Formation of sarcomeres in developing myotubes: role of mechanical stretch and contractile activation

2000 ◽  
Vol 279 (6) ◽  
pp. C1801-C1811 ◽  
Author(s):  
Patrick G. De Deyne
Keyword(s):  
Electrical Stimulation ◽  
Structural Alignment ◽  
Major Change ◽  
Mechanical Stretch ◽  
Experimental Conditions ◽  
Pharmacological Agents ◽  
Cultured Myotubes ◽  
Contractile Activation ◽  
Sarcomere Assembly ◽  
Passive Stretch

In a series of experiments, cultured myotubes were exposed to passive stretch or pharmacological agents that block contractile activation. Under these experimental conditions, the formation of Z lines and A bands (morphological structures, resulting from the specific structural alignment of sarcomeric proteins, necessary for contraction) was assessed by immunofluorescence. The addition of an antagonist of the voltage-gated Na+ channels [tetrodotoxin (TTX)] for 2 days in developing rat myotube cultures led to a nearly total absence of Z lines and A bands. When contractile activation was allowed to resume for 2 days, the Z lines and A bands reappeared in a significant way. The appearance of Z lines or A bands could not be inhibited nor facilitated by the application of a uniaxial passive stretch. Electrical stimulation of the cultures increased sarcomere assembly significantly. Antagonists of L-type Ca2+ channels (verapamil, nifedipine) combined with electrical stimulation led to the absence of Z lines and A bands to the same degree as the TTX treatment. Western blot analysis did not show a major change in the amount of sarcomeric α-actinin nor a shift in myosin heavy chain phenotype as a result of a 2-day passive stretch or TTX treatment. Results of experiments suggest that temporal Ca2+ transients play an important factor in the assembly and maintenance of sarcomeric structures during muscle fiber development.

Reflex Inhibition of Normal Cramp Following Electrical Stimulation of the Muscle Tendon

2007 ◽  
Vol 98 (3) ◽  
pp. 1102-1107 ◽  
Author(s):  
Serajul I. Khan ◽  
John A. Burne
Keyword(s):  
Electrical Stimulation ◽  
Maximum Voluntary Contraction ◽  
Voluntary Contraction ◽  
Muscle Cramp ◽  
Emg Activity ◽  
Reflex Inhibition ◽  
Experimental Conditions ◽  
Inhibitory Feedback ◽  
Two Phases ◽  
Stimulation Of

Muscle cramp was induced in one head of the gastrocnemius muscle (GA) in eight of thirteen subjects using maximum voluntary contraction when the muscle was in the shortened position. Cramp in GA was painful, involuntary, and localized. Induction of cramp was indicated by the presence of electromyographic (EMG) activity in one head of GA while the other head remained silent. In all cramping subjects, reflex inhibition of cramp electrical activity was observed following Achilles tendon electrical stimulation and they all reported subjective relief of cramp. Thus muscle cramp can be inhibited by stimulation of tendon afferents in the cramped muscle. When the inhibition of cramp-generated EMG and voluntary EMG was compared at similar mean EMG levels, the area and timing of the two phases of inhibition (I1, I2) did not differ significantly. This strongly suggests that the same reflex pathway was the source of the inhibition in both cases. Thus the cramp-generated EMG is also likely to be driven by spinal synaptic input to the motorneurons. We have found that the muscle conditions that appear necessary to facilitate cramp, a near to maximal contraction of the shortened muscle, are also the conditions that render the inhibition generated by tendon afferents ineffective. When the strength of tendon inhibition in cramping subjects was compared with that in subjects that failed to cramp, it was found to be significantly weaker under the same experimental conditions. It is likely that reduced inhibitory feedback from tendon afferents has an important role in generating cramp.

scholarly journals Regulation of Cyr61 gene expression by mechanical stretch through multiple signaling pathways

2001 ◽  
Vol 281 (5) ◽  
pp. C1524-C1532 ◽  
Author(s):  
Isao Tamura ◽  
Joel Rosenbloom ◽  
Edward Macarak ◽  
Brahim Chaqour
Keyword(s):  
Gene Expression ◽  
Signaling Pathways ◽  
Rho Kinase ◽  
Mechanical Stretch ◽  
Selective Inhibition ◽  
Early Gene ◽  
Actin Dynamics ◽  
Mrna Levels ◽  
Bladder Smooth Muscle ◽  
Pharmacological Agents

The cysteine-rich protein 61 (Cyr61) is a signaling molecule with functions in cell migration, adhesion, and proliferation. This protein is encoded by an immediate early gene whose expression is mainly induced by serum growth factors. Here we show that Cyr61 mRNA levels increase sharply in response to cyclic mechanical stretch applied to cultured bladder smooth muscle cells. Stretch-induced changes of Cyr61 transcripts were transient and accompanied by an increase of the encoded protein that localized mainly to the cytoplasm and nucleus of the cells. With the use of pharmacological agents that interfere with known signaling pathways, we show that transduction mechanisms involving protein kinase C and phosphatidylinositol 3-kinase activation partly blocked stretch-induced Cyr61 gene expression. Selective inhibition of Rho kinase pathways altered this stretch effect as well. Meanwhile, using inhibitors of the actin cytoskeleton, we show that Cyr61 gene expression is sensitive to mechanisms that sense actin dynamics. These results establish the regulation of Cyr61 gene by mechanical stretch and provide clues to the key signaling molecules involved in this process.

