Monkey Skeletal Muscle Research Articles

Tropomyosin Isoform Diversity in the Cynomolgus Monkey Heart and Skeletal Muscles Compared to Human Tissues.

Old world monkeys separated from the great apes, including the ancestor of humans, about 25 million years ago, but most of the genes in humans and various nonhuman primates are quite similar even though their anatomical appearances are quite different. Like other mammals, primates have four tropomyosin genes (TPM1, TPM2, TPM3, and TPM4) each of which generates a multitude of TPM isoforms via alternative splicing. Only TPM1 produces two sarcomeric isoforms (TPM1α and TPM1κ), and TPM2, TPM3, and TPM4 each generate one sarcomeric isoform. We have cloned and sequenced TPM1α, TPM1κ, TPM2α, TPM3α, and TPM4α with RNA from cynomolgus (Cyn) monkey hearts and skeletal muscle. We believe this is the first report of directly cloning and sequencing of these monkey transcripts. In the Cyn monkey heart, the rank order of TPM isoform expression is TPM1α > TPM2α > TPM1κ > TPM3α > TPM4α. In the Cyn monkey skeletal muscle, the rank order of expression is TPM1α > TPM2α > TPM3α > TPM1κ > TPM4α. The major differences in the human heart are the increased expression of TPM1κ, although TPM1α is still the dominant transcript. In the Cyn monkey heart, the only sarcomeric TPM isoform at the protein level is TPM1α. This is in contrast to human hearts where TPM1α is the major sarcomeric isoform but a lower quantity of TPM1κ, TPM2α, and TPM3α is also detected at the protein level. These differences of tropomyosin and/or other cardiac protein expression in human and Cyn monkey hearts may reflect the differences in physiological activities in daily life.

Single-nucleus profiling unveils a geroprotective role of the FOXO3 in primate skeletal muscle aging.

Age-dependent loss of skeletal muscle mass and function is a feature of sarcopenia, and increases the risk of many aging-related metabolic diseases. Here, we report phenotypic and single-nucleus transcriptomic analyses of non-human primate skeletal muscle aging. A higher transcriptional fluctuation was observed in myonuclei relative to other interstitial cell types, indicating a higher susceptibility of skeletal muscle fiber to aging. We found a downregulation of FOXO3 in aged primate skeletal muscle, and identified FOXO3 as a hub transcription factor maintaining skeletal muscle homeostasis. Through the establishment of a complementary experimental pipeline based on a human pluripotent stem cell-derived myotube model, we revealed that silence of FOXO3 accelerates human myotube senescence, whereas genetic activation of endogenous FOXO3 alleviates human myotube aging. Altogether, based on a combination of monkey skeletal muscle and human myotube aging research models, we unraveled the pivotal role of the FOXO3 in safeguarding primate skeletal muscle from aging, providing a comprehensive resource for the development of clinical diagnosis and targeted therapeutic interventions against human skeletal muscle aging and the onset of sarcopenia along with aging-related disorders.

Circular RNAs in monkey muscle: age-dependent changes.

Circular RNAs (circRNAs) have been identified in numerous species, including human, mouse, nematode, and coelacanth. They are believed to function as regulators of gene expression at least in part by sponging microRNAs. Here, we describe the identification of circRNAs in monkey (Rhesus macaque) skeletal muscle. RNA sequencing analysis was employed to identify and annotate ~12,000 circRNAs, including numerous circular intronic RNAs (ciRNAs), from skeletal muscle of monkeys of a range of ages. Reverse transcription followed by real-time quantitative (q)PCR analysis verified the presence of these circRNAs, including the existence of several highly abundant circRNAs, and the differential abundance of a subset of circRNAs as a function of age. Taken together, our study has documented systematically circRNAs expressed in skeletal muscle and has identified circRNAs differentially abundant with advancing muscle age. We propose that some of these circRNAs might influence muscle function.

