The relationships between cyclic AMP, calcium and prostaglandins as second messengers

There is considerable dissatisfaction with present second messenger hypotheses involving cyclic nucleotides and calcium. The recent findings that methyl xanthines and adenosine are prostaglandin antagonists casts doubt on much of the evidence in favour of the cyclic AMP hypothesis. There is evidence that allosteric sites may modify the binding of calcium and cyclic nucleotides to key cellular regulators and that a range of substances including steroids, adenosine and prostaglandins may occupy those sites. This concept introduces much needed flexibility into the second messenger concept, allows many experiments to be reinterpreted and has major implications throughout the biomedical sciences.     

Lipid raft redox signaling: molecular mechanisms in health and disease

Lipid rafts, the sphingolipid and cholesterol-enriched membrane microdomains, are able to form different membrane macrodomains or platforms upon stimulations, including redox signaling platforms, which serve as a critical signaling mechanism to mediate or regulate cellular activities or functions. In particular, this raft platform formation provides an important driving force for the assembling of NADPH oxidase subunits and the recruitment of other related receptors, effectors, and regulatory components, resulting, in turn, in the activation of NADPH oxidase and downstream redox regulation of cell functions. This comprehensive review attempts to summarize all basic and advanced information about the formation, regulation, and functions of lipid raft redox signaling platforms as well as their physiological and pathophysiological relevance. Several molecular mechanisms involving the formation of lipid raft redox signaling platforms and the related therapeutic strategies targeting them are discussed. It is hoped that all information and thoughts included in this review could provide more comprehensive insights into the understanding of lipid raft redox signaling, in particular, of their molecular mechanisms, spatial-temporal regulations, and physiological, pathophysiological relevances to human health and diseases.     

Membranes as messengers in T cell adhesion signaling

Talin and RapL are components of molecular pathways that regulate the avidity of the integrin lymphocyte function-associated antigen 1 (LFA-1) for its ligand, intercellular adhesion molecule 1. In this review, we discuss recent advances in our understanding of LFA-1 affinity regulation and signaling and discuss a scenario for how Talin and Rap1 might act in synergy to achieve regulation of LFA-1 that is tailored to the specific functional requirements of different situations. Speedy delivery of signals may be crucial, and membrane trafficking from endosomes and the Golgi apparatus seem to be essential in delivering the messages from spatially segregated surface receptors.     

Cholinoreceptor neurons of the snail: Identification,plasticity, and its regulation by opioids and second messengers

A review of the author's own data on the cholinoreceptors of identified neurons of the snail and the regulation of their plasticity. The following sections are presented: the identification of the cholinoreceptors; the coupling of the cholinoreceptors with ion channels; the identification of the opioid receptors; the modulation by opioids of the activity of the cholinoreceptors; the plasticity of the cholinoreceptors; the regulation of the plasticity of the cholinoreceptors by opioids; the regulation of the plasticity of the cholinoreceptors by second messengers; the second messengers involved in the regulation of the plasticity of the cholinoreceptors by the opiate kappa agonist, bremazocine. Translated from Zhurnal Vysshei Nervnoi Deyatel'nosti imeni I. P. Pavlova, Vol. 42, No. 6, pp. 1271–1286, November–December, 1992.     

Structural similarities among malaria toxins insulin second messengers, and bacterial endotoxin.

Malaria toxin causes hypoglycemia and induction of tumor necrosis factor. Extracts of parasitized erythrocytes which were coeluted and copurified with one of the two subtypes of mammalian insulin-mimetic inositolphosphoglycans similarly induced fibroblast proliferation in the absence of serum. In addition, induction of tumor necrosis factor in macrophages by malaria toxin and by lipopolysaccharide from Escherichia coli was enhanced by pretreatment of these toxins with alpha-galactosidase. Thus, parasitized erythrocytes contain both soluble inositolphosphoglycan-like insulin second messengers and endotoxin-like lipidic molecules.     

Phosphoinositide lipid second messengers: new paradigms for transepithelial signal transduction

Multiple forms of phosphatidylinositol are generated by differential phosphorylation of the inositol headgroup. These phosphoinositides, specifically PI(4,5)P₂, have been implicated as modulators in a variety of transport processes. The data indicate that phosphoinositides can modulate transporters directly or via the activation of down-stream signaling components. The phosphoinositide pathway has been linked to changes in transporter kinetics, intracellular signaling, membrane targeting and membrane stability. Recent results obtained for several of the well-characterized transport systems suggest the need to reassess the role of PI(4,5)P₂ and question whether lower abundance forms of the phosphoinositides, notably PI(3,4,5)P₃ (PIP₃) and PI(3,4)P₂, are the pertinent transport regulators. In contrast to PI(4,5)P₂, these latter forms represent a dynamic, regulated pool, the characteristics of which are more compatible with the nature of signaling intermediates. A recently described, novel transepithelial signaling pathway has been demonstrated for PIP₃ in which a signal initiated on the basolateral membrane is transduced to the apical membrane entirely within the planar face of the inner leaflet of the plasma membrane. The new paradigms emerging from recent studies may be widely applicable to transporter regulation in other cell types and are particularly relevant for signaling in polarized cells.     

