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Biology Open Jun 2024Regular spatial patterns are ubiquitous forms of organization in nature. In animals, regular patterns can be found from the cellular scale to the tissue scale, and from...
Regular spatial patterns are ubiquitous forms of organization in nature. In animals, regular patterns can be found from the cellular scale to the tissue scale, and from early stages of development to adulthood. To understand the formation of these patterns, how they assemble and mature, and how they are affected by perturbations, a precise quantitative description of the patterns is essential. However, accessible tools that offer in-depth analysis without the need for computational skills are lacking for biologists. Here, we present PatternJ, a novel toolset to analyze regular one-dimensional patterns precisely and automatically. This toolset, to be used with the popular imaging processing program ImageJ/Fiji, facilitates the extraction of key geometric features within and between pattern repeats in static images and time-lapse series. We validate PatternJ with simulated data and test it on images of sarcomeres from insect muscles and contracting cardiomyocytes, actin rings in neurons, and somites from zebrafish embryos obtained using confocal fluorescence microscopy, STORM, electron microscopy, and brightfield imaging. We show that the toolset delivers subpixel feature extraction reliably even with images of low signal-to-noise ratio. PatternJ's straightforward use and functionalities make it valuable for various scientific fields requiring quantitative one-dimensional pattern analysis, including the sarcomere biology of muscles or the patterning of mammalian axons, speeding up discoveries with the bonus of high reproducibility.
Topics: Animals; Axons; Image Processing, Computer-Assisted; Zebrafish; Sarcomeres; Somites; Software; Algorithms
PubMed: 38887972
DOI: 10.1242/bio.060548 -
Advances in Experimental Medicine and... 2024This chapter will describe basic structural and functional features of the contractile apparatus of muscle cells of the heart, namely, cardiomyocytes and smooth muscle... (Review)
Review
This chapter will describe basic structural and functional features of the contractile apparatus of muscle cells of the heart, namely, cardiomyocytes and smooth muscle cells. Cardiomyocytes form the contractile myocardium of the heart, while smooth muscle cells form the contractile coronary vessels. Both muscle types have distinct properties and will be considered with respect to their cellular appearance (brick-like cross-striated versus spindle-like smooth), arrangement of contractile proteins (sarcomeric versus non-sarcomeric organization), calcium activation mechanisms (thin-filament versus thick-filament regulation), contractile features (fast and phasic versus slow and tonic), energy metabolism (high oxygen versus low oxygen demand), molecular motors (type II myosin isoenzymes with high adenosine diphosphate [ADP]-release rate versus myosin isoenzymes with low ADP-release rates), chemomechanical energy conversion (high adenosine triphosphate [ATP] consumption and short duty ratio versus low ATP consumption and high duty ratio of myosin II cross-bridges [XBs]), and excitation-contraction coupling (calcium-induced calcium release versus pharmacomechanical coupling). Part of the work has been published (Neuroscience - From Molecules to Behavior", Chap. 22, Galizia and Lledo eds 2013, Springer-Verlag; with kind permission from Springer Science + Business Media).
Topics: Humans; Myocardial Contraction; Animals; Myocytes, Cardiac; Calcium; Energy Metabolism; Myocytes, Smooth Muscle; Excitation Contraction Coupling
PubMed: 38884723
DOI: 10.1007/978-3-031-44087-8_21 -
Scientific Reports Jun 2024Connectin (also known as titin) is a giant striated muscle protein that functions as a molecular spring by providing elasticity to the sarcomere. Novex-3 is a short...
Connectin (also known as titin) is a giant striated muscle protein that functions as a molecular spring by providing elasticity to the sarcomere. Novex-3 is a short splice variant of connectin whose physiological function remains unknown. We have recently demonstrated using in vitro analyses that in addition to sarcomere expression, novex-3 was also expressed in cardiomyocyte nuclei exclusively during fetal life, where it provides elasticity/compliance to cardiomyocyte nuclei and promotes cardiomyocyte proliferation in the fetus, suggesting a non-sarcomeric function. Here, we analyzed novex-3 knockout mice to assess the involvement of this function in cardiac pathophysiology in vivo. Deficiency of novex-3 compromised fetal cardiomyocyte proliferation and induced the enlargement of individual cardiomyocytes in neonates. In adults, novex-3 deficiency resulted in chamber dilation and systolic dysfunction, associated with Ca dysregulation, resulting in a reduced life span. Mechanistic analyses revealed a possible association between impaired proliferation and abnormal nuclear mechanics, including stiffer nuclei positioned peripherally with stabilized circumnuclear microtubules in knockout cardiomyocytes. Although the underlying causal relationships were not fully elucidated, these data show that novex-3 has a vital non-sarcomeric function in cardiac pathophysiology and serves as an early contributor to cardiomyocyte proliferation.
