Calcium sparks

{{Short description|Higher release of Calcium in cells}} A '''calcium spark''' is the microscopic release of calcium (Ca²⁺) from a store known as the sarcoplasmic reticulum (SR), located within muscle cells.<ref name=":4">{{cite journal | last1 = Cheng | first1 = H. | last2 = Lederer | first2 = W.J. | last3 = Cannell | first3 = M.B. | year = 1993 | title = Calcium sparks: elementary events underlying excitation-contraction coupling in heart muscle | journal = Science | volume = 262 | issue = 5134| pages = 740–744 | doi = 10.1126/science.8235594 | pmid = 8235594 | bibcode = 1993Sci...262..740C }}</ref> This release occurs through an ion channel within the membrane of the SR, known as a ryanodine receptor (RyR), which opens upon activation.<ref>Lanner, J.T., Georgiou, D.K., Joshi, A.D. and Hamilton, S.L. (2010) 'Ryanodine receptors: Structure, expression, molecular details, and function in calcium release', 2(11)</ref> 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,<ref>Cannell, M. and Kong, C. (2011) 'Local control in cardiac E-C coupling', Journal of Molecular and Cellular Cardiology, 52(2), pp. 298–303.</ref> as well as whole cell changes.

==Activation== As mentioned above, Ca²⁺ sparks depend on the opening of ryanodine receptors, of which there are three types: * Type 1 – found mainly in skeletal muscle * Type 2 – found mainly in the heart * Type 3 – found in smooth muscle and neurones Opening of the channel allows Ca²⁺ to pass from the SR, into the cell. This increases the local Ca²⁺ concentration around the RyR, by a factor of 10.<ref>Hoang-Trong, T.M., Ullah, A. and Jafri, S.M. (2015) 'Calcium sparks in the heart: Dynamics and regulation', 6</ref> Calcium sparks can either be evoked or spontaneous, as described below. thumb|440x440px|Figure 1: An evoked calcium spark, in a cardiac muscle cell.

===Evoked=== Electrical impulses, known as action potentials, travel along the cell membrane (sarcolemma) of muscle cells.<ref>Lodish, H., Berk, A., Zipursky, L.S., Matsudaira, P., Baltimore, D. and Darnell, J. (2000) The action potential and conduction of electric impulses. Available at: [https://www.ncbi.nlm.nih.gov/books/NBK21668/]{{dead link|date=July 2025|bot=medic}}{{cbignore|bot=medic}} (Accessed: 11 February 2017)</ref> Located in the sarcolemma of smooth muscle cells are receptors, called dihydropyridine receptors (DHPR). In skeletal and cardiac muscle cells, however, these receptors are located within structures known as T-tubules, that are extensions of the plasma membrane penetrating deep into the cell (see figure 1).<ref>{{cite journal | last1 = Brette | first1 = F. | last2 = Orchard | first2 = C. | year = 2003 | title = T-tubule function in mammalian cardiac myocytes | journal = Circulation Research | volume = 92 | issue = 11| pages = 1182–92 | doi=10.1161/01.res.0000074908.17214.fd| pmid = 12805236 | doi-access = free | citeseerx = 10.1.1.334.2517 }}</ref><ref>{{Cite journal|last1=Cheng|first1=Heping|last2=Lederer|first2=W. J.|date=2008-10-01|title=Calcium Sparks|journal=Physiological Reviews|language=en|volume=88|issue=4|pages=1491–1545|doi=10.1152/physrev.00030.2007|issn=0031-9333|pmid=18923188}}</ref> These DHPRs are located directly opposite to the ryanodine receptors, located on the sarcoplasmic reticulum<ref>{{cite journal | last1 = Scriven | first1 = D. R. L. | last2 = Dan | first2 = P. | last3 = Moore | first3 = E. D. W. | year = 2000 | title = Distribution of proteins implicated in excitation-contraction coupling in rat ventricular myocytes | journal = Biophys. J. | volume = 79 | issue = 5| pages = 2682–2691 | doi=10.1016/s0006-3495(00)76506-4| pmid = 11053140 | pmc = 1301148 | bibcode = 2000BpJ....79.2682S }}</ref> and activation, by the action potential causes the DHPRs to change shape.<ref>{{cite journal | last1 = Araya | first1 = R. | last2 = Liberona | first2 = J. | last3 = Cárdenas | first3 = J. | last4 = Riveros | first4 = N. | last5 = Estrada | first5 = M. | last6 = Powell | first6 = J. | last7 = Carrasco | first7 = M. | last8 = Jaimovich | first8 = E. | year = 2003 | title = Dihydropyridine receptors as voltage sensors for a depolarization-evoked, IP3R-mediated, slow calcium signal in skeletal muscle cells | journal = The Journal of General Physiology | volume = 121 | issue = 1| pages = 3–16 | doi=10.1085/jgp.20028671| pmid = 12508050 | pmc = 2217318 }}</ref>

