Intracellular calcium governs the driving potential that determines the depolarisa- tion phase of the MSMC AP [36, 40, 67]; numerous conductances are gated by intra- cellular calcium, and calcium is essential in excitation-contraction coupling [68, 69]. At rest, cytosolic calcium is kept at around 100 nM, while extracellular calcium is about 1 mM, which creates a steep concentration gradient from outside the cell to inside the cell [30]. Due to this concentration gradient, intracellular calcium can be raised quickly by opening plasma membrane calcium channels [36]. On the other hand, energy must be expended to keep the cytosolic calcium sufficiently low (in fact, prolonged elevation of intracellular calcium is toxic [70, 71]). Finally, tightly controlled mechanisms are required to control the influx and the removal of calcium. Cytosolic calcium increases via several pathways, which are depicted schematically in Figure 1.13 and include: calcium channels in the plasma membrane (voltage-gated calcium channels, as well as receptor-, second-messenger, and mechanically-operated channels); calcium release from internal stores such as the endoplasmic reticulum (ER), and mitochondria. This release is mediated by calcium channels or receptors: the ryanodine receptor (RyR), also called the calcium-induced calcium-release chan- nel (CICR), and the inositol (1,4,5)-triphosphate (IP3) receptor which responds to
from the cell mainly through calcium ATPase pumps (PMCA: this is a plasma mem- brane calcium pump, which used the energy in the form of ATP to displace calcium against its concentration gradient) and via sodium-calcium exchange (NCX) which exploits the energy stored in the Na+ gradient to remove calcium out of the cell at the expense of Na+ entry [30]. Moreover, calcium can be sequestered in inter- nal stores (ER, mitochondria) through ATPase pumps such as SERCA, a calcium pump located in the membrane of the ER whose function is to accumulate calcium in the internal stores using energy of ATP. Finally, cytosolic calcium is buffered by large cytosolic proteins. In fact, approximately 99% of total intracellular calcium is bound to buffers. An in-depth treatment is provided by Keener and Sneyd [30].
buffers ER/SR
cytosol
Plasma membrane
Pumps and exchangers Calcium channels
Ca2+
Ca2+
Ca2+
Figure 1.13: General scheme of the main processes involved in changes in cy- tosolic calcium.
Mathematical models in the literature can be based on a description of the influxes and effluxes of calcium through the plasma membrane and internal stores. Depending on the level of biological detail, the resulting model can range from simple two-variables models to complex models with thousands of variables [30]. A simplification that is often made is that the concentration of calcium is the same throughout the cell. This eliminates spatial-dependent variables and leaves only dependence on time. There are, however, important spatial patterns that are not accounted for in such models, such as calcium waves spreading through the MSMC (detailed discussion in Chapter 6).
1.4.1 The two-pool model
The two-pool model, proposed by Goldbeter et al [72], distinguishes two separate pools: one being IP3-sensitive and the other being Ca2+-sensitive. Agonists stim-
ulation results in a rise of IP3. The rise allows the release of a certain amount
of Ca2+ from an IP3-sensitive store. The release of Ca2+ then triggers further re-
lease of Ca2+ from a second store, sensitive to Ca2+ and insensitive to IP3, through
other receptors (CICR; calcium-induced, calcium release). The state variables are the calcium concentrations in the cytosol and in the Ca2+-sensitive pool. In the model, a steady Ca2+ flux into the cytosol is caused by IP3; this flux is constant
for constant intracellular IP3 concentration and is treated as a control parameter.
Thus, by varying this flux, the behaviour of the model can be studied at different constant IP3 concentrations [30]. The concentration of Ca2+ in the IP3-sensitive
store is assumed to be constant as the store refills quickly from the extracellular medium. A diagram of the model is shown in Figure 1.14. The model describes IP3-induced oscillations, but experimental evidence indicates that the role of Ca2+
is more complicated than is assumed in this model. Moreover, the model does not encompass all of the signalling pathways. Other models extend the two-pool to de- scribe additional pathways in Ca2+ signalling by adding a third pool to model the role of mitochondria. Ca2+-sensitive store Ca2+ extrusion Ca2+-induced Ca2+-released Ca2+ influx Ca2+ IP3-sensitive store Uptake leak IP3-dependent release
Figure 1.14: Schematic diagram of the two-pool model.
1.4.2 The De Young Keizer model
Higher-dimensional models were subsequently developed. The De Young Keizer approach [73] incorporates a detailed model of the IP3 receptors. De Young et
al [73] assumed that the IP3 receptor is composed of three independent and identical
subunits [73]. Each subunit has three binding sites: an IP3-activation binding site;
a Ca2+-activation binding site; and a Ca2+-inactivation binding site. Each of these binding sites can be either occupied or unoccupied. Hence, the subunits have eight different states. Ca2+ flux is induced by the IP3 receptor only if IP3 and Ca2+ are
both bound. Even though this model is based on more realistic assumptions, it does not agree as well with experimental data as does the two-pool model [30].
1.4.3 The Atri model
Atri et al [74] suggested another approach to modelling calcium release, based on the assumption that Ca2+ inactivates the IP3 receptors in a cooperative manner. Their
model assumes that the IP3receptor consists of three binding domains: domain 1 for
IP3 activation; domain 2 for Ca2+ activation; and domain 3 for Ca2+ inactivation.
Accordingly, for the receptor to conduct Ca2+ current, IP3 needs to be bound to
domain 1, Ca2+bound to domain 2, and Ca2+not bound to domain 3. Consequently, Ca2+ activates the IP3 receptor if bound to domain 2 and inactivates it if bound to
domain 3. The model assumes that the Ca2+concentration in the ER is high and so well buffered, that depletion of the ER has a negligible effect on intracellular Ca2+ dynamics [30].
1.4.4 The calcium-induced calcium-release model
Another way to model the calcium release from intracellular stores is through the RyR, which are similar to the IP3 receptors in that Ca2+can activate and inactivate
them, which trigger CICR from the SR/ER. Friel et al [75] developed a simple CICR model that provides a description of the behaviour of the Ca2+dynamics. A diagram of the model is shown in Figure 1.15. CICR is of particular importance in cardiac cells [30], where membrane depolarisation causes a small influx of Ca2+ through a plasma membrane voltage-dependent calcium channel, which, in turn, triggers the release of Ca2+ through the RyR on the membrane of the intracellular store. Subsequently, higher-dimensional models of CICR [17, 76] and considerably more complex ones [77] have been constructed. In the CICR models, it is not yet clear how Ca2+ inactivates the RyR and whether this inactivation plays a significant role [30].
ER/SR
Fluxes exchange with external medium
Ca2+
Fluxes exchange with cytoplasm
Figure 1: cc.
Figure 1.15: Schematic diagram of the CICR model.
1.4.5 A minimal model for the myometriocyte
In cardiac myocytes, calcium enters via voltage-gated calcium channels in the plasma membrane. This calcium entry activates SR calcium release via the RyR. Further- more, calcium is released via IP3-dependent calcium release. On the other hand, in
the myometrium cells of all species, the increase in cytosolic calcium triggered by the action potential arises mostly from the opening of L-type and T-type voltage-gated calcium channels in the plasma membrane. Although this rise can be augmented by release from the internal stores, calcium entry from the extracellular space is the major source of calcium-triggered contractions [29]. We developed a minimal model for MSMC calcium dynamics. The model only considers the increase of [Ca2+]i due
to the plasma membrane calcium channels, while ignoring the Ca2+release from the stores. Details of the minimal model will be shown in Chapter 4.