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Article: Eukaryotic chemotaxis at a glance.

Bagorda, Anna / Parent, Carole A

2008 Volume 121, Issue Pt 16, Page(s) 2621–2624

MeSH term(s)	Animals ; Chemotactic Factors/physiology ; Chemotaxis/physiology ; Cytoskeleton/physiology ; Eukaryotic Cells/physiology ; Humans ; Models, Biological ; Receptors, Cell Surface/physiology ; Signal Transduction/physiology
Chemical Substances	Chemotactic Factors ; Receptors, Cell Surface
Language	English
Publishing date	2008-08-06
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2993-2
ISSN	1477-9137 ; 0021-9533
ISSN (online)	1477-9137
ISSN	0021-9533
DOI	10.1242/jcs.018077
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Article: Eukaryotic chemotaxis at a glance

Bagorda, Anna / Parent, Carole A

Journal of cell science. 2008 Aug. 15, v. 121, no. 16

2008

Language	English
Dates of publication	2008-0815
Size	p. 2621-2624.
Publishing place	The Company of Biologists Limited
Document type	Article
ZDB-ID	2993-2
ISSN	1477-9137 ; 0021-9533
ISSN (online)	1477-9137
ISSN	0021-9533
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Direct biochemical measurements of signal relay during Dictyostelium development.

Das, Satarupa / Rericha, Erin C / Bagorda, Anna / Parent, Carole A

The Journal of biological chemistry

2011 Volume 286, Issue 44, Page(s) 38649–38658

Abstract: Upon starvation, individual Dictyostelium discoideum cells enter a developmental program that leads to collective migration and the formation of a multicellular organism. The process is mediated by extracellular cAMP binding to the G protein-coupled cAMP ...

Abstract	Upon starvation, individual Dictyostelium discoideum cells enter a developmental program that leads to collective migration and the formation of a multicellular organism. The process is mediated by extracellular cAMP binding to the G protein-coupled cAMP receptor 1, which initiates a signaling cascade leading to the activation of adenylyl cyclase A (ACA), the synthesis and secretion of additional cAMP, and an autocrine and paracrine activation loop. The release of cAMP allows neighboring cells to polarize and migrate directionally and form characteristic chains of cells called streams. We now report that cAMP relay can be measured biochemically by assessing ACA, ERK2, and TORC2 activities at successive time points in development after stimulating cells with subsaturating concentrations of cAMP. We also find that the activation profiles of ACA, ERK2, and TORC2 change in the course of development, with later developed cells showing a loss of sensitivity to the relayed signal. We examined mutants in PKA activity that have been associated with precocious development and find that this loss in responsiveness occurs earlier in these mutants. Remarkably, we show that this loss in sensitivity correlates with a switch in migration patterns as cells transition from streams to aggregates. We propose that as cells proceed through development, the cAMP-induced desensitization and down-regulation of cAMP receptor 1 impacts the sensitivities of chemotactic signaling cascades leading to changes in migration patterns.
MeSH term(s)	Adenylyl Cyclases/metabolism ; Biochemistry/methods ; Chemotaxis/physiology ; Cyclic AMP/metabolism ; Cyclic AMP-Dependent Protein Kinases/metabolism ; Dictyostelium/growth & development ; Fluorescence Resonance Energy Transfer/methods ; MAP Kinase Signaling System ; Models, Biological ; Mutation ; Phosphorylation ; Receptors, Cyclic AMP/metabolism ; Signal Transduction
Chemical Substances	Receptors, Cyclic AMP ; Cyclic AMP (E0399OZS9N) ; Cyclic AMP-Dependent Protein Kinases (EC 2.7.11.11) ; Adenylyl Cyclases (EC 4.6.1.1)
Language	English
Publishing date	2011-09-12
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.M111.284182
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Article: Chemotaxis: moving forward and holding on to the past.

Bagorda, Anna / Mihaylov, Vassil A / Parent, Carole A

Thrombosis and haemostasis

2006 Volume 95, Issue 1, Page(s) 12–21

Abstract: The ability of cells to sense external chemical cues and respond by directionally migrating towards them is a fundamental process called chemotaxis. This phenomenon is essential for many biological responses in the human body, including the invasion of ... ...