Topics: Animals; Myocytes, Cardiac; Cell Proliferation; Mice, Knockout; Mice; Cell Nucleus; Connectin; Sarcomeres; Muscle Proteins; Calcium
PubMed: 38877142
DOI: 10.1038/s41598-024-64608-1 -
Medicine and Science in Sports and... Jun 2024Increases in skeletal muscle size occur in response to prolonged exposure to resistance training that is typically ascribed to increased muscle fibre size. Whether...
INTRODUCTION
Increases in skeletal muscle size occur in response to prolonged exposure to resistance training that is typically ascribed to increased muscle fibre size. Whether muscle fibre number also changes remains controversial, and a paucity of data exists about myofibrillar structure. This cross-sectional study compared muscle fibre and myofibril characteristics in long-term resistance-trained (LRT) versus untrained (UNT) individuals.
METHODS
The maximal anatomical cross-sectional area (ACSAmax) of the biceps brachii muscle was measured by MRI in 16 LRT (5.9 ± 3.5 years' experience) and 13 UNT males. A muscle biopsy was taken from the biceps brachii to measure muscle fibre area, myofibril area and myosin spacing. Muscle fibre number, myofibril number in total and per fibre were estimated by dividing ACSAmax by muscle fibre area or myofibril area, and muscle fibre area by myofibril area, respectively.
RESULTS
Compared to UNT, LRT individuals had greater ACSAmax (+70%, P < 0.001), fibre area (+29%, P = 0.028), fibre number (+34%, P = 0.013), and myofibril number per fibre (+49%, P = 0.034) and in total (+105%, P < 0.001). LRT individuals also had smaller myosin spacing (-7%, P = 0.004; i.e. greater packing density) and a tendency towards smaller myofibril area (-16%, P = 0.074). ACSAmax was positively correlated with fibre area ( r = 0.526), fibre number ( r = 0.445) and myofibril number (in total r = 0.873 and per fibre r = 0.566), and negatively correlated with myofibril area ( r = -0.456) and myosin spacing ( r = -0.382) (all P < 0.05).
CONCLUSIONS
The larger muscles of LRT individuals exhibited more fibres in cross-section and larger muscle fibres, which contained substantially more total myofibrils and more packed myofilaments than UNT participants, suggesting plasticity of muscle ultrastructure.
PubMed: 38875487
DOI: 10.1249/MSS.0000000000003495 -
Circulation. Genomic and Precision... Jun 2024Hypertrophic cardiomyopathy (HCM) is caused by sarcomere gene mutations (genotype-positive HCM) in ≈50% of patients and occurs in the absence of mutations...
Integrating Clinical Phenotype With Multiomics Analyses of Human Cardiac Tissue Unveils Divergent Metabolic Remodeling in Genotype-Positive and Genotype-Negative Patients With Hypertrophic Cardiomyopathy.
BACKGROUND
Hypertrophic cardiomyopathy (HCM) is caused by sarcomere gene mutations (genotype-positive HCM) in ≈50% of patients and occurs in the absence of mutations (genotype-negative HCM) in the other half of patients. We explored how alterations in the metabolomic and lipidomic landscape are involved in cardiac remodeling in both patient groups.
METHODS
We performed proteomics, metabolomics, and lipidomics on myectomy samples (genotype-positive N=19; genotype-negative N=22; and genotype unknown N=6) from clinically well-phenotyped patients with HCM and on cardiac tissue samples from sex- and age-matched and body mass index-matched nonfailing donors (N=20). These data sets were integrated to comprehensively map changes in lipid-handling and energy metabolism pathways. By linking metabolomic and lipidomic data to variability in clinical data, we explored patient group-specific associations between cardiac and metabolic remodeling.
RESULTS
HCM myectomy samples exhibited (1) increased glucose and glycogen metabolism, (2) downregulation of fatty acid oxidation, and (3) reduced ceramide formation and lipid storage. In genotype-negative patients, septal hypertrophy and diastolic dysfunction correlated with lowering of acylcarnitines, redox metabolites, amino acids, pentose phosphate pathway intermediates, purines, and pyrimidines. In contrast, redox metabolites, amino acids, pentose phosphate pathway intermediates, purines, and pyrimidines were positively associated with septal hypertrophy and diastolic impairment in genotype-positive patients.
CONCLUSIONS
We provide novel insights into both general and genotype-specific metabolic changes in HCM. Distinct metabolic alterations underlie cardiac disease progression in genotype-negative and genotype-positive patients with HCM.
