The plasma membrane Ca2+-ATPase (PMCA) is a ubiquitously expressed, ATP-driven Ca2+ pump that is critical for maintaining low resting cytosolic Ca2+ ([Ca2+]i) in all eukaryotic cells. Since cytotoxic Ca2+ overload has such a central role in cell death, the PMCA represents an essential "linchpin" for the delicate balance between cell survival and cell death. In general, impaired PMCA activity and reduced PMCA expression leads to cytotoxic Ca2+ overload and Ca2+ dependent cell death, both apoptosis and necrosis, whereas maintenance of PMCA activity or PMCA overexpression is generally accepted as being cytoprotective. However, the PMCA has a paradoxical role in cell death depending on the cell type and cellular context. The PMCA can be differentially regulated by Ca2+-dependent proteolysis, can be maintained by a localised glycolytic ATP supply, even in the face of global ATP depletion, and can be profoundly affected by the specific phospholipid environment that it sits within the membrane. The major focus of this review is to highlight some of the controversies surrounding the paradoxical role of the PMCA in cell death and survival, challenging the conventional view of ATP-dependent regulation of the PMCA and how this might influence cell fate.

The dynamics of Ca²⁺ release and Ca²⁺‐activated Cl⁻ currents in two related, but functionally distinct exocrine cells, were studied to gain insight into how the molecular specialization of Ca²⁺ signalling machinery are utilized to produce different physiological endpoints: in this case, fluid or exocytotic secretion. Digital imaging and patch‐clamp methods were used to monitor the temporal and spatial properties of changes in cytosolic Ca²⁺ concentration ([Ca²⁺]_c) and Cl⁻ currents following the controlled photolytic release of caged‐InsP₃ or caged‐Ca²⁺. In parotid and pancreatic acinar cells, changes in [Ca²⁺]_c and activation of a Ca²⁺‐activated Cl⁻ current occurred with close temporal coincidence. In parotid, a rapid global Ca²⁺ signal was invariably induced, even with low‐level photolytic release of threshold amounts of InsP₃. In pancreas, threshold stimulation generated an apically delimited [Ca²⁺]_c signal, while a stronger stimulus induced a global [Ca²⁺]_c signal which exhibited characteristics of a propagating wave. InsP₃ was more effective in parotid, where [Ca²⁺]_c signals initiated with shorter latency and exhibited a faster time‐to‐peak than in pancreas. The increased potency of InsP₃ in parotid probably results from a four‐fold higher number of InsP₃ receptors as measured by radiolabelled InsP₃ binding and western blot analysis. The Ca²⁺ sensitivity of the Cl⁻ channels in parotid and pancreas was determined from the [Ca²⁺]‐current relationship measured during a dynamic ‘Ca²⁺ ramp’ produced by the continuous, low‐level photolysis of caged‐Ca²⁺. In addition to a greater number of InsP₃ receptors, the Cl⁻ current density of parotid acinar cells was more than four‐fold greater than that of pancreatic cells. Whereas activation of the current was tightly coupled to increases in Ca²⁺ in both cell types, local Ca²⁺ clearance was found to contribute substantially to the deactivation of the current in parotid. These data reveal specializations of common modules of Ca²⁺‐release machinery and subsequent effector activation that are specifically suited to the distinct functional roles of these two related cell types.

Cancer is caused by excessive cell proliferation and a propensity to avoid cell death, while the spread of cancer is facilitated by enhanced cellular migration, invasion, and vascularization. Cytosolic Ca²⁺ is central to each of these important processes, yet to date, there are no cancer drugs currently being used clinically, and very few undergoing clinical trials, that target the Ca²⁺ signalling machinery. The aim of this review is to highlight some of the emerging evidence that targeting key components of the Ca²⁺ signalling machinery represents a novel and relatively untapped therapeutic strategy for the treatment of cancer.

