48 MUSCLE CHARACTERISTICS OF BOVINE CLONE OFFSPRING F1 COMPARED WITH CLONES

2010 ◽  
Vol 22 (1) ◽  
pp. 181 ◽  
Author(s):  
I. Cassar-Malek ◽  
Y. Heyman ◽  
B. Picard ◽  
C. Richard ◽  
P. Chavatte-Palmer ◽  
...  

Information on clone offspring F1 is limited, especially in species with a long inter-generation interval such as cattle. As cloned cattle exhibit a slight delay in muscle maturation until puberty (Jurie et al. 2009 Animal 3, 244-250), the present study aimed to investigate the contractile and metabolic muscle characteristics of F1 at 8, 12, and 18 months of age. Repeated biopsies of the semitendinosus muscle were collected on 10 F1 heifers born after AI of cloned cows at the experimental farm of INRA. Muscle characteristics of these offspring were compared with those of 9 female clones and 8 AI control heifers previously biopsied at the same ages. All animals (clones, F1, and controls) were female Holstein, born and raised under the same conditions in the same farm. Biopsy samples were stored frozen at -80°C until analysis for contractile and metabolic characteristics. The type of contractile fibers was determined from the proportion of the different myosin heavy chain (MyHC) isoforms separated by electrophoresis. Oxidative metabolism was assessed by isocitrate dehydrogenase (ICDH) and cytochrome-c oxidase (COX) activities (μmol min-1 per gram of muscle). Data were analyzed separately for each time of biopsy using the GLM procedure of SAS (SAS Institute, Cary, NC, USA). The statistical model contained the group as fixed effect. When a significant effect was detected, differences between least squares means were further separated by the PDIFF option of SAS. Comparison of contractile characteristics from the 3 groups of animals is presented in Table 1. The proportion of MyHC I (slow oxidative isoform) and MyHC IIx (fast glycolytic isoform) in the muscles of F1 was not significantly different from those of controls at 8 and 12 months of age. F1 had different muscle contractile properties compared with clones at 12 months of age. At 8 months of age, F1 had greater ICDH activity than controls (1.39 ± 0.22 v. 0.54 ± 0.007; P ≤ 0.002) and greater COX activities (11.4 ± 1.6 v. 4.2 ± 0.9; P ≤ 0.003), but this was not observed later on. Altogether, these data indicate that the muscles of F1 were more oxidative than those of controls. This was not related to a higher percentage of type I fibers but rather to a higher percentage of type IIA fibers. A delay in muscle maturation was only partially found in F1. Table 1.Contractile characteristics (mean ± SE) of the semitendinosus muscle from controls (n = 8), clones (n = 9) and clone offspring (F1, n = 10) heifers


1992 ◽  
Vol 73 (1) ◽  
pp. 179-187 ◽  
Author(s):  
B. J. Petrof ◽  
A. M. Kelly ◽  
N. A. Rubinstein ◽  
A. I. Pack

Although the association between hypothyroidism and obstructive sleep apnea is well established, the effect of thyroid hormone deficiency on contractile proteins in pharyngeal dilator muscles responsible for maintaining upper airway patency is unknown. In the present study, the effects of hypothyroidism on myosin heavy chain (MHC) expression were examined in the sternohyoid, geniohyoid, and genioglossus muscles of adult rats (n = 20). The relative proportions of MHC isoforms present were determined using MHC-specific monoclonal antibodies and oligonucleotide probes. All control muscles showed a paucity of type I MHC fibers, with greater than 90% of fibers containing fast-twitch type II MHCs. In the genioglossus muscle, a population of non-IIa non-IIb fast-twitch type II fibers (putatively identified as type IIx MHC fibers) were detected. Hypothyroidism induced significant changes in MHC expression in all muscles studied. In the sternohyoid, type I fibers increased from 6.2 to 16.9%, whereas type IIa fibers increased from 25.9 to 30.7%. Type I fibers in the geniohyoid increased from 1.2 to 12.8%, whereas type IIa fibers increased from 34.1 to 42.7%. The genioglossus showed the smallest relative increase in type I expression but the greatest induction of type IIa MHC. None of the muscles examined demonstrated reinduction of embryonic or neonatal MHC in response to thyroid hormone deficiency. In summary, hypothyroidism alters the MHC profile of pharyngeal dilators in a muscle-specific manner. These changes may play a role in the pathogenesis of obstructive apnea in hypothyroid patients.



2021 ◽  
pp. 036354652110302
Author(s):  
Wen-Chung Tsai ◽  
Tung-Yang Yu ◽  
Gwo-Jyh Chang ◽  
Hsiang-Ning Chang ◽  
Li-Ping Lin ◽  
...  

