During postnatal development, hyperplastic and hypertrophic processes of skeletal muscle growth depend on the activation, proliferation, differentiation, and fusion of satellite cells (SC). Therefore, molecular and functional SC heterogeneity is an important component of muscle plasticity and will greatly affect long-term growth performance and muscle health. However, its regulation by cell intrinsic and extrinsic factors is far from clear. In particular, there is only minor information on the early postnatal period which is critical for muscle maturation and the establishment of adult SC pools. Here, we separated two SC subpopulations (P40/50, P50/70) from muscle of 4-day-old piglets. Our results characterize P40/50 as homogeneous population of committed (high expression of Myf5), fast-proliferating muscle progenitors. P50/70 constituted a slow-proliferating phenotype and contains high numbers of differentiated SC progeny. During culture, P50/70 is transformed to a population with lower differentiation potential that contains 40% Pax7-positive cells. A reversible state of low mitochondrial activity that results from active down-regulation of ATP-synthase is associated with the transition of some of the P50/70 cells to this more primitive fate typical for a reserve cell population. We assume that P40/50 and P50/70 subpopulations contribute unequally in the processes of myofiber growth and maintenance of the SC pool.
The male and the hermaphrodite forms of the nematode Caenorhabditis elegans (C. elegans) differ markedly in anatomy, nervous system and behavior at adulthood. Using the male mutants fog-2, him-5, and him-8, we compared body proportions and composition, and aspects of carbohydrate metabolism and gene expression between the C. elegans sexes in three adult stages. In all experiments, both sexes were grown on the same plate and separated using flow cytometry. The fat to fat-free mass ratio and the body volume-adjusted fat mass is similar between the sexes, although the body size is more than 50% smaller in adult males than in age-matched hermaphrodites. The volume-adjusted total RNA content is approximately 2-fold lower in males. Biochemical and NMR-based analyses reveal higher trehalose levels and much lower glucose levels in males than in hermaphrodites. The resulting trehalose-to-glucose ratio is 5.4-fold higher in males. These sex differences are reflected in gene expression data because the genes encoding key enzymes of the glycolysis and trehalose synthesis pathways are more highly expressed in males than in hermaphrodites. Notably, expression of the phosphofructokinase gene (C50F4.2) is 29-fold higher in males. Comparative analysis of gene expression data identifies 285 male-specific and 160 hermaphrodite-specific genes. These include transcription factor and C-type lectin-encoding genes. More than 35% of all C-type lectin genes are more highly expressed in males. The expression of many C-type lectin genes differs by a factor of >100 between the sexes. In conclusion, we found sex differences in carbohydrate metabolism that are linked to gene expression and identified certain lectin genes that are differentially expressed by the C. elegans sexes.
Low birth weight (LBW) can cause lifelong impairments in muscle development and growth. Satellite cells (SC) and their progeny are crucial contributors to myogenic processes. This study provides new data on LBW in piglets combining insights on energy metabolism, muscle capillarization and differences in SC presence and function. To this aim, muscle tissues as well as isolated myogenic cells of 4-day-old German Landrace piglets were analyzed. For the first time two heterogeneous SC subpopulations, which contribute differently to muscle development, were isolated from LBW pigs by Percoll density gradient centrifugation. The muscles of LBW piglets showed a reduced DNA, RNA, and protein content as well as lower activity of the muscle specific enzymes CK, ICDH, and LDH compared to their normal birth weight siblings. We assume that deficits in energy metabolism and capillarization are associated with reduced bioavailability of SC, possibly leading to early exhaustion of the SC reserve cell pool and the cells' premature differentiation. The pig remains one of the most important farm animals worldwide and nowadays also represents a highly appreciated model system for scientific studies. In the past decades pigs were mainly selected for economic traits like reproductive fitness, growth performance, or litter size. An analysis of wild boar litters in Europe showed a mean litter size between 4.75-6.28 piglets 1. In contrast, in domestic pigs a mean litter size of 10.9 piglets in 1992 was further increased by 12% to 12.2 piglets already in 2001 2. Varona and colleagues even reported a mean litter size of 14.23 piglets in Landrace pigs 3. In polytocous species uterus capacity is a limiting factor, becoming important from day 25 of gestation on 4,5 , and more than 14 embryos can be considered as intrauterine crowding 5 possibly leading to intrauterine competition and retardation of prenatal growth 5,6. Bigger litter sizes can lead to intrauterine growth retardation (IUGR) due to insufficient development of the placenta in relation to the number of embryos, which are not sufficiently supplied with oxygen and nutrients 5. Consequently, birth weight variation (and with this also weaning weight variability) increases in large litters; and with increasing litter birth weight more low birth weight (LBW) piglets are born 2,7,8. Several important factors like birth weight, weaning weight, and gender contribute to postnatal growth performance of pigs 9 , whereas the birth weight is the earliest and most easily accessible one. Up to 15-20% of pigs exhibit a low birth weight 10 associated with developmental disadvantages compared to their normal birth weight (NBW) litter mates. Low birth weight, for instance caused by IUGR, is also relevant for other species like sheep or cattle 10,11 and for humans, where for example in the U.S. approximately 5-15% of all children have to cope with IUGR 11-13. Some LBW pigs show compensatory growth but exhibit a higher risk for infection diseases and in general their survival rate is still reduced 9,14,...
