Ion channels versus ion pumps: the principal difference, in principle.

The incessant traffic of ions across cell membranes is controlled by two kinds of border guards: ion channels and ion pumps. Open channels let selected ions diffuse rapidly down electrical and concentration gradients, whereas ion pumps labour tirelessly to maintain the gradients by consuming energy to slowly move ions thermodynamically uphill. Because of the diametrically opposed tasks and the divergent speeds of channels and pumps, they have traditionally been viewed as completely different entities, as alike as chalk and cheese. But new structural and mechanistic information about both of these classes of molecular machines challenges this comfortable separation and forces its re-evaluation.

Physiology of Astroglia.

Astrocytes are principal cells responsible for maintaining the brain homeostasis. Additionally, these glial cells are also involved in homocellular (astrocyte-astrocyte) and heterocellular (astrocyte-other cell types) signalling and metabolism. These astroglial functions require an expression of the assortment of molecules, be that transporters or pumps, to maintain ion concentration gradients across the plasmalemma and the membrane of the endoplasmic reticulum. Astrocytes sense and balance their neurochemical environment via variety of transmitter receptors and transporters. As they are electrically non-excitable, astrocytes display intracellular calcium and sodium fluctuations, which are not only used for operative signalling but can also affect metabolism. In this chapter we discuss the molecules that achieve ionic gradients and underlie astrocyte signalling.

Cardiac optogenetics.

Optogenetics is an emerging technology for optical interrogation and control of biological function with high specificity and high spatiotemporal resolution. Mammalian cells and tissues can be sensitized to respond to light by a relatively simple and well-tolerated genetic modification using microbial opsins (light-gated ion channels and pumps). These can achieve fast and specific excitatory or inhibitory response, offering distinct advantages over traditional pharmacological or electrical means of perturbation. Since the first demonstrations of utility in mammalian cells (neurons) in 2005, optogenetics has spurred immense research activity and has inspired numerous applications for dissection of neural circuitry and understanding of brain function in health and disease, applications ranging from in vitro to work in behaving animals. Only recently (since 2010), the field has extended to cardiac applications with less than a dozen publications to date. In consideration of the early phase of work on cardiac optogenetics and the impact of the technique in understanding another excitable tissue, the brain, this review is largely a perspective of possibilities in the heart. It covers the basic principles of operation of light-sensitive ion channels and pumps, the available tools and ongoing efforts in optimizing them, overview of neuroscience use, as well as cardiac-specific questions of implementation and ideas for best use of this emerging technology in the heart.

Mechanism of osmoregulatory adaptation in tilapia.

The shortage of freshwater resource in many countries leads to a shift to develop aquaculture in brackish water and sea water. Tilapias are euryhaline that can thrive from freshwater to full sea water. They and their hybrids are the best candidate species for cultivation in brackish habitats. Thus, understanding their osmoregulatory mechanisms will help to breed or genetically engineer salt tolerant species. In this paper, we review recent progress in understanding the mechanisms of osmoregulatory adaptations in tilapia.

Ion transport proteins anchor and regulate the cytoskeleton.

Structurally diverse ion transport proteins anchor the spectrin-actin cytoskeleton to the plasma membrane by binding directly to linker proteins of the ankyrin and protein 4.1 families. Cytoskeletal anchoring regulates cell shape and restricts the activity of ion transport proteins to specialised membrane domains. New directions are being forged by recent findings that localised anchoring by ion transport proteins regulates the ordered assembly of actin filaments and the actin-dependent processes of cell adhesion and motility.

Mechanisms of action, physiological effects, and complications of hypothermia.