Mechanical loading regulates NOS expression and activity in developing and adult skeletal muscle

1998 ◽  
Vol 275 (1) ◽  
pp. C260-C266 ◽  
Author(s):  
James G. Tidball ◽  
Eliane Lavergne ◽  
Kim S. Lau ◽  
Melissa J. Spencer ◽  
James T. Stull ◽  
...  
Keyword(s):  
Cyclic Loading ◽  
Muscle Activation ◽  
Weight Bearing ◽  
No Production ◽  
Adult Skeletal Muscle ◽  
No Release ◽  
Cultured Myotubes ◽  
Passive Stretch ◽  
Expression And Activity

The hypothesis that changes in muscle activation and loading regulate the expression and activity of neuronal nitric oxide (NO) synthase (nNOS) was tested using in vitro and in vivo approaches. Removal of weight bearing from rat hindlimb muscles for 10 days resulted in a significant decrease in nNOS protein and mRNA concentration in soleus muscles, which returned to control concentrations after return to weight bearing. Similarly, the concentration of nNOS in cultured myotubes increased by application of cyclic loading for 2 days. NO release from excised soleus muscles was increased significantly by a single passive stretch of 20% or by submaximal activation at 2 Hz, although the increases were not additive when both stimuli were applied simultaneously. Increased NO release resulting from passive stretch or activation was dependent on the presence of extracellular calcium. Cyclic loading of cultured myotubes also resulted in a significant increase in NO release. Together, these findings show that activity of muscle influences NO production in the short term, by regulating NOS activity, and in the long term, by regulating nNOS expression.

scholarly journals Activation of the heat shock response in a primary cellular model of motoneuron neurodegeneration-evidence for neuroprotective and neurotoxic effects

2009 ◽  
Vol 14 (2) ◽  
Author(s):  
Bernadett Kalmar ◽  
Linda Greensmith
Keyword(s):  
Cell Death ◽  
Heat Shock ◽  
Heat Shock Response ◽  
Apoptotic Cell ◽  
Experimental Conditions ◽  
Pharmacological Agents ◽  
Shock Response ◽  
Induced Apoptosis ◽  
Shock Proteins ◽  
The Relationship

AbstractPharmacological up-regulation of heat shock proteins (hsps) rescues motoneurons from cell death in a mouse model of amyotrophic lateral sclerosis. However, the relationship between increased hsp expression and neuronal survival is not straightforward. Here we examined the effects of two pharmacological agents that induce the heat shock response via activation of HSF-1, on stressed primary motoneurons in culture. Although both arimoclomol and celastrol induced the expression of Hsp70, their effects on primary motoneurons in culture were significantly different. Whereas arimoclomol had survival-promoting effects, rescuing motoneurons from staurosporin and H2O2 induced apoptosis, celastrol not only failed to protect stressed motoneurons from apoptosis under same experimental conditions, but was neurotoxic and induced neuronal death. Immunostaining of celastrol-treated cultures for hsp70 and activated caspase-3 revealed that celastrol treatment activates both the heat shock response and the apoptotic cell death cascade. These results indicate that not all agents that activate the heat shock response will necessarily be neuroprotective.

Avoidance and approach learning motivated by stimulation of identical hypothalamic loci

1959 ◽  
Vol 197 (1) ◽  
pp. 153-157 ◽  
Author(s):  
George W. Brown ◽  
Bertram D. Cohen
Keyword(s):  
Electrical Stimulation ◽  
Lateral Hypothalamus ◽  
Opportunity To Learn ◽  
Hypothalamic Stimulation ◽  
Experimental Conditions ◽  
Approach Response ◽  
Lateral Hypothalamic ◽  
Stimulating Electrodes ◽  
Second Procedure ◽  
Stimulation Of

Cats with stimulating electrodes implanted in the lateral hypothalamus were subjected to two types of experimental procedures. In the first procedure the cats were given an opportunity to learn to avoid hypothalamic stimulation which produces a typical ‘hypothalamic rage’ response. The second procedure allows the same cats to learn to approach an area where the hypothalamic stimulus is administered. In both procedures, electrical stimulation was delivered through identical electrodes, yet each animal learned the appropriate avoidance or approach response, depending upon the experimental conditions. Therefore, lateral hypothalamic stimulation may act as an energizing, drive-arousing, operation to produce both avoidance and approach learning in cats.