Lipid raft redox signaling platforms in endothelial dysfunction

In response to various stimuli, membrane lipid rafts (LRs) are clustered to aggregate or recruit NADPH oxidase subunits and related proteins in vascular endothelial cells (ECs), forming redox signaling platforms. These LR signaling platforms may play important roles in the normal regulation of endothelial function and in the development of endothelial dysfunction or injury under pathological conditions. This LR-mediated mechanism now takes center stage in cell signaling for the regulation of many cellular activities or cell function such as ECs redox signaling, phagosomal activity of phagocytes, and cell apopotosis of lymphocytes. This brief review summarizes current evidence that relates to the formation of LR redox signaling platforms and their features in ECs, the functional significance of these signaling platforms in mediating death receptor activation-induced endothelial dysfunction, and the mechanisms initiating or promoting the formation of these platforms. It is expected that information provided here will help readers to understand this new signaling mechanism and perhaps extend the LR signaling platform concept to other research areas related to death receptors, redox signaling, endothelial biology, and cell/molecular biology of the cardiovascular system.     

Galanin receptors and their second messengers in the lumbar dorsal spinal cord

The ligand binding properties of galanin receptors were examined in crude synaptosomal fraction preparations of lumbar dorsal spinal cord, using chloramin-T mono-iodinated porcine Tyr²⁶ galanin as ligand. The equilibrium binding of [¹²⁵I]galanin showed the presence of a single population of high-affinity binding sites with K_D= 0.6±0.2 nM in a concentration of 55±15 fmol mg^-1 protein (B_max. The N-terminal fragments galanin (1–16) and galanin (1–12) fully displaced specific [¹²⁵1]galanin binding from membranes with IC₅₀ values 6 nM and 4 μM, respectively. The C-terminal fragment galanin (17–29) did not displace [¹²⁵I]galanin when applied in the concentration range 10^-11–10^-4 M. GTP inhibited the specific binding of [¹²⁵I] galanin in a concentration dependent manner, with 54% inhibition at 1 mM, suggesting that the galanin receptor found in lumbar dorsal spinal cord is G-protein coupled. Second messenger systems, through which the galanin receptor in lumbar dorsal spinal cord may exert its effect, were also studied. A galanin (10,μm) produced inhibition (58%) of the depolarization induced cGMP increase was found, whereas galanin (10 μM) did not inhibit the noradrenalin (100 μM) activated CAMP synthesis or phosphoinositide turnover in tissue slices of the spinal cord. Bilateral transection of the sciatic nerve at midthigh level 14 days prior to the binding experiment was performed, a treatment which is known to cause a dramatic increase of galanin-like immunoreactivity in the superficial layers of the dorsal spinal cord, dorsal root ganglion and in galanin mRNA levels, but no significant effect on B_max or K_D of the galanin receptor was found.     

Human nicotinic acetylcholine receptor: the influence of second messengers on activation and desensitization

The amplitude and time course of acetylcholine(ACh)-induced membrane current were determined in cells of the human medulloblastoma cell line TE 671. ACh was applied and washed out very rapidly (about 50 ms) by a shift of a cell between two streams of solution, one of which contained the transmitter. ACh-induced current was recorded in the whole-cell mode of patch clamping. The time course of activation of the ACh-induced current could not be resolved because the method of ACh application was still too slow. Desensitization started immediately with ACh application; it could be described by two time constants. With an ACh concentration of 3 μM, the fast time constant was about 0.5 s and the slow time constant was about 3.9 s. When the ACh concentration was raised in steps to 100 mM, the peak amplitude of the current increased, reached a maximum at 1 mM and decreased again. The rate of desensitization was directly correlated with increasing ACh concentration. Current amplitude and desensitization time constants were not affected by intracellular application of cAMP or of a catalytic subunit of a cAMP-dependent protein kinase. When the intracellular calcium concentration was raised at a constant magnesium concentration, desensitization time constants remained unaffected, but the current amplitude decreased. This decrease is not caused by a decrease in single-channel conductance, therefore it may represent a decrease in the number of activatable channels.     

