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Appearance, distribution, and amount of intramuscular fat (IMF), often referred to as marbling, are highly variable and depend on environmental and genetic factors. On the molecular level, the concerted action of several drivers, including hormones, receptors, transcription factors, etc., determines where clusters of adipocytes arise. Therefore, the aim of future studies remains to identify such factors as biological markers of IMF to increase the ability to identify animals that deposit IMF early in age to increase efficiency of high-quality meat production. In an attempt to unravel the cellular development of marbling, we investigated the abundance of markers for adipogenic differentiation during fattening of cattle and the transcriptome of muscle and dissected IMF. Markers of different stages of adipogenic differentiation are well known from cell culture experiments. They are usually transiently expressed, such as delta-like homolog 1 (DLK1) that is abundant in preadipocytes and absent during differentiation to mature adipocytes. It is even a greater challenge to detect those markers in live animals. Within skeletal muscles, hyperplasia and hypertrophy of adipocytes can be observed throughout life. Therefore, development of marbling requires, on the cellular level, recruitment, proliferation, and differentiation of adipogenic cells to store excess energy in the form of lipids in new cells. In a recent study, we investigated the localization and abundance of early markers of adipogenic differentiation, such as DLK1, in bovine muscle tissue. An inverse relationship between IMF content and number of DLK1-positive cells in bovine muscle was demonstrated. Considering the cellular environment of differentiating adipocytes in muscle and the secretory action of adipocytes and myocytes, it becomes obvious that cross talk between cells via adipokines and myokines may be important for IMF development. Secreted proteins can act on other cells, inhibiting or stimulating their function via autocrine and paracrine actions. Such factors with potential influence on IMF, among them, agouti signaling protein and thrombospondin 4, were identified in transcriptome analyses and further investigated. Furthermore, results from transcriptome analysis indicate involvement of genes that are not directly related to adipogenesis and lipid metabolism, providing new candidates for future research. [ABSTRACT FROM AUTHOR]

Muscle fiber development during gestation determines the muscle structure at birth and establishes the conditions for muscle development in growing cattle. Differences in muscle structure among beef cattle breeds and between beef and dairy cattle are obvious already shortly after birth. The objective of the study was to investigate the development of muscle fibers and muscle fiber bundle structure in semitendinosus muscle of divergent cattle breeds from 3 mo of gestation until birth. Fetuses of Gennan Angus (GA), Galloway (GW), Belgian Blue (BB), and Holstein Friesian (HF) were harvested at 3, 4.5, 6, or 9 mo of gestation. Muscle sections were analyzed for fiber size and types as well as for bundle structure. The results confirmed that primary muscle fiber development occurs mainly during the first trimester of gestation. All fibers were initially positive for fetal fast myosin. Slow myosin as a marker for fiber maturation was detected in primary fibers at 3 mo of gestation showing a weak immunostaining. During the second trimester, the intensity of immunostaining strongly increased indicating increased slow myosin protein expression. Concurrently, the shape of primary fibers changed from myotubes to myofibers whereas the size stayed nearly constant. The main increase in muscle mass during the second trimester was caused by secondary fiber development. As an example, the ratio between secondary and primary fibers increased in Holstein Friesian fetuses from 5.9 at 4.5 mo of gestation to 21.6 at 6 mo of gestation. Primary and secondary fibers continued to growth during the third trimester. Regional differences in the density of slow muscle fibers were detected leading to greater variation within the muscle than among breeds. Structural organization of muscle fibers in muscle fiber bundles developed early in fetal life. At first, large main bundles were visible. Smaller structural units defined as primary bundles were measurable at 6 mo of gestation when most fibers were developed. The size of primary bundles nearly doubled from 6 mo of gestation to birth in all breeds. In summary, differences among breeds in the early fetal muscle fiber development were detected in contractile differentiation and partly in muscle fiber bundle structure. A prolonged secondary fiber generation and altered contractile differentiation may be involved in breed differences of postnatal muscle development. [ABSTRACT FROM AUTHOR]

Skeletal muscle is a very heterogeneous tissue consisting of diverse cell types with specific transcription profiles. Therefore, the measured mRNA abundance of a certain cell type marker is influenced by the transcriptional activity as well as by the usually unknown number of contributing cells in the sample. In studies on the transcriptional activity of adipogenic genes, as indicators for the development of intramuscular adipocytes, an altered number of adipocytes or respective progenitor cells can mask changes in transcriptional activity. To overcome this problem, we started to use laser microdissection to isolate RNA of adipocytes and muscle fibers separately for downstream analysis. Even muscle fiber types can be collected and analyzed separately. Laser microdissection in combination with biopsy techniques enables gene expression studies of particular cell types during the life cycle of an animal. First experiences using laser microdissection for adipogenic gene expression studies in bovine skeletal muscle are described, and the influence of sample preparation and future challenges are discussed. [ABSTRACT FROM AUTHOR]

