The sarcoplasmic reticulum (SR) is a membrane-bound structure found within muscle cells that is similar to the smooth endoplasmic reticulum in other cells. The main function of the SR is to store calcium ions (Ca2+). Calcium ion levels are kept relatively constant, with the concentration of calcium ions within a cell being 10,000 times smaller than the concentration of calcium ions outside the cell.This means that small increases in calcium ions within the cell are easily detected and can bring about important cellular changes (the calcium is said to be a second messenger; see calcium in biology for more details). Calcium is used to make calcium carbonate (found in chalk) and calcium phosphate, two compounds that the body uses to make teeth and bones. This means that too much calcium within the cells can lead to hardening (calcification) of certain intracellular structures, including the mitochondria, leading to cell death. Therefore, it is vital that calcium ion levels are controlled tightly, and can be released into the cell when necessary and then removed from the cell.
The sarcoplasmic reticulum is a network of tubules that extend throughout muscle cells, wrapping around (but not in direct contact with) the myofibrils (contractile units of the cell). Cardiac and skeletal muscle cells contain structures called transverse tubules (T-tubules), which are extensions of the cell membrane that travel into the centre of the cell. T-tubules are closely associated with a specific region of the SR, known as the terminal cisternae in skeletal muscle, with a distance of roughly 12 nanometers, separating them. This is the primary site of calcium release.The longitudinal SR are thinner projects, that run between the terminal cisternae/junctional SR, and are the location where ion channels necessary for calcium ion absorption are most abundant. These processes are explained in more detail below and are fundamental for the process of excitation-contraction coupling in skeletal, cardiac and smooth muscle.
The SR contains ion channel pumps, within its membrane that are responsible for pumping Ca2+ into the SR. As the calcium ion concentration within the SR is higher than in the rest of the cell, the calcium ions won't freely flow into the SR, and therefore pumps are required, that use energy, which they gain from a molecule called adenosine triphosphate (ATP). These calcium pumps are called Sarco(endo)plasmic reticulum Ca2+ ATPases (SERCA). There are a variety of different forms of SERCA, with SERCA 2a being found primarily in cardiac and skeletal muscle.
SERCA consists of 13 subunits (labelled M1-M10, N, P and A). Calcium ions bind to the M1-M10 subunits (which are located within the membrane), whereas ATP binds to the N, P and A subunits (which are located outside the SR). When 2 calcium ions, along with a molecule of ATP, bind to the cytosolic side of the pump (i.e. the region of the pump outside the SR), the pump opens. This occurs because ATP (which contains three phosphate groups) releases a single phosphate group (becoming adenosine diphosphate). The released phosphate group then binds to the pump, causing the pump to change shape. This shape change causes the cytosolic side of the pump to open, allowing the two Ca2+ to enter. The cytosolic side of the pump then closes and the sarcoplasmic reticulum side opens, releasing the Ca2+ into the SR.
A protein found in cardiac muscle, called phospholamban (PLB) has been shown to prevent SERCA from working. It does this by binding to the SERCA and decreasing its attraction (affinity) to calcium, therefore preventing calcium uptake into the SR. Failure to remove Ca2+ from the cytosol, prevents muscle relaxation and therefore means that there is a decrease in muscle contraction too. However, molecules such as adrenaline and noradrenaline, can prevent PLB from inhibiting SERCA. When these hormones bind to a receptor, called a beta 1 adrenoceptor, located on the cell membrane, they produce a series of reactions (known as a cyclic AMP pathway) that produces an enzyme called protein kinase A (PKA). PKA can add a phosphate to PLB (this is known as phosphorylation), preventing it from inhibiting SERCA and allowing for muscle relaxation.
Located within the SR is a protein called calsequestrin. This protein can bind to around 50 Ca2+, which decreases the amount of free Ca2+ within the SR (as more is bound to calsequestrin).Therefore, more calcium can be stored (the calsequestrin is said to be a buffer). It is primarily located within the junctional SR/luminal space, in close association with the calcium release channel (described below).