BACKGROUND: Mild to moderate hypothermia (32-35 degrees C) is the first treatment with proven efficacy for postischemic neurological injury. In recent years important insights have been gained into the mechanisms underlying hypothermia's protective effects; in addition, physiological and pathophysiological changes associated with cooling have become better understood. OBJECTIVE: To discuss hypothermia's mechanisms of action, to review (patho)physiological changes associated with cooling, and to discuss potential side effects. DESIGN: Review article. INTERVENTIONS: None. MAIN RESULTS: A myriad of destructive processes unfold in injured tissue following ischemia-reperfusion. These include excitotoxicty, neuroinflammation, apoptosis, free radical production, seizure activity, blood-brain barrier disruption, blood vessel leakage, cerebral thermopooling, and numerous others. The severity of this destructive cascade determines whether injured cells will survive or die. Hypothermia can inhibit or mitigate all of these mechanisms, while stimulating protective systems such as early gene activation. Hypothermia is also effective in mitigating intracranial hypertension and reducing brain edema. Side effects include immunosuppression with increased infection risk, cold diuresis and hypovolemia, electrolyte disorders, insulin resistance, impaired drug clearance, and mild coagulopathy. Targeted interventions are required to effectively manage these side effects. Hypothermia does not decrease myocardial contractility or induce hypotension if hypovolemia is corrected, and preliminary evidence suggests that it can be safely used in patients with cardiac shock. Cardiac output will decrease due to hypothermia-induced bradycardia, but given that metabolic rate also decreases the balance between supply and demand, is usually maintained or improved. In contrast to deep hypothermia (

[The mechanism and control of neuropathic pain].

Neuropathic pain is a debilitating pain that occurs after nerve injury and is generally resistant to currently available treatments including morphine. Such pain involves aberrant excitability in dorsal horn neurons after nerve injury. Emerging evidence indicate that the enhanced activity of dorsal horn neurons requires a communication with microglia. Results of our laboratory have shown that activating P2X4R upregulated in spinal microglia after nerve injury contributes to neuropathic pain through a release of BDNF from microglia, which is a crucial factor to signal to dorsal horn neurons to cause neuronal hyperexcitability. Activated spinal microglia also express P2Y12R, and P2Y12R-KO mice display impaired neuropathic pain. The mechanisms of microglia activation are unknown, but our recent study shows that interferon-gamma (IFN-gamma) can be an important factor that causes spinal microglia activation after nerve injury. IFN-beta upregulates P2X4R in microglia and causes P2X4R-dependent allodynia. These findings suggest that purinoceptors in spinal microglia is crucial for pathological intractable pain.

Salinity tolerance in halophytes.

Halophytes, plants that survive to reproduce in environments where the salt concentration is around 200 mm NaCl or more, constitute about 1% of the world's flora. Some halophytes show optimal growth in saline conditions; others grow optimally in the absence of salt. However, the tolerance of all halophytes to salinity relies on controlled uptake and compartmentalization of Na+, K+ and Cl- and the synthesis of organic 'compatible' solutes, even where salt glands are operative. Although there is evidence that different species may utilize different transporters in their accumulation of Na+, in general little is known of the proteins and regulatory networks involved. Consequently, it is not yet possible to assign molecular mechanisms to apparent differences in rates of Na+ and Cl- uptake, in root-to-shoot transport (xylem loading and retrieval), or in net selectivity for K+ over Na+. At the cellular level, H+-ATPases in the plasma membrane and tonoplast, as well as the tonoplast H+-PPiase, provide the trans-membrane proton motive force used by various secondary transporters. The widespread occurrence, taxonomically, of halophytes and the general paucity of information on the molecular regulation of tolerance mechanisms persuade us that research should be concentrated on a number of 'model' species that are representative of the various mechanisms that might be involved in tolerance.

Acidocalcisomes - conserved from bacteria to man.

Recent work has shown that acidocalcisomes, which are electron-dense acidic organelles rich in calcium and polyphosphate, are the only organelles that have been conserved during evolution from prokaryotes to eukaryotes. Acidocalcisomes were first described in trypanosomatids and have been characterized in most detail in these species. Acidocalcisomes have been linked with several functions, including storage of cations and phosphorus, polyphosphate metabolism, calcium homeostasis, maintenance of intracellular pH homeostasis and osmoregulation. Here, we review acidocalcisome ultrastructure, composition and function in different trypanosomatids and other organisms.

Osmotic tolerance of avian spermatozoa: influence of time, temperature, cryoprotectant and membrane ion pump function on sperm viability.