Abstract	The ability of cells to sense external chemical cues and respond by directionally migrating towards them is a fundamental process called chemotaxis. This phenomenon is essential for many biological responses in the human body, including the invasion of neutrophils to sites of inflammation. Remarkably, many of the molecular mechanisms involved in controlling neutrophils chemotaxis arose millions of years ago in the simple eukaryotic organism Dictyostelium discoideum. Both neutrophils and Dictyostelium use G protein-coupled signaling cascades to mediate chemotactic responses, which are responsible for transducing external cues into highly organized cytoskeletal rearrangements that ultimately lead to directed migration. By using the genetically and biochemically tractable organism Dictyostelium as a model system, it has been possible to decipher many of the signal transduction events that are involved in chemotaxis.
MeSH term(s)	Actins/metabolism ; Animals ; Chemotactic Factors/metabolism ; Chemotaxis ; Chemotaxis, Leukocyte ; Cytoskeleton/metabolism ; Dictyostelium/enzymology ; Humans ; Models, Biological ; Myosin Type II/metabolism ; Neutrophils/enzymology ; PTEN Phosphohydrolase/metabolism ; Phosphatidylinositol 3-Kinases/metabolism ; Receptors, G-Protein-Coupled/metabolism ; Signal Transduction
Chemical Substances	Actins ; Chemotactic Factors ; Receptors, G-Protein-Coupled ; Phosphatidylinositol 3-Kinases (EC 2.7.1.-) ; PTEN Phosphohydrolase (EC 3.1.3.67) ; Myosin Type II (EC 3.6.1.-)
Language	English
Publishing date	2006-01
Publishing country	Germany
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	518294-3
ISSN	0340-6245
ISSN	0340-6245
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Article: Real-time measurements of cAMP production in live Dictyostelium cells

Bagorda, Anna / Das, Satarupa / Rericha, Erin C / Chen, David / Davidson, Jean / Parent, Carole A

Journal of cell science. 2009 Nov. 1, v. 122, no. 21

2009

Abstract: Cyclic AMP has a crucial role during the entire developmental program of the social amoebae Dictyostelium, acting both as an intracellular second messenger and, when secreted, as a directional cue that is relayed to neighboring cells during chemotaxis. ... ...

Abstract	Cyclic AMP has a crucial role during the entire developmental program of the social amoebae Dictyostelium, acting both as an intracellular second messenger and, when secreted, as a directional cue that is relayed to neighboring cells during chemotaxis. Although significant knowledge about cAMP production in chemotaxing cells has been derived from studies performed on cell populations, cAMP dynamics at the single cell level have not been investigated. To examine this, we used a FRET-based cAMP sensor that possesses high cAMP sensitivity and great temporal resolution. We show the transient profile of cAMP accumulation in live Dictyostelium cells and establish that chemoattractants control intracellular cAMP dynamics by regulating synthesis via the adenylyl cyclase ACA. aca⁻ cells show no significant change in FRET response following chemoattractant addition. Furthermore, cells lacking ACB, the other adenylyl cyclase expressed in chemotaxing cells, behave similarly to wild-type cells. We also establish that the RegA is the major phosphodiesterase that degrades intracellular cAMP in chemotaxis-competent cells. Interestingly, we failed to measure intracellular cAMP compartmentalization in actively chemotaxing cells. We conclude that cytosolic cAMP, which is destined to activate PKA, is regulated by ACA and RegA and does not compartmentalize during chemotaxis.
Language	English
Dates of publication	2009-1101
Size	p. 3907-3914.
Publishing place	The Company of Biologists Limited
Document type	Article
ZDB-ID	2993-2
ISSN	1477-9137 ; 0021-9533
ISSN (online)	1477-9137
ISSN	0021-9533
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Article ; Online: Real-time measurements of cAMP production in live Dictyostelium cells.

Bagorda, Anna / Das, Satarupa / Rericha, Erin C / Chen, David / Davidson, Jean / Parent, Carole A