Lipid second messengers derived from glycerolipids and sphingolipids,and their role in smooth muscle function*

The processes that link activation of an external receptor to the internal mechanisms that elicit a physiological response have been the subject of extensive investigation. It has been established that rather than just being an inert barrier to protect the cell from environmental damage, there are populations of phospholipids located within the plasma membrane that act as a reservoir for signalling molecules and when a receptor binds its appropriate activating ligand a chain of events is initiated which leads to the breakdown of these lipids and the release of second messengers. Such processes are rapid enough for physiological responses to be effected. The purpose of this review is to examine the profile of lipid second messengers derived from glycerophospholipids and sphingolipids. In the former class are included phosphoinositide and phosphatidylcholine and the latter includes sphingomyelin. Hydrolysis of such parent compounds is mediated by phospholipases and the profile of metabolites appears to be agonist specific and modulated by a number of mechanisms including heterotrimeric G-protein subunits, small G-proteins, alterations in intracellular calcium concentration, protein kinase C and tyrosine kinases. The recent interest in sphingolipids, particularly in vascular smooth muscle cells, has been provoked by the observation that ceramide and sphingoid base formation is observed in response to vasoconstrictor hormones.     

Galanin receptors and their second messengers in the lumbar dorsal spinal cord.

The ligand binding properties of galanin receptors were examined in crude synaptosomal fraction preparations of lumbar dorsal spinal cord, using chloramin-T mono-iodinated porcine Tyr26 galanin as ligand. The equilibrium binding of [125I]galanin showed the presence of a single population of high-affinity binding sites with KD = 0.6 +/- 0.2 nM in a concentration of 55 +/- 15 fmol mg-1 protein (Bmax). The N-terminal fragments galanin (1-16) and galanin (1-12) fully displaced specific [125I]galanin binding from membranes with IC50 values 6 nM and 4 microM, respectively. The C-terminal fragment galanin (17-29) did not displace [125I]galanin when applied in the concentration range 10(-11)-10(-4) M. GTP inhibited the specific binding of [125I] galanin in a concentration dependent manner, with 54% inhibition at 1 mM, suggesting that the galanin receptor found in lumbar dorsal spinal cord is G-protein coupled. Second messenger systems, through which the galanin receptor in lumbar dorsal spinal cord may exert its effect, were also studied. A galanin (10 microM) produced inhibition (58%) of the depolarization induced cGMP increase was found, whereas galanin (10 microM) did not inhibit the noradrenalin (100 microM) activated cAMP synthesis or phosphoinositide turnover in tissue slices of the spinal cord. Bilateral transection of the sciatic nerve at midthigh level 14 days prior to the binding experiment was performed, a treatment which is known to cause a dramatic increase of galanin-like immunoreactivity in the superficial layers of the dorsal spinal cord, dorsal root ganglion and in galanin mRNA levels, but no significant effect on Bmax or KD of the galanin receptor was found.     

Sphingosine 1-phosphate: a prototype of a new class of second messengers

Sphingosine 1-phosphate (SPP) is an important sphingolipid-derived second messenger in mammalian cells that acts to promote proliferation and to inhibit apoptosis. Various growth factors increase the intracellular concentration of SPP by activating sphingosine kinase, the molecular cloning of which has revealed that it defines a new type of lipid kinase. Cell fate is influenced by the balance between the intracellular concentration of SPP and that of ceramide, a pro-apoptotic sphingolipid metabolite. The observation that a similar "rheostat" is a determinant of cell survival in yeast cells exposed to heat shock indicates that it is an evolutionarily conserved mechanism of stress regulation. SPP also acts extracellularly to inhibit cell motility and to influence cell morphology, effects that appear to be mediated by the G protein-coupled receptor EDG1. These observations indicate that SPP is the prototype of a new class of lipid mediators that exert both intracellular and extracellular actions.     

Co-stimulation and counter-stimulation: lipid raft clustering controls TCR signaling and functional outcomes

T cell receptor (TCR) antigen recognition induces the formation of a specialized 'immunological synapse' at the T cell : antigen presenting cell (APC) junction. This junction is generated by the recruitment and exclusion of particular proteins from the contact area and is required for T cell activation. We and others have hypothesized that lipid raft/non-raft partitioning provides a molecular basis for protein sorting which organizes the TCR, co-stimulators, signal transducers and the actin cytoskeleton at the T cell : APC interface. Here we discuss the emerging paradigm that co-stimulators induce the directional transport and clustering of lipid rafts at the T cell : APC interface, thus generating platform(s) specialized for processive and sustained TCR signal transduction and T cell activation. We also discuss recent data implicating the involvement of 'counter-stimulators' and other negative regulators which prevent optimal raft clustering at the TCR contact site and, thus, facilitate T cell inactivation and tolerance induction.     

