scholarly journals Cyclic Adenosine Monophosphate Regulates Calcium Channels in the Plasma Membrane of Arabidopsis Leaf Guard and Mesophyll Cells

2004 ◽  
Vol 279 (34) ◽  
pp. 35306-35312 ◽  
Author(s):  
Fouad Lemtiri-Chlieh ◽  
Gerald A. Berkowitz
Keyword(s):  
Plasma Membrane ◽  
Calcium Channels ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Mesophyll Cells

scholarly journals Effects of integrin-mediated cell adhesion on plasma membrane lipid raft components and signaling

2011 ◽  
Vol 22 (18) ◽  
pp. 3456-3464 ◽  
Author(s):  
Andrés Norambuena ◽  
Martin A. Schwartz
Keyword(s):  
Plasma Membrane ◽  
Cell Adhesion ◽  
Lipid Raft ◽  
Rho Gtpases ◽  
Cancer Metastasis ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Membrane Lipid ◽  
Cell Detachment ◽  
Suspended Cells

Anchorage dependence of cell growth, which is mediated by multiple integrin-regulated signaling pathways, is a key defense against cancer metastasis. Detachment of cells from the extracellular matrix triggers caveolin-1–dependent internalization of lipid raft components, which mediates suppression of Rho GTPases, Erk, and phosphatidylinositol 3-kinase in suspended cells. Elevation of cyclic adenosine monophosphate (cAMP) following cell detachment is also implicated in termination of growth signaling in suspended cells. Studies of integrins and lipid rafts, however, examined mainly ganglioside GM1 and glycosylphosphatidylinositol-linked proteins as lipid raft markers. In this study, we examine a wider range of lipid raft components. Whereas many raft components internalized with GM1 following cell detachment, flotillin2, connexin43, and Gαs remained in the plasma membrane. Loss of cell adhesion caused movement of many components from the lipid raft to the nonraft fractions on sucrose gradients, although flotillin2, connexin43, and H-Ras were resistant. Gαs lost its raft association, concomitant with cAMP production. Modification of the lipid tail of Gαs to increase its association with ordered domains blocked the detachment-induced increase in cAMP. These data define the effects of that integrin-mediated adhesion on the localization and behavior of a variety of lipid raft components and reveal the mechanism of the previously described elevation of cAMP after cell detachment.

scholarly journals Visualization of β-adrenergic receptor dynamics and differential localization in cardiomyocytes

2021 ◽  
Vol 118 (23) ◽  
pp. e2101119118
Author(s):  
Marc Bathe-Peters ◽  
Philipp Gmach ◽  
Horst-Holger Boltz ◽  
Jürgen Einsiedel ◽  
Michael Gotthardt ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Adrenergic Receptor ◽  
Cardiac Contractility ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Receptor Subtypes ◽  
Specific Cell ◽  
Adult Cardiomyocytes ◽  
Outer Plasma Membrane

A key question in receptor signaling is how specificity is realized, particularly when different receptors trigger the same biochemical pathway(s). A notable case is the two β‐adrenergic receptor (β‐AR) subtypes, β1 and β2, in cardiomyocytes. They are both coupled to stimulatory Gs proteins, mediate an increase in cyclic adenosine monophosphate (cAMP), and stimulate cardiac contractility; however, other effects, such as changes in gene transcription leading to cardiac hypertrophy, are prominent only for β1‐AR but not for β2-AR. Here, we employ highly sensitive fluorescence spectroscopy approaches, in combination with a fluorescent β‐AR antagonist, to determine the presence and dynamics of the endogenous receptors on the outer plasma membrane as well as on the T-tubular network of intact adult cardiomyocytes. These techniques allow us to visualize that the β2‐AR is confined to and diffuses within the T-tubular network, as opposed to the β1‐AR, which is found to diffuse both on the outer plasma membrane as well as on the T-tubules. Upon overexpression of the β2‐AR, this compartmentalization is lost, and the receptors are also seen on the cell surface. Such receptor segregation depends on the development of the T-tubular network in adult cardiomyocytes since both the cardiomyoblast cell line H9c2 and the cardiomyocyte-differentiated human-induced pluripotent stem cells express the β2‐AR on the outer plasma membrane. These data support the notion that specific cell surface targeting of receptor subtypes can be the basis for distinct signaling and functional effects.