Caloric restriction delays aging-induced cellular phenotypes in rhesus monkey skeletal muscle

Sarcopenia is the age-related loss of skeletal muscle mass and function and is characterized by a reduction in muscle mass and fiber cross-sectional area, alterations in muscle fiber type and mitochondrial functional changes. In rhesus monkeys, calorie restriction (CR) without malnutrition improves survival and delays the onset of age-associated diseases and disorders including sarcopenia. We present a longitudinal study on the impact of CR on early stage sarcopenia in the upper leg of monkeys from ~ 16 years to ~ 22 years of age. Using dual-energy X-ray absorptiometry we show that CR delayed the development of maximum muscle mass and, unlike Control animals, muscle mass of the upper leg was preserved in CR animals during early phase sarcopenia. Histochemical analyses of vastus lateralis muscle biopsies revealed that CR opposed age-related changes in the proportion of Type II muscle fibers and fiber cross-sectional area. In contrast the number of muscle fibers with mitochondrial electron transport system enzyme abnormalities (ETS ab) was not significantly affected by CR. Laser capture microdissection of ETS ab fibers and subsequent PCR analysis of the mitochondrial DNA revealed large deletion mutations in fibers with abnormal mitochondrial enzyme activities. CR did not prevent stochastic mitochondrial deletion mutations in muscle fibers but CR may have contributed to the maintenance of affected fibers.

Molecular analyses of mtDNA deletion mutations in microdissected skeletal muscle fibers from aged rhesus monkeys

Mitochondrial DNA (mtDNA) deletion mutations co-localize with electron transport system (ETS) abnormalities in rhesus monkey skeletal muscle fibers. Using laser capture microdissection in conjunction with PCR and DNA sequence analysis, mitochondrial genomes from single sections of ETS abnormal fibers were characterized. All ETS abnormal fibers contained mtDNA deletion mutations. Deletions were large, removing 20-78% of the genome, with some to nearly all of the functional genes lost. In one-third of the deleted genomes, the light strand origin was deleted, whereas the heavy strand origin of replication was conserved in all fibers. A majority (27/39) of the deletion mutations had direct repeat sequences at their breakpoints and most (36/39) had one breakpoint within or in close proximity to the cytochrome b gene. Several pieces of evidence support the clonality of the mtDNA deletion mutation within an ETS abnormal region of a fiber: (a) only single, smaller than wild-type, PCR products were obtained from each ETS abnormal region; (b) the amplification of mtDNA from two regions of the same ETS abnormal fiber identified identical deletion mutations, and (c) a polymorphism was observed at nucleotide position 16103 (A and G) in the wild-type mtDNA of one animal (sequence analysis of an ETS abnormal region revealed that mtDNA deletion mutations contained only A or G at this position). Species-specific differences in the regions of the genomes lost as well as the presence of direct repeat sequences at the breakpoints suggest mechanistic differences in deletion mutation formation between rodents and primates.

Selenoprotein W accumulates primarily in primate skeletal muscle, heart, brain and tongue.

The human selenoprotein W coding region with the selenocysteine codon (TGA) changed to a cysteine codon (TGT) was fused to six histidine codons (at its 3' end), cloned into a prokaryotic expression vector (pTrc99a), and the corresponding mutated selenoprotein W was expressed in bacteria. The protein was purified by Ni-NTA agarose column and reverse phase HPLC. Polyclonal antibodies raised against this protein were used in Western blots to determine tissue distribution of selenoprotein W from rhesus monkeys fed a commercial chow. Selenoprotein W was found in several tissues with highest amounts in skeletal muscle and heart (muscle 6 fold greater than liver) and lowest levels in liver, but selenium concentrations were highest in kidneys (10 fold greater than muscle) and lowest in skeletal muscle. Northern blots using a human selenoprotein W cDNA probe indicated that mRNA levels were highest in monkey skeletal muscle and heart (2-2.5 fold greater than in liver), which is similar to the pattern found with a human multiple tissue Northern blot. However, as in the monkey, selenium concentrations were highest in human kidney and lowest in skeletal muscle and heart. Thus, selenoprotein W protein levels correlated with selenoprotein W mRNA levels but not with tissue selenium concentrations.

Effect of long-term caloric restriction on GLUT4, phosphatidylinositol-3 kinase p85 subunit, and insulin receptor substrate-1 protein levels in rhesus monkey skeletal muscle.

