Role Of Stretch-activated Channels Research Articles

The Role of Stretch-Activated Channels in Mediating mTORC1 Signaling in Isolated Rat Soleus Muscle in Response to Mechanical Stimulation after Functional Unloading

The aim of the present work was to identify the role of stretch-activated ion channels (SAC) in the transmission of mechanical signals to the key mTORC1/p70S6k anabolic pathways in the soleus muscle in rats after seven days of functional unloading of the hindlimbs. Functional unloading of the hindlimb muscles was by antiorthostatic suspension (hindlimb suspension, HS). The rats were divided into four groups: 1) group C (animal-house control; 2) C + Gd (controls whose muscles were incubated with the SAC inhibitor GdCl3; 3) HS (functional unloading for seven days); and 4) HS + Gd (a group with seven days of antiorthostatic suspension followed by incubation of muscles with GdCl3). After completion of antiorthostatic suspension, isolated soleus muscles were subjected to a series of eccentric contractions (EC). Muscles from intact animals responded to EC with increases in the phosphorylation of key markers of mTORC1 signaling: p70S6k and rpS6. The extent of mechanically induced increases in the levels of phosphorylation of these protein markers were significantly reduced to essentially identical extents in the “pure” suspension (HS) group and the HS group given the SAC blocker (HS + Gd) as compared with the increase in muscles from intact control group. These results lead to the conclusion that decreases in the mechanically dependent responses of the mTORC1 signal pathway in the soleus muscle in rats after seven days of functional unloading may be linked with inactivation of stretch-activated ion channels.

PO-064 A potential role of stretch-activated channels in anabolic mechanotransduction in the atrophied rat soleus muscle

Objective Prolonged immobilization or unloading of skeletal muscle causes muscle disuse atrophy, which is characterized by a reduction in muscle cross-sectional area and compromised contractile function. To date, the mechanisms of anabolic mechanotransduction in the atrophied mammalian skeletal muscle remain poorly understood. The aim of the present study was to assess a possible role of stretch-activated ion channels (SAC) in the propagation of a mechanical signal to anabolic signaling and protein synthesis (PS) in an isolated rat soleus muscle following mechanical unloading.
 Methods The mechanical unloading was performed via hindlimb suspension (HS). Twenty-eight male Wistar rats weighing were randomly assigned to the following 4 groups (n=7/group): 1) vivarium control (C), 2) control + SAC inhibitor (gadolinium) (C+Gd3+), 3) 7-day HS (HS), 4) 7-day HS + SAC inhibitor (HS+Gd3+). Following unloading, an isolated rat soleus was placed in an organ culture medium and subjected to a bout of eccentric contractions (EC). Upon completion of the EC, muscles were collected for Western blot analyses to determine the content of the key anabolic markers. The rate of PS was measured by SUnSET technique.
 Results EC-induced increase in PS was significantly less in the HS and HS+ Gd3+ groups vs. the C group. There was no statistically significant difference between the HS and HS+ Gd3+ groups in terms of EC-induced increase in muscle PS. A decrease in EC-induced phosphorylation of p70S6K, 4E-BP1, RPS6 and GSK-3beta in the 7-day unloaded soleus treated with SAC inhibitor did not differ from that of the 7-day unloaded soleus without SAC blockade. Thus, the inhibition of SAC with gadolinium did not lead to further decline in EC-induced phosphorylation of the key anabolic markers and muscle PS.
 Conclusions The results of the study suggest that attenuation of mTORC1-signaling and PS in response to EC in unloaded soleus muscle may be associated with inactivation of SAC. The study was supported by the RFBR grant # 16-34-60055.

Mechano-electric finite element model of the left atrium

Mechanical stretch plays a major role in modulating atrial function, being responsible for beat-by-beat responses to changes in chamber preload, enabling a prompt regulation of cardiac function. Mechano-electric coupling (MEC) operates through many mechanisms and has many targets, making it experimentally difficult to isolate causes and effects especially under sinus conditions where effects are more transient and subtle. Therefore, modelling is a powerful tool to help understand the role of MEC with respect to the atrial electromechanical interaction. We propose a cellular-based computational model of the left atrium that includes a strongly coupled MEC component and mitral flow component to account for correct pressure generation in the atrial chamber as a consequence of blood volume and contraction. The method was applied to a healthy porcine left atrium. Results of the strongly coupled simulation show that strains are higher in the areas adjacent to the mitral annulus, the rim of the appendage, around the pulmonary venous trunks and at the location of the Bachmann's bundle, approximately between the mitral annulus and the region where the venous tissue transitions into atrial. These are regions where arrhythmias are likely to originate. The role of stretch-activated channels was very small for sinus rhythm for the single cardiac beat simulation, although tension development was very sensitive to stretch. The method could be applied to investigate potential therapeutic interventions acting on the mechano-electrical properties of the left atrium.

