31 results on '"Ferkey, Denise M."'
Search Results
2. Structural Domains Required for Caenorhabditis elegans G Protein-coupled Receptor Kinase 2 (GRK-2) Function in Vivo
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Wood, Jordan F., Wang, Jianjun, Benovic, Jeffrey L., and Ferkey, Denise M.
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- 2012
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3. Caenorhabditis elegans TRPV channels function in a modality-specific pathway to regulate response to aberrant sensory signaling
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Ezak, Meredith J., Hong, Elizabeth, Chaparro-Garcia, Angela, and Ferkey, Denise M.
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Caenorhabditis elegans -- Physiological aspects ,Caenorhabditis elegans -- Genetic aspects ,Gene expression -- Research ,Gene mutations -- Research ,Protein-protein interactions -- Analysis ,Biological sciences - Published
- 2010
4. The Prop1-like homeobox gene unc-42 specifies the identity of synaptically connected neurons.
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Berghoff, Emily G., Glenwinkel, Lori, Bhattacharya, Abhishek, HaoSheng Sun, Varol, Erdem, Mohammadi, Nicki, Antone, Amelia, Yi Feng, Nguyen, Ken, Cook, Steven J., Wood, Jordan F., Masoudi, Neda, Cros, Cyril C., Ramadan, Yasmin H., Ferkey, Denise M., Hall, David H., and Hobert, Oliver
- Published
- 2021
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5. Chemosensory signal transduction in Caenorhabditis elegans.
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Ferkey, Denise M., Sengupta, Piali, and L'Etoile, Noelle D.
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CAENORHABDITIS elegans , *SENSORY receptors , *SENSORY perception , *CELL receptors , *CELLULAR signal transduction , *SMELL , *GENETIC techniques , *TASTE , *ODORS , *PROMPTS (Psychology) - Abstract
Chemosensory neurons translate perception of external chemical cues, including odorants, tastants, and pheromones, into information that drives attraction or avoidance motor programs. In the laboratory, robust behavioral assays, coupled with powerful genetic, molecular and optical tools, have made Caenorhabditis elegans an ideal experimental system in which to dissect the contributions of individual genes and neurons to ethologically relevant chemosensory behaviors. Here, we review current knowledge of the neurons, signal transduction molecules and regulatory mechanisms that underlie the response of C. elegans to chemicals, including pheromones. The majority of identi- fied molecules and pathways share remarkable homology with sensory mechanisms in other organisms. With the development of new tools and technologies, we anticipate that continued study of chemosensory signal transduction and processing in C. elegans will yield additional new insights into the mechanisms by which this animal is able to detect and discriminate among thousands of chemical cues with a limited sensory neuron repertoire. [ABSTRACT FROM AUTHOR]
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- 2021
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6. INX-18 and INX-19 play distinct roles in electrical synapses that modulate aversive behavior in Caenorhabditis elegans.
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Voelker, Lisa, Upadhyaya, Bishal, Ferkey, Denise M., Woldemariam, Sarah, L'Etoile, Noelle D., Rabinowitch, Ithai, and Bai, Jihong
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CAENORHABDITIS elegans ,SYNAPSES ,ANIMAL behavior ,AVERSIVE stimuli ,SMALL molecules ,SENSORY neurons ,INSULAR cortex - Abstract
In order to respond to changing environments and fluctuations in internal states, animals adjust their behavior through diverse neuromodulatory mechanisms. In this study we show that electrical synapses between the ASH primary quinine-detecting sensory neurons and the neighboring ASK neurons are required for modulating the aversive response to the bitter tastant quinine in C. elegans. Mutant worms that lack the electrical synapse proteins INX-18 and INX-19 become hypersensitive to dilute quinine. Cell-specific rescue experiments indicate that inx-18 operates in ASK while inx-19 is required in both ASK and ASH for proper quinine sensitivity. Imaging analyses find that INX-19 in ASK and ASH localizes to the same regions in the nerve ring, suggesting that both sides of ASK-ASH electrical synapses contain INX-19. While inx-18 and inx-19 mutant animals have a similar behavioral phenotype, several lines of evidence suggest the proteins encoded by these genes play different roles in modulating the aversive quinine response. First, INX-18 and INX-19 localize to different regions of the nerve ring, indicating that they are not present