Journal of Muscle Research and Cell Motility最新文献

{"title":"Optimizing 2D in vitro differentiation conditions for C2C12 murine myoblasts on gelatin hydrogel.","authors":"Veronica Sian, Per Harald Jonson, Anna Vainio, Helena Luque, Swethaa Natraj Gayathri, Peter Hackman, Bjarne Udd, Marco Savarese, Jaakko Sarparanta","doi":"10.1007/s10974-025-09711-0","DOIUrl":"https://doi.org/10.1007/s10974-025-09711-0","url":null,"abstract":"<p><p>Optimizing in vitro differentiation protocols for skeletal muscle cells is essential for producing mature, functional myotubes suitable for disease modeling and therapeutic screening. While C2C12 murine myoblasts are a widely used model, achieving consistent and advanced differentiation remains challenging. In this study, we systematically evaluated conditions that improve myotube formation and maturation in a 2D culture system using ultra-compliant gelatin hydrogels. We compared standard and commercial differentiation media and identified that supplementation of DMEM with 2% horse serum and 10% Opti-MEM (DMO) supported robust myotube formation, with thin and aligned fibers. Insulin supplementation significantly increased expression of myosin heavy chain (MyHC) and calsequestrin, while pyruvate provided additional benefit by further enhancing myotube maturation. Media change frequency was also critical: daily replacement was necessary to maintain optimal differentiation, although the addition of insulin and pyruvate partly mitigated the effects of less frequent changes. Application of electrical pulse stimulation (EPS) improved sarcomeric α-actinin organization without significantly altering MyHC isoform expression. RNA sequencing confirmed transcriptional reprogramming consistent with myogenic progression, including early upregulation of key muscle-specific genes. Our findings present a cost-effective, reproducible protocol that supports advanced C2C12 differentiation in a scalable 2D system, offering practical guidance for generating mature, functional myotubes in vitro for both basic and translational muscle research.</p>","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145251582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Festschrift for Professor Jennifer Morgan.","authors":"Michelle Peckham","doi":"10.1007/s10974-025-09709-8","DOIUrl":"https://doi.org/10.1007/s10974-025-09709-8","url":null,"abstract":"","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145225503","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Regulation of Autonomic Motility in Human Gastric Muscle and the Gastroepiploic Artery by Stretch and pH.","authors":"Dae Hoon Kim, Seung Myoung Son, Woong Choi, Chan Hyung Kim, Hun Sik Kim, Ki Bae Kim, Wen-Xie Xu, Sang Jin Lee, Young Chul Kim, Hyo-Yung Yun","doi":"10.1007/s10974-025-09699-7","DOIUrl":"https://doi.org/10.1007/s10974-025-09699-7","url":null,"abstract":"<p><p>Human gastric motility is regulated by both slow wave activity and membrane excitability. Regulation of gastric function involves adapting motility through repetitive stretches during feeding and digestion. Alongside gastric motility, gastric vascular motility must also be accurately regulated. The physiological function of stretch-activated K⁺ channels has been demonstrated in the relaxation mechanisms of the uterus and bladder. For these reasons, this study was designed to investigate whether stretch-activated K⁺ channels are involved in the functional regulation of human gastric muscle and vessels. We examined human gastric body tissues and gastroepiploic arteries from patients who underwent gastrectomy using a conventional contractile measurement system and Western immunoblot. High concentrations of K⁺ (50 mM) induced tonic contraction (4 g) in human gastric circular muscle from the body. Acetylcholine (ACh, 10 µM) also induced an initial peak (3 g), tonic (1.1 g), and phasic contractions (1.5 g; 2.5 cycles/min). L-methionine, known to block TWIK (two-pore domain weak inward rectifying K_2P channel)-related K⁺ channels (TREK-1), produced sustained contraction (2 g) in gastric smooth muscle in the presence of a cocktail of K⁺ channel blockers. Additionally, channel inhibitors such as extracellular acidosis (MES ([pH]_o = 6.4)), quinidine, bupivacaine, and lidocaine enhanced spontaneous contractions by 224%, 183%, 138%, and 127% of control, respectively, in the presence of L-methionine. Concurrently, we analyzed the physiological role of TREK-1 and TASK-2 in the human gastroepiploic artery. The ring of the human gastroepiploic artery produced tonic contraction (2.8 g) under high K⁺ (50 mM). Following stimulation with high K⁺, the artery exhibited spontaneous vasoconstriction known as vasomotion (2.7 g; 0.13 