In cardiac and smooth muscle, activation of the DHPR results in it forming an ion channel.<ref>{{cite journal | last1 = Kotlikoff | first1 = M | year = 2003 | title = Calcium-induced calcium release in smooth muscle: The case for loose coupling | journal = Progress in Biophysics and Molecular Biology | volume = 83 | issue = 3| pages = 171–91 | doi=10.1016/s0079-6107(03)00056-7| pmid = 12887979 | doi-access = }}</ref> This allows Ca²⁺ to pass into the cell, increasing the local Ca²⁺ concentration, around the RyR. When four Ca²⁺ molecules bind to the RyR, it opens, resulting in a larger release of Ca²⁺, from the SR . This process, of using Ca²⁺ to activate release of Ca²⁺ from the SR is known as calcium-induced calcium release (CICR).<ref>{{cite journal | last1 = Fabiato | first1 = A | year = 1983 | title = Calcium-induced release of calcium from the cardiac sarcoplasmic reticulum | journal = Am. J. Physiol. | volume = 245 | issue = 1 | pages = C1–C14 | doi=10.1152/ajpcell.1983.245.1.c1 | pmid = 6346892}}</ref>

However, in skeletal muscle the DHPR touches the RyR. Therefore, the shape change of the DHPR activates the RyR directly, without the need for Ca²⁺ to flood into the cell first. This causes the RyR to open, allowing Ca²⁺ to be released from the SR.<ref>{{cite journal | last1 = Meissner | first1 = G. | last2 = Lu | first2 = X. | year = 1995 | title = Dihydropyridine receptor-ryanodine receptor interactions in skeletal muscle excitation-contraction coupling | journal = Bioscience Reports | volume = 15 | issue = 5| pages = 399–408 | doi=10.1007/bf01788371| pmid = 8825041 | s2cid = 32810845 }}</ref>

===Spontaneous === Ca²⁺ sparks can also occur in cells at rest (i.e. cells that have not been stimulated by an action potential). This occurs roughly 100 times every second in each cell<ref name=":0" /> and is a result of the resting Ca²⁺ concentration being high enough to start local CICR. An increase in Ca²⁺ within the SR is also.thought to bind to Ca²⁺ sensitive sites on the inside of the RyR causing the channel to be more likely to open. As well as this, a protein called calsequestrin (found within the SR) detaches from the RyR, when calcium concentration is high, again allowing the channel to open (see sarcoplasmic reticulum for more details). Similarly, a decrease in Ca²⁺ concentration within the SR has also proven to lower RyR sensitivity. This is thought to be due to the calsequestrin binding more strongly to the RyR, preventing it from opening and decreasing the likelihood of a spontaneous spark.<ref>{{Cite journal|last1=Bassani|first1=J. W.|last2=Yuan|first2=W.|last3=Bers|first3=D. M.|date=1995-05-01|title=Fractional SR Ca release is regulated by trigger Ca and SR Ca content in cardiac myocytes|journal=American Journal of Physiology. Cell Physiology|language=en|volume=268|issue=5|pages=C1313–C1319|issn=0363-6143|pmid=7762626|doi=10.1152/ajpcell.1995.268.5.c1313}}</ref>