in the same synapses. Second, removing inx-18 disrupts the distribution of INX-19, while removing inx-19 does not alter INX-18 localization. Finally, by using a fluorescent cGMP reporter, we find that INX-18 and INX-19 have distinct roles in establishing cGMP levels in ASK and ASH. Together, these results demonstrate that electrical synapses containing INX-18 and INX-19 facilitate modulation of ASH nociceptive signaling. Our findings support the idea that a network of electrical synapses mediates cGMP exchange between neurons, enabling modulation of sensory responses and behavior. Animals are constantly adjusting their behavior to respond to changes in the environment or to their internal state. This behavior modulation is achieved by altering the activity of neurons and circuits through a variety of neuroplasticity mechanisms. Chemical synapses are known to impact neuroplasticity in several different ways, but the diversity of mechanisms by which electrical synapses contribute is still being investigated. Electrical synapses are specialized sites of connection between neurons where ions and small signaling molecules can pass directly from one cell to the next. By passing small molecules through electrical synapses, neurons may be able to modify the activity of their neighbors. In this study we identify two genes that contribute to electrical synapses between two sensory neurons in C. elegans. We show that these electrical synapses are crucial for proper modulation of sensory responses, as without them animals are overly responsive to an aversive stimulus. In addition to pinpointing their sites of action, we present evidence that they may be contributing to neuromodulation by facilitating passage of the small molecule cGMP between neurons. Our work provides evidence for a role of electrical synapses in regulating animal behavior. [ABSTRACT FROM AUTHOR]
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- 2019
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7. GRK Roles in C. elegans.
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Wood, Jordan F. and Ferkey, Denise M.
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- 2016
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8. Aversive Behavior in the Nematode C. elegans Is Modulated by cGMP and a Neuronal Gap Junction Network.
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Krzyzanowski, Michelle C., Woldemariam, Sarah, Wood, Jordan F., Chaubey, Aditi H., Brueggemann, Chantal, Bowitch, Alexander, Bethke, Mary, L’Etoile, Noelle D., and Ferkey, Denise M.
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AVERSIVE stimuli ,SENSE organs ,NOCICEPTORS ,NOCICEPTIVE pain ,NEURAL circuitry - Abstract
All animals rely on their ability to sense and respond to their environment to survive. However, the suitability of a behavioral response is context-dependent, and must reflect both an animal’s life history and its present internal state. Based on the integration of these variables, an animal’s needs can be prioritized to optimize survival strategies. Nociceptive sensory systems detect harmful stimuli and allow for the initiation of protective behavioral responses. The polymodal ASH sensory neurons are the primary nociceptors in C. elegans. We show here that the guanylyl cyclase ODR-1 functions non-cell-autonomously to downregulate ASH-mediated aversive behaviors and that ectopic cGMP generation in ASH is sufficient to dampen ASH sensitivity. We define a gap junction neural network that regulates nociception and propose that decentralized regulation of ASH signaling can allow for rapid correlation between an animal’s internal state and its behavioral output, lending modulatory flexibility to this hard-wired nociceptive neural circuit. [ABSTRACT FROM AUTHOR]
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- 2016
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9. The protein arginine methyltransferase PRMT5 promotes D2-like dopamine receptor signaling.
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Krzyzanowski, Michelle C., Likhite, Neah, Jackson, Christopher A., Wood, Jordan F., Yu, Michael C., Ferkey, Denise M., Birkaya, Barbara, Michaels, Kerry L., Mao-Shih Liang, Lei, Pedro, Andreadis, Stelios T., and Clark, Stewart D.
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- 2015
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10. The C. elegans cGMP-Dependent Protein Kinase EGL-4 Regulates Nociceptive Behavioral Sensitivity.
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Krzyzanowski, Michelle C., Brueggemann, Chantal, Ezak, Meredith J., Wood, Jordan F., Michaels, Kerry L., Jackson, Christopher A., Juang, Bi-Tzen, Collins, Kimberly D., Yu, Michael C., L'Etoile, Noelle D., and Ferkey, Denise M.