cycles/min), which was completely inhibited by nifedipine, a voltage-dependent L-type Ca²⁺ channel (VDCC_L) blocker. BayK 8644, an activator of VDCC_L, induced vasomotion, which was also inhibited by nifedipine. In the human artery, L-methionine induced a vascular tonic contraction (0.15 g) and enhanced vasomotion by 179%. Additionally, lidocaine induced peak and tonic contractions of 1 g and 0.7 g, respectively. Both L-methionine and lidocaine also enhanced vasomotion induced by BayK 8644. The molecular presence of TREK-1 and TASK-2 was confirmed via Western blot in human gastric muscle, gastric mucosa, and artery, respectively. These findings suggest that TREK-1 and TASK-2 may be significant regulators of human gastric muscle and vascular motility.</p>","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145149337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"LncRNA Xist acts as a miR-486-5p sponge to modulate myoblast proliferation by recruiting Stk4.","authors":"Wenlun Wang, Huang Yan, Li Wenbin, Yu Chaohang, Wu Shengcai, Cai Xioqing, Zhou Nanqing, Yuan Qing, Hu Qianmin, Zhang Feng, Zhu Lingyun","doi":"10.1007/s10974-025-09708-9","DOIUrl":"https://doi.org/10.1007/s10974-025-09708-9","url":null,"abstract":"<p><p>Cell proliferation plays a crucial role in muscle atrophy-regeneration. However, the functional roles of Long non-coding RNA X-inactive specific transcript (lncRNA Xist) in C2C12 cell proliferation remain poorly characterized. We identified lncRNA Xist as a regulator of cell proliferation kinetics by acting as a competing endogenous RNA (ceRNA) in C2C12 cells. Downregulation or overexpression of lncRNA Xist correlated with enhanced or impaired cell proliferation, respectively. Mechanistically, lncRNA Xist sponges miR-486-5p to regulate serine/threonine kinase 4 (Stk4) expression, thereby modulating C2C12 cell proliferation in vitro. The lncRNA Xist/miR-486-5p/Stk4 ceRNA network plays an essential role in C2C12 proliferation, suggesting lncRNA Xist as a potential therapeutic target for muscle atrophy.</p>","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":""},"PeriodicalIF":1.7,"publicationDate":"2025-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145091863","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Theoretical treatment of tension transients in muscle following sudden changes in orthophosphate concentration: implications for energy transduction.","authors":"Alf Månsson","doi":"10.1007/s10974-025-09698-8","DOIUrl":"10.1007/s10974-025-09698-8","url":null,"abstract":"<p><p>The relative timing of the force-generating power stroke and release of the ATP-hydrolysis product orthophosphate (Pi) in actomyosin energy transduction is debated. It may be explored by studying the tension response to sudden changes in [Pi] during isometric muscle contraction (Pi-transients; rate constant k_Pi) and by the rate of redevelopment of isometric force (k_tr) after a period of unloaded shortening at varied [Pi]. Most studies of these types are interpreted using simple kinetic schemes that ignore the range of elastic strains of actin-attached myosin cross-bridges. We found that the only simple scheme which accounts for the experimental findings of single exponential Pi-transients with k_Pi ≈ k_tr has force-generation coincident with actin-myosin attachment. This characteristics could compromise the high power output of muscle. We therefore turned to a mechanokinetic model, allowing consideration of the varying elastic cross-bridge strains. Our model assumes Pi-release between cross-bridge attachment and the force-generating power stroke. However, power strokes only occur if cross-bridges attach in a pre-power-stroke state with zero or negative elastic strain (counteracting shortening). The model suggests two components of the Pi-transients. One is attributed to slow cross-bridge detachment from the pre-power-stroke state at positive elastic strain upon Pi-binding. The other is due to Pi-induced shifts in equilibrium with rapid power stroke reversal. The slow component dominates for all parameter values tested but the fast component is ubiquitous, predicting a biphasic Pi-transient in disagreement with experiments. Strikingly, however, the mechanokinetic model gives different predictions than apparently similar simple kinetic schemes and we do not rule out the existence of parameter values leading to a negligible fast component. We also show that the assumption of secondary Pi-binding sites on myosin outside the active site removes the fast component albeit without predicting that k_tr ≈ k_Pi. Additional studies are required to finally corroborate that k_tr ≈ k_Pi in experiments but also to further develop mechanokinetic models combined with multistep Pi-release.