Abstract: The objective of this study was to investigate the effect of long-term heat stress on performance traits and plasma enzyme activities in Naked-neck (Na) chickens and commercial breeds (New Hampshire, NH and Lohmann white, LW) with their F1 crosses (NaxNH and NaxLW). A total of 240 female chickens were randomly assigned to a completely randomized design in a 5×2 factorial arrangement (five genetic groups and two ambient temperatures: high=30–32°C; normal=18–20°C). Body weights were measured at 20 and 68weeks of age. Eggs were collected once daily while egg weight and feed intake were determined at 28-d intervals. Blood samples were taken from 12 randomly selected birds per genotype and ambient temperature (12 out of 24 hens) at 22, 38, 51 and 65weeks age. Levels of glutamic–pyruvic transaminase (GPT), glutamic–oxaloacetic transaminase (GOT) and creatine kinase (CK) were determined in blood plasma. Compared with controls, the commercial hens showed significant (p<0.05) performance reductions under thermal stress with respect to egg production (33%), egg mass (24.2%), feed intake (15%) and shell thickness (24.3%). At high ambient temperature, the NaxLW cross outperformed all other genotypes with respect to heat tolerance in body weight, egg number, egg weight, feed intake and shell thickness; and thus, appeared to be suitable genetic combinations. Compared with controls, heat stress significantly (p<0.05) increased CK, GPT and GOT activities in all genotypes. However, the response of heat stressed genotypes at different ages was inconsistent and variable. In conclusion, the present study showed that the Naked-neck chickens and their F1 crosses were more thermo-tolerant compared with commercial layer hens. Although plasma enzyme activities uniformly increased due to heat stress, the response of genotypes with age was inconsistent. [Copyright &y& Elsevier]

Nuclear transcription factor Y, beta (NFYB) was evaluated as candidate gene for congenital splay leg in piglets based on data from differential display and QTL analysis. We mapped NFYB to pig chromosome 5 (SSC5). By assigning further five porcine genes from the corresponding region on human chromosome (HSA) 12q23.3→ q24.11 to SSC5 and 14 we could confine an evolutionary breakpoint from an interval of more than 10 Mb to less than 400 kb. Comparative sequence analysis of the coding region of NFYB in healthy and splay leg piglets revealed no polymorphism. Inter-species conservation of the codons ranges from 87% to 95% between pig, human, cow, dog, rat and mouse, respectively. The expression of NFYB in M. biceps femoris was not different between healthy and splay leg piglets. However, healthy male piglets had a significantly higher expression than females. Our results exclude NFYB as candidate gene for congenital splay leg but provide a basis for selection of further candidates for the disease from SSC5. Copyright © 2007 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Abstract: The myogenic factors (MYF) 5 and 6 are integral to the initiation and development of skeletal muscle and to the maintenance of its phenotype. Thus, they are candidate genes for growth- and meat quality-related traits. We performed a comparative sequence analysis of the MYF5/MYF6 locus in swine, cattle, dog, chicken and zebrafish on the basis of structural and functional information from human and mouse. Beside the characterization of upstream regulatory elements recently identified in mice, we demonstrate the existence of further highly conserved elements (E1 to E4) which may play a role in the regulation of MYF5 and MYF6 expression. Comparative sequence analysis of putative regulatory sequences in swine revealed a total of 21 single nucleotide polymorphisms (SNP) including 1 and 6 SNPs new for the promoters of MYF5 and MYF6, respectively. The conserved organization of the locus in vertebrates indicates a common basic mechanism of muscle development. However, the existence of numerous regulatory elements at large distances to MYF5 and MYF6 points to a very complex pattern of the gene regulation with significant differences between species. [Copyright &y& Elsevier]

The melanocortin 4 receptor is expressed in virtually all brain regions of mammals and plays an important role in energy homeostasis. Polymorphisms in this gene may thus be related to growth and obesity. In pigs, a non-synonymous polymorphic site was described (Asp298Asn) and demonstrated to affect cAMP production and to alter adenylyl cyclase signalling. Association studies revealed significant linkage of this mutation with production trait in pigs. In this study, 207 Lithuanian White pigs were genotyped at the MC4R locus and analysed on relationships between genotype and breeding values for several performance traits. The observed allele and genotype frequencies did not deviate significantly from Hardy–Weinberg equilibrium (wildtype allele 0.59; mutant allele 0.41) and are comparable with those described in other Large White populations. The mutant Asn298 allele of the MC4R gene was significantly associated with increased test daily gain, higher lean meat percentage and lower backfat thickness. There was a trend towards an improved feed conversion ratio (p = 0.065) in animals with the mutant allele whereas no significant effect was found on lifetime daily gain. These results indicate that the MC4R polymorphism should be integrated in selection programmes in the Lithuanian White to improve carcass composition. [ABSTRACT FROM AUTHOR]