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NEURONS ,PROTEIN kinases ,G proteins ,PHOSPHORYLATION ,QUININE - Abstract
Signaling levels within sensory neurons must be tightly regulated to allow cells to integrate information from multiple signaling inputs and to respond to new stimuli. Herein we report a new role for the cGMP-dependent protein kinase EGL-4 in the negative regulation of G protein-coupled nociceptive chemosensory signaling. C. elegans lacking EGL-4 function are hypersensitive in their behavioral response to low concentrations of the bitter tastant quinine and exhibit an elevated calcium flux in the ASH sensory neurons in response to quinine. We provide the first direct evidence for cGMP/PKG function in ASH and propose that ODR-1, GCY-27, GCY-33 and GCY-34 act in a non-cell-autonomous manner to provide cGMP for EGL-4 function in ASH. Our data suggest that activated EGL-4 dampens quinine sensitivity via phosphorylation and activation of the regulator of G protein signaling (RGS) proteins RGS-2 and RGS-3, which in turn downregulate Gα signaling and behavioral sensitivity. [ABSTRACT FROM AUTHOR]
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- 2013
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11. A Functional Nuclear Localization Sequence in the C. elegans TRPV Channel OCR-2.
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Ezak, Meredith J. and Ferkey, Denise M.
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NUCLEOTIDE sequence , *CAENORHABDITIS elegans , *TRP channels , *GENETIC transcription , *GENE expression , *CYTOSOL , *NEURONS , *CALCIUM channels - Abstract
The ability to modulate gene expression in response to sensory experience is critical to the normal development and function of the nervous system. Calcium is a key activator of the signal transduction cascades that mediate the process of translating a cellular stimulus into transcriptional changes. With the recent discovery that the mammalian Cav1.2 calcium channel can be cleaved, enter the nucleus and act as a transcription factor to control neuronal gene expression, a more direct role for the calcium channels themselves in regulating transcription has begun to be appreciated. Here we report the identification of a nuclear localization sequence (NLS) in the C. elegans transient receptor potential vanilloid (TRPV) cation channel OCR-2. TRPV channels have previously been implicated in transcriptional regulation of neuronal genes in the nematode, although the precise mechanism remains unclear. We show that the NLS in OCR-2 is functional, being able to direct nuclear accumulation of a synthetic cargo protein as well as the carboxy-terminal cytosolic tail of OCR-2 where it is endogenously found. Furthermore, we discovered that a carboxy-terminal portion of the full-length channel can localize to the nucleus of neuronal cells. These results suggest that the OCR-2 TRPV cation channel may have a direct nuclear function in neuronal cells that was not previously appreciated. [ABSTRACT FROM AUTHOR]
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- 2011
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12. Remote control of ion channels and neurons through magnetic-field heating of nanoparticles.
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Heng Huang, Delikanli, Savas, Hao Zeng, Ferkey, Denise M., and Pralle, Arnd
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TISSUES ,MAGNETIC fields ,TEMPERATURE ,CELLS ,PROTEINS ,THERMOMETERS - Abstract
Recently, optical stimulation has begun to unravel the neuronal processing that controls certain animal behaviours. However, optical approaches are limited by the inability of visible light to penetrate deep into tissues. Here, we show an approach based on radio-frequency magnetic-field heating of nanoparticles to remotely activate temperature-sensitive cation channels in cells. Superparamagnetic ferrite nanoparticles were targeted to specific proteins on the plasma membrane of cells expressing TRPV1, and heated by a radio-frequency magnetic field. Using fluorophores as molecular thermometers, we show that the induced temperature increase is highly localized. Thermal activation of the channels triggers action potentials in cultured neurons without observable toxic effects. This approach can be adapted to stimulate other cell types and, moreover, may be used to remotely manipulate other cellular machinery for novel therapeutics. [ABSTRACT FROM AUTHOR]
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- 2010
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13. The C. elegans D2-Like Dopamine Receptor DOP-3 Decreases Behavioral Sensitivity to the Olfactory Stimulus 1-Octanol.