</p>","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":"193-213"},"PeriodicalIF":1.7,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12454480/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144626590","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"miR-2400 promotes proliferation of bovine skeletal muscle-derived satellite cells by regulating MAGED1 genes expression.","authors":"Li Yang, Hai-Jing Luo, Zhi-An Gong, Wen-Tian Zhang, Jing-Xuan Cui, Xue-Peng Fu, Wei-Wei Zhang","doi":"10.1007/s10974-025-09695-x","DOIUrl":"10.1007/s10974-025-09695-x","url":null,"abstract":"<p><p>microRNAs play a crucial role in the intricate process of muscle satellite cells proliferation and differentiation. Previous studies have demonstrated that miR-2400 can regulate bovine skeletal muscle satellite cell (MuSCs) proliferation, yet the underlying mechanism remains incompletely elucidated. In this study, we employed bioinformatics prediction and dual luciferase reporter assays to establish that miR-2400 directly targets the 3' untranslated regions (UTRs) of melanoma antigen family D1 (MAGED1) mRNA, thereby suppressing its expression. To ascertain whether miR-2400 affects the proliferation of MuSCs through MAGED1, we constructed the MAGED1 interference vector using RNA interference technology (RNAi) and assessed changes in MuSCs proliferation subsequent to MAGED1 interference. The experimental data indicate that the cell viability and the rate of EdU-positive cells of MuSCs were increased after interference with MAGED1. The proportion of S-phase cells and the expression level of cell cycle-associated proteins CCND2 and CCNB1 increased. These findings align with miR-2400's role in promoting cell proliferation and suggest that miR-2400 exerts its effects by directly targeting MAGED1.</p>","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":"183-192"},"PeriodicalIF":1.7,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144025714","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The impact of mitokine MOTS-c administration on the soleus muscle of rats subjected to a 7-day hindlimb suspension.","authors":"Daria A Sidorenko, Irina D Lvova, Sergey A Tyganov, Boris S Shenkman, Kristina A Sharlo","doi":"10.1007/s10974-025-09700-3","DOIUrl":"10.1007/s10974-025-09700-3","url":null,"abstract":"<p><p>The aim of the study was to investigate the effect of MOTS-c on the key functional alterations in the rat soleus muscle during 7-day unloading - the transformation of slow fibers into fast ones, atrophy and increased fatigue. We daily intraperitoneally injected male Wistar rats with a short mitochondrial peptide MOTS-c during 7-day unloading of their hind limbs. After the end of the experiment, we conducted an ex vivo fatigue test of soleus muscle and showed that the MOTS-c administration prevents increased fatigue during 7-day hind limb unloading. Also, using immunohistochemical analysis, we showed that MOTS-c prevents the transformation of slow fibers into fast ones, mitigates the slow muscle atrophy fibers (but not fast ones) of the soleus muscle. In the group receiving MOTS-c, the decrease in Akt and GSK3β phosphorylation was prevented, and the 18 S and 28 S rRNA levels were at the control level. The ubiquitin ligases MuRF and Atrogin-1 mRNA were also reduced compared to the hindlimb unloading group with placebo. In addition, MOTS-c prevented a decrease in the expression of a few mitochondrial biogenesis parameters and the level of ACC phosphorylation (AMPK target). Thus, the MOTS-C injections during hind limb unloading lead to the normalization of several protein synthesis and degradation processes and support the expression of genes that ensure muscle resistance to fatigue.</p>","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":"215-229"},"PeriodicalIF":1.7,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144553780","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Skeletal muscle alterations in Marfan syndrome: a systematic review.","authors":"Audrei R Santos, Rita M S Gutierrez, Tatiana E Koike, Talita C Conte, Caroline C Real, Nicolas A Dumont, Elen H Miyabara","doi":"10.1007/s10974-025-09706-x","DOIUrl":"10.1007/s10974-025-09706-x","url":null,"abstract":"<p><p>Marfan syndrome is an autosomal dominant multisystemic connective tissue disorder caused by mutations in the FBN1 gene. Although clinical changes in the cardiovascular, ocular, and skeletal systems have been described in detail in Marfan syndrome patients, investigations about skeletal muscle alterations are still incipient. This systematic review describes cellular, molecular, and functional changes in skeletal muscles of patients and