Cyclic nucleotides, phosphorylated proteins, and the nervous system.

Some postsynaptic effects of several classes of neurotransmitters appear to be mediated or modulated through the cyclic nucleotides, cyclic AMP and cyclic GMP. Available evidence suggests that the molecular mechanism by which the cyclic nucleotides carry out this second messenger role in nerve cells involves regulation of the state of phosphorylation of specific neuronal proteins. Phosphorylated proteins also appear to be involved in mediating certain of the actions of several other classes of regulatory agents, including calcium and the steroid hormones.     

Extracellular ATP and other nucleotides—ubiquitous triggers of intercellular messenger release

Extracellular nucleotides, and ATP in particular, are cellular signal substances involved in the control of numerous (patho)physiological mechanisms. They provoke nucleotide receptor-mediated mechanisms in select target cells. But nucleotides can considerably expand their range of action. They function as primary messengers in intercellular communication by stimulating the release of other extracellular messenger substances. These in turn activate additional cellular mechanisms through their own receptors. While this applies also to other extracellular messengers, its omnipresence in the vertebrate organism is an outstanding feature of nucleotide signaling. Intercellular messenger substances released by nucleotides include neurotransmitters, hormones, growth factors, a considerable variety of other proteins including enzymes, numerous cytokines, lipid mediators, nitric oxide, and reactive oxygen species. Moreover, nucleotides activate or co-activate growth factor receptors. In the case of hormone release, the initially paracrine or autocrine nucleotide-mediated signal spreads through to the entire organism. The examples highlighted in this commentary suggest that acting as ubiquitous triggers of intercellular messenger release is one of the major functional roles of extracellular nucleotides. While initiation of messenger release by nucleotides has been unraveled in many contexts, it may have been overlooked in others. It can be anticipated that additional nucleotide-driven messenger functions will be uncovered with relevance for both understanding physiology and development of therapy.     

光周期诱导棉铃虫滞育的生化基础

滞育,是有规律地发生在昆虫的某一世代中某一虫态的一种生理生态特性,是昆虫种群适应不良环境的一种生理过程。由于其重要性而得到了生态学家、生理学家、物候学家们的极大重视。通过大量研究,对环境因素诱导、保持和终止滞育的作用,对激素调节滞育的作用,已经有了基本的认识。肯定了光周期和温度是诱导、保持和终止滞育的主要环境因素,     

Changes in the lymphoid tissue and metabolic shifts in the liver in thymus hormone deficiency]

The investigation has been performed on 44 Wistar rats, some of them at the age of 2 months have been subjected to thymectomy. In 3, 6 and 12 months in the spleen, axillary and popliteal lymph nodes density of cell populations, position of lymphatic nodules have been determined, and in the liver--activity of key enzymes of gluconeogenesis. In the animals with the deficiency of the thymic hormones the most manifested changes are seen in 3 and 12 months: decrease in cell population density in the lymph nodes and in their T-dependent zones, decrease in amount of small lymphocytes, in density of the lymphoid nodules arrangement. The dynamics of processes is also followed at analysis of the key enzymes activity of gluconeogenesis in the liver: glucose-6-phosphatase activity is decreased in 3 months after thymectomy, phosphoenolpyruvate carboxycinase activity--in 12 months. Fructose-1,6-diphosphotase activity is not changed. The thymic hormones deficiency is supposed to disturb the ratio of cyclic nucleotides and in this way participate both in inhibition of the immune function and in decrease of metabolic activity mediated by cyclic adenosine monophosphate.     

Pathobiology of biliary epithelia

Cholangiocytes are epithelial cells that line the intra- and extrahepatic biliary tree. They serve predominantly to mediate the content of luminal biliary fluid, which is controlled via numerous signaling pathways influenced by endogenous (e.g., bile acids, nucleotides, hormones, neurotransmitters) and exogenous (e.g., microbes/microbial products, drugs etc.) molecules. When injured, cholangiocytes undergo apoptosis/lysis, repair and proliferation. They also become senescent, a form of cell cycle arrest, which may prevent propagation of injury and/or malignant transformation. Senescent cholangiocytes can undergo further transformation to a senescence-associated secretory phenotype (SASP), where they begin secreting pro-inflammatory and pro-fibrotic signals that may contribute to disease initiation and progression. These and other concepts related to cholangiocyte pathobiology will be reviewed herein. This article is part of a Special Issue entitled: Cholangiocytes in Health and Disease edited by Jesus Banales, Marco Marzioni, Nicholas LaRusso and Peter Jansen.     

