S. G. Velleman

Extrinsic regulation of domestic animal-derived satellite cells

Satellite cells are the postnatal myogenic cells, as they provide myonuclei to support skeletal muscle hypertrophy and are principal cells responsible for myofiber repair and regeneration. Even though research with satellite cells from meat animals is new, considerable data exist to suggest that these cells are regulated through both intrinsic and extrinsic mechanisms. This review covers the present status of the extrinsic factors known or postulated to modulate meat animal satellite cell growth and development.

Heterogeneity in growth and differentiation characteristics in male and female satellite cells isolated from turkey lines with different growth rates

The effects of growth- and gender-related differences on satellite cell proliferation and differentiation were investigated using satellite cells isolated from the pectoralis major muscle of a turkey line selected for increased 16-week body weight (F-line) and its unselected randombred control (RBC2-line). Proliferation rates within the F- and RBC2-lines did not differ between sexes. The F-line male and female satellite cells when compared to the RBC2-line male and female satellite cells proliferated at a faster rate. Differentiation rates were increased for the F-line male cells compared to both the F-line female and RBC2-line male satellite cells. No difference in differentiation rate was noted within the RBC2-line satellite cells. For satellite cells from females, the RBC2-line differentiated faster than the F-line. Morphological data on myotube length and the number of nuclei per myotube supported the differentiation data in that F-line male satellite cells had the longest myotubes with the most nuclei, there was no significant difference between myotubes within the RBC2-line, and female-derived myotubes from the RBC2-line were longer than those of the F-line by 96 h of fusion. These data are suggestive of both growth- and gender- related differences in satellite cell proliferation and differentiation.

Histopathologic and Myogenic Gene Expression Changes Associated with Wooden Breast in Broiler Breast Muscles

SUMMARY The wooden breast condition is a myopathy affecting the pectoralis major (p. major) muscle in fast-growing commercial broiler lines. Currently, wooden breast–affected birds are phenotypically detected by palpation of the breast area, with affected birds having a very hard p. major muscle that is of lower value. The objective of this study was to compare the wooden breast myopathy in two fast-growing broiler lines (Lines A and B) with incidence of wooden breast to a slower growing broiler Line C with no phenotypically observable wooden breast. One of the characteristics of the wooden breast condition is fibrosis of the p. major muscle. Morphologic assessment of Lines A and B showed significant fibrosis in both lines, but the collagen distribution and arrangement of the collagen fibrils was different. In Line A, the collagen fibrils were tightly packed, whereas in Line B the collagen fibrils were diffuse. This difference in collagen organization may be due to the expression of the extracellular matrix proteoglycan decorin. Decorin is a regulator of collagen crosslinking and is expressed at significantly higher levels in Line A wooden breast–affected p. major muscle, which would lead to tightly packed collagen fibers due to high levels of collagen crosslinking. Furthermore, expression of the muscle-specific transcriptional regulatory factors for proliferation and differentiation of muscle cells leading to the regeneration of muscle in response to muscle damage was significantly elevated in Line A, and only the factor for differentiation, myogenin, was increased in Line B. The results from this study provide initial evidence that the etiology of the wooden breast myopathy may vary between fast-growing commercial broiler lines.

The role of syndecan-4 and attached glycosaminoglycan chains on myogenic satellite cell growth

The syndecans are a family of cell-surface heparan sulfate proteoglycans consisting of a core protein with covalently attached glycosaminoglycan (GAG) chains. Syndecan-4 expression in skeletal muscle is increased in growth-selected animals during proliferation. Previous studies have suggested that cell-surface heparan sulfate proteoglycans like syndecan-4 are involved in fibroblast growth factor 2 (FGF2) signaling by FGF2 binding to the heparan sulfate chains. Fibroblast growth factor 2 is a potent stimulator of muscle proliferation and an intense inhibitor of differentiation. To investigate the functional contribution of the attached syndecan-4 GAG chains, a turkey syndecan-4 full length cDNA was cloned and mutated at two or all three potential GAG attachment sites at Ser38, Ser65, and Ser67 resulting in all possible one-chain mutants and a no-chain mutant. Turkey satellite cells were transfected with the wild-type syndecan-4, one-chain mutants, no-chain mutant, or the empty vector and assayed for cell proliferation, differentiation, and FGF2 responsiveness. The wild-type syndecan-4, one-chain mutants, and no-chain mutant inhibited cell proliferation and delayed initial differentiation but did not alter FGF2 responsiveness or function through the mitogen-activated protein kinase pathway. There was no difference between the wild-type syndecan-4, one-chain, and no-chain mutants during these stages. These data suggest that syndecan-4 functions in an FGF2-independent manner and the GAG chains attached to the syndecan-4 core protein are not required for syndecan-4 to affect turkey satellite cell proliferation and the initial differentiation.