Potential factors influencing sperm survival under hypertonic conditions were evaluated in the Sandhill crane (Grus canadensis) and turkey (Meleagridis gallopavo). Sperm osmotolerance (300-3000 mOsm/kg) was evaluated after: (1) equilibration times of 2, 10, 45 and 60 min at 4 degrees C versus 21 degrees C; (2) pre-equilibrating with dimethylacetamide (DMA) or dimethylsulfoxide (Me2SO) at either 4 degrees C or 21 degrees C; and (3) inhibition of the Na+/K+ and the Na+/H+ antiporter membrane ionic pumps. Sperm viability was assessed using the eosin-nigrosin live/dead stain. Species-specific differences occurred in response to hypertonic conditions with crane sperm remaining viable under extreme hypertonicity (3000 mOsm/kg), whereas turkey sperm viability was compromised with only slightly hypertonic (500 mOsm/kg) conditions. The timing of spermolysis under hypertonic conditions was also species-specific, with a shorter interval for turkey (2 min) than crane (10 min) sperm. Turkey sperm osmotolerance was slightly improved by lowering the incubation temperature from 21 to 4 degrees C. Pre-equilibrating sperm with DMA reduced the incidence of hypertonic spermolysis only in the crane, at both room and refrigeration temperature. Inhibiting the Na+/K+ and the Na+/H+ antiporter membrane ion pumps did not impair resistance of crane and turkey spermatozoa to hypertonic stress; pump inhibition actually increased turkey sperm survival compared to control sperm. Results demonstrate marked species specificity in osmotolerance between crane and turkey sperm, as well as in the way temperature and time of exposure affect sperm survival under hypertonic conditions. Differences are independent of the role of osmotic pumps in these species.

Pancreatic duct secretion: experimental methods, ion transport mechanisms and regulation.

The pancreatic ductal tree conveys enzymatic acinar products to the duodenum and secretes the fluid and ionic components of pancreatic juice. The physiology of pancreatic duct cells has been widely studied, but many questions are still unanswered concerning their mechanisms of ionic transport. Differences in the transport mechanisms operating in the ductal epithelium has been described both among different species and in the different regions of the ductal tree. In this review we summarize the methods developed to study pancreatic duct secretion both in vivo and in vitro, the different mechanisms of ionic transport that have been reported to date in the basolateral and luminal membranes of pancreatic ductal cells and the regulation of pancreatic duct secretion by nervous, endocrine and paracrine influences.

[Advances in the study of the microbial efflux pumps and its inhibitors development].

Drug resistant bacteria is an increasingly urgent challenge to public health. Bacteria adaptation and extensive abuse of antibiotics contribute to this dilemma. Active efflux of antibiotics is employed by the bacteria to survive the antibiotic pressure. Efflux pump is one of the hot spots of current drug related studies and ideal targets for the improvement of treatment. The efflux pumps and related mechanisms of action, regulation of expression and methodologies were summarized. Comparative genomics analyses were employed to elucidate the underlying mechanisms of action and evolution of efflux pump as exemplified by the Mycobacterium in our lab, which is a crucial re-emerging threat to global public health. The pathway and state-of-art drug development of efflux pump related drugs are included too.

Mechanisms of high salinity tolerance in plants.

Among abiotic stresses, high salinity stress is the most severe environmental stress, which impairs crop production on at least 20% of irrigated land worldwide. In response to high salinity stress, various genes get upregulated, the products of which are involved either directly or indirectly in plant protection. Some of the genes encoding osmolytes, ion channels, receptors, components of calcium signaling, and some other regulatory signaling factors or enzymes are able to confer salinity-tolerant phenotypes when transferred to sensitive plants. Overall, the susceptibility or tolerance to high salinity stress in plants is a coordinated action of multiple stress responsive genes, which also cross talk with other components of stress signal transduction pathways. High salinity exerts its negative impact mainly by disrupting the ionic and osmotic equilibrium of the cell. In saline soils, high levels of sodium ions lead to plant growth inhibition and even death; therefore, mechanisms of salinity tolerance involve sequestration of Na(+) and Cl(-) in vacuoles of the cells, blocking of Na(+) entry into the cell, Na(+) exclusion from the transpiration stream, and some other mechanisms that help in salinity tolerance. Understanding these mechanisms of stress tolerance, along with a plethora of genes involved in the stress signaling network, is important to improve high salinity stress tolerance in crops plants. This chapter first describes the adverse effect of salinity stress and general pathway for the plant stress response, followed by roles of various ion pumps, calcium, SOS pathways, ABA, transcription factors, mitogen-activated protein kinases, glycine betaine, proline, reactive oxygen species, and DEAD-box helicases in salinity stress tolerance. The cross-tolerance between stresses is also mentioned.