The spatial genetic structure of common hamsters (Cricetus cricetus) was investigated using three partial mitochondrial (mt) genes and 11 nuclear microsatellite loci. All marker systems revealed significant population differentiation across Europe. Hamsters in central and western Europe belong largely to two allopatric mitochondrial lineages south and northwest of the Carpathian and Sudetes. The southern group,‘Pannonia’, comprises populations inside the Carpathian basin (Czech Republic, Hungary) while the second group,‘North’, includes hamsters from Belgium, the Netherlands, France, and Germany. Isolation of the lineages is maintained by a combination of geographical and ecological barriers. Both main phylogeographical groups show signs of further subdivision. North is separated into highly polymorphic central German and less polymorphic western populations, which most likely split during late glacial expansion (15 000–10 000 bp). Clock estimates based on haplotype distributions predict a divergence of the two major lineages 85 000–147 000bp. Expansion times fall during the last glaciation (115 000–10 000 bp) corroborating fossil data, which identifyCricetus cricetusas characteristic of colder climatic phases. Despite the allopatry of mt haplotypes, there is an overlap of nuclear microsatellite alleles between phylogeographical units. Although there are strong evidence that Pannonian hamsters have persisted inside the Carpathian basin over the last 50 000 years, genetic differentiation among European hamsters has mainly been caused by immigration from different eastern refugia. Possible source populations are likely to be found in the Ukrainian and the southern Russian plains— core areas of hamster distribution. From there, hamsters have repeatedly expanded during the Quaternary. [ABSTRACT FROM AUTHOR]

We have isolated and characterized 18 microsatellite loci in the Peking duck ( Anas platyrhynchos). The average number of alleles per locus was 3.5, ranging from one to six in domestic Peking ducks ( n = 40). All of the markers were polymorphic in a sample of five mallards ( A. platyrhynchos; two to eight alleles). Seventeen of the 18 markers amplified in Muscovy ducks ( Cairina moschata) with 11 being polymorphic in our sample ( n = 14). Amplification of the markers in different species comprising the subfamilies Anatinae and Anserinae indicates their potential value for population genetic applications in a wide range of waterfowl species. [ABSTRACT FROM AUTHOR]

Reports on the development and characterization of the first nine polymorphic dinucleotide repeat loci in the Mongolian gerbil. Examination of inheritance and linkage of microsatellites in seven gerbil families; Estimation of the level of genetic polymorphism; Comparison of genetic variability of laboratory and wild gerbils.

Introduces articles concerning animal genetics. Dinucleotide repeat polymorphism at the ovine loci; Analysis of porcine melanocortin-5 receptor genes; Identification of polymorphisms in head and hinge region of feline cardiac Beta-myosin heavy chain gene; Linkage and physical mapping of sheep perforin gene.

Birth weight differences have been shown to affect body composition in pigs and other species. However, data on effects of birth weight differences on morphological and cellular traits of carcass in bulls with a longer fattening period are scarce. Consequently, we have analyzed 2 groups of bulls from an experimental F2 cross (Charolais × Holstein) with low birth weight (LBW; 35.6 ± 0.7 kg; n = 20) and high birth weight (HBW; 59.4 ± 0.9 kg; n = 20). All animals were kept in a tight stall barn from d 121; were fed a standardized, concentrate-based diet ad libitum; and were slaughtered at an age of 547 d. The feed intake was higher in the HBW group but feed conversion ratio was not different. Live weights were not different between the 4th and 15th month of age but then diverged, resulting in significantly higher weights at slaughter at 18 mo in the HBW group (717 ± 13 vs. 661 ± 10 kg; P < 0.05). Bulls of the HBW group accreted more protein (14.9 ± 0.15 vs. 14.4 ± 0.17%; P = 0.017) and LBW bulls accreted more fat (17.2 ± 0.81 vs. 14.3 ± 0.79%; P = 0.016) in the carcass. The absolute and relative weights of omental, perirenal, and visceral adipose depots were significantly higher in LBW bulls. Weight of longissimus (MLD) was higher in the HBW bulls (8.6 ± 0.15 vs. 7.7 ± 0.13 kg; P < 0.001) compared with LBW animals. However, there were no significant differences in muscle fiber traits in MLD. The intramuscular fat (IMF) content of MLD from LBW bulls was significantly increased compared with that of HBW animals (3.9 ± 0.5 vs. 2.5 ± 0.3%; P = 0.012). Analysis of cellular parameters of the IMF revealed increased diameters (94.4 ± 1.7 vs. 81.4 ± 2.4 μm; P < 0.01) of adipocytes and larger but not more marbling flecks as cause for higher IMF in LBW bulls. Our results demonstrate long-lasting effects of large differences in birth weight on carcass traits. Crossbred bulls could not compensate low birth weights even under ad libitum feeding regimen within a period of 18 mo in this experiment. Our data indicate that metabolic differences resulted in different partitioning of nutrients with a shift to lipid accumulation in bulls with lower birth weight in this crossbreed population. [ABSTRACT FROM AUTHOR]

