine monophosphate (cAMP) pathways signaling pathways were affected by CLA treatment and 98 of the 775 genes were found to be regulated by hepatocyte nuclear factor 4alpha, a transcription factor important in controlling liver metabolic status.

Feed intake, average daily gain, feed efficiency, and real-time ultrasound traits in Duroc pigs: II. Genomewide association

Efficient use of feed resources has become a clear challenge for the U.S. pork industry as feed costs continue to be the largest variable expense. The availability of the Illumina Porcine60K BeadChip has greatly facilitated whole-genome association studies to identify chromosomal regions harboring genes influencing those traits. The current study aimed at identifying genomic regions associated with variation in feed efficiency and several production traits in a Duroc terminal sire population, including ADFI, ADG, feed conversion ratio, residual feed intake (RFI), real-time ultrasound back fat thickness (BF), ultrasound muscle depth, intramuscular fat content (IMF), birth weight (BW at birth), and weaning weight (BW at weaning). Single trait association analyses were performed using Bayes B models with 35,140 SNP on 18 autosomes after quality control. Significance of nonoverlapping 1-Mb length windows (n = 2,380) were tested across 3 QTL inference methods: posterior distribution of windows variances from Monte Carlo Markov Chain, naive Bayes factor, and nonparametric bootstrapping. Genes within the informative QTL regions for the traits were annotated. A region ranging from166 to 140 Mb (4-Mb length) on SSC 1, approximately 8 Mb upstream of the MC4R gene, was significantly associated with ADFI, ADG, and BF, where SOCS6 and DOK6 are proposed as the most likely candidate genes. Another region affecting BW at weaning was identified on SSC 4 (84-85 Mb), harboring genes previously found to influence both human and cattle height: PLAG1, CHCHD7, RDHE2 (or SDR16C5), MOS, RPS20, LYN, and PENK. No QTL were identified for RFI, IMF, and BW at birth. In conclusion, we have identified several genomic regions associated with traits affecting nutrient utilization that could be considered for future genomic prediction to improve feed utilization.

Effects of reduced panel, reference origin, and genetic relationship on imputation of genotypes in Hereford cattle.

The objective of this study was to investigate alternative methods of designing and using reduced SNP panels for imputing SNP genotypes. Two purebred Hereford populations, an experimental population known as Line 1 Hereford (L1, n = 240) and registered Hereford with American Hereford Association (AHA, n = 311), were used. Using different reference samples of 62 to 311 animals with 39,497 SNP on 29 autosomes and study samples of 57 or 62 animals for which genotypes were available for ~2,600 SNP (reduced panels), imputations were performed to predict the other ~36,900 loci that had been masked. An imputation package, including LinkPHASE and DAGPHASE, was used for imputation. Four reduced panels differing in minor allele frequency (MAF) and marker spacing were evaluated. Reduced panels included every 15th SNP across the genome (SNP_space), commercial Illumina Bovine3K Beadchip (SNP_3K), SNP with the highest MAF (SNP_MAF), and SNP with high MAF that were also evenly spaced across the genome (SNP_MS). Imputation accuracy was defined as the correlation of imputed genotypes and real genotypes. Reference samples were either from L1 or AHA. Among animals with genotypes, genetic relationships were estimated based on molecular marker genotypes or pedigree. Reduced panel design, number of animals in the reference sample, reference origin and genetic relationship between animals in the reference, and study samples all affected imputation accuracy (P < 0.001). Across genotyping schemes, imputed genotypes from SNP_MS had the greatest accuracy. A 0.1 increase in average pedigree relationship or average molecular relationship between reference and study samples increased imputation accuracy 10 to 20%. Using reference samples from the L1 population resulted in lower imputation accuracy than using reference samples from the admixed population AHA (P < 0.001). Increasing the number of animals in the reference panel by 100 individuals increased imputation accuracy by 8% when pedigree relationship was used as a covariate and 6% when molecular relationship was used as a covariate. We concluded that imputation accuracy would be increased through optimization of reduced panel design and genotyping strategy.