Caloric restriction (CR), a reduction in calorie intake without malnutrition, improves insulin sensitivity in various species, including mice, rats, rhesus and cynomolgus monkeys, and humans. Skeletal muscle is quantitatively the most important tissue for blood glucose clearance. Therefore, we assessed the effect of 6 years of CR (30% reduction in calorie intake) in male rhesus monkeys (14-20 years old) on muscle expression of several proteins involved in insulin action. Whole body insulin sensitivity (assessed by Modified Minimal Model) was significantly increased in CR relative to Control monkeys. CR did not alter the expression of GLUT4 glucose transporter or phosphatidylinositol-3 kinase p85 subunit (PI3K). Insulin receptor substrate-1(IRS-1) abundance tended to be greater for CR compared to Control monkeys (p = .051), but correlational analysis revealed no association between IRS-1 and insulin sensitivity (r2 = .075, p = .271). These findings indicate that the CR-induced increase in insulin sensitivity in rhesus monkeys is unrelated to alterations in GLUT4, P13K, and IRS-1 abundance.

Purification, Characterization, and Glutathione Binding to Selenoprotein W From Monkey Muscle

Selenoprotein W was purified from monkey skeletal muscle to investigate its binding of glutathione. The purification was accomplished by concentration of the cytosol with an Amicon cell, gel filtration using Sephadex G-50, cation-exchange chromatography with CM-Sephadex, and reverse-phase high-pressure liquid chromatography using a C-18 Vydac column. Selenoprotein W was monitored during purification by slot blots. These steps resulted in an electrophoretically pure selenoprotein W preparation that was estimated by gel filtration to have molecular weight of about 10 kDa. N-terminal amino acid sequencing was used to confirm that the pure proteins were selenoprotein W. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI) revealed that the proteins existed in three masses of 9635 ± 7, 9371 ± 11, and 9330 ± 5 Da. The theoretical mass of the protein predicted from the cDNA sequence is 9330 Da. The 9635-Da form of the protein was shown to contain bound glutathione (306 Da), which could be released by reduction with dithiothreitol at 50°C. The form with a mass of 9371 Da is assumed to result from binding of an unidentified 41-Da moiety to the 9330-Da form of the protein. MALDI peptide mapping with endoproteinase Glu-C suggested that glutathione is bound to the 36th amino acid (cysteine) of selenoprotein W.

Velocity, force, power, and Ca2+ sensitivity of fast and slow monkey skeletal muscle fibers.

In this study, we determined the contractile properties of single chemically skinned fibers prepared from the medial gastrocnemius (MG) and soleus (Sol) muscles of adult male rhesus monkeys and assessed the effects of the spaceflight living facility known as the experiment support primate facility (ESOP). Muscle biopsies were obtained 4 wk before and immediately after an 18-day ESOP sit, and fiber type was determined by immunohistochemical techniques. The MG slow type I fiber was significantly smaller than the MG type II, Sol type I, and Sol type II fibers. The ESOP sit caused a significant reduction in the diameter of type I and type I/II (hybrid) fibers of Sol and MG type II and hybrid fibers but no shift in fiber type distribution. Single-fiber peak force (mN and kN/m2) was similar between fiber types and was not significantly different from values previously reported for other species. The ESOP sit significantly reduced the force (mN) of Sol type I and MG type II fibers. This decline was entirely explained by the atrophy of these fiber types because the force per cross-sectional area (kN/m2) was not altered. Peak power of Sol and MG fast type II fiber was 5 and 8.5 times that of slow type I fiber, respectively. The ESOP sit reduced peak power by 25 and 18% in Sol type I and MG type II fibers, respectively, and, for the former fiber type, shifted the force-pCa relationship to the right, increasing the Ca2+ activation threshold and the free Ca2+ concentration, eliciting half-maximal activation. The ESOP sit had no effect on the maximal shortening velocity (Vo) of any fiber type. Vo of the hybrid fibers was only slightly higher than that of slow type I fibers. This result supports the hypothesis that in hybrid fibers the slow myosin heavy chain would be expected to have a disproportionately greater influence on Vo.