Abstract Congenital splay leg is a hereditary disease observed in newborn piglets. The etiology and pathogenetic mechanism of the disorder are still unknown. The gene for cyclin-dependent protein kinase inhibitor 3 (CDKN3) was identified as a potential candidate gene in a differential display experiment. We analyzed the gene on sequence variations and compared its expression in M. biceps femoris between healthy and affected piglets. Comparative sequencing of the coding region of three healthy and four splay leg piglets revealed twelve single nucleotide polymorphisms (SNP) resulting in six amino acid exchanges in the deduced sequences. However, all polymorphisms were observed in healthy as well as in splay leg piglets thus excluding structural differences of the gene as a cause of the disease. Besides full length transcripts, we found a variety of aberrantly transcribed cDNA in clones derived from M. biceps femoris of healthy as well as of splay leg piglets. All alternative transcripts coexist with normal cDNA. Expression analysis revealed a trend towards higher values in M. biceps femoris of splay leg piglets supporting the results obtained from a differential display.

Studies the nucleotide sequence and physical mapping of the porcine cyclin-dependent kinase inhibitor 3 (CDKN3) gene. Genus and species; Origin of clone; Comparison with related sequence.

Focuses on the chromosomal localization and partial cDNA sequence of the porcine adenocine triphosphate synthase subunit e gene. Origin of the clone; Comparison with related sequences; Chromosomal location.

Abstract: The experiment was conducted to study the development of intramuscular fat in Japanese Black (JB) compared to Holstein (HS) steers and to find breed differences for fat depot development and distribution in the carcass under equal feeding conditions. Additional to slaughter samples, biopsy samples of longissimus muscle (LM) and subcutaneous fat, taken at 10, 14, 18, and 22months of age, were used for histological and molecular investigations. Japanese Black steers stored about 14% more fat in the LM (P=0.001), resulting in larger marbling flecks (P<0.001). Muscle fibers and intramuscular adipocytes in both breeds responded to the high energy feeding with significant enlargement, which was faster in JB. Histograms of intramuscular adipocytes size showed a shift toward larger cells during growth, but also the abundance of small, developing adipocytes. This development was accompanied by a correlated up-regulation of adipogenic genes until 22months of age. [Copyright &y& Elsevier]

Abstract: The incidence of hyper-contracted giant fibres in pig postmortem skeletal muscle is closely related to poor meat quality in terms of pale, soft, and exudative pork. Detection of a predisposition to develop giant fibres in live pigs could help to predict pork quality and to exclude affected pigs from genetic selection. The abundance and proportion of giant fibres in longissimus muscle were highest in Piétrain followed by Landrace, Large White, and Leicoma pigs of market weight. The postmortem development of giant fibres could be successfully simulated in vitro incubating biopsy samples from longissimus muscle at 37°C for 60min. For repeated measurements on three samples the intraclass correlation coefficient for the number of giant fibres/cm2 was for biopsy and for carcass samples. “Simulated” giant fibres exhibited ultrastructural changes in plasma membrane, myofibrils, mitochondria, and sarcoplasmatic reticulum as shown previously for giant fibres in carcass samples. [Copyright &y& Elsevier]

