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Phosphoinositol/Ca2+ pathway in the cardiac k-opioid receptor: physiological role and alternations upontolerance盛建中, Sheng, Jianzhong. January 1997 (has links)
published_or_final_version / Physiology / Doctoral / Doctor of Philosophy
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Investigation on pre- and postsynaptic Ca²⁺ signaling in neuronal model systems /Krjukova, Jelena, January 2004 (has links)
Diss. (sammanfattning) Uppsala : Univ., 2004. / Härtill 4 uppsatser.
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Effector regulation domains on G[alpha]16 and their role in the activation of phospholipase C[Beta] and other effectors /Yu, Yan Mei. January 2004 (has links)
Thesis (M. Phil.)--Hong Kong University of Science and Technology, 2004. / Includes bibliographical references (leaves 94-103). Also available in electronic version. Access restricted to campus users.
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DNF2 et SYMCRK : deux gènes impliqués dans le contrôle symbiotique des réactions de défense chez Medicago truncatula / DNF2 and SYMCRK : two genes involved in the symbiotic control of defense reaction in Medicago truncatulaBourcy, Marie 21 March 2013 (has links)
Medicago truncatula forme une association symbiotique avec Sinorhizobium meliloti qui conduit à la formation de nodosités fixatrices d’azote. Les cellules symbiotiques végétales accueillent des centaines de bactéries qui restent viables dans la nodosité et se différencient en bactéroïdes fixateurs d’azote. Dans le but de mieux comprendre les mécanismes moléculaires nécessaires à la mise en place de cette interaction, nous avons recherché de nouveaux gènes de plante requis pour une symbiose effective en utilisant des approches de génétique directe et inverse. Des méthodes de biologie cellulaire et moléculaire ont été utilisées pour caractériser le phénotype des mutants et mieux comprendre la fonction biologique de ces gènes.Le gène symbiotique DNF2 code une phosphatidylinositol phospholipase C putative. Les nodosités formées par le mutant dnf2 contiennent une zone de fixation qui est réduite et dans laquelle les rhizobia ne se différencient pas complètement en bactéroïdes. De plus ces nodosités sénescent rapidement et présentent des réactions similaires à des réponses de défense. Sous certaines conditions d’expérimentation, le phénotype sauvage peut être restauré chez ce mutant ce qui montre le caractère conditionnel du phénotype.Le gène symbiotique SYMCRK code un récepteur kinase riche en cystéine. Le phénotype du mutant symCRK est similaire à celui de dnf2, ce qui suggère que ces deux gènes sont impliqués dans des processus aboutissant à des réponses similaires, probablement la persistance des bactéries dans les cellules végétales ou l’inhibition des réactions de défense de la plante. Les phénotypes Fix- atypiques des mutants dnf2 et symCRK suggèrent que les gènes correspondants sont impliqués dans les processus de répression des défenses de la plante et de persistance des bactéroïdes. / Medicago truncatula and Sinorhizobium meliloti form a symbiotic association resulting in the formation of nitrogen-fixing nodules. In the nodules, symbiotic plant cells home and maintain hundreds of viable bacteria which are differentiated into bacteroids, the nitrogen-fixing form of rhizobia. In order to better understand the molecular mechanism sustaining this phenomenon, we used a combination of forward and reverse genetics approaches to identify genes required for nitrogen fixation. In addition we have used cell and molecular biology to characterize the phenotype of the corresponding mutants and to gain an insight into the genes functions.The symbiotic gene DNF2 encodes a putative phosphatidylinositol phospholipase C-like protein. Nodules formed by the mutant contain a zone of infected cells reduced to a few cell layers. In this zone, bacteria do not differentiate properly into bacteroids. Mutant nodules senesce rapidly and they exhibit defense-like reactions. The dnf2 symbiotic phenotype has been shown to be dependent on the experimental conditions.The symbiotic gene SYMCRK encodes a cystein-rich receptor kinase. The symCRK phenotype is similar to dnf2 suggesting that the two genes SYMCRK and DNF2 are participating in similar processes. This atypical phenotype amongst Fix- mutants unravels DNF2 and SYMCRK as new actors of bacteroid persistence inside symbiotic plant cells and repression of plant defense.