Effects of surface electrostimulation on the structure and metabolic properties in monkey skeletal muscle

Adaptative changes in skeletal muscle following surface electrical stimulation (SES) were investigated in rhesus monkeys. SES was performed on the triceps brachialis muscle (TB) according to an intermittent pattern. The procedure was carried out for 3 wk, using a current with a medium frequency of 60 Hz normally observed in fast motor axons. The histochemical assays performed on biopsies taken from proximal and distal parts of the TB muscle, before and after the SES program, showed that the distribution of fibers typed by ATPase was unaffected. On the other hand, SES led to an overall increase in the mean fiber cross-sectional area (FCSA); P < 0.01 (+13.7%, NS, in proximal portion, vs +31%, P < 0.01 in distal portion). This increase in size occurred in all fiber types. SES was shown to induce an overall increase in capillary to fiber ratio (C/F; +11.06%, NS, in proximal portion, vs +25.93%, P < 0.05 in distal portion). The number of capillaries surrounding fiber Type II (CAFII) was significantly increased by SES (P < 0.05): +3.21%, NS, in proximal portion, versus +21.47%, P < 0.05 in distal portion. Moreover, the number of capillaries surrounding fiber Type I (CAFI) was statistically unaffected by SES. These results suggest that a stimulation of capillary growth may occur following SES-training. Citrate synthase activity was significantly increased after SES. This enhancement in oxidative potential was shown to occur in all fiber types (NADH-diaphorase staining).(ABSTRACT TRUNCATED AT 250 WORDS)

Protein sequence determination by ESI-MS and LSI-MS tandem mass spectrometry: parvalbumin primary structures from cat, gerbil and monkey skeletal muscle

Protein sequence determination by ESI-MS and LSI-MS tandem mass spectrometry: parvalbumin primary structures from cat, gerbil and monkey skeletal muscle

Occurrence of cathepsin D isozymes with different specificities in monkey skeletal muscle

Cathepsin D was highly purified from the skeletal muscle of Japanese monkey (Macaca fuscata fuscata) by a procedure including affinity chromatography on concanavalin A-Sepharose and pepstatin-Sepharose, and then resolved into ten isozymes (A through J) by isoelectric focusing. When examined for specificity toward oxidized insulin B chain, isozyme A was highly specific and cleaved exclusively the Leu15-Tyr16 bond, whereas isozyme F was less specific, cleaving the Leu15-Tyr16 and Glu13-Ala14 bonds, with slower cleavages at several other bonds. These results demonstrate for the first time the occurrence of isozymes with different specificities among cathepsin D isozymes obtained from a single source.

Purification and characterization of the parvalbumin from monkey skeletal muscle

1. 1. A high affinity Ca 2+ binding and low mol. wt protein, parvalbumin, was purified from monkey skeletal muscle. 2. 2. As compared with other animals, only one component and a lower content of monkey parvalbumin were found. 3. 3. This may suggest that both the component and the content of parvalbumin decreases with biological evolution. 4. 4. The parvalbumin was found to have a mol. wt of 11,400, a pI of 5.1, a high aspartic acid and lysine content, maximum absorption at around 260nm, a blocked amino-terminal, an immunological distinction, 2 mol Ca 2+ binding/mol, and a conformational change by Ca 2+ binding. 5. 5. Parvalbumin was shown to have alpha type properties.

Responsiveness of Monkey Skeletal Muscle Arteries to Vasoconstrictor Substances before and after Cold Storage