The present study was conducted to assess the effects of the probiotic Enterococcus faecium AL41 ( EF ) and of the enteric pathogen Salmonella Enteritidis PT4 ( SE ) on the development of posthatch pectoralis major muscle ( PM ) of broiler chicks. The four experimental groups were control ( CON ), EF, SE, and EF + SE (EFSE). EF and SE were given per os from days 1 to 7 and at day 4 posthatch, respectively. Muscle samples from 6 chicks per group were taken at day 8 (D8) and day 11 (D11) to evaluate PM myofiber growth, capillarization, DNA, RNA, and protein content, as well as enzyme activities (isocitrate dehydrogenase, lactate dehydrogenase, creatine kinase). PM growth rate was 7.45 ± 2.7 g/d in non-SE groups (CON, EF) and 5.10 ± 1.82 g/d in SE-infected groups ( P < 0.02). Compared with group CON, application of bacteria (groups EF and SE) reduced the fiber cross-sectional area (246 and 262 vs. 347 ± 19 μm 2 ) and the number of myonuclei per fiber (0.66 and 0.64 vs. 0.79 ± 0.03). At D11, hypertrophic myofiber growth normalized in the EF group, but negative effects persisted in SE and EFSE birds contributing to lower daily PM gain. In addition, SE infection strongly disturbed PM capillarization. Negative effects on capillary cross-sectional area and on the area (%) covered by capillaries persisted until D11 in the SE group, whereas pre-feeding of EF restored capillarization in the EFSE group to control levels. We conclude that supplementation of the probiotic bacteria EF AL41 had positive effects on PM capillarization and, thus, on delivery of O 2 , supply of nutrients, and removal of metabolites. Supplementation of probiotic bacteria might therefore reduce energetic stress and improve muscle health and meat quality during SE infection.
Muscle stem cells, termed satellite cells (SC), and SC-derived myogenic progenitor cells (MPC) are involved in postnatal muscle growth, regeneration, and muscle adaptability. They can be released from their natural environment by mechanical disruption and tissue digestion. The literature contains several isolation protocols for porcine SC/MPC including various digestion procedures, but comparative studies are missing. In this report, classic trypsinization and a more complex trypsin, collagenase, and DNase (TCD) digestion were performed with skeletal muscle tissue from 4- to 5-d-old piglets. The two digestion procedures were compared regarding cell yield, viability, myogenic purity, and in vitro cell function. The TCD digestion tended to result in higher cell yields than digestion with solely trypsin (statistical trend p = 0.096), whereas cell size and viability did not differ. Isolated myogenic cells from both digestion procedures showed comparable proliferation rates, expressed the myogenic marker Desmin, and initiated myogenic differentiation in vitro at similar levels. Thus, TCD digestion tended to liberate slightly more cells without changes in the tested in vitro properties of the isolated cells. Both procedures are adequate for the isolation of SC/MPC from juvenile porcine muscles but the developmental state of the animal should always be considered.
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