Background: The increasing use of platelet-rich plasma (PRP) to treat muscle injuries raises concerns because transforming growth factor–beta (TGF-β) in PRP may promote fibrosis in the injured muscle and thus impair muscle regeneration. Purpose: To investigate whether suramin (a TGF-β inhibitor) can reduce muscle fibrosis to improve healing of the injured muscle after PRP treatment and identify the underlying molecular mechanism. Study Design: Controlled laboratory study. Methods: Myoblasts isolated from the gastrocnemius muscle of Sprague Dawley rats were treated with PRP or PRP plus suramin. MTT assays were performed to evaluate cell viability. The expression of fibrosis-associated proteins (such as type I collagen and fibronectin), Smad2, and phosphorylated Smad2 was determined using Western blot analysis and immunofluorescent staining. An anti–TGF-β antibody was employed to verify the role of TGF-β in fibronectin expression. Gastrocnemius muscles were injured through a partial transverse incision and then treated using PRP or PRP plus suramin. Hematoxylin and eosin staining was conducted to evaluate the healing process 7 days after the injury. Immunofluorescent staining was performed to evaluate fibronectin expression. Muscle contractile properties—fast-twitch and tetanic strength—were evaluated through electric stimulation. Results: PRP plus 25 μg/mL of suramin promoted myoblast proliferation. PRP induced fibronectin expression in myoblasts, but suramin reduced this upregulation. The anti–TGF-β antibody also reduced the upregulation of fibronectin expression in the presence of PRP. The upregulation of phosphorylated Smad2 by PRP was reduced by either the anti–TGF-β antibody or suramin. In the animal study, no significant difference was discovered in muscle healing between the PRP versus PRP plus suramin groups. However, the PRP plus suramin group had reduced fibronectin expression at the injury site. Fast-twitch strength and tetanic strength were significantly higher in the injured muscle treated using PRP or PRP plus suramin. Conclusion: Simultaneous PRP and suramin use reduced fibrosis in the injured muscle and promoted healing without negatively affecting the muscle’s contractile properties. The underlying molecular mechanism may be associated with the phosphorylated Smad2 pathway. Clinical Relevance: Simultaneous PRP and suramin use may reduce muscle fibrosis without compromising muscle contractile properties and thus improve muscle healing.



2020 ◽  
Vol 98 (4) ◽  
Author(s):  
Cadra L Van Bibber-Krueger ◽  
Ashley M Collins ◽  
Kelsey J Phelps ◽  
Travis G O’Quinn ◽  
Terry A Houser ◽  
...  

Abstract The objective of this study was to determine the impact of quality grade and steak location on color stability of semitendinosus (ST) steaks during a 9-d refrigerated study. Twenty-one ST muscles (12 Choice and 9 Select) were purchased from a commercial beef packing plant and fabricated into twelve 2.54-cm thick steaks per muscle. Steaks 1, 6, and 12 were designated for immunohistochemistry while remaining steak locations of proximal (steaks 2 to 4), middle (5 to 8), and distal (9 to 11) were randomly assigned to 0, 4, or 9 d of simulated retail display. Surface color attributes of day-9 steaks were recorded daily by a visual color panel and spectrophotometer. On days 0, 4, and 9 of display, steaks were analyzed for metmyoglobin reducing ability (MRA) and oxygen consumption (OC). Grade × day of display (DOD) interactions were detected for L*, a*, surface oxymyoglobin (OMb) and metmyoglobin (MMb) percentages, and visual panel surface redness and discoloration scores (P ≤ 0.02); however, no Grade × DOD interactions were observed for MRA or OC (P > 0.17). There were location main effect (LOC) × DOD interactions for L*, a*, surface MMb, visual panel surface redness and discoloration, and MRA (P ≤ 0.02). Distal steaks had lower L* values compared with the other locations (P < 0.01), which coincided with steaks being rated visually darker red (P < 0.01). Proximal steaks had greater a* values and had less surface discoloration than middle steaks (P < 0.05), which had an increased percentage of surface MMb (P ≤ 0.04). Distal and proximal steaks had increased MRA compared with middle steaks on days 0 and 4 (P < 0.05), and distal steaks had greater OC than the other locations throughout display (P < 0.01). There were fewer type I fibers at the proximal end with a greater percentage located at the middle and distal ends, and an increased percentage of type IIX fibers at the middle and proximal locations (P ≤ 0.01). Less type IIA fibers were detected at the middle LOC compared with the other two locations (P < 0.10). Larger type I, IIA, and IIX fibers were located at the proximal and middle locations compared with the distal LOC (P < 0.01). ST color and color-stability characteristics were influenced by DOD and LOC, which may partially be explained by differences in fiber types among locations.