scholarly journals Cyclic AMP modulates the rate of ‘constitutive’ exocytosis of apical membrane proteins in Madin-Darby canine kidney cells

1995 ◽  
Vol 108 (5) ◽  
pp. 1931-1943 ◽  
Author(s):  
M. Brignoni ◽  
O.P. Pignataro ◽  
M.L. Rodriguez ◽  
A. Alvarez ◽  
D.E. Vega-Salas ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Adenylate Cyclase ◽  
Apical Membrane ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Apical Plasma Membrane ◽  
Madin Darby Canine Kidney ◽  
Cell Contacts ◽  
Darby Canine Kidney ◽  
Canine Kidney

Madin-Darby canine kidney and other epithelial cell lines (e.g. Caco-2, MCF-10A and MCF-7) develop intracellular vacuoles composed of apical membrane displaying microvilli (VACs) when impaired from forming normal cell-to-cell contacts. In a previous publication, we showed that VACs are rapidly exocytosed upon treatment with 8-Br-3′,5′-cyclic adenosine monophosphate (8-Br-cAMP), a membrane-permeable analog of cAMP, and that this exocytosis correlates with variations in the cellular cAMP concentration in response to the cell-cell contacts. In the present work, we tested the hypothesis that cAMP may be a positive modulator of the ‘constitutive’ exocytic pathway. To mimic conditions in cells with incomplete intercellular contacts, the intracellular levels of cAMP were decreased by means of two independent approaches: (i) pores were induced in the plasma membrane with the polypeptidic antibiotic subtilin, thus allowing small molecules (including cAMP) to permeate and move out of the cytoplasm; and (ii) adenylate cyclase and protein kinase A were blocked with specific inhibitors. In all cases, the intracellular levels of cAMP were measured and, in porated cells, equilibrated to simulate the corresponding physiological intracellular concentrations. The decrease in cAMP within the physiological range resulted in a decreased rate of transport of an apical marker of the constitutive pathway (influenza virus hemagglutinin) from the trans-Golgi network to the apical plasma membrane. Likewise, the delivery of a number of cellular apical proteins to the plasma membrane was retarded at low cAMP concentrations. The inhibitors of adenylate cyclase failed to block basolateral delivery of vesicular stomatitis virus G protein. This differential modulatory effect may represent a differentiation-dependent control of the insertion of apical membrane in epithelial cells.

Effect of dopamine on adenylate cyclase activity, polyphosphoinositide metabolism and cytosolic calcium concentrations in frog pituitary melanotrophs

1993 ◽  
Vol 136 (3) ◽  
pp. 421-429 ◽  
Author(s):  
L. Desrues ◽  
M. Lamacz ◽  
B. G. Jenks ◽  
H. Vaudry ◽  
M. C. Tonon
Keyword(s):  
Calcium Channels ◽  
Inositol Trisphosphate ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Cytosolic Calcium ◽  
Inhibitory Effect ◽  
Intermediate Lobe ◽  
Camp Production ◽  
Membrane Phospholipids ◽  
Voltage Dependent

