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Signal transduction in the brain : modulation of receptor-mediated inositol phospholipid breakdown by potassium and fluoride ionsTiger, Gunnar January 1990 (has links)
Neurotransmitter receptor types mediating the generation of intracellular signals are of two types; ligand-gated ion channels and G protein coupled receptors. The effector enzyme phosphoinositide-specific phospholipase C (PLC) is modulated by stimulation of G protein coupled receptors, leading to an increased breakdown of inositol phospholipids ("Ptdlns breakdown").In recent years, the receptors in the brain coupled to PLC and modulation of such receptor-mediated Ptdlns breakdown have been characterised. One such modulation is the "potassium effect", whereby an increase in the assay [K+] from 6 to 18 mM potentiates the Ptdlns breakdown response to the muscarinic agonist carbachol in the rat brain. It has been speculated that this effect is one way of enhancing the signal :noise ratio of muscarinic neurotransmission. The mechanisms responsible for the potassium effect have been studied in this thesis.Initial methodological studies indicated that the temperature of the Krebs buffer used after tissue dissection was an important factor regulating the Ptdlns response to receptor stimulation. Expressing the Ptdlns breakdown response as a fraction of the total labelled phosphoinositides was more useful than other ways of expressing the data. Acid extraction of the Lipid fraction was also superior to neutral extraction.Miniprismspreparedfrompig striatum and hippocampus showed qualitative (but not quantitative) similarities with the rat with respect to stimulation by carbachol, noradrenaline and the potassium effect. Dopamine also stimulated Ptdlns breakdown, though probably via a noradrenergic mechanism.The enhancing actions of potassium appeared to be selective for muscarinic Ml-type receptors. Thus glutamate, quisqualate and NaF-stimulated Ptdlns breakdown are not affected by raised [K+].The potassium effect is brought about by two mechanisms. In calcium-free Krebs buffer, the effect could be mimicked by the calcium channel agonist BAY K-8644 and partially antagonised by verapamil. At an assay [Ca2*] of 2.52 mM, however, modulation of calcium uptake had little effect on carbachol-stimulated Ptdlns breakdown at either normal or raised [K+]. The synergy between potassium and carbachol at252 mM Ca?+ is not dependent upon tissue depolarisation perse, since other ways of depolarising the tissue did not enhance the response to carbachol. It is suggested that potassium might have a direct effect on the muscarinic Ml-type receptor - G protein - PLC complex.In order to investigate this possibility, the effect of fluoride ions (which activate G proteins via formation of AlF4) on basal and carbachol-stimulated Ptdlns breakdown was investigated. Fluoride ions inhibited the enhanced breakdown response to carbachol found at raised [K+]. However, this effect is secondary to effects of fluoride on PLC substrate availibility rather than on G protein function. / digitalisering@umu
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Structural Studies of Arabidopsis Thaliana Inositol Polyphosphate Multi-KinaseEndo-Streeter, Stuart Tamotsu January 2009 (has links)
<p>Inositol Polyphosphate Multi-Kinase (IPMK, also known as ArgRIII, Arg82, and IPK2) is a central component of the inositol signaling system, catalyzing the phosphorylation of at least four different inositol polyphosphate species in vivo with in vitro activity observed for three more. Each of these IP species is sterically unique and the phosphorylation target varies between the 6'-, 3'-, or 5'-hydroxyls, classifying IPMK as a 6/3/5-kinase. The products of IPMK have been linked to multiple processes including cell cycle regulation, transcriptional control, telomere length regulation, mRNA export and various phenotypes including mouse embryonic and fly larvae development, and stress responses in plants and yeast. Linking specific IP species and cellular processes has been complicated by the inability to distinguish between the different effects of the various IP species generated by IPMK. Deletion of IPMK affects the IP populations of all its various substrates and products and therefore the role of a single IP species cannot be tracked. The goals of this work were to address the question of substrate selectivity and develop new tools to probe inositol signaling in vivo through a combination of structural, enzymatic, and genomic techniques.</p><p>The structure of Arabidopsis thaliana IPMK is reported at 2.9Å resolution and in conjunction with a new model of inositol selectivity has been used to design constructs with altered substrate profiles. In vitro and in vivo experiments have confirmed that IPMK identifies substrate inositol polyphosphate species through a recognition surface that requires phosphate groups occupy specific pockets and rejects those with axial phosphate groups in specific regions. In vivo experiments have linked specific inositol polyphosphate species to nitrogen metabolism and temperature sensitivity in yeast and established the potential for these constructs to be used to probe signaling in other organisms.</p> / Dissertation
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The Development Of The Novel Synthesis For ConduritolsCantekin, Seda 01 September 2006 (has links) (PDF)
Conduritols have attracted a great deal of attention in recent years owing to biological activities exhibited by them and also their usefulness in the synthesis of other natural products and pharmaceuticals. Conduritols show interesting inhibitor activity for some glycosidases and serve as important intermediates in the synthesis of inositols. In this study, novel synthetic strategies leading to highly functionalized cyclohexene derivative containing an aromatic ring, namely phenyl-substituted conduritol was achieved successfully for the first time. As the starting material, substituted benzoquinone derivative, biphenyl-2-ol was used. Bromination of one double bond followed by the reduction of the carbonyl groups served the desired skeleton. Further substitution of the bromine atoms and functionalization of the remaining double bond opened up an entry for the synthesis of substituted conduritol and inositol derivatives. Moreover, by the use of hydroquinone as a starting material we developed new synthetic methodologies for the synthesis of another conduritol derivative, bromo-substituted conduritol for the first time. Oxidation of hydroquinone to the corresponding quinone followed by the bromination of one double bond gave the desired product. After the reduction of the carbonyl groups and protection of the hydroxyl groups, the key compound for the synthesis of bromo-substituted conduritol was obtained. As a result, we enabled to synthesize new conduritol derivatives and we had considerable advance for the synthesis of other conduritol derivatives, inositols and quercitols.