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Molecular Mechanisms Underlying Phosphatidylinositol-Specific Phospholipase C Mediated Regulation Of Lipid MetabolismRupwate, Sunny Dinkar 05 1900 (has links) (PDF)
Phosphoinositide-specific phospholipase C (PLC) is involved in Ca2+ mediated signalling events that lead to altered cellular status. PLC activation causes hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) and generates two second messengers, inositol 1,4,5-trisphosphate (IP3) and 1,2-diacylglycerol. Each has distinct role in depending on the cell type in mammalian cells, IP3 binds to intracellular receptors, stimulating the release of sequestered Ca2+. DAG remains in the membrane, where it can activate members of the protein kinase C (PKC) family. In plant absence of PKC keeps the question open as to what is the role of DAG in plants. The role of IP3 apart form triggering calcium release is not known, although the phosphorylated product of IP3 by groups of kinases has been implicated in certain nuclear signalling pathway.
Using various sequence-analysis methods on plant PLC sequences, we identified two conserved motifs in known PLC sequences. The identified motifs are located in the C2 domain of plant PLCs and are not found in any other protein. These motifs are specifically found in the Ca2+ binding loops and form adjoining beta strands. Further, we identified certain conserved residues that are highly distinct from corresponding residues of animal PLCs. The motifs reported here could be used to annotate plant-specific phospholipase C sequences. Furthermore, we demonstrated that the C2 domain alone is capable of targeting PLC to the membrane in response to a Ca2+ signal. We also showed that the binding event results from a change in the hydrophobicity of the C2 domain upon Ca2+ binding. Bioinformatic analyses revealed that all PLCs from Arabidopsis and rice lack a transmembrane domain, myristoylation and GPI-anchor protein modifications. Our bioinformatic study indicates that plant PLCs are located in the cytoplasm, the nucleus and the mitochondria. Our results suggest that there are no distinct isoforms of plant PLCs, as have been proposed to exist in the soluble and membrane associated fractions. The same isoform could potentially be present in both subcellular fractions, depending on the calcium level of the cytosol.
we have used Saccharomyces cerevisiae as a model system to investigate physiological function of PLC in regulation of lipid metabolism. S. cerevisiae synthesizes membrane phospholipids via a pathway which appears to be similar to that of higher eukaryotes. The synthesis of glycerolipid begins with the formation of phosphatidic acid which is quantitatively a minor lipid but is responsible for the repression of UNAINO-containing phospholipid biosynthetic gene by governing localization of Opi1. When the levels of phosphatidic acid are lowered which causes translocation of Opi1 from endoplasmic reticulum membrane to nucleus, where it binds to INO2 of the INO2-INO4 activator complex thereby attenuating transcriptional activation. The expression of phospholipid biosynthetic gene is affected by many conditions which include carbon source, nutrient availability, growth stage, pH and temperature. The well studied conditions which regulate phospholipid biosynthetic genes transcription are through exogenous supplementation of inositol, which is achieved by lowering of phosphatidic acid levels by its utilization for the synthesis of phosphatidylinositol. Since inositol was able to change regulates phospholipid biosynthetic gene we proposed to investigate inositol triphosphate role in such regulation. We overexpressed a plant phospholipase C in yeast to study its effect on lipid biosynthesis. The overexpressed yeast cells were subjected to microarray analysis and the result were confirmed by Q-PCR. The result obtained indicated that there was decrease in the expression of UNAINO-containing genes. To further validate our observation we carried out an in vivo assay to determined activity of enzyme involved in phospholipid biosynthesis. These results were in accordance with our expression analysis further supporting our hypothesis. Our study indicates that phospholipase c regulates phospholipid biosynthesis at transcription level in response to various stimuli.