Introduction: Porcine congenital splay leg syndrome (PCS) is a major birth defect in piglets, resulting in lameness and high mortality rates. The multifactorial pathogenesis of PSC is not well understood but includes a polygenic inheritance., Methods: Here, in addition to morphological investigations, we characterized the expression of myogenic genes and functional (proliferation and differentiation) properties of myogenic precursor/satellite cells (SATCs) in 1 day-old PCS piglets, non-affected littermates (LCs), and piglets from PCS-free healthy litters (HCs). In addition, PCS phenotypes were related to the SNP Homer1&#95;rs325197091 within the Homer1 locus, which has been identified as a potential hereditary cause of PCS., Results and Discussion: Samples from musculus semitendinosus (ST) of PCS piglets had a higher proportion of type II fibers, reflecting myofiber immaturity. In addition, myofiber atrophy, a lower number of myonuclei per fiber (ST), and a higher apoptotic activity (in ST and longissimus dorsi muscle; LD) were found in the PCS group. A higher proportion of cycling committed myoblasts (Pax7 + /Ki67 + cells) occurred in samples from PCS-affected piglets, and on the other hand, the mRNA expression of genes involved in differentiation (muscle differentiation 1; MyoD, myogenin; MyoG) was repressed compared with HCs. Cultured SATCs from PCS-affected animals showed a temporal shift in peak expression of Pax7, MyoD, and MyoG toward days 3 and 4 of their 7 days differentiation regime. In vitro experiments with isolated SATCs confirmed the lower differentiation potential and the delayed progression of the myogenic processes in cells from piglets with PCS phenotype. In addition, Pax7 and desmin were differently expressed in Homer1&#95;rs325197091 genotype variants (GG, GA, and AA). Both genes showed the lowest expression in the homozygous AA-variant, which was most frequently found in PCS-affected animals. The homozygous AA-variant was also associated with lower expression of the truncated Homer1-subtype 205. Thus, we hypothesize that in PCS, the balance between Homer1 proteins and its signaling functions is changed in a way detrimental to the myogenic differentiation program. Our results demonstrated direct negative effects of the Homer1 AA genotype on Pax7 expression, but the exact mode of action still needs to be elucidated., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Schumacher, Reyer, Maak and Röntgen.)

Background: Combining the results of within-population genome-wide association studies (GWAS) based on whole-genome sequences into a single meta-analysis (MA) is an accurate and powerful method for identifying variants associated with complex traits. As part of the H2020 BovReg project, we performed sequence-level MA for beef production traits. Five partners from France, Switzerland, Germany, and Canada contributed summary statistics from sequence-based GWAS conducted with 54,782 animals from 15 purebred or crossbred populations. We combined the summary statistics for four growth, nine morphology, and 15 carcass traits into 16 MA, using both fixed effects and z-score methods., Results: The fixed-effects method was generally more informative to provide indication on potentially causal variants, although we combined substantially different traits in each MA. In comparison with within-population GWAS, this approach highlighted (i) a larger number of quantitative trait loci (QTL), (ii) QTL more frequently located in genomic regions known for their effects on growth and meat/carcass traits, (iii) a smaller number of genomic variants within the QTL, and (iv) candidate variants that were more frequently located in genes. MA pinpointed variants in genes, including MSTN, LCORL, and PLAG1 that have been previously associated with morphology and carcass traits. We also identified dozens of other variants located in genes associated with growth and carcass traits, or with a function that may be related to meat production (e.g., HS6ST1, HERC2, WDR75, COL3A1, SLIT2, MED28, and ANKAR). Some of these variants overlapped with expression or splicing QTL reported in the cattle Genotype-Tissue Expression atlas (CattleGTEx) and could therefore regulate gene expression., Conclusions: By identifying candidate genes and potential causal variants associated with beef production traits in cattle, MA demonstrates great potential for investigating the biological mechanisms underlying these traits. As a complement to within-population GWAS, this approach can provide deeper insights into the genetic architecture of complex traits in beef cattle., (© 2023. ’Institut National de Recherche en Agriculture, Alimentation et Environnement (INRAE).)

In contracting muscles, carbohydrates and fatty acids serve as energy substrates; the predominant utilization depends on the workload. Here, we investigated the contribution of non-mitochondrial and mitochondrial metabolic pathways in response to repeated training in a polygenic, paternally selected marathon mouse model (DUhTP), characterized by exceptional running performance and an unselected control (DUC), with both lines descended from the same genetic background. Both lines underwent three weeks of high-speed treadmill training or were sedentary. Both lines' muscles and plasma were analyzed. Muscle RNA was sequenced, and KEGG pathway analysis was performed. Analyses of muscle revealed no significant selection-related differences in muscle structure. However, in response to physical exercise, glucose and fatty acid oxidation were stimulated, lactate dehydrogenase activity was reduced, and lactate formation was inhibited in the marathon mice compared with trained control mice. The lack of lactate formation in response to exercise appears to be associated with increased lipid mobilization from peripheral adipose tissue in DUhTP mice, suggesting a specific benefit of lactate avoidance. Thus, results from the analysis of muscle metabolism in born marathon mice, shaped by 35 years (140 generations) of phenotype selection for superior running performance, suggest increased metabolic flexibility in male marathon mice toward lipid catabolism regulated by lactate dehydrogenase.