Extracellular matrix proteoglycan decorin-mediated myogenic satellite cell responsiveness to transforming growth factor-β1 during cell proliferation and differentiation

Transforming growth factor-beta1 (TGF-beta1) is a potent inhibitor of muscle cell proliferation and differentiation. Decorin, a small proteoglycan in the extracellular matrix, binds to TGF-beta1 and modulates the activity of TGF-beta1 during muscle cell growth and development. However, its interaction with TGF-beta1 and involvement in myogenesis is not well characterized. In the present study, chicken myogenic satellite cells, myogenic precursors for muscle growth and repair, were isolated from the pectoralis major muscle and used to investigate the biological function of TGF-beta1 and decorin during myogenesis. The over-expression of decorin in satellite cells significantly increased cell proliferation, compared to the control cells. Consistent with this result, reducing decorin expression decreased cell proliferation, which suggests a decorin-mediated mechanism is involved in the regulation of myogenic satellite cell proliferation. Satellite cells over-expressing decorin were less sensitive to TGF-beta1 during proliferation, which indicates that decorin may sequester TGF-beta1 leading to increased proliferation. During satellite cell differentiation, the over-expression of decorin induced differentiation by increasing the muscle specific creatine kinase concentration. However, the addition of TGF-beta1 diminished decorin-mediated cell responsiveness to TGF-beta1 during differentiation. Taken together, these results suggest that decorin induces myogenic satellite cell proliferation and differentiation by regulating cellular responsiveness to TGF-beta1. An alternative TGF-beta1-independent pathway may be involved in the regulation of satellite cells by decorin.

Expression and localization of the proteoglycan decorin during the progression of cholesterol induced atherosclerosis in Japanese quail: implications for interaction with collagen type I and lipoproteins

The temporal and spatial distribution, and relative levels of the proteoglycan decorin and collagen type I were examined during the progression of atherosclerosis in the dorsal aortas of Japanese quail selected for cholesterol induced atherosclerosis. The quail were placed on either a control or 0.5% added cholesterol diet at approximately 16 weeks of age. Dorsal aortas were collected at 1- or 2-week intervals over a 15-week period after initiating cholesterol feeding. Biochemical analysis for decorin and collagen type I showed that both increased in the cholesterol-fed birds compared to control-fed birds beginning at 9 weeks and continued through the duration of the study. Through immunohistochemical staining for decorin and collagen type I, the spatial localization of decorin and collagen type I in control and less severe plaques in cholesterol-fed birds was most prominent in the arterial adventitia. However, in severe atherosclerotic plaques, decorin was localized in foam cell regions and collagen type I was found surrounding the foam cell regions where decorin accumulated. These results demonstrated a localization of decorin in the core of the atherosclerotic plaque foam cell region with collagen type I being located on the plaque surface.

Adipogenic differentiation state-specific gene expression as related to bovine carcass adiposity

Genetic regulation of the site of fat deposition is not well defined. The objective of this study was to investigate adipogenic differentiation state-specific gene expression in feedlot cattle (>75% Angus; <25% Simmental parentage) of varying adipose accretion patterns. Four groups of 4 steers were selected via ultrasound for the following adipose tissue characteristics: low subcutaneous-low intramuscular (LSQ-LIM), low subcutaneous-high intramuscular (LSQ-HIM), high subcutaneous-low intramuscular (HSQ-LIM), and high subcutaneous-high intramuscular (HSQ-HIM). Adipose tissue from the subcutaneous (SQ) and intramuscular (IM) depots was collected at slaughter. The relative expression of adipogenic genes was evaluated using quantitative PCR. Data were analyzed using the mixed model of SAS, and gene expression data were analyzed using covariate analysis with ribosomal protein L19 as the covariate. No interactions (P > 0.10) were observed between IM and SQ adipose tissue depots for any of the variables measured. Therefore, only the main effects of high and low accretion within a depot and the effects of depot are reported. Steers with LIM had smaller mean diameter IM adipocytes (P < 0.001) than HIM steers. Steers with HSQ had larger mean diameter SQ adipocytes (P < 0.001) than LSQ. However, there were no differences (P > 0.10) in any of the genes measured due to high or low adipose accretion. Preadipogenic delta-like kinase1 mRNA was greater in the IM than the SQ adipose tissue; conversely, differentiating and adipogenic genes, lipoprotein lipase, PPARγ, fatty acid synthetase, and fatty acid binding protein 4 were greater (P < 0.001) in the SQ than the IM depot. Intramuscular adipocytes were smaller than SQ adipocytes and had greater expression of the preadipogenic gene, indicating that more hyperplasia was occurring. Meanwhile, SQ adipose tissue contained much larger (P < 0.001) adipocytes that had a greater expression (P < 0.001) of differentiating and adipogenic genes than did the IM adipose tissue, indicating more cells were undergoing differentiation and hypertrophy. Adipogenic differentiation state-specific gene expression was not different in cattle with various phenotypes, but adipogenesis in the SQ and IM adipose tissues seems to occur independently.