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CA²⁺-selective TRPM channels regulate IP₃-dependent CA²⁺ oscillations in the C. elegans intestineXing, Juan, January 2009 (has links)
Thesis (Ph. D. in Pharmacology)--Vanderbilt University, Dec. 2009. / Title from title screen. Includes bibliographical references.
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The modulating effect of myo-inositol and other antidepressants on the mRNA levels and protein expression of selected subcellular enzymes / Marina van RooyenVan Rooyen, Marina January 2005 (has links)
myo-lnositol (mIns), a natural component of the human diet and essential precursor of
several signalling pathways, including that of G protein-coupled receptors, has also been
shown to be effective in the treatment of psychiatric disorders such as depression, obsessive
compulsive disorder and panic disorder. Most likely since mlns is a simple isomer of
glucose, no serious side effects have been reported with its use, even at high oral doses of
mlns. Previous studies suggest that the therapeutic action of mlns may include reduced
serotonin 5HTzA and muscarinic acetylcholine receptor function. An important signal
transduction system that may possibly be involved in the mechanism of action of
antidepressants is phosphoinositide (PI) turnover. In this signalling system PI-phospholipase
C (PLCpl), that is implicated in the in the mechanism of action of antidepressants and
anxiolytics, is activated.
The mechanism of action of mlns, however, still remains elusive and needs further
investigation. In this study a possible modulatory role of 24-hour pre-treatment of human
neuroblastoma cell line (SH-SY5Y) with mlns on mRNA levels and protein expression of
phospholipase C-p1 (PLCP1) and glycogen synthase kinase 3P (GSK3p) was investigated.
The effects of mlns were also compared to that of other prototype antidepressants, such as
fluoxetine (a selective serotonin reuptake inhibitor), imipramine (a tricyclic antidepressant),
lithium and another drug with potential antidepressant effects, sildenafil (phosphodiesterase
5-type (PDE5) inhibitor). Real-time reverse transcription Polymerase Chain Reaction (RTPCR)
was performed in order to investigate the mRNA levels, while protein expression in
membranes and the cytosol fraction of cells were quantified with Western blots.
The expression of PLCPl was decreased after pre-treatments with imipramine or myoinositol
in combination with fluoxetine. In addition, sildenafil alone or in combination with
myo-inositol, also decreased the expression of membrane-bound PLCp1. However, a 24-
hour pre-treatment with lithium did not alter PLCPl expression significantly. Determined
mRNA levels for the expression of PLCPl were consistent in these findings, except for the
inhibition of the mRNA for the expression of PLCPl also after lithium treatment. The reduced
PLCpl mRNA levels after lithium pre-treatment may suggest the involvement of posttranscriptional
modification (or delayed translational effects) of PLCpl after lithium treatment.
The data from the current study suggest that antidepressant action may include
downregulation of PLCPl expression and that modulators of the nitric oxidecGMP pathway
(e.g. sildenafil as a PDE5 inhibitor) may exhibit similar properties. / Thesis (M.Sc. (Pharmacology))--North-West University, Potchefstroom Campus, 2005.
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Castor Bean (Ricinus Communis L.) Genes Involved in Phytic Acid Biosynthetic Pathways: Expression Analysis in Response to Phosphate and Characterization of Inositol 1,3,4,5,6-Pentakisphosphate 2-KinaseYu, Jaeju 18 January 2013 (has links)
During seed development, myo-inositol (Ins) hexakisphosphate or phytic acid (PA) is stored in the form of phytin with mineral cations, and is mobilized following germination, releasing these nutrients that are required for seedling growth. Outside its role in seeds, PA and other phosphoylated Ins derivatives play critical roles in biological processes in many eukaryotes. PA also has negative influences on nutrition in both non-ruminant animals and humans due to its lack of digestibility.