Calcium ion release from the SR, occurs in the junctional SR/terminal cisternae through a ryanodine receptor (RyR) and is known as a calcium spark.There are three types of ryanodine receptor, RyR1 (in skeletal muscle), RyR2 (in cardiac muscle) and RyR3 (in the brain). Calcium release through ryanodine receptors in the SR is triggered differently in different muscles. In cardiac and smooth muscle an electrical impulse (action potential) triggers calcium ions to enter the cell through an L-type calcium channel located in the cell membrane (smooth muscle) or T-tubule membrane (cardiac muscle). These calcium ions bind to and activate the RyR, producing a larger increase in intracellular calcium. In skeletal muscle, however, the L-type calcium channel is bound to the RyR. Therefore, activation of the L-type calcium channel, via an action potential, activates the RyR directly, causing calcium release (see calcium sparks for more details). Also, caffeine (found in coffee) can bind to and stimulate RyR. Caffeine makes the RyR more sensitive to either the action potential (skeletal muscle) or calcium (cardiac or smooth muscle), thereby producing calcium sparks more often (this is partially responsible for caffeine's effect on heart rate).
Triadin and Junctin are proteins found within the SR membrane, that are bound to the RyR. The main role of these proteins is to anchor calsequestrin (see above) to the ryanodine receptor. At ‘normal’ (physiological) SR calcium levels, calsequestrin binds to the RyR, Triadin and Junctin, which prevents the RyR from opening.If calcium concentration within the SR falls too low, there will be less calcium bound to the calsequestrin. This means that there is more room on the calsequestrin, to bind to the junctin, triadin and ryanodine receptor, therefore it binds tighter. However, if calcium within the SR rises too high, more calcium binds to the calsequestrin and therefore it binds to the junctin-triadin-RyR complex less tightly. The RyR can therefore open and release calcium into the cell.
In addition to the effects that PKA had on phospholamban (see above) that resulted in increased relaxation of the cardiac muscle, PKA (as well as another enzyme called calmodulin kinase II) can also phosphorylate ryanodine receptors. When phosphorylated, RyRs are more sensitive to calcium, therefore they open more often and for longer periods of time. This increases calcium release from the SR, increasing the rate of contraction.Therefore, in cardiac muscle, activation of PKA, through the cyclic AMP pathway, results in increased muscle contraction (via RyR2 phosphorylation) and increased relaxation (via phospholamban phosphorylation), which increases heart rate.
The mechanism behind the termination of calcium release through the RyR is still not fully understood. Some researchers believe it is due to the random closing of ryanodine receptors (known as stochastic attrition), or the ryanodine receptors becoming inactive after a calcium spark,while others believe that a decrease in SR calcium, triggers the receptors to close (see calcium sparks for more details).
The breakdown of the sarcoplasmic reticulum, along with the resultant release of calcium, is an important contributor to rigor mortis, the stiffening of muscles after death.
If the concentration of calcium increases in the sarcoplasm then it can also cause muscles stiffness.
A sarcomere is the smallest functional unit of striated muscle tissue. It is the repeating unit between two Z-lines. Skeletal muscles are composed of tubular muscle cells which are formed during embryonic myogenesis. Muscle fibers contain numerous tubular myofibrils. Myofibrils are composed of repeating sections of sarcomeres, which appear under the microscope as alternating dark and light bands. Sarcomeres are composed of long, fibrous proteins as filaments that slide past each other when a muscle contracts or relaxes. The costamere is a different component that connects the sarcomere to the sarcolemma.
SERCA, or sarco/endoplasmic reticulum Ca2+-ATPase, or SR Ca2+-ATPase, is a calcium ATPase-type P-ATPase. Its major function is to transport calcium from the cytosol into the sarcoplasmic reticulum.
Muscle contraction is the activation of tension-generating sites within muscle cells. In physiology, muscle contraction does not necessarily mean muscle shortening because muscle tension can be produced without changes in muscle length, such as when holding something heavy in the same position. The termination of muscle contraction is followed by muscle relaxation, which is a return of the muscle fibers to their low tension-generating state.