Previous attempts to increase the level of flaxseed in hens' diet for the production of n-3 polyunsaturated fatty acids (n-3 PUFAs)-enriched eggs have been commonly associated with undesirable effects on production efficiency, lipid health indices, and oxidative stability of eggs, requiring adequate research attention. This study investigated the effects of feeding a moderate level of flaxseed (FS) and plant polyphenol extracts (PPEs) on fatty acid content, oxidative stability, and lipid health indices in eggs of slow-growing Sasso T451A laying hens. One hundred and five hens were assigned to five groups (seven replicates of three) and fed on FS (75 g flaxseed and no antioxidants), VE8 (75 g flaxseed and 800 mg vitamin E), TS8 (75 g flaxseed and 800 mg Thymus schimperi ), DA8 (75 g flaxseed and 800 mg Dodonaea angustifolia ), and CD8 (75 g flaxseed and 800 mg Curcuma domestica ) extract per kg diets. The egg yolk content of eicosapentaenoic acid (EPA, C20:5 n-3) in the DA8, TS8, and CD8 diets and docosahexaenoic acid (DHA, C22:6 n-3) in TS8 and CD8 diets significantly ( p < 0.05) increased compared with the FS diet. The FS diet significantly increased the malondialdehyde (MDA) content in egg yolks, whereas the TS8 diet decreased it by 67% ( p < 0.05). Little difference was observed in yolk fatty acid content between cooked and raw eggs. Production of n-3 PUFA-enriched eggs with favorable lipid health indices was possible through inclusion of PPEs extracted from local plant species grown in Ethiopia and a moderate dose of flaxseed in the diet of laying hens., Competing Interests: The authors declare no conflict of interest.

One way to improve the growth of low-birth-weight (LBW) piglets can be stimulation of the cellular development of muscle by optimized amino acid supply. In the current study, it was investigated how glutamine (Gln) supplementation affects muscle tissue of LBW and normal-birth-weight (NBW) piglets. Longissimus and semitendinosus muscles of 96 male piglets, which were supplemented with 1 g Gln/kg body weight or alanine, were collected at slaughter on day 5 or 26 post natum (dpn), one hour after injection with Bromodeoxyuridine (BrdU, 12 mg/kg). Immunohistochemistry was applied to detect proliferating, BrdU-positive cells in muscle cross-sections. Serial stainings with cell type specific antibodies enabled detection and subsequent quantification of proliferating satellite cells and identification of further proliferating cell types, e.g., preadipocytes and immune cells. The results indicated that satellite cells and macrophages comprise the largest fractions of proliferating cells in skeletal muscle of piglets early after birth. The Gln supplementation somewhat stimulated satellite cells. We observed differences between the two muscles, but no influence of the piglets' birth weight was observed. Thus, Gln supplements may not be considered as effective treatment in piglets with low birth weight for improvement of muscle growth.

Phosphorus (P) inclusion in broiler diets needs to meet the physiological demands at a specific developmental stage to ensure the performance, health, and welfare of the birds and minimize nutrient losses. Toward a more efficient utilization of P in broiler husbandry, a timed nutritional conditioning strategy might enhance the endogenous mechanisms of mineral homeostasis and thus reduce dietary P supply of mineral sources. In this study, following a variable P supply in the starter phase, the effects of a dietary P depletion of broiler chickens were investigated at different developmental stages. Physiological adaptation mechanisms were elucidated based on zootechnical performance, endocrine parameters, regulation of intestinal P transport, bone characteristics, and health aspects. The results revealed a marked response to P depletion at the earliest developmental phase, after which indications of effective compensatory mechanism were detectable with advancing ages. Potential mechanisms that enable broilers to maintain mineral homeostasis primarily include endocrine control mediated by calcitriol actions, as well as intestinal P uptake and mineral mobilization from the bone. Conclusively, the precise timing, duration, and extent of a P depletion strategy in the broiler chicken might be considered for optimized nutrient utilization., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Adding flaxseed was found to decrease oxidative stability in feed and increase the antioxidant needs of chicken. This has also been associated with a decrease in the nutritional value and oxidative stability of meat if sufficient dietary antioxidants are not included. Furthermore, dietary flaxseed has been explored in fast-growing chickens as such studies are limited with slow-growing chickens. Thus, this study aimed to evaluate the effects of feeding plant polyphenol extracts as an antioxidant alongside flaxseed on fatty acid content, oxidative stability, and lipid health indices in breast muscle of slow-growing Sasso T451A dual-purpose chicken. A total of 126 chickens assigned to six groups (seven replicates of three) were fed on NC (control and no antioxidants), FS (75 g flaxseed and no antioxidants), VE8 (75 g flaxseed and 800 mg vitamin E), TS8 (75 g flaxseed and 800 mg Thymus schimperi), DA8 (75 g flaxseed and 800 mg Dodonaea angustifolia) and CD8 (75 g flaxseed and 800 mg Curcuma domestica) extract per kg diet. Feeding on CD8 and VE8 in raw and TS8, CD8 and VE8 diets in cooked breast muscle increased (p < 0.05) the C22:6n − 3 (DHA) and C20:5n − 3 (EPA) contents compared to the FS diet. Feeding FS increased (p < 0.05) the malondialdehyde (MDA) content in breast muscle, whereas TS8 in cooked and raw and CD8 and DA8 diets in raw breast muscle decreased it (p < 0.05). No added benefit was observed in feeding VE8 over plant extracts in terms of improving fatty acid composition and lipid health indices and reducing lipid oxidation in breast meat.