Topics: Humans; Cardiomyopathy, Hypertrophic; Male; Female; Middle Aged; Genotype; Adult; Phenotype; Myocardium; Metabolomics; Proteomics; Lipidomics; Lipid Metabolism; Sarcomeres; Energy Metabolism; Aged; Multiomics
PubMed: 38853772
DOI: 10.1161/CIRCGEN.123.004369 -
Preparative Biochemistry & Biotechnology Jun 2024Recombinant myofibril-bound serine proteinase (rMBSP) was successfully expressed in GS115 in our laboratory. However, low production of rMBSP in shake flask...
Recombinant myofibril-bound serine proteinase (rMBSP) was successfully expressed in GS115 in our laboratory. However, low production of rMBSP in shake flask constraints further exploration of properties. A 5-L high cell density fermentation was performed and the fermentation medium was optimized. Response surface methodology (RSM) was used to optimize the culture condition through modeling three selected parameter. Under the optimized culture medium (LBSM, 1% yeast powder and 1% peptone) and culture conditions (induction pH 5.5, temperature 29 °C, time 40 h), the yield of rMBSP was 420 mg/L in a 5-L fermenter, which was a 6-fold increase over thar, expressed in flask cultivation. The desired enzyme was purified by two-step, which yielded a 33.7% recovery of a product that had over 85% purity. The activity of purified rMBSP was significantly inhibited by Ca, Mg, SDS, guanidine hydrochloeide, acetone, isopropanol, chloroform, -hexane and -heptane. Enzymatic analysis revealed a of 2.89 ± 0.09 μM and a of 14.20 ± 0.12 nM•min for rMBSP. LC-MS/MS analysis demonstrated the specific cleavage of bovine serum albumin by rMPSP. These findings suggest that rMPSP has potential as a valuable enzyme for protein science research.
PubMed: 38847777
DOI: 10.1080/10826068.2024.2361159 -
Meat Science Sep 2024This study investigated the effect of ultrasound (US) combined with pre- and post-addition of κ-carrageenan (KC) on the gelling properties, structural characteristics...
Potential mechanisms and effects of ultrasound treatment combined with pre- and post-addition of κ-carrageenan on the gelling properties and rheological behavior of myofibrillar proteins under low-salt condition.
This study investigated the effect of ultrasound (US) combined with pre- and post-addition of κ-carrageenan (KC) on the gelling properties, structural characteristics and rheological behavior of myofibrillar proteins (MP) under low-salt conditions. The results showed that US combined with either pre- or post-addition of KC rendered higher gel strength and water holding capacity (WHC) of MP gels than those treated with US alone and added with KC alone (P < 0.05). US combined with pre-addition of KC facilitated the binding between MP and KC, which enhanced the gel strength and WHC of the mixed MP gels and significantly improved the rheological behavior of MP. This was also confirmed by the highest surface hydrophobicity, disulfide bonds and β-sheet content of the MP gels with US combined with pre-addition of KC. Moreover, microstructural results reflected a denser structure for the pre-addition of KC in combination with US. However, US combined with post-addition of KC resulted in limited MP unfolding and relatively weak hydrophobic interactions in the composite gels, which were less effective in improving the gel properties of the MP gels. This study provides potential strategies for enhancing the gelling properties of low-salt meat products via application of US and KC.
Topics: Carrageenan; Rheology; Animals; Gels; Meat Products; Hydrophobic and Hydrophilic Interactions; Food Handling; Muscle Proteins; Swine; Myofibrils
PubMed: 38838569
DOI: 10.1016/j.meatsci.2024.109554 -
ELife Jun 2024Styxl2, a poorly characterized pseudophosphatase, was identified as a transcriptional target of the Jak1-Stat1 pathway during myoblast differentiation in culture. Styxl2...
Styxl2, a poorly characterized pseudophosphatase, was identified as a transcriptional target of the Jak1-Stat1 pathway during myoblast differentiation in culture. Styxl2 is specifically expressed in vertebrate striated muscles. By gene knockdown in zebrafish or genetic knockout in mice, we found that Styxl2 plays an essential role in maintaining sarcomere integrity in developing muscles. To further reveal the functions of Styxl2 in adult muscles, we generated two inducible knockout mouse models: one with being deleted in mature myofibers to assess its role in sarcomere maintenance, and the other in adult muscle satellite cells (MuSCs) to assess its role in de novo sarcomere assembly. We find that Styxl2 is not required for sarcomere maintenance but functions in de novo sarcomere assembly during injury-induced muscle regeneration. Mechanistically, Styxl2 interacts with non-muscle myosin IIs, enhances their ubiquitination, and targets them for autophagy-dependent degradation. Without Styxl2, the degradation of non-muscle myosin IIs is delayed, which leads to defective sarcomere assembly and force generation. Thus, Styxl2 promotes de novo sarcomere assembly by interacting with non-muscle myosin IIs and facilitating their autophagic degradation.