== Calcium after release == There are roughly 10,000 clusters of ryanodine receptors within a single cardiac cell, with each cluster containing around 100 ryanodine receptors.<ref name=":0">{{cite journal | last1 = Cheng | first1 = H. | last2 = Lederer | first2 = W. | year = 2008 | title = Calcium sparks | journal = Physiological Reviews | volume = 88 | issue = 4| pages = 1491–545 | doi=10.1152/physrev.00030.2007| pmid = 18923188 }}</ref> During a single spontaneous spark, when Ca²⁺ is released from the SR, the Ca²⁺ diffuses throughout the cell. As the RyRs in the heart are activated by Ca²⁺, the movement of the Ca²⁺ released during a spontaneous spark, can activate the neighbouring RyRs within the same cluster. However, there usually isn't enough Ca²⁺ present in a single spark to reach a neighbouring cluster of receptors.<ref name=":0"/> The calcium can, however, signal back to the DHPR causing it to close and preventing further influx of calcium as a form of negative feedback for long term stability <ref>{{Cite journal |last=Grantham |first=C.J. |last2=Cannell |first2=M.B. |date=1996 |title=Ca2+ Influx During the Cardiac Action Potential in Guinea Pig Ventricular Myocytes |url=https://www.ahajournals.org/doi/10.1161/01.RES.79.2.194 |journal=Circulation Research |language=en |volume=79 |issue=2 |pages=194–200 |doi=10.1161/01.RES.79.2.194 |issn=0009-7330|url-access=subscription }}</ref>.

An increase in Ca²⁺ concentration within the cell or the production of a larger spark, can lead to a large enough calcium released that the neighbouring cluster can be activated by the first. This is known as spark-induced spark activation and can lead to a Ca²⁺ wave of calcium release spreading across the cell.<ref name=":0"/>

During evoked Ca²⁺ sparks, all clusters of ryanodine receptors, throughout the cell are activated at almost exactly the same time <ref>{{Cite journal |last=Cannell |first=M.B. |last2=Cheng |first2=H. |last3=Lederer |first3=W.J. |date=1994 |title=Spatial non-uniformities in [Ca2+]i during excitation-contraction coupling in cardiac myocytes |url=https://linkinghub.elsevier.com/retrieve/pii/S0006349594806770 |journal=Biophysical Journal |language=en |volume=67 |issue=5 |pages=1942–1956 |doi=10.1016/S0006-3495(94)80677-0 |pmc=1225569 |pmid=7858131}}</ref>. This produces an increase in Ca²⁺ concentration across the whole cell (not just locally) and is known as a whole cell Ca²⁺ transient. This Ca²⁺ then binds to a protein, called troponin, initiating contraction, through a group of proteins known as myofilaments.<ref>{{cite conference | last1 = Herzberg | first1 = O. | last2 = Moult | first2 = J. | last3 = James | first3 = M. | chapter = Calcium Binding to Skeletal Muscle Troponin C and the Regulation of Muscle Contraction | year = 1986 | title = Ciba Foundation Symposium 122 ‐ Calcium and the Cell | conference = Ciba Foundation Symposium of 1985 | volume = 122 | pages = 120–44 | pmid = 3792134 | doi = 10.1002/9780470513347.ch8 | series = Novartis Foundation Symposia | isbn = 9780470513347 | editor1=David Evered| editor2=Julie Whelan }}</ref>

In smooth muscle cells, the Ca²⁺ released during a spark is used for muscle relaxation. This is because, the Ca²⁺ that enters the cell via the DHPR in response to the action potential, stimulates both muscle contraction and calcium release from the SR. The Ca²⁺ released during the spark, then activates two other ion channels on the membrane. One channel allows potassium ions to exit the cell, whereas the other allows chloride ions to leave the cell. The result of this movement of ions, is that the membrane voltage becomes more negative. This deactivates the DHPR (which was activated by the positive membrane potential produced by the action potential), causing it to close and stopping the flow of Ca²⁺into the cell, leading to relaxation.<ref>{{cite journal | last1 = Webb | first1 = R | year = 2003 | title = Smooth muscle contraction and relaxation | journal = Advances in Physiology Education | volume = 27 | issue = 4| pages = 201–6 | doi=10.1152/advances.2003.27.4.201| pmid = 14627618 }}</ref>

==Termination== The mechanism by which SR Ca²⁺ release terminates is still not fully understood. Current main theories are outlined below:<ref name=":3" />