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Ezak, Meredith J. and Ferkey, Denise M.
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DOPAMINE receptors , *NEUROTRANSMITTER receptors , *DOPAMINE agents , *ROTIGOTINE , *DOPAMINE agonists , *OLFACTORY receptors , *ESCHERICHIA coli , *CELL receptors , *HORMONE receptors - Abstract
We previously found that dopamine signaling modulates the sensitivity of wild-type C. elegans to the aversive odorant 1-octanol. C. elegans lacking the CAT-2 tyrosine hydroxylase enzyme, which is required for dopamine biosynthesis, are hypersensitive in their behavioral avoidance of dilute concentrations of octanol. Dopamine can also modulate the context-dependent response of C. elegans lacking RGS-3 function, a negative regulator of Gα signaling. rgs-3 mutant animals are defective in their avoidance of 100% octanol when they are assayed in the absence of food (E. coli bacterial lawn), but their response is restored when they are assayed in the presence of food or exogenous dopamine. However, it is not known which receptor might be mediating dopamine's effects on octanol avoidance. Herein we describe a role for the C. elegans D2-like receptor DOP-3 in the regulation of olfactory sensitivity. We show that DOP-3 is required for the ability of food and exogenous dopamine to rescue the octanol avoidance defect of rgs-3 mutant animals. In addition, otherwise wild-type animals lacking DOP-3 function are hypersensitive to dilute octanol, reminiscent of cat-2 mutants. Furthermore, we demonstrate that DOP-3 function in the ASH sensory neurons is sufficient to rescue the hypersensitivity of dop-3 mutant animals, while dop-3 RNAi knockdown in ASH results in octanol hypersensitivity. Taken together, our data suggest that dopaminergic signaling through DOP-3 normally acts to dampen ASH signaling and behavioral sensitivity to octanol. [ABSTRACT FROM AUTHOR]
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- 2010
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14. C. elegans G Protein Regulator RGS-3 Controls Sensitivity to Sensory Stimuli
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Ferkey, Denise M., Hyde, Rhonda, Haspel, Gal, Dionne, Heather M., Hess, Heather A., Suzuki, Hiroshi, Schafer, William R., Koelle, Michael R., and Hart, Anne C.
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CELLULAR signal transduction , *SENSORY receptors , *PROTEINS , *SENSORY neurons - Abstract
Summary: Signal transduction through heterotrimeric G proteins is critical for sensory response across species. Regulator of G protein signaling (RGS) proteins are negative regulators of signal transduction. Herein we describe a role for C. elegans RGS-3 in the regulation of sensory behaviors. rgs-3 mutant animals fail to respond to intense sensory stimuli but respond normally to low concentrations of specific odorants. We find that loss of RGS-3 leads to aberrantly increased G protein-coupled calcium signaling but decreased synaptic output, ultimately leading to behavioral defects. Thus, rgs-3 responses are restored by decreasing G protein-coupled signal transduction, either genetically or by exogenous dopamine, by expressing a calcium-binding protein to buffer calcium levels in sensory neurons or by enhancing glutamatergic synaptic transmission from sensory neurons. Therefore, while RGS proteins generally act to downregulate signaling, loss of a specific RGS protein in sensory neurons can lead to defective responses to external stimuli. [Copyright &y& Elsevier]
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- 2007
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15. G Protein-Coupled Receptor Kinase Function Is Essential for Chemosensation in C. elegans
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Fukuto, Hana S., Ferkey, Denise M., Apicella, Alfonso J., Lans, Hannes, Sharmeen, Tahira, Chen, Wei, Lefkowitz, Robert J., Jansen, Gert, Schafer, William R., and Hart, Anne C.