Ryanodine receptors form a class of intracellular calcium channels in various forms of excitable animal tissue like muscles and neurons. There are three major isoforms of the ryanodine receptor, which are found in different tissues and participate in different signaling pathways involving calcium release from intracellular organelles. The RYR2 ryanodine receptor isoform is the major cellular mediator of calcium-induced calcium release (CICR) in animal cells.
Phospholamban, also known as PLN or PLB, is a micropeptide protein that in humans is encoded by the PLN gene. Phospholamban is a 52-amino acid integral membrane protein that regulates the calcium (Ca2+) pump in cardiac muscle cells.
T-tubules are extensions of the cell membrane that penetrate into the centre of skeletal and cardiac muscle cells. With membranes that contain large concentrations of ion channels, transporters, and pumps, T-tubules permit rapid transmission of the action potential into the cell, and also play an important role in regulating cellular calcium concentration.
Myocardial contractility represents the innate ability of the heart muscle (cardiac muscle or myocardium) to contract. The ability to produce changes in force during contraction result from incremental degrees of binding between different types of tissue, that is, between filaments of myosin (thick) and actin (thin) tissue. The degree of binding depends upon the concentration of calcium ions in the cell. Within an in vivo intact heart, the action/response of the sympathetic nervous system is driven by precisely timed releases of a catecholamine, which is a process that determines the concentration of calcium ions in the cytosol of cardiac muscle cells. The factors causing an increase in contractility work by causing an increase in intracellular calcium ions (Ca++) during contraction.
Calcium-induced calcium release (CICR) describes a biological process whereby calcium is able to activate calcium release from intracellular Ca2+ stores (e.g., endoplasmic reticulum or sarcoplasmic reticulum). Although CICR was first proposed for skeletal muscle in the 1970s, it is now known that CICR is unlikely to be the primary mechanism for activating SR calcium release. Instead, CICR is thought to be crucial for excitation-contraction coupling in cardiac muscle. It is now obvious that CICR is a widely occurring cellular signaling process present even in many non-muscle cells, such as in the insulin-secreting pancreatic beta cells, epithelium, and many other cells. Since CICR is a positive-feedback system, it has been of great interest to elucidate the mechanism(s) responsible for its termination.
Calsequestrin is a calcium-binding protein that acts as a calcium buffer within the sarcoplasmic reticulum. The protein helps hold calcium in the cisterna of the sarcoplasmic reticulum after a muscle contraction, even though the concentration of calcium in the sarcoplasmic reticulum is much higher than in the cytosol. It also helps the sarcoplasmic reticulum store an extraordinarily high amount of calcium ions. Each molecule of calsequestrin can bind 18 to 50 Ca2+ ions. Sequence analysis has suggested that calcium is not bound in distinct pockets via EF-hand motifs, but rather via presentation of a charged protein surface. Two forms of calsequestrin have been identified. The cardiac form Calsequestrin-2 (CASQ2) is present in cardiac and slow skeletal muscle and the fast skeletal form Calsequestrin-1(CASQ1) is found in fast skeletal muscle. The release of calsequestrin-bound calcium (through a calcium release channel) triggers muscle contraction. The active protein is not highly structured, more than 50% of it adopting a random coil conformation. When calcium binds there is a structural change whereby the alpha-helical content of the protein increases from 3 to 11%. Both forms of calsequestrin are phosphorylated by casein kinase 2, but the cardiac form is phosphorylated more rapidly and to a higher degree. Calsequestrin is also secreted in the gut where it deprives bacteria of calcium ions..
Catecholaminergic polymorphic ventricular tachycardia (CPVT) is an inherited genetic disorder that predisposes those affected to potentially life-threatening abnormal heart rhythms or arrhythmias. The arrhythmias seen in CPVT typically occur during exercise or at times of emotional stress, and classically take the form of bidirectional ventricular tachycardia or ventricular fibrillation. Those affected may be asymptomatic, but they may also experience blackouts or even sudden cardiac death.