ALTERATIONS OF ECCENTRICALLY EVOKED PROTEIN SYNTHESIS IN ISOLATED RAT SOLEUS FOLLOWING GRAVITATIONAL UNLOADING: ROLE OF FAK

ALTERATIONS OF ECCENTRICALLY EVOKED PROTEIN SYNTHESIS IN ISOLATED RAT SOLEUS FOLLOWING GRAVITATIONAL UNLOADING: ROLE OF FAK

Intracellular calcium accumulation following eccentric contractions in rat skeletal muscle in vivo: role of stretch-activated channels

Although the accumulation of intracellular calcium ions ([Ca2+]i) is associated with muscle damage, little is known regarding the temporal profile of muscle [Ca2+]i under in vivo conditions, and, specifically, the effects of different contraction types [e.g., isometric (ISO); eccentric (ECC)] on [Ca2+]i remain to be determined. The following hypotheses were tested. 1) For 90 min at rest, an in vivo vs. in vitro preparation would better maintain initial [Ca2+]i. 2) Compared with ISO, ECC contractions (50 contractions, 10 sets, 5-min interval) would lead to a greater increase of [Ca2+]i. 3) Elevated [Ca2+]i during ECC would be reduced or prevented by the stretch-activated ion channel blockers streptomycin and gadolinium (Gd3+). Spinotrapezius muscles of Wistar rats were exteriorized (in vivo) or excised (in vitro). [Ca2+]i was evaluated by loading the muscle with fura 2-AM using fluorescence imaging. [Ca2+]i rose progressively beyond 40 min at rest under in vitro but not in vivo conditions during the 90-min protocol. In vivo [Ca2+]i increased more rapidly during ECC (first set) than ISO (fifth set) (P < 0.05 vs. precontraction values). The peak level of [Ca2+]i was increased by 21.5% (ISO) and 42.8% (ECC) after 10 sets (both P < 0.01). Streptomycin and Gd3+ abolished the majority of [Ca2+]i increase during ECC (69 and 86% reduction, respectively; P < 0.01 from peak [Ca2+]i of ECC). In conclusion, in vivo quantitative analyses demonstrated that ECC contractions elevate [Ca2+]i significantly more than ISO contractions and that stretch-activated channels may play a permissive role in this response.

Mechanotransduction pathways of low-intensity ultrasound in C-28/I2 human chondrocyte cell line

Low-intensity ultrasound (LIUS) has recently been considered to be an effective method to induce cartilage repair and/or regeneration after injury. Nevertheless, there is no study to provide a cellular mechanism or signal pathways of LIUS stimulation. The current study is designed to investigate the effects of LIUS on the mechanotransduction pathways in C-28/I2, an immortalized human chondrocyte cell line. C-28/I2 cells were treated with LIUS at an intensity of 200 mW/cm2 using Noblelife from Duplogen. The role of stretch-activated channels (SAC) and integrins that are most well-known mechanoreceptors on the chondrocyte cell surface was first examined in mediating the LIUS effects on the expression of type II collagen and aggrecan. When analysed by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry, gadolinium (a specific inhibitor of SACs) or GRGDSP (a peptide inhibitor of integrins) specifically reduced the LIUS-induced elevation of type II collagen and aggrecan expressions depending on the incubation time. In addition, the LIUS treatment of C-28/I2 cells induced the phosphorylation of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) but not p38 kinase among the members of the mitogen-activated protein kinases (MAPKs). The phosphorylation of ERK by LIUS was repressed by a specific inhibitor of the ERK pathway and integrin function. These results suggest that the LIUS signal might be mediated via canonical mechanoreceptors of SACs and integrins and subsequently through JNK and ERK pathways. The present study provides the first evidence for the activation of the mechanotransduction pathways by LIUS in human chondrocytes.