There have been two parallel PA biosynthetic pathways proposed, yet, the pathway is still poorly understood in terms of its regulation and enzymes involved. Here, genes encoding enzymes putatively implicated in castor bean PA biosynthetic pathways were identified in the genome and expression followed. Isolated castor bean embryos have the ability to resynthesize PA following germination if exogenous phosphate is available. It was found that the genes purported to act in PA synthesis were constitutively expressed in the embryos regardless of the availability of phosphate.
Castor bean Ins 1,3,4,5,6-pentakisphosphate 2-kinase (RcIPK1), catalyzing the final reaction in PA biosynthesis, regardless of pathway, was chosen for further study.
Even though only one copy of the RcIPK1 gene was present in the genome, numerous RNA variants were found, most likely due to alternative splicing events. These variants encoded six closely related protein isoforms based on in silico analysis. Functional analyses using yeast mutant strains lacking the IPK1 gene revealed that only three of the mRNA variants could rescue the temperature-sensitive growth phenotype, and it was demonstrated by HPLC analysis of Ins phosphates that their ability to complement the missing yeast IPK1 enzyme was associated with enzyme activity. Only these three isoforms possessed conserved motif III and motif IV important for IPK1 catalytic activity.
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Structural Analysis of the N-terminal Acetyltransferase A ComplexNeubauer, Julie January 2012 (has links)
<p>NatA binds inositol hexakisphosphate and other ligands, and exhibits conformational flexibility dependent on the ligand bound.</p> / Dissertation
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Effets du myo-inositol sur la perméabilité à l'eau d'ovocytes de Xenopus laevis exprimant les formes native et mutée D150E de l'aquaporine-2Lussier, Yoann January 2007 (has links)
Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal
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INORGANIC AND ORGANIC PHOSPHORUS INTERACTIONS WITH HYDROXY-INTERLAYERED SOIL MINERALSShumaker II, Paul D. 01 January 2008 (has links)
Phosphorus (P), a necessary plant and animal nutrient, can also lead to eutrophication of fresh waters when in excess. Appropriate P management is necessary to prevent fresh water pollution. Mineralogy of soil clays has been shown to affect P adsorption, desorption, and movement through soils. Specifically, hydroxy-interlayered minerals have been shown to adsorb and retain inorganic P in soil systems. This study was designed to determine the sorption and desorption characteristics of inorganic, organic, and mixed forms of P interacting with soil hydroxy-interlayered vermiculites (HIV) and smectites (HIS), and compare the findings to sorption and desorption processes of natural aluminum (Al) and Iron (Fe) hydroxide minerals. Results indicate natural Al and Fe hydroxide minerals sorbed and retained P more strongly than hydroxy-interlayered minerals in our samples and inositol hexakisphosphate was more highly sorbed and retained than inorganic P.
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Organic phosphorus speciation in environmental samples : Method development and applicationsParaskova, Julia V. January 2014 (has links)
This thesis investigates the development of new methodology for the identification and quantification of organic phosphorus compounds in environmental samples. Phosphorus is a vital element for primary production and one of the factors contributing to eutrophication. Eutrophication of aquatic systems leads to algal blooms, changes in ecological balance and deteriorating water quality. Difficulties in studying organic phosphorus stem from the fact that organic phosphorus is present in the environment in a variety of forms and each form may have different degradation and turnover time, having very different effects on eutrophication. New methods for the quantification of phosphorus derived from three groups of organic phosphorus compounds were developed. For the determination of phosphorus derived from DNA and phospholipids selective extraction was combined with digestion and colorimetric determination of the extracted phosphate. For quantification of inositol phosphates high performance liquid chromatography was coupled with tandem mass spectrometry using electrospray ionization. The methods were applied to studying the distribution of these compounds in a small catchment and in the case of DNA-P and phospholipid-P, the degradation of the fractions in lake sediments. The studies showed that phosphorus bound to DNA, phospholipids and inositol phosphates constitute a sizeable part of the total phosphorus in different environmental samples. The phospholipid-P fraction was the smallest one, accounting for, on average, only a few percent of the total phosphorus in the sample. Inositol phosphates were most prevalent in the soils, with inositol hexakisphosphate accounting for over 10% of the total phosphorus content. The highest content of DNA-P was found in sediments and it was shown that DNA-P degrades more rapidly than phospholipid-P and therefore plays a more critical role in internal loading.
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