Overall, these data suggest that the C2 domain of plant PLC plays a vital role in calcium signalling. Further it can be inferred from this study that PI-PLC regulates lipid metabolism in S. cerevisiae.
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A Peptide Selectively Uncoupling BDNF Receptor TrkB from Phospholipase C gamma 1 Prevents Epilepsy and Anxiety-like DisorderGu, Bin January 2015 (has links)
<p>Temporal lobe epilepsy is a common and devastating disorder that features recurrent seizures and is often associated with pathologic anxiety and hippocampal sclerosis. An episode of prolonged seizures (status epilepticus) is thought to promote development of human temporal lobe epilepsy years later. A chemical-genetic approach established proof of concept that transiently inhibiting the receptor tyrosine kinase, TrkB, following status epilepticus prevented epilepsy, anxiety-like behavior and hippocampal damage in a mouse model, providing rationale for developing a therapeutic targeting TrkB signaling. To circumvent the undesirable consequence that global inhibition of TrkB exacerbates neuronal degeneration following status epilepticus, we sought to identify both the TrkB-activated signaling pathway mediating these pathologies and a compound that uncouples TrkB from the responsible signaling effector. To accomplish these goals, we used genetically modified mice and a model of seizures and epilepsy induced by a chemoconvulsant. Genetic inhibition of TrkB-mediated phospholipase C gamma 1 (PLC gamma 1) signaling suppressed seizures induced by a chemoconvulsant, leading to design of a peptide (pY816) that inhibited the interaction of TrkB with PLC gamma 1. We demonstrate that pY816 selectively inhibits TrkB-mediated activation of PLC gamma 1 both in vitro and in vivo. Treatment with pY816 prior to administration of a chemoconvulsant suppressed seizures in a dose- and time-dependent manner. Treatment with pY816 initiated after chemoconvulsant-evoked status epilepticus and continued for just three days suppressed seizure-induction of epilepsy, anxiety-like behavior and hippocampal damage assessed months later. This study elucidates the signaling pathway by which TrkB activation produces diverse neuronal activity-driven pathologies and demonstrates therapeutic benefits of an inhibitor of this pathway in an animal model in vivo. A strategy of uncoupling a receptor tyrosine kinase from a signaling effector may prove useful in diverse diseases in which excessive receptor tyrosine kinase signaling contributes.</p> / Dissertation
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Spatiotemporale Organisation der Interaktion von Gq Protein-Untereinheiten und der Phospholipase Cβ3 / Spatiotemporal patterns of interaction of Gq protein subunits and phospholipase Cβ3Pollinger, Thomas January 2012 (has links) (PDF)
Die G-Protein vermittelte Aktivierung der Phospholipase Cβ (PLCβ) stellt einen primären Mechanismus dar, um eine Vielzahl von physiologischen Ereignissen zu regulieren, z.B. die Kontraktion glatter Muskelzellen, Sekretion oder die Modulation der synaptischen Transmission. Sowohl Gαq- als auch Gβγ-Untereinheiten sind dafür bekannt mit PLCβ Enzymen zu interagieren und diese zu aktivieren. Über die Dynamik dieser Interaktion und den relative Beitrag der G-Protein Untereinheiten ist jedoch nur wenig bekannt. Unter Verwendung Fluoreszenz Resonanz Energie Transfer (FRET)- basierter Methoden in lebenden Zellen, wurde die Kinetik der Rezeptor-induzierten Interaktion zwischen Gβγ und Gαq Untereinheiten, die Interaktion von sowohl der Gαq als auch der Gβγ-Untereinheit mit der PLCβ3 und die Interaktion des regulator of G-Protein signaling 2 (RGS2) mit Gαq-Untereinheiten untersucht. Um die Untersuchung der Protein-Protein-Interaktion auf die Zellmembran zu beschränken, wurde die Total-Internal Reflection Fluorescence (TIRF) Mikroskopie angewandt. Zeitlich hoch auflösendes, ratiometrisches FRET-Imaging offenbarte eine