Control of ribosomal protein phosphorylation in HeLa cells

The effects of a large series of hormones, cyclic nucleotides and metabolic inhibitors on phosphorylation of ribosomal protein S6 in HeLa cells suggest that at least two metabolic pathways are involved. One responds to insulin and epidermal growth factor; the other responds to adenosine 3′, 5′-cyclic monophosphate. Some phosphodiesterase inhibitors can suppress the phosphorylation of S6 that ordinarily is stimulated by insulin.     

THE INFLUENCE OF SECRETIN, GLUCAGON AND OTHER PEPTIDES, OF AMINO ACIDS, PROSTAGLANDIN ENDOPEROXIDE ANALOGUES AND DIAZEPAM ON THE LEVEL OF ADENOSINE 3'',5''-CYCLIC MONOPHOSPHATE IN NEUROBLASTOMA GLIOMA HYBRID CELLS

Abstract— Neuroblastoma glioma hybrid cells display many properties of neurons. A series of compounds, among them a number of amino acids, peptides and peptide hormones were tested for their ability to influence the level of adenosine 3',5'-cyclic monophosphate (cyclic AMP) in the hybrid cells. Two prostaglandin endoperoxide analogues exhibit a weak stimulatory action, if applied in at least micromolar concentrations. At nanomolar concentrations, only the gastrointestinal hormones secretin and glucagon stimulate the formation of cyclic AMP, as detected in the presence of a phosphodiesterase inhibitor. The effect of secretin but not that of glucagon is antagonized by secretin-(5–27), suggesting that secretin and glucagon act on the cells via different receptors. These results appear to be noteworthy since (a) an effect of secretin or glucagon on a cell with neuronal characteristics has not yet been described, and (b) many peptide hormones have been detected both in the gastrointestinal tract and the nervous system.     

Bromination of guanosine and cyclic GMP confers resistance to metabolic processing by B cells

The metabolic fates of 8-bromoguanosine (8BrGuo) and 8-bromoguanosine-3'5'-cyclic monophosphate (8Br-cGMP) were examined in cultures of murine B lymphocytes. These compounds exert striking immunostimulatory effects upon bone marrow-derived lymphoid cells in vitro. Both 8BrGuo and 8Br-cGMP were resistant to metabolic processing by these cells. That purine metabolic pathways are intact and operant in B cells was demonstrated by the ready degradation and phosphorylation of native guanosine and cyclic GMP. Inaccessibilty of the substrate to the relevant enzymes was ruled out as an explanation by the observation that the brominated compounds also were resistant to processing in broken cell preparations. Moreover, 8BrGuo did not interfere with the cellular machinery for metabolizing native guanosine. The implications of these observations for studying the actions of purine nucleotides, cyclic nucleotides, and their enzymatic processing in B cells are discussed.     

Observations on prostaglandins in normal and leukemic human lymphocytes

Prostaglandins E (PGE) and F2 alpha (PGF2 alpha) were measured in lymphocytes of normal subjects, children with acute lymphocytic leukemia (ALL), and adults with chronic lymphocytic leukemia (CLL). In ALL lymphocytes PGE increased from a normal value of 25 pgrams to 270 pgrams/10(6) cells, and PGF 2 alpha increased from a normal value of 31 pgrams to 482 pgrams/10(6) cells. In CLL lymphocytes, levels of PGE and PGF2 alpha were normal or low. When normal lymphocytes were stimulated with phytohemagglutinin (PHA), the level of PGE and PGF2 alpha fluctuated, followed by corresponding changes in the level of cyclic nucleotides. In cultured ALL lymphocytes, the level of PGE remained high, while cyclic 3':5'-adenosine monophosphate (c-AMP) level was constantly low, and the initial level of PGF2 alpha fluctuated in relation to similar oscillations of cyclic 3':5'-guanosine monophosphate (c-GMP). These values were lower, although not significantly, when ALL lymphocytes were stimulated with PHA. When CLL lymphocytes were stimulated with PHA, the level of PGE remained low (20 pgrams), as did that of c-AMP. The level of PGF2 alpha, after a brief initial increase (130 pgrams), returned to and remained at a lower level (60 pgrams) while the level of c-GMP was persistently high. These results suggest: (1) prostaglandins may indirectly influence the cell cycle, possibly through modulation of cyclase activity and levels of cyclic nucleotides; and (2) some derangement of this regulatory mechanism may be present in leukemic lymphocytes.     