A genome‐wide association study of direct gestation length in US Holstein and Italian Brown populations

Direct gestation length influences economically important traits in dairy cattle that are related to birth and peri-natal survival of the calf. The objective of this study was to identify single nucleotide polymorphisms (SNPs) that are significantly associated with direct gestation length through a genome-wide association study. Data used in the analysis included 7,308,194 cow gestation lengths from daughters of 4743 United States Holstein sires in the Cooperative Dairy DNA Repository population and 580,157 gestation lengths from 749 sires in the Italian Brown population. Association analysis included 36,768 and 35,082 SNPs spanning all autosomes for Holstein and Brown Swiss, respectively. Multiple shrinkage Bayesian was employed. Estimates of heritability for both populations were moderate, with values of 0.32 (±0.03) and 0.29 (±0.02) for Holstein and Brown Swiss, respectively. A panel of SNPs was identified, which included SNPs that have significant effects on direct gestation length, of which the strongest candidate region is located on chromosome 18. Two regions not previously linked to direct calving ease and calf survival were identified on chromosome 7 and 28, corresponding to regions that contain genes related to embryonic development and foetal development. SNPs were also identified in regions that have been previously mapped for calving difficulty and longevity. This study identifies target regions for the investigation of direct foetal effects, which are a significant factor in determining the ease of calving.

Estimates of variance components for genetic correlations among swine estrus traits.

Variance components and genetic correlations were estimated among estrus, puberty, growth, and composition traits in Landrace-Large White gilts (n = 1,225; Genetic Improvement Services, Newton Grove, NC) from 59 sires and 330 dams. Four groups of gilts entered the North Carolina Swine Evaluation Station in Clayton at an average age of 162 d and were checked daily for estrus. Once 70% of gilts had reached puberty, recording of estrus symptoms occurred every 12 h for 30 d, using fence-line boar contact. Subjective estrus traits were maximum strength of standing reflex with or without a boar present, total strength of standing reflex with or without a boar present, and strength of vulva reddening and swelling. Objective estrus traits consisted of vulva redness, vulva width, length of estrus in consecutive days based on 12-h observations, and age at puberty (AGEPUB). Growth and composition traits included puberty weight, days to 114 kg (DYS), 10th-rib backfat, and 10th-rib LM area at 114 kg (BF, LMA) and puberty. Variance components were estimated using AIREMLF90 with an animal model. All models included gilt development diet class and breed composition as fixed effects, entry age as a covariate (except DYS, BF, and LMA), a random common litter effect, and a random animal genetic effect. Heritability estimates for length of estrus, maximum strength of the standing reflex with a boar, total strength of the standing reflex with a boar, maximum strength of the standing reflex without a boar, total strength of the standing reflex without a boar, vulva redness, strength of vulva reddening and swelling, and vulva width were 0.21, 0.13, 0.26, 0.42, 0.42, 0.26, 0.45, and 0.58, respectively. Heritability estimates for AGEPUB, puberty weight, 10th-rib backfat at puberty, 10th-rib LM area at puberty, DYS, BF, and LMA were 0.29, 0.39, 0.41, 0.38, 0.24, 0.47, and 0.39, respectfully. Common litter effect estimates ranged from 0.01 to 0.09. The estimated genetic correlation between length of estrus and maximum strength of standing reflex with a boar was 0.99. Genetic correlations between AGEPUB and length of estrus, maximum strength of standing reflex with a boar, and vulva redness were -0.23, -0.32, and 0.20, respectively. Length of estrus had positive genetic associations with DYS and BF (0.30 and 0.29, respectively). It was concluded that past selection for lean BW gain may have weakened the strength of the standing reflex and that sufficient genetic variation exists to make selection for improved swine estrus traits effective.

Changes in chondrocyte gene expression following in vitro impaction of porcine articular cartilage in an impact injury model

Our objective was to monitor chondrocyte gene expression at 0, 3, 7, and 14 days following in vitro impaction to the articular surface of porcine patellae. Patellar facets were either axially impacted with a cylindrical impactor (25 mm/s loading rate) to a load level of 2,000 N or not impacted to serve as controls. After being placed in organ culture for 0, 3, 7, or 14 days, total RNA was isolated from full thickness cartilage slices and gene expression measured for 17 genes by quantitative real‐time RT‐PCR. Targeted genes included those encoding proteins involved with biological stress, inflammation, or anabolism and catabolism of cartilage extracellular matrix. Some gene expression changes were detected on the day of impaction, but most significant changes occurred at 14 days in culture. At 14 days in culture, 10 of the 17 genes were differentially expressed with col1a1 most significantly up‐regulated in the impacted samples, suggesting impacted chondrocytes may have reverted to a fibroblast‐like phenotype.