Spike-Wave Complexes and Fast Components of Cortically Generated Seizures. IV. Paroxysmal Fast Runs in Cortical and Thalamic Neurons

1998 ◽  
Vol 80 (3) ◽  
pp. 1495-1513 ◽  
Author(s):  
Igor Timofeev ◽  
François Grenier ◽  
Mircea Steriade
Keyword(s):  
Electrical Stimulation ◽  
High Frequency ◽  
Cortical Neurons ◽  
Field Potential ◽  
Thalamic Nuclei ◽  
Experimental Conditions ◽  
Thalamic Neurons ◽  
Single Action ◽  
Spike Bursts ◽  
Spike Wave

Timofeev, Igor, François Grenier, and Mircea Steriade. Spike-wave complexes and fast components of cortically generated seizures. IV. Paroxysmal fast runs in cortical and thalamic neurons. J. Neurophysiol. 80: 1495–1513, 1998. In the preceding papers of this series, we have analyzed the cellular patterns and synchronization of neocortical seizures occurring spontaneously or induced by electrical stimulation or cortical infusion of bicuculline under a variety of experimental conditions, including natural states of vigilance in behaving animals and acute preparations under different anesthetics. The seizures consisted of two distinct components: spike-wave (SW) or polyspike-wave (PSW) at 2–3 Hz and fast runs at 10–15 Hz. Because the thalamus is an input source and target of cortical neurons, we investigated here the seizure behavior of thalamic reticular (RE) and thalamocortical (TC) neurons, two major cellular classes that have often been implicated in the generation of paroxysmal episodes. We performed single and dual simultaneous intracellular recordings, in conjunction with multisite field potential and extracellular unit recordings, from neocortical areas and RE and/or dorsal thalamic nuclei under ketamine-xylazine and barbiturate anesthesia. Both components of seizures were analyzed, but emphasis was placed on the fast runs because of their recent investigation at the cellular level. 1) The fast runs occurred at slightly different frequencies and, therefore, were asynchronous in various cortical neuronal pools. Consequently, dorsal thalamic nuclei, although receiving convergent inputs from different neocortical areas involved in seizure, did not express strongly synchronized fast runs. 2) Both RE and TC cells were hyperpolarized during seizure episodes with SW/PSW complexes and relatively depolarized during the fast runs. As known, hyperpolarization of thalamic neurons deinactivates a low-threshold conductance that generates high-frequency spike bursts. Accordingly, RE neurons discharged prolonged high-frequency spike bursts in close time relation with the spiky component of cortical SW/PSW complexes, whereas they fired single action potentials, spike doublets, or triplets during the fast runs. In TC cells, the cortical fast runs were reflected as excitatory postsynaptic potentials appearing after short latencies that were compatible with monosynaptic activation through corticothalamic pathways. 3) The above data suggested the cortical origin of these seizures. To further test this hypothesis, we performed experiments on completely isolated cortical slabs from suprasylvian areas 5 or 7 and demonstrated that electrical stimulation within the slab induces seizures with fast runs and SW/PSW complexes, virtually identical to those elicited in intact-brain animals. The conclusion of all papers in this series is that complex seizure patterns, resembling those described at the electroencephalogram level in different forms of clinical seizures with SW/PSW complexes and, particularly, in the Lennox-Gastaut syndrome of humans, are generated in neocortex. Thalamic neurons reflect cortical events as a function of membrane potential in RE/TC cells and degree of synchronization in cortical neuronal networks.

scholarly journals The effect of electrical stimulation on the incorporation of l-[U-14C]valine into the protein of chopped tissue from guinea-pig cerebral cortex

1970 ◽  
Vol 118 (5) ◽  
pp. 791-800 ◽  
Author(s):  
C. T. Jones ◽  
P. Banks
Keyword(s):  
Cerebral Cortex ◽  
Electrical Stimulation ◽  
Guinea Pig ◽  
High Energy ◽  
Initial Increase ◽  
Nuclear Fraction ◽  
High Energy Phosphate ◽  
Experimental Conditions ◽  
Relative Contribution ◽  
Energy Dependent

1. Chopped tissue from guinea-pig cerebral cortex carried out an energy-dependent incorporation of [14C]valine into protein. 2. At all times studied the nuclear fraction of the homogenized tissue accounted for about 25% of the total labelled protein. 3. Electrical stimulation at first increased, but subsequently decreased, the rate of incorporation of [14C]valine into protein of the chopped tissue. 4. The initial increase in the incorporation of [14C]valine into protein occurred in the nuclear fraction. At later times electrical stimulation decreased the incorporation into all the subcellular fractions, but the relative contribution of the nuclear fraction to the total labelled protein increased. 5. These changes are discussed in relation to the changes in the rates of respiration, glycolysis, high-energy phosphate content and intracellular Na+ and K+ concentrations, which were measured under the same experimental conditions as those used to study protein synthesis.