The effect of nutritional status and muscle fiber type on myogenic satellite cell fate and apoptosis

Satellite cells (SC) are multipotential stem cells that can be induced by nutrition to alter their cellular developmental fate, which may vary depending on their fiber type origin. The objective of the current study was to determine the effect of restricting protein synthesis on inducing adipogenic transdifferentiation and apoptosis of SC originating from fibers of the fast glycolytic pectoralis major (p. major) and fast oxidative and glycolytic biceps femoris (b. femoris) muscles of the chicken. The availability of the essential sulfur amino acids Met and Cys was restricted to regulate protein synthesis during SC proliferation and differentiation. The SC were cultured and treated with 1 of 6 Met/Cys concentrations: 60/192, 30/96 (control), 7.5/24, 3/9.6, 1/3.2, or 0/0 mg/L. Reductions in Met/Cys concentrations from the control level resulted in increased lipid staining and expression of the adipogenic marker genes peroxisome proliferator-activated receptor gamma and stearoyl-CoA desaturase during differentiation in the p. major SC. Although b. femoris SC had increased lipid staining at lower Met/Cys concentrations, there was no increase in expression of either adipogenic gene. For both muscle types, SC Met/Cys, concentration above the control increased the expression of peroxisome proliferator-activated receptor gamma and stearoyl-CoA desaturase during differentiation. As Met/Cys concentration was decreased during proliferation, a dose-dependent decline in all apoptotic cells occurred except for early apoptotic cells in the p. major, which had no treatment effect (P < 0.05). During differentiation, decreasing Met/Cys concentration caused an increase in early apoptotic cells in both fiber types and no effect on late apoptotic cells except for an increase in the p. major 7.5/24 mg/L of Met/Cys treatment. In general, the viability of the SC was unaffected by the Met/Cys concentration except during proliferation in the p. major 0/0 mg/L of Met/Cys treatment, which increased SC viability. These data demonstrate the effect of nutrition on SC transdifferentiation to an adipogenic lineage and apoptosis, and the effect of fiber type on this response in an in vitro context.

Genetics of Growth and Reproduction in the Turkey. 17. Changes in Genetic Parameters Over Forty Generations of Selection for Increased Sixteen-Week Body Weight

A line (F) of turkeys was selected over 40 generations for increased 16-wk BW. The base population for the F line was a randombred control population that was maintained without conscious selection and used to remove yearly environmental variation in the F line. Selection was effective in increasing 16-wk BW in the F line. Selection differentials based on the mean of the selected parents minus the mean of the entire population (intended) and intended selection differentials weighted for number of offspring produced (actual) did not differ consistently, indicating that natural selection was not opposing artificial selection during the reproduction of the F line. The realized heritability of 16-wk BW in the F line, based on the linear regression of the selection response on accumulated actual selection differential, declined with selection. For both sexes combined, the realized heritability was 0.309 +/- 0.022 (SE), 0.268 +/- 0.033, 0.268 +/- 0.026, 0.166 +/- 0.016, and 0.242 +/- 0.004, respectively, for generations 1 to 10, 11 to 20, 21 to 30, 31 to 40, and 1 to 40. Genetic increases in 16-wk BW in the F line over 40 generations of selection were positively associated with BW at other ages (8 and 20 wk of age and at 50% production), shank length and width at 16 wk of age, days from stimulatory lighting to production of the first egg, and egg weight but were negatively associated with egg production, intensity of lay (maximum and average clutch length and rate of lay), and walking ability. Over the 40 generations of selection, genetic increases in BW in the F line were not associated with changes in broodiness or mortality to 8 wk of age. During generations 31 to 40, BW at 8 and 20 wk of age continued to increase in the F line, but there was no significant change in adult BW, and the only significant change in reproduction traits was for average clutch length (-0.030). Because the genetic changes in some correlated traits were not consistent in all generation intervals studied, the genetic correlation between the selected trait (16-wk BW) and the correlated trait apparently changed with selection.

Effects of glypican‐1 on turkey skeletal muscle cell proliferation, differentiation and fibroblast growth factor 2 responsiveness

The heparan sulfate proteoglycan, glypican‐1, is a low affinity receptor for fibroblast growth factor 2 (FGF2). Fibroblast growth factor 2 is a potent stimulator of skeletal muscle cell proliferation and an inhibitor of differentiation. Heparan sulfate proteoglycans like glypican‐1 are required for FGF2 to transduce an intracellular signal. Understanding the role of glypican‐1 in the regulation of FGF2‐mediated signaling is important in furthering the understanding of the biological processes involved in muscle development and growth. In the current study, a turkey glypican‐1 expression vector construct was transfected into turkey myogenic satellite cells resulting in the overexpression of glypican‐1. The proliferation, differentiation, and responsiveness to FGF2 were measured in control and transfected cell cultures. The overexpression of glypican‐1 in turkey myogenic satellite cells increased both satellite cell proliferation and FGF2 responsiveness, but decreased the rate of differentiation. The current data support glypican‐1 modulation of both proliferation and differentiation through an FGF2‐mediated pathway.