Previously, microRNA-100 (miR-100) and its putative mRNA target, insulin-like growth factor receptor-1 ( IGF1R ) were identified as differentially and inversely expressed in bovine longissimus dorsi (LD) muscles with divergent intramuscular fat (IMF) content by our group. While IGF1R signaling is implicated in myogenesis and muscle lipid metabolism, the underlying regulatory mechanisms are poorly understood. In the present study, we aimed to investigate the regulation of IGF1R by miR-100 during bovine muscle satellite cell (BMSC) myogenesis and lipid deposition. MiR-100 was confirmed to target the IGF1R 3'-untranslated region (3'-UTR) by luciferase reporter assay. Furthermore, expression of miR-100 and IGF1R was reciprocal during BMSC differentiation, suggesting a crosstalk between the two. Correspondingly, miR-100 mimic (agomiR) suppressed the levels of IGF1R , PI3K/AKT pathway signaling, myogenic gene MYOG , muscle structural components MYH7 and MYH8, whereas the inhibitor (antagomiR) had no clear stimulating effects. The IGF1R inhibitor (BMS-754807) curtailed receptor levels and triggered atrophy in muscle myotubes but did not influence miR-100 expression. AgomiR increased oleic acid-induced lipid deposition in BMSC myotubes supporting its involvement in intramuscular fat deposition, while antagomiR had no effect. Moreover, mitochondrial beta-oxidation and long-chain fatty acid synthesis-related genes were modulated by agomiR addition. Our results demonstrate modulatory roles of miR-100 in BMSC development, lipid deposition, and metabolism and suggest a role of miR-100 in marbling characteristics of meat animals and fat oxidation in muscle.

In 2002, a transmembrane protein-now known as FNDC5-was discovered and shown to be expressed in skeletal muscle, heart, and brain. It was virtually ignored for 10 years, until a study in 2012 proposed that, in response to exercise, the ectodomain of skeletal muscle FNDC5 was cleaved, traveled to white adipose tissue, and induced browning. The wasted energy of this browning raised the possibility that this myokine, named irisin, might mediate some beneficial effects of exercise. Since then, more than 1000 papers have been published exploring the roles of irisin. A major interest has been on adipose tissue and metabolism, following up the major proposal from 2012. Many studies correlating plasma irisin levels with physiological conditions have been questioned for using flawed assays for irisin concentration. However, experiments altering irisin levels by injecting recombinant irisin or by gene knockout are more promising. Recent discoveries have suggested potential roles of irisin in bone remodeling and in the brain, with effects potentially related to Alzheimer's disease. We discuss some discrepancies between research groups and the mechanisms that are yet to be determined. Some important questions raised in the initial discovery of irisin, such as the role of the mutant start codon of human FNDC5 and the mechanism of ectodomain cleavage, remain to be answered. Apart from these specific questions, a promising new tool has been developed-mice with a global or tissue-specific knockout of FNDC5. In this review, we critically examine the current knowledge and delineate potential solutions to resolve existing ambiguities., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Endocrine Society.)

Muscle growth of low birth weight (LBW) piglets may be improved with adapted nutrition. This study elucidated effects of glutamine (Gln) supplementation on the cellular muscle development of LBW and normal birth weight (NBW) piglets. Male piglets (n = 144) were either supplemented with 1 g Gln/kg body weight or an isonitrogeneous amount of alanine (Ala) between postnatal day 1 and 12 (dpn). Twelve piglets per group were slaughtered at 5, 12 and 26 dpn, one hour after injection with Bromodeoxyuridine (BrdU, 12 mg/kg). Muscle samples were collected and myogenic cells were isolated and cultivated. Expression of muscle growth related genes was quantified with qPCR. Proliferating, BrdU-positive cells in muscle sections were detected with immunohistochemistry indicating different cell types and decreasing proliferation with age. More proliferation was observed in muscle tissue of LBW-GLN than LBW-ALA piglets at 5 dpn, but there was no clear effect of supplementation on related gene expression. Cell culture experiments indicated that Gln could promote cell proliferation in a dose dependent manner, but expression of myogenesis regulatory genes was not altered. Overall, Gln supplementation stimulated cell proliferation in muscle tissue and in vitro in myogenic cell culture, whereas muscle growth regulatory genes were barely altered.