Calcium Dependence of Contractile Activation of Isolated Sheep Urethra I: Responses to Electrical Stimulation

1991 ◽  
Vol 69 (4) ◽  
pp. 263-269 ◽  
Author(s):  
Angeles Garcia-Pascual ◽  
Gonzalo Costa ◽  
Albino Garcia-Sacristan ◽  
Karl-Erik Andersson
Keyword(s):  
Electrical Stimulation ◽  
Calcium Dependence ◽  
Contractile Activation

scholarly journals Acute Neuromuscular Electrical Stimulation (NMES) With Blood Flow Restriction: The Effect of Restriction Pressures

2020 ◽  
pp. 1-9
Author(s):  
Paul Head ◽  
Mark Waldron ◽  
Nicola Theis ◽  
Stephen David Patterson
Keyword(s):  
Blood Flow ◽  
Electrical Stimulation ◽  
Isometric Contraction ◽  
Perceived Exertion ◽  
Neuromuscular Electrical Stimulation ◽  
Muscle Thickness ◽  
Ratings Of Perceived Exertion ◽  
Experimental Conditions ◽  
Maximal Voluntary Isometric Contraction ◽  
Flow Restriction

Context: Neuromuscular electrical stimulation (NMES) combined with blood flow restriction (BFR) has been shown to improve muscular strength and size better than NMES alone. However, previous studies used varied methodologies not recommended by previous NMES or BFR research. Objective: The present study investigated the acute effects of NMES combined with varying degrees of BFR using research-recommended procedures to enhance understanding and the clinical applicability of this combination. Design: Randomized crossover. Setting: Physiology laboratory. Participants: A total of 20 healthy adults (age 27 [4] y; height 177 [8] cm; body mass 77 [13] kg). Interventions: Six sessions separated by at least 7 days. The first 2 visits served as familiarization, with the experimental conditions performed in the final 4 sessions: NMES alone, NMES 40% BFR, NMES 60% BFR, and NMES 80% BFR. Main Outcome Measures: Maximal voluntary isometric contraction, muscle thickness, blood pressure, heart rate, rating of perceived exertion, and pain were all recorded before and after each condition. Results: The NMES 80% BFR caused greater maximal voluntary isometric contraction decline than any other condition (−38.9 [22.3] N·m, P < .01). Vastus medialis and vastus lateralis muscle thickness acutely increased after all experimental conditions (P < .05). Pain and ratings of perceived exertion were higher after NMES 80% BFR compared with all other experimental conditions (P < .05). No cardiovascular effects were observed between conditions. Conclusion: The NMES combined with 80% BFR caused greater acute force decrement than the other conditions. However, greater perceptual ratings of pain and ratings of perceived exertion were observed with NMES 80% BFR. These acute observations must be investigated during chronic interventions to corroborate any relationship to changes in muscle strength and size in clinical populations.

scholarly journals Molecular and Biomechanical Adaptations to Mechanical Stretch in Cultured Myotubes

2021 ◽  
Vol 12 ◽  
Author(s):  
Dapeng Ren ◽  
Jing Song ◽  
Ran Liu ◽  
Xuemin Zeng ◽  
Xiao Yan ◽  
...  
Keyword(s):  
Mechanical Stretch ◽  
Biomechanical Properties ◽  
Structural Element ◽  
The Past ◽  
Future Challenges ◽  
Cultured Myotubes ◽  
Myotube Hypertrophy ◽  
And Oxidative Stress ◽  
Made In

Myotubes are mature muscle cells that form the basic structural element of skeletal muscle. When stretching skeletal muscles, myotubes are subjected to passive tension as well. This lead to alterations in myotube cytophysiology, which could be related with muscular biomechanics. During the past decades, much progresses have been made in exploring biomechanical properties of myotubes in vitro. In this review, we integrated the studies focusing on cultured myotubes being mechanically stretched, and classified these studies into several categories: amino acid and glucose uptake, protein turnover, myotube hypertrophy and atrophy, maturation, alignment, secretion of cytokines, cytoskeleton adaption, myotube damage, ion channel activation, and oxidative stress in myotubes. These biomechanical adaptions do not occur independently, but interconnect with each other as part of the systematic mechanoresponse of myotubes. The purpose of this review is to broaden our comprehensions of stretch-induced muscular alterations in cellular and molecular scales, and to point out future challenges and directions in investigating myotube biomechanical manifestations.

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