Journal of cell science

2009 Volume 122, Issue Pt 21, Page(s) 3907–3914

Abstract	Cyclic AMP has a crucial role during the entire developmental program of the social amoebae Dictyostelium, acting both as an intracellular second messenger and, when secreted, as a directional cue that is relayed to neighboring cells during chemotaxis. Although significant knowledge about cAMP production in chemotaxing cells has been derived from studies performed on cell populations, cAMP dynamics at the single cell level have not been investigated. To examine this, we used a FRET-based cAMP sensor that possesses high cAMP sensitivity and great temporal resolution. We show the transient profile of cAMP accumulation in live Dictyostelium cells and establish that chemoattractants control intracellular cAMP dynamics by regulating synthesis via the adenylyl cyclase ACA. aca(-) cells show no significant change in FRET response following chemoattractant addition. Furthermore, cells lacking ACB, the other adenylyl cyclase expressed in chemotaxing cells, behave similarly to wild-type cells. We also establish that the RegA is the major phosphodiesterase that degrades intracellular cAMP in chemotaxis-competent cells. Interestingly, we failed to measure intracellular cAMP compartmentalization in actively chemotaxing cells. We conclude that cytosolic cAMP, which is destined to activate PKA, is regulated by ACA and RegA and does not compartmentalize during chemotaxis.
MeSH term(s)	Adenylyl Cyclases/genetics ; Adenylyl Cyclases/metabolism ; Chemotaxis ; Cyclic AMP/metabolism ; Cytosol/metabolism ; Dictyostelium/cytology ; Dictyostelium/enzymology ; Dictyostelium/genetics ; Dictyostelium/metabolism ; Fluorescence Resonance Energy Transfer ; Protozoan Proteins/genetics ; Protozoan Proteins/metabolism
Chemical Substances	Protozoan Proteins ; Cyclic AMP (E0399OZS9N) ; Adenylyl Cyclases (EC 4.6.1.1)
Language	English
Publishing date	2009-10-06
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2993-2
ISSN	1477-9137 ; 0021-9533
ISSN (online)	1477-9137
ISSN	0021-9533
DOI	10.1242/jcs.051987
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Article: Chemotaxis: moving forward and holding on to the past

Bagorda, Anna / Mihaylov, Vassil A. / Parent, Carole A.

Thrombosis and Haemostasis

2006 Volume 95, Issue 01, Page(s) 12–21

Abstract	The ability of cells to sense external chemical cues and respond by directionally migrating towards them is a fundamental process called chemotaxis. This phenomenon is essential for many biological responses in the human body, including the invasion of neutrophils to sites of inflammation. Remarkably, many of the molecular mechanisms involved in controlling neutrophils chemotaxis arose millions of years ago in the simple eukaryotic organism Dictyostelium discoideum. Both neutrophils and Dictyostelium use G protein-coupled signaling cascades to mediate chemotactic responses, which are responsible for transducing external cues into highly organized cytoskeletal rearrangements that ultimately lead to directed migration. By using the genetically and biochemically tractable organism Dictyostelium as a model system, it has been possible to decipher many of the signal transduction events that are involved in chemotaxis.
Keywords	Dictyostelium ; neutrophils ; chemotactic signaling
Language	English
Publishing date	2006-01-01
Publisher	Schattauer GmbH
Publishing place	Stuttgart ; New York
Document type	Article
ZDB-ID	518294-3
ISSN	2567-689X ; 0340-6245
ISSN (online)	2567-689X
ISSN	0340-6245
DOI	10.1160/TH05-07-0483
Database	Thieme publisher's database

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Article: Protein kinase A gating of a pseudopodial-located RhoA/ROCK/p38/NHE1 signal module regulates invasion in breast cancer cell lines.

Cardone, Rosa A / Bagorda, Anna / Bellizzi, Antonia / Busco, Giovanni / Guerra, Lorenzo / Paradiso, Angelo / Casavola, Valeria / Zaccolo, Manuela / Reshkin, Stephan J

Molecular biology of the cell

2005 Volume 16, Issue 7, Page(s) 3117–3127

Abstract: Metastasis results from a sequence of selective events often involving interactions with elements of the tumor-specific physiological microenvironment. The low-serum component of this microenvironment confers increased motility and invasion in breast ... ...