Even stressed cells are individuals: second messengers of free radicals in pathophysiology of cancer

Pathophysiological processes associated with disturbances in cell and tissue oxidative homeostasis, are associated with self-catalyzed process of lipid peroxidation. The end products of lipid peroxidation are reactive aldehydes such as 4-hydroxy-2-nonenal (HNE), acting as "second messengers of free radicals." Although reactive aldehydes were first recognized only as cytotoxic, new evidence has come to light, related to their cell growth regulatory functions achieved through cell signaling. The variable appearance of HNE in several organs indicates that its mode of action might be related to an individual cell stress adaptation. The underlying mechanism could be that specific mutations and epigenetic changes on one hand interfere with hormesis on the other. The precise role of oxidative stress and lipid peroxidation in these processes still needs more clarification at molecular level. Finally, an individual approach to each patient, based on the individual cell response to stress, opens a new possibility of integrative medicine in cancer treatment and strongly supports modern concepts of personalized medicine.     

Lipid raft compartmentalization of urokinase receptor signaling in human neutrophils

Urokinase plasminogen activator (uPA) receptors (uPAR) can be engaged for activation signaling either by aggregation or by binding exogenous uPA. These signaling mechanisms require uPAR to associate with two distinct adhesion proteins, L-selectin and complement receptor 3 (CR3), respectively. uPAR contains a glycosylphosphatidylinositol anchor, suggesting that it is concentrated within glycosphingolipid-enriched microdomains, or "lipid rafts". This study was undertaken to determine the extent to which uPAR-mediated signaling is compartmentalized to lipid rafts. Human neutrophil uPAR was cross-linked or stimulated with uPA after pretreatment with the lipid raft-disrupting agents, methyl-beta-cyclodextrin or filipin III. Both agents suppressed increases in intracellular Ca(2+) concentrations ([Ca(2+)](i)) triggered by cross-linking, but did not affect [Ca(2+) ](i) in response to uPA. Neutrophil membranes were separated into lipid raft and non-raft fractions, revealing the presence of uPAR and L-selectin, but the virtual absence of CR3 alpha chain in lipid rafts, either constitutively or in response to uPAR aggregation. Fluorescence resonance energy transfer experiments confirmed close proximity of a lipid raft marker to both uPAR and L-selectin, but not CR3. We conclude that uPAR can engage distinct signaling pathways involving different partner proteins that are functionally and physically segregated from one another in both lipid raft and non-raft domains of the plasma membrane.     

Lipids in dendritic cell biology: messengers, effectors, and antigens

Dendritic cells (DC) are the most professional APC, which induce and coordinate immune responses. The principal task of DC is T cell activation, although DC also interact with and regulate other cell types. The present review serves to illustrate the increasing evidence that lipids play an important role in DC biology. In addition to being fuel stores and structural components of cellular membranes such as in other cell types, lipids act as second messengers and as effectors throughout all steps of DC differentiation and regulate important DC functions. The recent finding that DC synthesize lipid antigens in response to bacterial stimulation and induce antibacterial, CD1-restricted T cells through antigenic mimicry further emphasizes the important role of lipids and DC at the blurring boundaries of innate and adaptive immunity.     

Effect of nitroglycerine on content of second messengers in myocytes of rat thoracic aorta

We evaluated the concentration dependence and time dependence of the effect of nitroglycerine on intracellular content of cAMP, cGMP, and free Ca²⁺. It was shown that after norepinephrine stimulation, nitroglycerine exhibited calcium-blocking activity in lower concentrations (starting from 10⁻⁷ M). Under conditions of experimental nitrate tolerance the dose-dependent effect of nitroglycerine on intracellular cGMP and Ca²⁺ was less pronounced. Calcium-blocking activity of nitroglycerine decreased most significantly upon stimulation of myocytes with norepinephrine. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 141, No. 5, pp. 526–529, May, 2006     

A novel hypothesis for the psycho-modulating effects of lithium: the role of essential fatty acids, eicosanoids and sub-cellular second messengers

Evidence is presented for a novel proposal for the mechanism of action of lithium in manic depressive psychosis. Lithium has well established effects on catecholaminergic--and hormone--stimulated adenyl cyclase activity and on cyclic AMP formation. Although there is conflicting evidence in the literature concerning the effects of the ion on cyclic nucleotide phosphodiesterase, not much is known of the effects of lithium on cyclic GMP. These two second messengers have been proposed to be mutually antagonistic in their actions but that a physiological balance between the two is essential for maintaining homeostasis of the human psyche. An in vivo animal study was undertaken to determine the effects of chronic lithium treatment on the dynamics and kinetics of these two cyclic nucleotides and phosphodiesterase in rat cerebral cortex. From these results, a possible functional coupling mechanism between the two second messenger systems and the effects of lithium are proposed. Lithium by means of its specific site of action, is unique among psychoactive drugs in that it can control both phases of bipolar illness. This point of action is proposed to be the metabolism of free fatty acids where lithium, by altering the availability of precursors for eicosanoid metabolism, is able to modulate both noradrenergic- and cholinergic-dependent pathways. By doing this, the ion is able to reestablish lost control over adrenergic and cholinergic balance critical for thought process and mood stability.