2005 ◽  
Vol 99 (4) ◽  
pp. 1494-1499 ◽  
Author(s):  
Hui Zhong ◽  
Roland R. Roy ◽  
Boonclaire Siengthai ◽  
V. Reggie Edgerton

The effects of short-term (4 days) and long-term (60 days) neuromuscular inactivity on myonuclear number, size, and myosin heavy chain (MHC) composition of isolated rat soleus fibers were determined using confocal microscopy and gel electrophoresis. Inactivity was produced via spinal cord isolation (SI), i.e., complete spinal cord transections at a midthoracic and a high sacral level and bilateral deafferentation between the transection sites. Compared with control, there was an increase in the percentage of fibers containing the faster MHC isoforms after 60, but not 4, days of SI. The mean sizes of type I and type I+IIa fibers were 41 and 27% and 66 and 56% smaller after 4 and 60 days of SI, respectively. Thus atrophy occurred earlier than the shift in myosin heavy chain (MHC) profile. The number of myonuclei was ∼30% higher in type I than type I+IIa fibers in control soleus, but after 60 days of SI these values were similar. The number of myonuclei per millimeter in type I fibers was significantly lower than control after 60 days of SI, whereas there was no change in type I+IIa fibers. Thus myonuclei were eliminated from fibers containing only type I MHC. Because the magnitude of the loss of myonuclei was less than the level of atrophy, the myonuclear domains of both type I and type I+IIa fibers were significantly lower than control. Thus chronic (60 days) inactivity results in smaller, faster fibers that contain a higher than normal amount of DNA per unit of cytoplasm. The absence of activation of muscle fibers that are normally the most active (pure type I fibers) resulted in most, but not all, fibers expressing some fast MHC isoforms. The results also indicate that a loss of myonuclei is not a prerequisite for sustained muscle fiber atrophy.



2007 ◽  
Vol 116 (9) ◽  
pp. 643-646 ◽  
Author(s):  
Melinda V. Davis ◽  
Albert L. Merati ◽  
Safwan S. Jaradeh ◽  
Joel H. Blumin

Objectives: Cricopharyngeal achalasia (CA) can be defined as inadequate opening of the cricopharyngeus muscle (CPM) resulting in dysphagia. Myosin heavy chain (MHC) isoform fiber type composition and size are key determinants of muscle function. These parameters have not been described in CA. It is hypothesized that there is a difference between the MHC isoform composition of the CPM in patients with the clinical diagnosis of CA and that in normal subjects. Methods: Patients who had received prior botulinum were excluded. The MHC fiber type composition and size in patients and cadaveric controls were determined by adenosine triphosphatase staining and image analysis. Results: The CPMs of 12 CA patients (6 male, 6 female; mean age, 61 years) and 5 control cadaveric subjects (3 male, 2 female; mean age, 67 years) were analyzed. There were relatively fewer type I fibers (67%) in patients with CA than in controls (81%), but the difference was not significant (p = 18). Type I fibers were slightly smaller in CA patients (38.7 μm) than in controls (47.2 μm), but this was not significantly different (p > 05). Of the 12 CA patients, 3 had type II predominance, a feature not seen in normal subjects. Conclusions: Patients with CA had relatively fewer type I fibers, although the difference was not statistically significant. The MHC isoform composition and fiber size were not different between CA patients and normal subjects. This is the first report to characterize the CPM in patients with CA.



1993 ◽  
Vol 265 (3) ◽  
pp. C842-C850 ◽  
Author(s):  
V. J. Caiozzo ◽  
S. Swoap ◽  
M. Tao ◽  
D. Menzel ◽  
K. M. Baldwin

The objectives of this study were to 1) examine the effect of hypo- and hyperthyroidism (triiodothyronine treatment) on the distribution of type IIA myosin heavy chain (MHC) in the soleus at the single fiber level and 2) correlate changes in the single fiber distribution of type IIA MHC with the maximal shortening velocity of whole skeletal muscle. The presence of the type IIA MHC in single fibers was determined using a monoclonal antibody reactive to the type IIA MHC and quantified with a Meridian ACAS 570 interactive laser cytometer. The findings of this study demonstrate that 1) hyperthyroidism significantly increases the relative number of muscle fibers that express type IIA MHC, 2) not all type I fibers are capable of expressing fast type IIA MHC under hyperthyroid conditions, and 3) there is a high correlation between maximal shortening velocity and the relative number of type IIA fibers. This latter observation suggests that the maximal shortening velocity of whole skeletal muscle may not be solely determined by its fastest fiber(s) but rather by the relative proportion of fibers expressing fast type IIA MHC.





Author(s):  
G. Scarlato ◽  
G. Pellegrini ◽  
C. Cerri ◽  
G. Meola ◽  
A. Veicsteinas

SUMMARY:Two cases of systemic carnitine deficiency are described. In both patients, carnitine concentration was lower than normal in serum and muscle tissue. In the first case, the illness began at age 35; the clinical manifestations were only muscular. In the second case, the illness began in childhood; there were intermittent episodes of hepatic enlargement and coma. An excessive lipid content was present in muscle tissue, especially in type I fibers, of both cases, and in the liver of the second patient. Ultrastructural studies of muscle tissue revealed important changes of mitochondria.During muscular exercise, aerobic and anaerobic metabolism were in vestigated. For a given relative work intensity, these patients showed abnormally high blood lactic acid concentration and lactic acid/pyruvic acid ratios. These data, together with the morphological alterations observed in mitochondria, suggest an impaired function of the respiratory chain, leading to a shift of the red/ox potential of the tissue towards a non reduced state.





Sign in / Sign up

Export Citation Format

Share Document