Abstract	Metastasis results from a sequence of selective events often involving interactions with elements of the tumor-specific physiological microenvironment. The low-serum component of this microenvironment confers increased motility and invasion in breast cancer cells by activating the Na+/H+ exchanger isoform 1 (NHE1). The present study was undertaken to characterize the signal transduction mechanisms underlying this serum deprivation-dependent activation of both the NHE1 and the concomitant invasive characteristics such as leading edge pseudopodia development and penetration of matrigel in breast cancer cell lines representing different stages of metastatic progression. Using pharmacological and genetic manipulation together with transport and kinase activity assays, we observe that the activation of the NHE1 and subsequent invasion by serum deprivation in metastatic human breast cells is coordinated by a sequential RhoA/p160ROCK/p38MAPK signaling pathway gated by direct protein kinase A phosphorylation and inhibition of RhoA. Fluorescence resonance energy transfer imaging of RhoA activity and immunofluorescence analysis of phospho-RhoA and NHE1 show that serum deprivation dynamically remodels the cell, forming long, leading edge pseudopodia and that this signal module is preferentially compartmentalized in these leading edge pseudopodia, suggesting a tight topographic relation of the signaling module to an invasion-specific cell structure.
MeSH term(s)	Breast Neoplasms/pathology ; Cation Transport Proteins/metabolism ; Cell Line, Tumor ; Collagen/metabolism ; Culture Media, Serum-Free/pharmacology ; Cyclic AMP-Dependent Protein Kinases/metabolism ; Disease Progression ; Down-Regulation ; Drug Combinations ; Enzyme Activation ; Fluorescence Resonance Energy Transfer ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Hydrogen-Ion Concentration ; Intracellular Signaling Peptides and Proteins ; Laminin/metabolism ; Membrane Proteins/metabolism ; Microscopy, Fluorescence ; Models, Biological ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Phosphorylation ; Protein Serine-Threonine Kinases/metabolism ; Proteoglycans/metabolism ; Pseudopodia/metabolism ; Serine/chemistry ; Signal Transduction ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers/metabolism ; Subcellular Fractions ; Time Factors ; Up-Regulation ; p38 Mitogen-Activated Protein Kinases/metabolism ; rho-Associated Kinases ; rhoA GTP-Binding Protein/metabolism
Chemical Substances	Cation Transport Proteins ; Culture Media, Serum-Free ; Drug Combinations ; Intracellular Signaling Peptides and Proteins ; Laminin ; Membrane Proteins ; Proteoglycans ; SLC9A1 protein, human ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers ; matrigel (119978-18-6) ; Serine (452VLY9402) ; Collagen (9007-34-5) ; Protein Serine-Threonine Kinases (EC 2.7.11.1) ; rho-Associated Kinases (EC 2.7.11.1) ; Cyclic AMP-Dependent Protein Kinases (EC 2.7.11.11) ; p38 Mitogen-Activated Protein Kinases (EC 2.7.11.24) ; rhoA GTP-Binding Protein (EC 3.6.5.2)
Language	English
Publishing date	2005-04-20
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1098979-1
ISSN	1939-4586 ; 1059-1524
ISSN (online)	1939-4586
ISSN	1059-1524
DOI	10.1091/mbc.e04-10-0945
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Article ; Online: The Na+-H+ exchanger-1 induces cytoskeletal changes involving reciprocal RhoA and Rac1 signaling, resulting in motility and invasion in MDA-MB-435 cells.

Paradiso, Angelo / Cardone, Rosa Angela / Bellizzi, Antonia / Bagorda, Anna / Guerra, Lorenzo / Tommasino, Massimo / Casavola, Valeria / Reshkin, Stephan J

Breast cancer research : BCR

2004 Volume 6, Issue 6, Page(s) R616–28

Abstract: Introduction: An increasing body of evidence shows that the tumour microenvironment is essential in driving neoplastic progression. The low serum component of this microenvironment stimulates motility/invasion in human breast cancer cells via activation ...

Abstract	Introduction: An increasing body of evidence shows that the tumour microenvironment is essential in driving neoplastic progression. The low serum component of this microenvironment stimulates motility/invasion in human breast cancer cells via activation of the Na+-H+ exchanger (NHE) isoform 1, but the signal transduction systems that underlie this process are still poorly understood. We undertook the present study to elucidate the role and pattern of regulation by the Rho GTPases of this serum deprivation-dependent activation of both NHE1 and subsequent invasive characteristics, such as pseudopodia and invadiopodia protrusion, directed cell motility and penetration of normal tissues. Methods: The present study was performed in a well characterized human mammary epithelial cell line representing late stage metastatic progression, MDA-MB-435. The activity of RhoA and Rac1 was modified using their dominant negative and constitutively active mutants and the activity of NHE1, cell motility/invasion, F-actin content and cell shape were measured. Results: We show for the first time that serum deprivation induces NHE1-dependent morphological and cytoskeletal changes in metastatic cells via a reciprocal interaction of RhoA and Rac1, resulting in increased chemotaxis and invasion. Deprivation changed cell shape by reducing the amount of F-actin and inducing the formation of leading edge pseudopodia. Serum deprivation inhibited RhoA activity and stimulated Rac1 activity. Rac1 and RhoA were antagonistic regulators of both basal and stimulated tumour cell NHE1 activity. The regulation of NHE1 activity by RhoA and Rac1 in both conditions was mediated by an alteration in intracellular proton affinity of the exchanger. Interestingly, the role of each of these G-proteins was reversed during serum deprivation; basal NHE1 activity was regulated positively by RhoA and negatively by Rac1, whereas RhoA negatively and Rac1 positively directed the stimulation of NHE1 during serum deprivation. Importantly, the same pattern of RhoA and Rac1 regulation found for NHE1 activity was observed in both basal and serum deprivation dependent increases in motility, invasion and actin cytoskeletal organization. Conclusion: Our findings suggest that the reported antagonistic roles of RhoA and Rac1 in cell motility/invasion and cytoskeletal organization may be due, in part, to their concerted action on NHE1 activity as a convergence point.
MeSH term(s)	Actins/metabolism ; Breast Neoplasms/enzymology ; Breast Neoplasms/metabolism ; Breast Neoplasms/pathology ; Cation Transport Proteins/metabolism ; Cell Line, Tumor ; Cell Movement/physiology ; Culture Media, Serum-Free ; Cytoskeleton/metabolism ; Cytoskeleton/pathology ; Enzyme Activation ; Humans ; Membrane Proteins/metabolism ; Neoplasm Invasiveness ; Signal Transduction/physiology ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers/metabolism ; rac1 GTP-Binding Protein/metabolism ; rhoA GTP-Binding Protein/metabolism
Chemical Substances	Actins ; Cation Transport Proteins ; Culture Media, Serum-Free ; Membrane Proteins ; SLC9A1 protein, human ; Sodium-Hydrogen Exchanger 1 ; Sodium-Hydrogen Exchangers ; rac1 GTP-Binding Protein (EC 3.6.5.2) ; rhoA GTP-Binding Protein (EC 3.6.5.2)
Language	English
Publishing date	2004
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2015059-3
ISSN	1465-542X ; 1465-5411
ISSN (online)	1465-542X
ISSN	1465-5411
DOI	10.1186/bcr922
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Article: Postnatal maturation of cortical serotonin lateral asymmetry in gerbils is vulnerable to both environmental and pharmacological epigenetic challenges.