The stainless steel cannula inserting method was used to observe effects of α-adrenoceptor agonists, 5-HT and KCl before and after cold storage (3–5 days, at 4°C) in skeletal muscle branches of the simian deep femoral artery. Epinephrine (EPI), norepinephrine (NE), phenylephrine (PE), methoxamine (MT) and 5-hydroxytryptamine (5-HT) induced marked monophasic vasoconstrictions in a dose-dependent manner. 5-HT induced a greater vasoconstriction in larger diameter vessels (old animals) than that in smaller ones (young animals), suggesting age-related responses. A selective α2-stimulant, Clonidine (CLO) or xylazine (XYL), produced only a slight vasoconstriction. Tyramine (TYR) also produced only a slight vasoconstrictor response. The order of potencies for inducing vasoconstrictions was EPI≥5-HT≥NE>MT=PE≫KCI>CLO=XYL=TYR. The vasoconstrictor responses to all used adrenergic agonists and 5-HT were not significantly influenced by the prolonged cold storage. However, KCI-induced constrictions were significantly suppressed by the cold storage. These results suggest that the postjunctional α-adrenoceptor in simian skeletal muscle arteries is mainly of the α1 -type. Since cold storage caused a significant suppression of the KCI-induced response but not those of adrenoceptor agonists and 5-HT, it was considered that the mechanism of calcium entry to the vascular smooth muscle of skeletal muscle arteries might be significantly damaged by the cold storage.

Responsiveness of monkey skeletal muscle arteries to vasoconstrictor substances before and after cold storage.

The stainless steel cannula inserting method was used to observe effects of alpha-adrenoceptor agonists, 5-HT and KCl before and after cold storage (3-5 days, at 4 degrees C) in skeletal muscle branches of the simian deep femoral artery. Epinephrine (EPI), norepinephrine (NE), phenylephrine (PE), methoxamine (MT) and 5-hydroxytryptamine (5-HT) induced marked monophasic vasoconstrictions in a dose-dependent manner. 5-HT induced a greater vasoconstriction in larger diameter vessels (old animals) than that in smaller ones (young animals), suggesting age-related responses. A selective alpha 2-stimulant, clonidine (CLO) or xylazine (XYL), produced only a slight vasoconstriction. Tyramine (TYR) also produced only a slight vasoconstrictor response. The order of potencies for inducing vasoconstrictions was EPI greater than or equal to 5-HT greater than or equal to NE greater than MT = PE much greater than KCl greater than CLO = XYL = TYR. The vasoconstrictor responses to all used adrenergic agonists and 5-HT were not significantly influenced by the prolonged cold storage. However, KCI-induced constrictions were significantly suppressed by the cold storage. These results suggest that the postjunctional alpha-adrenoceptor in simian skeletal muscle arteries is mainly of the alpha 1-type. Since cold storage caused a significant suppression of the KCl-induced response but not those of adrenoceptor agonists and 5-HT, it was considered that the mechanism of calcium entry to the vascular smooth muscle of skeletal muscle arteries might be significantly damaged by the cold storage.

Localization and inhibition of calcium-activated neutral protease (CANP) in primate skeletal muscle and peripheral nerve

Localization of calcium-activated neutral protease in normal monkey ( Cebus apella) skeletal muscle and peripheral nerve was studied by application of the indirect immunofluorescent and peroxidase-antiperoxidase techniques for light and electron microscopy. In muscle, the protease was demonstrated in association with endomysial collagen fibrils, basal lamina, sarcolemma, and Z-bands and also at neuromuscular junctions. In nerve, the enzyme was demonstrated in association with endoneurial collagen fibrils, basal lamina, axolemma, and neurofilaments of both myelinated and unmyelinated axons. Intramuscular injections of the thiol protease inhibitor, leupeptin, abolished Ca-activated neutral protease immunoreactivity in both muscle and peripheral nerve. These data suggest that the protease is localized both intracellularly and extracellularly in normal primate muscle and peripheral nerve. Inhibition of basal lamina associated neutral protease by leupeptin after denervation may hold significance for protecting this structure against degradation and preserving it as a neurotrophic lattice for axon regeneration.

Functional properties of palmaris longus muscles of rhesus monkeys transplanted as index finger flexors.

This experiment with skeletal muscle autografts in monkeys was designed to retest previous findings that transplanted skeletal muscle can regenerate to a functional degree in primates without predenervation and to test a new hypothesis that increased functional demands on regenerated muscle grafts in monkeys may result in improved functional capacity of the grafts. Rhesus monkey index flexors were replaced with free palmaris longus muscle autografts with microneural anastomoses between the graft motor nerve and the severed profundus motor nerve. One monkey was taught selective index flexion before grafting and continued with this program after grafting to test the effect of training on the graft. Mature grafts were evaluated for in vivo contractile properties and by histology and histochemistry and were compared with a group of normal Rhesus palmaris longus muscles. The results reconfirm the capacity of nonpredenervated monkey skeletal muscle grafts to regenerate and to achieve some contractile ability and suggest that training of free muscle grafts may enhance recovery of their functional and structural properties.