===Local depletion of SR Ca²⁺=== This theory suggests that during a calcium spark, as calcium flows out of the SR, the concentration of Ca²⁺ within the local region of the SR becomes too low to support continued release. However, this was not thought to be the case for spontaneous sparks as the total release during a Ca²⁺ spark is small compared to total SR Ca²⁺ content and researchers have produced sparks lasting longer than 200 milliseconds, therefore showing that there is still Ca²⁺ left within the SR after a 'normal' (200ms) spark.<ref name="Bers, D.M. 2002 pp. 198">{{cite journal | last1 = Bers | first1 = D.M. | year = 2002 | title = Cardiac excitation-contraction coupling | journal = Nature | volume = 415 | issue = 6868| pages = 198–205 | doi = 10.1038/415198a | bibcode = 2002Natur.415..198B | pmid = 11805843 | s2cid = 4337201 }}</ref> However local depletion in the junctional SR may be much larger than previously thought (see <ref name ="Kong et al. 2013">{{cite journal | author = Kong CHT, Laver DR, Cannell MB | title = Extraction of Sub-microscopic Ca Fluxes from Blurred and Noisy Fluorescent Indicator Images with a Detailed Model Fitting Approach | journal = PLOS Comput Biol | year = 2013 | volume = 9 | issue = 2 | pages = e1002931–7 | doi = 10.1371/journal.pcbi.1002931 | pmid = 23468614 | pmc = 3585382 | bibcode = 2013PLSCB...9E2931K | doi-access = free }}</ref><ref name=":5">{{Cite journal |last=Laver |first=D.R. |last2=Kong |first2=C.H.T. |last3=Imtiaz |first3=M.S. |last4=Cannell |first4=M.B. |date=2013 |title=Termination of calcium-induced calcium release by induction decay: An emergent property of stochastic channel gating and molecular scale architecture |url=https://linkinghub.elsevier.com/retrieve/pii/S002228281200377X |journal=Journal of Molecular and Cellular Cardiology |volume=54 |pages=98–100 |doi=10.1016/j.yjmcc.2012.10.009 |issn=0022-2828|url-access=subscription }}</ref>) and this leads to the local Ca²⁺ flux being able to support local CICR. This loss of local CICR strength is the reversal of the normal activation process in CICR and effectively snowballs until the release flux is reduced to the point where only a few RyRs remain open at any one time and stochastic attrition (see below) can take place<ref name=":5" /><ref>{{Cite journal |last=Gillespie |first=Dirk |last2=Fill |first2=Michael |date=2013 |title=Pernicious attrition and inter-RyR2 CICR current control in cardiac muscle |url=https://linkinghub.elsevier.com/retrieve/pii/S0022282813000382 |journal=Journal of Molecular and Cellular Cardiology |language=en |volume=58 |pages=53–58 |doi=10.1016/j.yjmcc.2013.01.011 |pmc=3628281 |pmid=23369697}}</ref> During the activation of a large number of ryanodine receptors however, as is the case during electrically evoked Ca²⁺ release , the entire SR is about 50% depleted of Ca²⁺ and this mechanism will play an important role in repriming of release.

===Stochastic attrition=== Despite the complicated name, this idea simply suggests that all ryanodine receptors in a cluster (and the associated dihydropyridine receptors) happen to randomly close at the same time. This would not only prevent calcium release from the SR, but it would also stop the stimulus for restarting calcium release (i.e. the flow of calcium through the DHPR).<ref name="ReferenceA">{{cite journal | last1 = Sham | first1 = J. S. K. |display-authors=etal | year = 1998| title = Termination of Ca2+ release by a local inactivation of ryanodine receptors in cardiac myocytes | journal = Proc. Natl. Acad. Sci. USA | volume = 95 | issue = 25| pages = 15096–15101 | doi=10.1073/pnas.95.25.15096| pmid = 9844021 | pmc = 24581 | bibcode = 1998PNAS...9515096S | doi-access = free }}</ref> However, due to number of RyRs in a cluster, this mechanism seems unlikely to be able to start the termination of release, as there is a very small probability that they would all close together at exactly the same time.<ref name=":3">{{Cite journal |last=Cannell |first=Mark B. |last2=Kong |first2=Cherrie H.T. |date=2017-09-04 |title=Quenching the spark: Termination of CICR in the submicroscopic space of the dyad |url=https://rupress.org/jgp/article/149/9/837/53274/Quenching-the-spark-Termination-of-CICR-in-the |journal=Journal of General Physiology |language=en |volume=149 |issue=9 |pages=837–845 |doi=10.1085/jgp.201711807 |issn=0022-1295 |pmc=5583711 |pmid=28798280}}</ref>