Importance of the efflux pump systems in the resistance of Mycobacterium tuberculosis to fluoroquinolones and linezolid.

OBJECTIVE: Our aim was to study the influence of efflux pump systems in the resistance of Mycobacterium tuberculosis to fluoroquinolones and linezolid. METHODS: We studied the mutations in gyrA and gyrB genes and the influence of efflux pump systems with 2 inhibitors (reserpine and MC 207.110). RESULTS: The effect of the active efflux system on the decrease in sensitivity to ciprofloxacin, moxifloxacin, levofloxacin, ofloxacin, gatifloxacin and linezolid was studied by investigating the variation in the in vitro activity of these compounds when assayed in association with reserpine and MC 207.110. These inhibitors exhibit activity both in strains that are resistant and in strains that are susceptible to these antibiotics. However, they are seen to be most active in resistant strains, since the minimum inhibitory concentration of the antibiotics studied in these strains was reduced between 2- and 6-fold. CONCLUSIONS: Therefore, these mechanisms are involved in the resistance to both compounds. It would be of interest to carry out further studies to determine to what extent these active efflux systems influence resistance to the different groups of drugs used in the treatment of tuberculosis, with a view to the possibility of using the inhibitors of these systems in future therapeutic applications.

Steady-state solutions in mathematical models of atrial cell electrophysiology and their stability.

The steady states of the Fenton-Karma, the Courtemanche and the Nygren cell models were studied by determining the fixed points of the dynamical system describing their cell kinetics. The linear stability of the fixed points was investigated, as well as their response to external stimuli. Symbolic calculations were carried out as far as possible in order to prove the existence of these fixed points. In the Fenton-Karma model, a unique stable fixed point was found, namely the resting state. In contrast, the Courtemanche model had an infinite number of fixed points. A bifurcation diagram was constructed by classifying these fixed points according to a conservation law. Initial conditions were identified, for which the dynamical behavior of the cell was auto-oscillatory. In its original formulation, the Nygren model had no fixed point. After having restored charge conservation, the system was found to have an infinite number of fixed points, resulting in a bifurcation diagram similar to that of the Courtemanche model. The approach proposed in this paper assists in the exploration of the high-dimensional parameter space of the cell models and the identification of the conditions leading to spontaneous pacemaker activity.

A preferential role for glycolysis in preventing the anoxic depolarization of rat hippocampal area CA1 pyramidal cells.

During brain anoxia or ischemia, a decrease in the level of ATP leads to a sudden decrease in transmembrane ion gradients [anoxic depolarization (AD)]. This releases glutamate by reversing the operation of glutamate transporters, which triggers neuronal death. By whole-cell clamping CA1 pyramidal cells, we investigated the energy stores that delay the occurrence of the AD in hippocampal slices when O2 and glucose are removed. With glycolytic and mitochondrial ATP production blocked in P12 slices, the AD occurred in approximately 7 min at 33 degrees C, reflecting the time needed for metabolic activity to consume the existing ATP and phosphocreatine, and for subsequent ion gradient decrease. Allowing glycolysis fueled by glycogen, in the absence of glucose, delayed the AD by 5.5 min, whereas superfused glucose prevented the AD for >1 h. With glycolysis blocked, the latency to the AD was 6.5 min longer when mitochondria were allowed to function, demonstrating that metabolites downstream of glycolysis (pyruvate, citric acid cycle intermediates, and amino acid oxidation) provide a significant energy store for oxidative phosphorylation. With glycolysis blocked but mitochondria functioning, superfusing lactate did not significantly delay the AD, showing that ATP production from lactate is much less than that from endogenous metabolites. These data demonstrate a preferential role for glycolysis in preventing the AD. They also define a hierarchy of pool sizes for hippocampal energy stores and suggest that brain ATP production from glial lactate may not be significant in conditions of energy deprivation.