Topics: Animals; Mice; Mice, Knockout; Proteolysis; Sarcomeres; Zebrafish; Phosphoprotein Phosphatases
PubMed: 38829202
DOI: 10.7554/eLife.87434 -
Food Research International (Ottawa,... Jul 2024Myofibrillar proteins are crucial for gel formation in processed meat products such as sausages and meat patties. Freeze-thaw cycles can alter protein properties,...
Myofibrillar proteins are crucial for gel formation in processed meat products such as sausages and meat patties. Freeze-thaw cycles can alter protein properties, impacting gel stability and product quality. This study aims to investigate the potential of thawed drip and its membrane-separated components as potential antifreeze agents to retard denaturation, oxidation and gel deterioration of myofibrillar proteins during freezing-thawing cycles of pork patties. The thawed drip and its membrane-separated components of > 10 kDa and < 10 kDa, along with deionized water, were added to minced pork at 10 % mass fraction and subjected to increasing freeze-thaw cycles. Results showed that the addition of thawed drip and its membrane separation components inhibited denaturation and structural changes of myofibrillar proteins, evidenced by reduced surface hydrophobicity and carbonyl content, increased free sulfhydryl groups, protein solubility and α-helix, as compared to the deionized water group. Correspondingly, improved gel properties including water-holding capacity, textural parameters and denser network structure were observed with the addition of thawed drip and its membrane separation components. Denaturation and oxidation of myofibrillar proteins were positively correlated with gel deterioration during freezing-thawing cycles. We here propose a role of thawed drip and its membrane separation components as cryoprotectants against myofibrillar protein gel deterioration during freeze-thawing cycles.
Topics: Freezing; Animals; Gels; Swine; Muscle Proteins; Myofibrils; Food Handling; Protein Denaturation; Meat Products; Hydrophobic and Hydrophilic Interactions; Solubility; Water; Oxidation-Reduction
PubMed: 38823861
DOI: 10.1016/j.foodres.2024.114461 -
Food Research International (Ottawa,... Jul 2024Different emulsion gel systems are widely applied to deliver functional ingredients. The effects and mechanisms of ultrasound-assisted emulsification (UAE) treatment and...
Effects and mechanisms of ultrasound-assisted emulsification treatment on the curcumin delivery and digestive properties of myofibrillar protein-carboxymethyl cellulose complex emulsion gel.
Different emulsion gel systems are widely applied to deliver functional ingredients. The effects and mechanisms of ultrasound-assisted emulsification (UAE) treatment and carboxymethyl cellulose (CMC) modifying the curcumin delivery properties and in vitro digestibility of the myofibrillar protein (MP)-soybean oil emulsion gels were investigated. The rheological properties, droplet size, protein and CMC distribution, ultrastructure, surface hydrophobicity, sulfhydryl groups, and zeta potential of emulsion gels were also measured. Results indicate that UAE treatment and CMC addition both improved curcumin encapsulation and protection efficiency in MP emulsion gel, especially for the UAE combined with CMC (UAE-CMC) treatment which encapsulation efficiency, protection efficiency, the release rate, and bioaccessibility of curcumin increased from 86.75 % to 97.67 %, 44.85 % to 68.85 %, 18.44 % to 41.78 %, and 28.68 % to 44.93 % respectively. The protein digestibility during the gastric stage was decreased after the CMC addition and UAE treatment, and the protein digestibility during the intestinal stage was reduced after the CMC addition. The fatty acid release rate was increased after CMC addition and UAE treatment. Apparent viscosity, storage modulus, and loss modulus were decreased after CMC addition while increased after UAE and UAE-CMC treatment especially the storage modulus increased from 0.26 Pa to 41 Pa after UAE-CMC treatment. The oil size was decreased, the protein and CMC concentration around the oil was increased, and a denser and uniform emulsion gel network structure was formed after UAE treatment. The surface hydrophobicity, free SH groups, and absolute zeta potential were increased after UAE treatment. The UAE-CMC treatment could strengthen the MP emulsion gel structure and decrease the oil size to increase the curcumin delivery properties, and hydrophobic and electrostatic interaction might be essential forces to maintain the emulsion gel.
Topics: Curcumin; Emulsions; Carboxymethylcellulose Sodium; Gels; Digestion; Rheology; Hydrophobic and Hydrophilic Interactions; Muscle Proteins; Soybean Oil; Viscosity; Particle Size; Myofibrils; Ultrasonic Waves
PubMed: 38823850
DOI: 10.1016/j.foodres.2024.114531