cGMP and cyclic nucleotide-gated channels participate in mouse sperm capacitation

During capacitation of mammalian sperm intracellular [Ca(2+)] and cyclic nucleotides increase, suggesting that CNG channels play a role in the physiology of sperm. Here we study the effect of capacitation, 8Br-cAMP (8-bromoadenosine 3',5'-cyclic monophosphate) and 8Br-cGMP (8-bromoguanosine 3',5'-cyclic monophosphate) on the macroscopic ionic currents of mouse sperm, finding the existence of different populations of sperm, in terms of the recorded current and its response to cyclic nucleotides. Our results show that capacitation and cyclic nucleotides increase the ionic current, having a differential sensitivity to cGMP (cyclic guanosine monophosphate) and cAMP (cyclic adenosine monophosphate). Using a specific inhibitor we determine the contribution of CNG channels to macroscopic current and capacitation.     

Elevation of cyclic 3'5' adenosine monophosphate levels by cholera toxin inhibits the generation of interleukin 2 activity

Several molecules can interact with membrane receptors on mononuclear cells to increase intracellular levels of cyclic adenosine monophosphate (cAMP). We used the cholera toxin (CT), a cAMP elevating agent, to study the influence of this nucleotide on the production of interleukin 2 (IL-2) by human peripheral blood mononuclear cells stimulated by phytohemagglutinin and phorbol myristate acetate. Stimulated generation of IL-2 activity was inhibited by CT but not by its B subunit. The inhibition was potentiated by addition of theophylline. Therefore the synthesis and/or release of IL-2 is controlled by intracellular cAMP levels and may be modulated by agents active on this nucleotide system, such as bacterial toxins, glycoprotein hormones, or neurotransmitters.     

Cyclic nucleotide content of tobacco BY-2 cells

The cyclic nucleotide content of cultured tobacco bright yellow-2 (BY-2) cells was determined, after freeze-killing, perchlorate extraction and sequential chromatography, by radioimmunoassay. The identities of the putative cyclic nucleotides, adenosine 3',5'-cyclic monophosphate (cyclic AMP), guanosine 3',5'-cyclic monophosphate (cyclic GMP) and cytidine 3',5'-cyclic monophosphate (cyclic CMP) were unambiguously confirmed by tandem mass spectrometry. The potential of BY-2 cell cultures as a model system for future investigations of cyclic nucleotide function in higher plants is discussed.     

Studies on cyclic adenosine 3' ,5'-monophosphate levels, Adenylate cyclase and phosphodiesterase activities in the dimorphic fungus Mucor rouxii.

The germination of spores of Mucor rouxii into hyphae was inhibited by 2 mm dibutyryl cyclic adenosine 3′,5′-monophosphate or 7 mm cyclic adenosine 3′,5′-monophosphate; under these conditions spores developed into budding spherical cells instead of filaments, provided that glucose was present in the culture medium. Removal of the cyclic nucleotides resulted in the conversion of yeast cells into hyphae. Dibutyryl cyclic adenosine 3′,5′-monophosphate (2 mm) also inhibited the transformation of yeast to mycelia after exposure of yeast culture to air.Since in all living systems so far studied adenylate cyclase and cyclic adenosine 3′,5′-monophosphate phosphodiesterase are involved in maintaining the intracellular cyclic adenosine monophosphate level, the activity of both enzymes and the intracellular concentration of cyclic adenosine monophosphate were investigated in yeast and mycelium extracts. Cyclic adenosine monophosphate phosphodiesterase and adenylate cyclase activities could be demonstrated in extracts of M. rouxii. The specific activity of adenylate cyclase did not vary appreciably with the fungus morphology. On the contrary, cyclic adenosine monophosphate phosphodiesterase activity was four- to sixfold higher in mycelial extracts than in yeast extracts and reflected quite accurately the observed changes in intracellular cyclic adenosine monophosphate levels; these were three to four times higher in yeast cells than in mycelium.     