mice with Marfan syndrome. Study selection (from EMBASE, MEDLINE, and Web of Science databases), data extraction, and quality appraisal were performed by two independent reviewers. A total of 2634 articles were identified; 26 were included in the analysis based on the selection criteria. The risk of bias was evaluated using the Critical Appraisal Skills Programme and Joanna Briggs Institute Critical Appraisal tool for human studies and the Systematic Review Centre for Laboratory Animal Experimentation RoB tool for animal studies. The findings indicate that skeletal muscle alterations in Marfan syndrome are characterized by fibrosis, reduced muscle mass and myofiber size, compromised muscle regeneration, and impaired muscle function. Future studies are warranted to investigate the mechanisms involved in the development of this muscle phenotype to help develop effective strategies to improve skeletal muscle function and the quality of life of individuals with Marfan syndrome.</p>","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":"253-272"},"PeriodicalIF":1.7,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144957651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The influences of ARHGEF9 on myoblasts migration and differentiation.","authors":"Shuang Li, Jin Xu, Wenjia Zhang, Yongze Liu, Huili Tong, Bingchen Liu","doi":"10.1007/s10974-025-09692-0","DOIUrl":"10.1007/s10974-025-09692-0","url":null,"abstract":"<p><p>Rho guanine nucleotide exchange factor 9 (ARHGEF9), as a protein that assists small GTPases, is widely present in various tissues. It has been reported to play an important role mainly in neurological diseases and gliomas. However, there have been no reports on its impact on skeletal muscle regeneration after injury. This study first demonstrated a significant increase in ARHGEF9 protein expression during the regeneration of skeletal muscle post-injury in mice. Secondly, during the differentiation of mouse C2C12 myoblasts, ARHGEF9 significantly increased and co-localized with actin filaments. Inhibition of ARHGEF9 significantly downregulated the migration rate and actin filaments polymerization of mouse C2C12 myoblasts, and significantly reduced the expression of proteins related to cell migration. Finally, inhibition of ARHGEF9 significantly reduced the differentiation ability of mouse C2C12 myoblasts. In summary, ARHGEF9 impacting on myoblasts migration and differentiation suggests that targeting ARHGEF9 could be beneficial for promoting skeletal muscle regeneration and repair.</p>","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":"153-165"},"PeriodicalIF":1.7,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143483175","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Residual force enhancement following hindlimb unloading and exercise prehabilitation.","authors":"Hyo-Seong Yeo, Jeong-Hun Bae, Jae-Young Lim","doi":"10.1007/s10974-025-09703-0","DOIUrl":"10.1007/s10974-025-09703-0","url":null,"abstract":"<p><p>This study was performed to investigate the effect of exercise prehabilitation (EPH) on passive force, residual force enhancement (RFE), and muscle protein expression in rat skeletal muscle. A total of 24 Sprague-Dawley rats (12 weeks old) were randomly divided into three groups: control (CON, n = 8), hindlimb unloading (HLU, n = 8), and exercise prehabilitation (EPH, n = 8). The HLU group underwent hindlimb unloading for 14 days, and the EPH group exercised for 14 days before HLU. Passive force and RFE were measured in the isolated soleus (SOL) muscle. Muscle protein expression and fiber CSA were analyzed by western blot and immunofluorescence. Body weight (BW) was higher in the CON group than in the HLU or EPH groups. Muscle weight (MW)/BW of SOL was lower in HLU than CON. Muscle fiber and physiological CSA were lower in both HLU and EPH groups than in CON. Peak passive force and passive force normalized to physiological CSA of the SOL were lower in treatment groups than CON. RFE (%) was lower in HLU compared to CON, while EPH had higher RFE than HLU in SOL. Myostatin expression was upregulated by HLU but downregulated by EPH. These results suggest that exercise prehabilitation before surgery can enhance muscle function and suppress myostatin expression. EPH may help preserve muscle function by influencing RFE and regulating muscle protein expression, highlighting the potential of exercise prehabilitation in pre-surgical care and rehabilitation.</p>","PeriodicalId":16422,"journal":{"name":"Journal of Muscle Research and Cell Motility","volume":" ","pages":"239-251"},"PeriodicalIF":1.7,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144753620","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}