Bioengineered human corneal endothelium for transplantation.

OBJECTIVE: To investigate whether the bioengineered human corneal endothelial cell (HCEC) monolayers harvested from thermoresponsive culture supports could be used as biological tissue equivalents. METHODS: Untransformed adult HCECs derived from eye bank corneas were cultivated on a thermoresponsive poly-N-isopropylacrylamide-grafted surface for 3 weeks at 37 degrees C. Confluent cell cultures with a phenotype and cell density similar to HCECs in vivo were detached as a laminated sheet by lowering the culture temperature to 20 degrees C. In vitro characteristics of the HCEC sheets were determined evaluating their viability and by scanning electron microscopy, immunohistochemistry, and histological studies. RESULTS: After separation from culture surfaces via a thermal stimulus, the HCEC sheets remained viable. Polygonal cell morphology and multiple cellular interconnections were observed throughout the HCEC sheets. Immunolocalization of zonula occludens-1 and Na+,K+-adenosine triphosphatase (ATPase) indicated the formation of tight junctions and the distribution of ionic pumps at the cell boundary. In addition, we ascertained that cultured HCECs have a monolayered architecture that mimics native corneal endothelium. CONCLUSION: These data suggest that a well-organized and functional HCEC monolayer can feasibly be used as tissue equivalents for replacing compromised endothelium.Clinical Relevance Bioengineered human corneal endothelium fabricated from thermoresponsive supports can potentially offer a new therapeutic strategy for corneal endothelial cell loss.

Ion transporters for fluid reabsorption in the rooster (Gallus domesticus) epididymal region.

Testicular fluid is highly condensed during its passage through the epididymal region in the avian species. In the present study, major ion transporters that are responsible for condensation mainly by water resorption in the reproductive tract as identified in the mammalian epididymis were localized within the rooster (Gallus domesticus) epididymis by immunohistochemistry. The results show that the efferent ductule epithelium expressed sodium-potassium ATPase (Na(+),K(+)-ATPase), carbonic anhydrase II (CAII) and sodium hydrogen exchanger isoform 3 (NHE3) and that the connecting ductule and epididymal duct epithelia expressed Na(+),K(+)-ATPase and CAII. These data suggest that a model proposed for reabsorption in mammalian efferent ductules can be applied to avian efferent ductules.

Calcium uptake by plant cells--channels and pumps acting in concert.

How do plant cells accomplish a net uptake of Ca(2+) but keep the membrane potential under control? Consideration of the voltage dependence of the depolarization-activated calcium channel and hyperpolarization-activated calcium channel types, and two other major transporters in the plasma membrane, the H(+)-ATPase and I(K,out), suggests that one channel is well suited for both nutritive and signalling Ca(2+) uptake whereas the other could be limited to a signalling function.

[Effect of plasma membrane ion permeability modulators on respiration and heat output of wheat roots].

A study was made of changes in the rates of respiration, heat production, and membrane characteristics in cells of excised roots of wheat seedlings under the modulation of plasma membrane ion permeability by two membrane active compounds: valinomycin (20 microM (V50)) and chlorpromazine (50 microM (CP50) and 100 microM (CP100)). Both compounds increased the loss of potassium ions, which correlated with the lowering of membrane potential, rate of respiration, and heat production after a 2 h exposure. The differences in alteration of these parameters were due to specific action of either compound on the membrane and to the extent of ion homeostasis disturbance. V20 had a weak effect on the studied parameters. V50 caused an increase of the rate of respiration and heat production, which enhanced following a prolonged action (5 h) and were associated with ion homeostatis restoration. The extent of alteration of membrane characteristics (an increase of potassium loss by roots, and lowering of cell membrane potential) as well as energy expense under the action of CP50 during the first period were more pronounced than in the presence of V50. During a prolonged action of CP50, the increase of respiration intensity and heat production correlated with partial recovery of ion homeostatis in cells. Essential lowering of membrane potential and substantial loss of potassium by cells, starting from the early stages of their response reaction, were followed by inhibition of respiration rate and heat production. Alterations of the structure and functional characteristics of excised root cells indicate the intensification of the membrane-tropic effect of a prolonged action of CP100, and the lack of cell energy resources.