Plant purine nucleotide cyclases

Cyclic nucleotides (cAMP and cGMP) play an essential role in many important cellular processes in prokaryotic and eukaryotic organisms. They are produced by purine nucleotide cyclases: adenylyl and guanylyl cyclases. They are classified as one of two distinct forms: soluble and bound to membranes. Beside the differences in enzyme localization, the domain structure and regulation of enzymes activity are also diverse. However, all cyclases possess three groups of important residues: substrate specifying residue, metal binding residues and transition state stabilization residues. The natural occurrence of cyclic nucleotides in plants is now established. It was shown that in higher plants cNMPs act as a second messengers in a large number of (patho)physiological responses. However, it is only recently that the first plant enzymes with AC and GC activity of the unique structure have been identified and functionally characterized. In this study a systematic analysis of all the known prokaryotic, fungal and animal cyclases was done and direct evidences for the presence AC and GC in plant cells were shown.     

Identification of Adenosine-3′,5′-Monophosphate as the Bacterial Attractant for Myxamoebae of Dictyostelium discoideum

Adenosine-3',5'-cyclic monophosphate was shown to be the compound found in Escherichia coli responsible for the attraction of the amoebae of the cellular slime mold Dictyostelium discoideum. A number of other nucleotides were tested and the following were active: tubercidin-3',5'-cyclic monophosphate, N(6)-2'-O-dibutyryl-adenosine-3',5'-cyclic monophosphate, 5'-methylene adenosine-3',5'-cyclic monophosphonate, guanosine-3',5'-cyclic monophosphate, uridine-3',5'-cyclic monophosphate, cytidine-3',5'-cyclic monophosphate, inosine-3',5'-cyclic monophosphate, and thymidine-3',5'-cyclic monophosphate. They were less active than adenosine-3',5'-cyclic monophosphate. It is suggested that cyclic adenosine monophosphate secreted by the bacteria is used by the amoebae as a means of sensing and orienting towards food.     

Observations on prostaglandins in normal and leukemic human lymphocytes

Prostaglandins E (PGE) and F₂ (PGF₂) were measured in lymphocytes of normal subjects, children with acute lymphocytic leukemia (ALL), and adults with chronic lymphocytic leukemia (CLL). In ALL lymphocytes PGE increased from a normal value of 25 pgrams to 270 pgrams/10⁶ cells, and PGF₂ increased from a normal value of 31 pgrams to 482 pgrams/10⁶ cells. In CLL lymphocytes, levels of PGE and PGF₂ were normal or low. When normal lymphocytes were stimulated with phytohemagglutinin (PHA), the level of PGE and PGF₂ fluctuated, followed by corresponding changes in the level of cyclic nucleotides. In cultured ALL lymphocytes, the level of PGE remained high, while cyclic 3′:5′-adenosine monophosphate (c-AMP) level was constantly low, and the initial high level of PGF₂ fluctuated in relation to similar oscillations of cyclic 3′:5′-guanosine monophosphate (c-GMP). These values were lower, although not significantly, when ALL lymphocytes were stimulated with PHA. When CLL lymphocytes were stimulated with PHA, the level of PGE remained low (20 pgrams), as did that of c-AMP. The level of PGF₂, after a brief initial increase (130 pgrams), returned to and remained at a lower level (60 pgrams) while the level of c-GMP was persistently high. These results suggest: (1) prostaglandins may indirectly influence the cell cycle, possibly through modulation of cyclase activity and levels of cyclic nucleotides; and (2) some derangement of this regulatory mechanism may be present in leukemic lymphocytes.     

Molecular pathways of cyclic nucleotide-induced inhibition of arterial smooth muscle cell proliferation

Cyclic adenosine 3',5'-monophosphate (cAMP) and cyclic guanosine 3',5'-monophosphate (cGMP) are second messengers involved in the intracellular signal transduction of a wide variety of extracellular stimuli. These signals regulate many biological processes including cell proliferation, differentiation, migration, and apoptosis. Recently, significant progress has been achieved in the molecular basis underlying cyclic nucleotide regulation of cell proliferation. This review summarizes our knowledge of the signaling pathways regulated by cyclic nucleotides in arterial smooth muscle cells.