ABSTRACT It has previously been shown that dopamine plays a pivotal role in the regulation of α-melanocyte-stimulating hormone (α-MSH) secretion from the intermediate lobe of the pituitary. In the present study, we have investigated the various intracellular mechanisms that are associated with the action of dopamine on frog pituitary melanotrophs. Dopamine reduced forskolin-stimulated cyclic adenosine monophosphate (cAMP) production and the inhibitory effect of dopamine was blocked by the dopaminergic D2 receptor antagonist sulpiride. The D2 receptor agonist apomorphine inhibited incorporation of [3H]inositol into membrane phospholipids. Dopamine also inhibited the formation of inositol trisphosphate and provoked accumulation of phosphatidylinositol bisphosphate. The inhibitory effect of dopamine on inositol trisphosphate production was mimicked by D2 receptor agonists and blocked by sulpiride. Using a double-wavelength microfluorimetric approach, we found that dopamine caused a rapid and transient decrease in K+-evoked stimulation of intracellular calcium concentration. The timecourses of the responses of the various intracellular messengers indicate that blockage of voltagedependent calcium channels is the primary event associated with activation of dopamine D2 receptors, while inhibition of polyphosphoinositide breakdown, related to blockage of voltage-dependent calcium channels, and reduction of cAMP production are secondary events which may contribute to the sustained inhibitory effect of dopamine on α-MSH release. Journal of Endocrinology (1993) 136, 421–429

scholarly journals PACmn for improved optogenetic control of intracellular cAMP

BMC Biology ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Shang Yang ◽  
Oana M. Constantin ◽  
Divya Sachidanandan ◽  
Hannes Hofmann ◽  
Tobias C. Kunz ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Signaling Pathways ◽  
Hippocampal Neurons ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Cell Types ◽  
Camp Signaling ◽  
Intracellular Camp ◽  
Extracellular Signals ◽  
Starting Point

Abstract Background Cyclic adenosine monophosphate (cAMP) is a ubiquitous second messenger that transduces extracellular signals in virtually all eukaryotic cells. The soluble Beggiatoa photoactivatable adenylyl cyclase (bPAC) rapidly raises cAMP in blue light and has been used to study cAMP signaling pathways cell-autonomously. But low activity in the dark might raise resting cAMP in cells expressing bPAC, and most eukaryotic cyclases are membrane-targeted rather than soluble. Our aim was to engineer a plasma membrane-anchored PAC with no dark activity (i.e., no cAMP accumulation in the dark) that rapidly increases cAMP when illuminated. Results Using a streamlined method based on expression in Xenopus oocytes, we compared natural PACs and confirmed bPAC as the best starting point for protein engineering efforts. We identified several modifications that reduce bPAC dark activity. Mutating a phenylalanine to tyrosine at residue 198 substantially decreased dark cyclase activity, which increased 7000-fold when illuminated. Whereas Drosophila larvae expressing bPAC in mechanosensory neurons show nocifensive-like behavior even in the dark, larvae expressing improved soluble (e.g., bPAC(R278A)) and membrane-anchored PACs exhibited nocifensive responses only when illuminated. The plasma membrane-anchored PAC (PACmn) had an undetectable dark activity which increased >4000-fold in the light. PACmn does not raise resting cAMP nor, when expressed in hippocampal neurons, affect cAMP-dependent kinase (PKA) activity in the dark, but rapidly and reversibly increases cAMP and PKA activity in the soma and dendrites upon illumination. The peak responses to brief (2 s) light flashes exceed the responses to forskolin-induced activation of endogenous cyclases and return to baseline within seconds (cAMP) or ~10 min (PKA). Conclusions PACmn is a valuable optogenetic tool for precise cell-autonomous and transient stimulation of cAMP signaling pathways in diverse cell types.

scholarly journals Iontophoretic release of cyclic AMP and dispersion of melanosomes within a single melanophore.