Purification and characterization of cathepsin B from monkey skeletal muscle.

Cathepsin B was purified about 11,000-fold from monkey skeletal muscle by ammonium sulfate fractionation and sequential column chromatographies monitored by assaying of Z-Phe-Arg-MCA hydrolase activity. The purified enzyme gave a single protein band on SDS-polyacrylamide gel electrophoresis, and its molecular weight was estimated to be 24,000 by gel filtration. It had a pH optimum of 6.5, required a thiol reducing agent for activation, and was inhibited by various thiol protease inhibitors. These properties were similar to those reported for cathepsins B from other sources. Although the enzyme scarcely hydrolyzed ordinary proteins, such as casein, hemoglobin, and bovine serum albumin, it degraded myosin and actin among various myofibrillar proteins. These results strongly suggested that skeletal muscle cathepsin B may participate in the degradation of muscle proteins in vivo. In addition, cathepsin B was shown to hydrolyze various neuropeptides such as Leu-enkephalin, beta-neoendorphin, alpha-neoendorphin, dynorphin(1-13), and substance P. It appeared to act on these peptides mainly as a dipeptidyl carboxypeptidase, although not so rigorously, presumably due to its endopeptidase activity.

Anion activation of monkey muscle creatine kinase

1. 1. Creatine kinase from rhesus monkey skeletal muscle is activated by acetate and other short chain fatty acids. 2. 2. Activation is associated with lower K m and higher V max values at less than saturating substrate concentrations but does not occur when both substrates are saturating. 3. 3. No co-operativity between subunits is evident in the activation process. 4. 4. It appears that acetate promotes the mutual enhancement by substrates in their binding by inducing the optimum enzyme conformation normally associated with substrate saturation. 5. 5. Conservation of this activation effect through the evolution of the phosphagen kinases implies that it may well be of physiological significance.

Effects of temperature on responses of fresh and refrigerated perfused blood vessels.

ARTICLESEffects of temperature on responses of fresh and refrigerated perfused blood vesselsJC Murphy, O Carrier Jr, and J SahadiJC Murphy, O Carrier Jr, and J SahadiPublished Online:01 Nov 1973https://doi.org/10.1152/ajplegacy.1973.225.5.1187MoreSectionsPDF (1 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformation Cited ByStereolithography 3D Printing of a Heat Exchanger for Advanced Temperature Control in Wire Myography25 January 2022 | Polymers, Vol. 14, No. 3Prolonged Cold Storage Diminishes the 5-Hydroxytryptamine- and Potassium Chloride-Mediated Contractions of Rat Thoracic AortaThe Tohoku Journal of Experimental Medicine, Vol. 217, No. 2Effects of prolonged cold storage on double peaked vasoconstrictor responses to periarterial nerve stimulation in isolated canine splenic arteries29 January 2009 | British Journal of Pharmacology, Vol. 126, No. 8Vascular Tissue Preservation TechniquesResponsiveness of Monkey Skeletal Muscle Arteries to Vasoconstrictor Substances before and after Cold StorageJapanese Journal of Pharmacology, Vol. 46, No. 3Vascular Responsiveness of Simian Digital Artery to Various Vasoactive SubstancesJournal of Investigative Dermatology, Vol. 86, No. 6Physical Factors of Regulation1 January 2011The effect of cold storage on the sensitivity to alpha and beta agonists in the isolated rabbit kidneyGeneral Pharmacology: The Vascular System, Vol. 7, No. 2-3 More from this issue > Volume 225Issue 5November 1973Pages 1187-1191 Copyright & PermissionsCopyright © 1973 by American Physiological Societyhttps://doi.org/10.1152/ajplegacy.1973.225.5.1187PubMed4745220History Published online 1 November 1973 Published in print 1 November 1973 Metrics