Acute pancreatitis (AP) is serious inflammatory disease of the pancreas. Accumulating evidence links diabetes with severity of AP, suggesting that endogenous insulin may be protective. We investigated this putative protective effect of insulin during cellular and in vivo models of AP in diabetic mice (Ins2^Akita) and Pancreatic Acinar cell-specific Conditional Insulin Receptor Knock Out mice (PACIRKO). Caerulein and palmitoleic acid (POA)/ethanol-induced pancreatitis was more severe in both Ins2^Akita and PACIRKO vs control mice, suggesting that endogenous insulin directly protects acinar cells in vivo. In isolated pancreatic acinar cells, insulin induced Akt-mediated phosphorylation of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 2 (PFKFB2) which upregulated glycolysis thereby preventing POA-induced ATP depletion, inhibition of the ATP-dependent plasma membrane Ca²⁺ ATPase (PMCA) and cytotoxic Ca²⁺ overload. These data provide the first mechanistic link between diabetes and severity of AP and suggest that phosphorylation of PFKFB2 may represent a potential therapeutic strategy for treatment of AP.

Knowledge of the diversity of ion channel form and function has increased enormously over the last 25 years. The initial impetus in channel discovery came with the introduction of the patch clamp method in 1981. Functional data from patch clamp experiments have subsequently been augmented by molecular studies which have determined channel structures. Thus the introduction of patch clamp methods to study ion channel expression in the choroid plexus represents an important step forward in our knowledge understanding of the process of CSF secretion. Two K⁺ conductances have been identified in the choroid plexus: Kv1 channel subunits mediate outward currents at depolarising potentials; Kir 7.1 carries an inward-rectifying conductance at hyperpolarising potentials. Both K⁺ channels are localised at the apical membrane where they may contribute to maintenance of the membrane potential while allowing the recycling of K⁺ pumped in by Na⁺-K⁺ ATPase. Two anion conductances have been identified in choroid plexus. Both have significant HCO₃ ^- permeability, and may play a role in CSF secretion. One conductance exhibits inward-rectification and is regulated by cyclic AMP. The other is carried by an outward-rectifying channel, which is activated by increases in cell volume. The molecular identity of the anion channels is not known, nor is it clear whether they are expressed in the apical or basolateral membrane. Recent molecular evidence indicates that choroid plexus also expresses the non-selective cation channels such as transient receptor potential channels (TRPV4 and TRPM3) and purinoceptor type 2 (P2X) receptor operated channels. In conclusion, good progress has been made in identifying the channels expressed in the choroid plexus, but determining the precise roles of these channels in CSF secretion remains a challenge for the future.

Acute pancreatitis is a serious and sometimes fatal inflammatory disease where the pancreas digests itself. The non-oxidative ethanol metabolites palmitoleic acid (POA) and POA-ethylester (POAEE) are reported to induce pancreatitis caused by impaired mitochondrial metabolism, cytosolic Ca²⁺ ([Ca²⁺]_i) overload and necrosis of pancreatic acinar cells. Metabolism and [Ca²⁺]_i are linked critically by the ATP-driven plasma membrane Ca²⁺-ATPase (PMCA) important for maintaining low resting [Ca²⁺]_i. The aim of the current study was to test the protective effects of insulin on cellular injury induced by the pancreatitis-inducing agents, ethanol, POA, and POAEE. Rat pancreatic acinar cells were isolated by collagenase digestion and [Ca²⁺]_i was measured by fura-2 imaging. An in situ [Ca²⁺]_i clearance assay was used to assess PMCA activity. Magnesium green (MgGreen) and a luciferase-based ATP kit were used to assess cellular ATP depletion. Ethanol (100 mm) and POAEE (100 μm) induced a small but irreversible Ca²⁺ overload response but had no significant effect on PMCA activity. POA (50–100 μm) induced a robust Ca²⁺ overload, ATP depletion, inhibited PMCA activity, and consequently induced necrosis. Insulin pretreatment (100 nm for 30 min) prevented the POA-induced Ca²⁺ overload, ATP depletion, inhibition of the PMCA, and necrosis. Moreover, the insulin-mediated protection of the POA-induced Ca²⁺ overload was partially prevented by the phosphoinositide-3-kinase (PI3K) inhibitor, LY294002. These data provide the first evidence that insulin directly protects pancreatic acinar cell injury induced by bona fide pancreatitis-inducing agents, such as POA. This may have important therapeutic implications for the treatment of pancreatitis.