Neddens, Jörg / Dawirs, Ralph R / Bagorda, Francesco / Busche, Andrea / Horstmann, Sven / Teuchert-Noodt, Gertraud

Brain research

2004 Volume 1021, Issue 2, Page(s) 200–208

Abstract: Long-term effects of postnatal differential rearing conditions and/or early methamphetamine (MA) application on serotonin (5-HT) fibre density were investigated in several cortical areas of both hemispheres of gerbils. The aim of this study was twofold: ( ...

Abstract	Long-term effects of postnatal differential rearing conditions and/or early methamphetamine (MA) application on serotonin (5-HT) fibre density were investigated in several cortical areas of both hemispheres of gerbils. The aim of this study was twofold: (1) Is the 5-HT fibre innervation of the cerebral cortex lateralised, and (2) if so, do postnatal environmental conditions and/or an early drug challenge interfere with development of 5-HT cerebral asymmetries? For that purpose, male gerbils were reared either under semi-natural or restricted environmental and social conditions, under both conditions once (on postnatal day 14) being treated with either a single dose of MA (50 mg/kg, i.p.) or saline. On postnatal day 110, 5-HT fibres were immunohistochemically stained and innervation densities quantified in prefrontal cortex, insular cortex, frontal cortex, parietal cortex, and entorhinal cortex. It was found that (1) 5-HT innervation in the cerebral cortex was clearly lateralised; (2) direction and extent of this asymmetry were not uniformly distributed over the different areas investigated; (3) both early methamphetamine challenge and rearing condition differentially interfered with adult 5-HT cerebral asymmetry; (4) combining MA challenge with subsequent restricted rearing tended to reverse the effects of MA on 5-HT cerebral asymmetry in some of the cortical areas investigated; and (5) significant responses in 5-HT cerebral asymmetry only occurred in prefrontal and entorhinal association cortices. The present findings suggest that the ontogenesis of cortical laterality is influenced by epigenetic factors and that disturbances of the postnatal maturation of lateralised functions may be associated with certain psychopathological behaviours.
MeSH term(s)	Adrenergic Agents/pharmacology ; Animals ; Cell Count ; Cerebral Cortex/drug effects ; Cerebral Cortex/physiology ; Epigenesis, Genetic ; Functional Laterality/drug effects ; Gerbillinae ; Image Processing, Computer-Assisted ; Immunohistochemistry ; Male ; Methamphetamine/pharmacology ; Nerve Fibers/drug effects ; Nerve Fibers/physiology ; Serotonin/metabolism ; Social Isolation
Chemical Substances	Adrenergic Agents ; Serotonin (333DO1RDJY) ; Methamphetamine (44RAL3456C)
Language	English
Publishing date	2004-09-24
Publishing country	Netherlands
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1200-2
ISSN	1872-6240 ; 0006-8993
ISSN (online)	1872-6240
ISSN	0006-8993
DOI	10.1016/j.brainres.2004.06.050
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