deutlich schnellere Dissoziation von Gαq und PLCβ3 nach Entzug purinerger Agonisten verglichen mit der Deaktivierung von Gq Proteinen in der Abwesenheit der PLCβ3. Dieser offensichtliche Unterschied in der Kinetik kann durch die GTPase-aktivierende Eigenschaft der PLCβ3 in lebenden Zellen erklärt werden. Weiterhin zeigte es sich, dass PLCβ3 die Gq Protein Kinetik in einem ähnlich Ausmaß beeinflusst wie RGS2, welches in vitro deutlich effizienter darin ist, die intrinsische GTPase Aktivität der Gαq-Untereinheit zu beschleunigen. Als Antwort auf die Rezeptorstimulation wurde sowohl eine Interaktion von Gαq-Untereinheiten als auch von Gq-abstammende Gβγ-Untereinheiten mit der PLCβ3 beobachtet. Darüber hinaus zeigte sich auch eine Agonist-abhängige Interaktion von Gαq und RGS2. In Abwesenheit einer Rezeptorstimulation konnte kein spezifisches FRET-Signal zwischen Gq Proteinen und der PLCβ3 oder RGS2 detektiert werden. Zusammengefasst ermöglichte das ratiometrische FRET-Imaging in der TIRF Mikroskopie neue Einsichten in die Dynamik und Interaktionsmuster des Gq-Signalwegs. / G protein-mediated activation of phospholipase Cβ (PLCβ) represents a primary mechanism to regulate many physiological events such induce smooth muscle contraction, secretion and modulation of synaptic transmission. Both Gαq- and Gβγ-subunits are known to interact and activate PLCβ enzymes, however little is known about the dynamics of this interactions and the relative contribution of the G protein subunits in intact cells. Using fluorescence resonance energy transfer- (FRET-) based assays in single intact cells we studies kinetics of receptor-induced interactions between Gβγ- and Gαq-subunits, interactions of both Gαq and Gβγ with PLCβ3 as well as interactions of regulator of G proteins signalling 2 (RGS2) with Gαq- and Gβγ-subunits. In order to restrict the protein/protein interaction studies to the cell membrane we applied total internal reflection (TIRF) microscopy. High temporal resolution ratiometric FRET imaging uncovered a markedly faster dissociation of Gαq and PLC upon withdrawal of purinergic agonists compared to the deactivation of Gq proteins in the absence of PLCβ3. This apparent difference in kinetics could be contributed to the GTPase-activating property of PLCβ3 in living cells. Furthermore we found that PLCβ3 modulated Gq protein kinetics to a similar extent compared to RGS2, which in vitro is about 100 fold more efficient in activating Gq-GTPase activity. We observed that both Gαq subunits and Gq-derived Gβγ-subunits interact with PLCβ3 in response to receptor stimulation. In the absence of receptor stimulation we did neither detect any specific FRET signals between Gq protein subunits and PLCβ3 nor did we detect any interactions between RGS2 and Gαq subunits. Finally we could not detect agonist- dependent FRET between RGS2 and Gβγ-subunits. Taken together, ratiometric FRET-imaging under conditions of TIRF allowed new insights into dynamics and interaction patterns within the Gq signalling pathway.
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Properties of phospholipase C-[Beta]-mediated signaling in H9c2 cardiac myoblasts /Kwon, Sun Hyung. January 1900 (has links)
Thesis (M.S.)--Oregon State University, 2008. / Printout. Includes bibliographical references (leaves 61-67). Also available on the World Wide Web.
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Regulation of phospholipase C and plasma membrane phosphatidylinositol 4,5-bisphosphate in insulin-secreting cells /Thore, Sophia, January 2006 (has links)
Diss. (sammanfattning) Uppsala : Uppsala universitet, 2006. / Härtill 3 uppsatser.
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Regulation of fibronectin assembly by PLC-[gamma]1Crooke, Cornelia. January 2009 (has links)
Thesis (Ph. D. in Biochemistry)--Vanderbilt University, May 2009. / Title from title screen. Includes bibliographical references.
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