Piglets with low birth weight (LBW) usually have reduced muscle mass and increased lipid deposition compared with their normal-birth-weight (NBW) littermates. Supplementation of piglets with amino acids during the first days of life may improve muscle growth and simultaneously alter the intramuscular lipid deposition. The aim of the current study was to investigate the influence of glutamine (Gln) supplementation during the early suckling period on lipid deposition in the longissimus muscle (MLD) and the role of different perilipin (PLIN) family members in this process. Four groups were generated consisting of 72 male LBW piglets and 72 NBW littermates. Piglets were supplemented with either 1 g Gln/kg body weight or an isonitrogenous amount of alanine (Ala) between days post natum (dpn) 1 and 12. Twelve piglets per group were slaughtered at 5, 12, and 26 dpn, and muscle tissue was collected. Perilipins were localized by immunohistochemistry in muscle sections. The mRNA and protein abundances of PLIN family members and related lipases were quantified by quantitative RT-PCR (qPCR) and western blots, respectively. While PLIN1 was localized around lipid droplets in mature and developing adipocytes, PLIN2 was localized at intramyocellular lipid droplets, PLIN3 and 4 at cell membranes of muscle fibers and adipocytes, and PLIN5 in the cytoplasm of undefined cells. The western blot results indicated higher protein abundances of PLIN2, 3, 4, and 5 in LBW piglets ( p < 0.05) at 5 dpn compared with their NBW littermates independent of supplementation, while not directly reflecting the mRNA expression levels. The mRNA abundance of PLIN2 was lower while PLIN4 was higher in piglets at 26 dpn in comparison with piglets at 5 dpn ( p < 0.01). Relative mRNA expression of LPL and CGI-58 was lowest in piglets at 5 dpn ( p < 0.001). However, ATGL mRNA was not influenced by birth weight or supplementation, but the Spearman correlation coefficient analysis revealed close correlations with PLIN2 , 4, and 5 mRNA at 5 and 26 dpn ( r > 0.5, p < 0.001). The results indicated the importance of birth weight and age for intramuscular lipid deposition and different roles of PLIN family members in this process, but no clear modulating effect of Gln supplementation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zhao, Albrecht, Li, Schregel, Sciascia, Metges and Maak.)

The porcine congenital splay leg syndrome (PCS), even though being of transient nature, is still one of the most important causes for piglet losses due to its high incidence and mortality. Although, described decades ago, the pathogenetic mechanism is still elusive. Numerous, mostly descriptive studies characterized the syndrome at clinical, histological and cellular levels but resulted in a highly diverse picture of the syndrome. Broad variability in phenotypical expression and, in case of proper care, the rapid recovery of affected animals complicated a systematical analysis of the underlying pathogenesis. Although, several environmental factors were discussed as potential causes of PCS, most of the evidence points to a hereditary basis of PCS. Nevertheless, only few of the suggested candidate genes from transcriptome and mapping analyses, like F-box protein 32 (FBXO32), could be confirmed so far. Only recently, a genome wide association study revealed genomic regions on five porcine chromosomes and named a number of potential candidate genes, among them homer scaffold protein 1 (HOMER1). This new candidate-a cellular scaffold protein-plays a role in a plethora of cellular signaling cascades, and is not only involved in skeletal muscle differentiation but also critical for muscular function. In this review, we critically elucidate the current state of knowledge in the field and evaluate current achievements in the identification of the pathogenetic mechanism for the syndrome., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Schumacher, Röntgen and Maak.)

Adapted nutrition can improve the growth of low birth weight (LBW) piglets. Since maternal milk is thought to provide insufficient glutamine (Gln) for LBW piglets, the current study investigated the influence of Gln supplementation during the early suckling period on development and lipid deposition in skeletal muscle. The weight differences between LBW and normal birth weight (NBW) littermates persisted from birth to slaughter ( p < 0.001). However, intramuscular Gln and Ala concentrations were altered in piglets according to the supplementation ( p < 0.01). There were larger muscle fibers ( p = 0.048) in Gln-supplemented piglets. Capillarization or nuclei number per muscle fiber was not influenced by birth weight (BiW) or Gln supplementation. Abundance of myosin heavy chain (MYH) isoforms was slightly altered by Gln supplementation. LBW piglets had more lipid droplets than NBW piglets at day 5 of life in both muscles ( p < 0.01). The differences decreased with age. Adipocyte development increased with age, but was not influenced by BiW or supplementation. The results indicate that BiW differences were accompanied by differences in lipid deposition and muscle fiber structure, suggesting a delayed development in LBW piglets. Supplementation with Gln may support piglets to overcome those disadvantages.