1977 ◽  
Vol 74 (3) ◽  
pp. 928-939 ◽  
Author(s):  
I I Geschwind ◽  
J M Horowitz ◽  
G M Mikuckis ◽  
R D Dewey
Keyword(s):  
Plasma Membrane ◽  
Cyclic Amp ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Cyclic Guanosine Monophosphate ◽  
Guanosine Monophosphate ◽  
Melanocyte Stimulating Hormone ◽  
Single Target ◽  
A Cell ◽  
Mean Time

Selective dispersion of melanosomes was often observed after iontophoretic injection of cyclic adenosine monophosphate (AMP) from a glass microelectrode positioned in a target melanophore in frog skin (as viewed from above through a microscope), with other melanophores in the field serving as controls. Because the skin has orderly arrays of several types of closely spaced cells, it is probable that at times the microelectrode also impales cells other than melanophores. When cyclic AMP injection inside a cell resulted in dispersion of melanosomes from a perinuclear position into dendritic processes, the onset of dispersion was relatively rapid, in many cases less than 4 min (mean time of onset, 5.3 +/- 2.9 [SD] min). A much slower dispersion (mean time of onset, 19.0 +/- 5.0 min) of melanosomes was observed when the microelectrode was positioned adjacent to a melanophore, and much larger quantities of cyclic AMP were released. In addition, no changes were observed for injections of 5'-AMP or cyclic guanosine monophosphate (GMP) through electrodes positioned inside or adjacent to melanophores. Potential measurements showed that after impaling a clell, a constant transmembrane potential could often be recorded over many minutes, indicating that the membrane tends to seal around the microelectrode. The results indicate that cyclic AMP acts more rapidly on the inside of a cell than when applied outside a cell and allowed to diffuse through the plasma membrane. This study introduces a model system whereby the properties of the plasma membrane and melanocyte-stimulating hormone (MSH) receptors can be studies within a single target cell.

scholarly journals Acylation of the incretin peptide exendin-4 directly impacts GLP-1 receptor signalling and trafficking

2021 ◽  
Author(s):  
Maria Lucey ◽  
Tanyel Ashik ◽  
Amaara Marzook ◽  
Yifan Wang ◽  
Joëlle Goulding ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Insulin Secretion ◽  
G Protein ◽  
Membrane Lipids ◽  
Acyl Chain ◽  
Cyclic Adenosine Monophosphate ◽  
Adenosine Monophosphate ◽  
Albumin Binding ◽  
Biased Agonism ◽  
Glucagon Like Peptide 1

AbstractThe glucagon-like peptide-1 receptor (GLP-1R) is a class B G protein-coupled receptor and mainstay therapeutic target for the treatment of type 2 diabetes and obesity. Recent reports have highlighted how biased agonism at the GLP-1R affects sustained glucose-stimulated insulin secretion through avoidance of desensitisation and downregulation. A number of GLP-1R agonists (GLP-1RAs) feature a fatty acid moiety to promote albumin binding in order to prolong their pharmacokinetics, but the potential for these ligand changes to influence GLP-1R signalling has rarely been investigated beyond potency assessments for cyclic adenosine monophosphate (cAMP). In this work we directly compare the prototypical GLP-1RA exendin-4 with its C-terminally acylated analogue, exendin-4-C16, for their relative propensities to recruit and activate G proteins and β-arrestins, endocytic and post-endocytic trafficking profiles, and interactions with model and cellular membranes. Both ligands had similar cAMP potency but the exendin-4-C16 showed ∼2.5-fold bias towards G protein recruitment and a ∼60% reduction in β-arrestin-2 recruitment efficacy compared to exendin-4, as well as reduced GLP-1R endocytosis and preferential targeting towards recycling pathways. These effects were associated with a reduced ability to promote the movement of the GLP-1R extracellular domain, as determined using a conformational biosensor approach, and a ∼70% increase in insulin secretion. Interactions with plasma membrane lipids were enhanced by the acyl chain. Exendin-4-C16 showed extensive albumin binding and was highly effective for lowering of blood glucose in mice over at least 72 hours. Overall, our study highlights the importance of a broad approach to the evaluation of GLP-1RA pharmacology.Significance statementAcylation is a common strategy to enhance the pharmacokinetics of peptide-based drugs. Our work shows how acylation can also affect various other pharmacological parameters, including biased agonism, receptor trafficking and interactions with the plasma membrane, which may be therapeutically important.

Sign in / Sign up

Export Citation Format

Share Document