In the histology of skeletal muscle, a triad is the structure formed by a T tubule with a sarcoplasmic reticulum (SR) known as the terminal cisterna on either side. Each skeletal muscle fiber has many thousands of triads, visible in muscle fibers that have been sectioned longitudinally. In mammals, triads are typically located at the A-I junction; that is, the junction between the A and I bands of the sarcomere, which is the smallest unit of a muscle fiber.
A calcium spark is the microscopic release of calcium (Ca2+) from a store known as the sarcoplasmic reticulum (SR), located within muscle cells. This release occurs through an ion channel within the membrane of the SR, known as a ryanodine receptor (RyR), which opens upon activation. This process is important as it helps to maintain Ca2+ concentration within the cell. It also initiates muscle contraction in skeletal and cardiac muscles and muscle relaxation in smooth muscles. Ca2+ sparks are important in physiology as they show how Ca2+ can be used at a subcellular level, to signal both local changes, known as local control, as well as whole cell changes.
Ryanodine receptor 2 (RYR2) is one of a class of ryanodine receptors and a protein found primarily in cardiac muscle. In humans, it is encoded by the RYR2 gene. In the process of cardiac calcium-induced calcium release, RYR2 is the major mediator for sarcoplasmic release of stored calcium ions.
Triadin, also known as TRDN, is a human gene associated with the release of calcium ions from the sarcoplasmic reticulum triggering muscular contraction through calcium-induced calcium release. Triadin is a multiprotein family, arising from different processing of the TRDN gene on chromosome 6. It is a transmembrane protein on the sarcoplasmic reticulum due to a well defined hydrophobic section and it forms a quaternary complex with the cardiac ryanodine receptor (RYR2), calsequestrin (CASQ2) and junctin proteins. The luminal (inner compartment of the sarcoplasmic reticulum) section of Triadin has areas of highly charged amino acid residues that act as luminal Ca2+ receptors. Triadin is also able to sense luminal Ca2+ concentrations by mediating interactions between RYR2 and CASQ2. Triadin has several different forms; Trisk 95 and Trisk 51, which are expressed in skeletal muscle, and Trisk 32 (CT1), which is mainly expressed in cardiac muscle.
Sarcolipin is a micropeptide protein that in humans is encoded by the SLN gene.
Terminal cisternae are enlarged areas of the sarcoplasmic reticulum surrounding the transverse tubules.
CXL 1020 is an experimental drug that is being investigated as a treatment for acute decompensated heart failure. CXL 1020 functions as a nitroxyl donor; nitroxyl is the reduced, protonated version of nitric oxide. Nitroxyl is capable of enhancing left ventricular contractility without increasing heart rate by modifying normal Ca2+ cycling through the sarcoplasmic reticulum as well as increasing the sensitivity of cardiac myofilaments to Ca2+.
The dyadic space is the name for the volume of cytoplasm between pairs (dyads) of areas where the cell membrane and an organelle such as the endoplasmic reticulum come into close contact of each other, creating what are known as dyadic clefts.
Cardiac excitation-contraction coupling (CardiacEC coupling) describes the series of events, from the production of an electrical impulse (action potential) to the contraction of muscles in the heart. This process is of vital importance as it allows for the heart to beat in a controlled manner, without the need for conscious input. EC coupling results in the sequential contraction of the heart muscles that allows blood to be pumped, first to the lungs (pulmonary circulation) and then around the rest of the body (systemic circulation) at a rate between 60 and 100 beats every minute, when the body is at rest. This rate can be altered, however, by nerves that work to either increase heart rate (sympathetic nerves) or decrease it (parasympathetic nerves), as the body's oxygen demands change. Ultimately, muscle contraction revolves around a charged atom (ion), calcium (Ca2+), which is responsible for converting the electrical energy of the action potential into mechanical energy (contraction) of the muscle. This is achieved in a region of the muscle cell, called the transverse-tubule during a process known as calcium induced calcium release.
Intrepicalcin (ViCaTx1) is a short peptide toxin found in the venom of scorpion Vaejovis intrepidus. It is one of a group of short, basic peptides called calcins, which bind to ryanodine receptors (RyRs) and thereby trigger calcium release from the sarcoplasmic reticulum.