===Inactivation/adaptation=== This theory suggests that after activation of the RyR and the subsequent release of Ca²⁺, the channel closes briefly to recover. During this time, either the channel cannot be reopened, even if calcium is present (i.e. the RyR is inactivated) or the channel can be reopened, however more calcium is required to activate it than usual (i.e. the RyR is in an adaptation phase)<ref>{{cite journal | last1 = Sham | first1 = J. S. K. |display-authors=etal | year = 1998 | title = Termination of Ca2+ release by a local inactivation of ryanodine receptors in cardiac myocytes | journal = Proc. Natl. Acad. Sci. USA | volume = 95 | issue = 25| pages = 15096–15101 | doi=10.1073/pnas.95.25.15096| pmid = 9844021 | pmc = 24581 | bibcode = 1998PNAS...9515096S | doi-access = free }}</ref>. This would mean that one-by-one the RyRs would close, thus ending the spark.<ref name="ReferenceA"/>

=== Sticky cluster theory === This theory suggests that the RyRs can comminicate their open and closed states across the cluster, perhaps via their physical conformation <ref>Sobie, E.A., Dilly, K.W., Cruz, J. dos S., Lederer, J.W. and Jafri, S.M. (2002) 'Termination of cardiac ca(2+) sparks: An investigative mathematical model of calcium-induced calcium release', 83(1)</ref>. Physical comminication now seems unlikely after electron micrographs showed that few RyRs actually touch each other. <ref>{{Cite journal |last=Asghari |first=Parisa |last2=Scriven |first2=David R.L. |last3=Sanatani |first3=Shubhayan |last4=Gandhi |first4=Sanjiv K. |last5=Campbell |first5=Andrew I.M. |last6=Moore |first6=Edwin D.W. |date=2014-07-07 |title=Nonuniform and Variable Arrangements of Ryanodine Receptors Within Mammalian Ventricular Couplons |url=https://www.ahajournals.org/doi/10.1161/CIRCRESAHA.115.303897 |journal=Circulation Research |language=en |volume=115 |issue=2 |pages=252–262 |doi=10.1161/CIRCRESAHA.115.303897 |issn=0009-7330}}</ref>

==Discovery== Spontaneous Ca²⁺ sparks were discovered in cardiac muscle cells, of rats, in 1992 by Peace Cheng and Mark B. Cannell in Jon Lederer's laboratory at the University of Maryland, Baltimore, U.S.A.<ref name=":4" />

Initially the idea was rejected by the scientific journal, Nature, who reviewers believed that the sparks were a consequence of the methodology (i.e. they were artifacts), and so wouldn't occur naturally within the body. However they were quickly recognised as being of fundamental importance to muscle physiology, playing a central role in excitation-contraction coupling.

The discovery was made possible due to improvements in confocal microscopes. This allowed for the detection of the release of Ca²⁺, which were highlighted using a substance known as fluo-3, which caused the Ca²⁺ to be detectable by fluorescence. Ca²⁺ “sparks” were so called because of the spontaneous, localised nature of the Ca²⁺ release as well as the fact that they are the initiation event of excitation-contraction coupling.

==Detection and analysis== Because of the importance of Ca²⁺ sparks in explaining the gating properties of ryanodine receptors in situ (within the body), many studies have focused on improving their detectability <ref name=":1">{{cite journal|date=February 1999|title=Amplitude distribution of Ca²⁺ sparks in confocal images: theory and studies with an automatic detection method|journal=Biophysical Journal|volume=76|issue=2|pages=606–17|doi=10.1016/S0006-3495(99)77229-2|pmc=1300067|pmid=9929467|vauthors=Cheng H, Song LS, Shirokova N|display-authors=etal}}</ref><ref name=":2">{{cite journal|date=January 2005|title=Ca²⁺ sparks in muscle cells: interactive procedures for automatic detection and measurements on line-scan confocal images series|journal=Computer Methods and Programs in Biomedicine|volume=77|issue=1|pages=57–70|doi=10.1016/j.cmpb.2004.06.004|pmid=15639710|vauthors=Sebille S, Cantereau A, Vandebrouck C|display-authors=etal}}</ref> in the hope that by accurately and reliably detecting all Ca²⁺ spark events, their properties can help us explain how muscle cell contraction is regulated.

==See also== *Calcium-induced calcium release *Confocal microscopy *Ryanodine receptor

==References== {{Reflist}}

==External links== '''Software''' * [http://sparkmasterhome.googlepages.com SparkMaster - Automated Ca²⁺ Spark Analysis with ImageJ] - Free software for Ca²⁺ spark analysis in confocal linescan images

Category:Cell biology