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G proteins , *MEMBRANE proteins , *PROTEINS , *NEURONS - Abstract
G protein-coupled receptors (GPCRs) mediate diverse signaling processes, including olfaction. G protein-coupled receptor kinases (GRKs) are important regulators of G protein signal transduction that specifically phosphorylate activated GPCRs to terminate signaling. Despite previously described roles for GRKs in GPCR signal downregulation, animals lacking C. elegans G protein-coupled receptor kinase-2 (Ce-grk-2) function are not hypersensitive to odorants. Instead, decreased Ce-grk-2 function in adult sensory neurons profoundly disrupts chemosensation, based on both behavioral analysis and Ca2+ imaging. Although mammalian arrestin proteins cooperate with GRKs in receptor desensitization, loss of C. elegans arrestin-1 (arr-1) does not disrupt chemosensation. Either overexpression of the C. elegans Gα subunit odr-3 or loss of eat-16, which encodes a regulator of G protein signaling (RGS) protein, restores chemosensation in Ce-grk-2 mutants. These results demonstrate that loss of GRK function can lead to reduced GPCR signal transduction and suggest an important role for RGS proteins in the regulation of chemosensation. [Copyright &y& Elsevier]
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- 2004
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16. Tcf4 can specifically recognize β-catenin using alternative conformations.
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Graham, Thomas A., Ferkey, Denise M., Mao, Feng, Kimelman, David, and Xu, Wenqing
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CANCER , *TRANSCRIPTION factors , *CRYSTALLOGRAPHY , *AMINO acids - Abstract
Accumulation of the Wnt pathway effector β-catenin is a hallmark of a number of cancers, including colon cancer. As β-catenin accumulates in the cell, it forms a complex with Tcf family transcription factors and activates the transcription of several critical genes involved in cell proliferation. Because Tcf4 is the predominant Tcf factor present in colon cancer cells, drugs that specifically disrupt the β-catenin?Tcf4 complex could be useful in treating colon cancers. Earlier structural and biochemical studies demonstrated that the central region of the β-catenin binding domain of Tcf is essential for anchoring Tcf to β-catenin via two conserved lysines in β-catenin (called the charged 'buttons'). Here we report the crystal structure of a β-catenin?Tcf4 complex at 2.0 Å resolution. Our structural and mutagenesis studies show that Tcf4 docks specifically to β-catenin using several distinct conformations in its essential central region. These conformations allow different glutamate residues in the central region of Tcf4 to form a salt bridge with the same critical charged button, Lys 312 of β-catenin. We propose that this interaction may be the first event in β-catenin?Tcf4 recognition. [ABSTRACT FROM AUTHOR]
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- 2001
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17. The Caenorhabditis elegans innexin INX-20 regulates nociceptive behavioral sensitivity.
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Chaubey, Aditi H., Sojka, Savannah E., Onukwufor, John O., Ezak, Meredith J., Vandermeulen, Matthew D., Bowitch, Alexander, Vodičková, Anežka, Wojtovich, Andrew P., and Ferkey, Denise M.
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PROTEIN metabolism , *NEURONS , *SENSORY receptors , *ANIMAL experimentation , *CAENORHABDITIS elegans , *PAIN threshold , *CELLULAR signal transduction , *ALLERGIES , *NOCICEPTIVE pain - Abstract
Organisms rely on chemical cues in their environment to indicate the presence or absence of food, reproductive partners, predators, or other harmful stimuli. In the nematode Caenorhabditis elegans, the bilaterally symmetric pair of ASH sensory neurons serves as the primary nociceptors. ASH activation by aversive stimuli leads to backward locomotion and stimulus avoidance. We previously reported a role for guanylyl cyclases in dampening nociceptive sensitivity that requires an innexin-based gap junction network to pass cGMP between neurons. Here, we report that animals lacking function of the gap junction component INX-20 are hypersensitive in their behavioral response to both soluble and volatile chemical stimuli that signal through G protein-coupled receptor pathways in ASH. We find that expressing inx-20 in the ADL and AFD sensory neurons is sufficient to dampen ASH sensitivity, which is supported by new expression analysis of endogenous INX-20 tagged with mCherry via the CRISPR-Cas9 system. Although ADL does not form gap junctions directly with ASH, it does so via gap junctions with the interneuron RMG and the sensory neuron ASK. Ablating either ADL or RMG and ASK also resulted in nociceptive hypersensitivity, suggesting an important role for RMG/ASK downstream of ADL in the ASH modulatory circuit. This work adds to our growing understanding of the repertoire of ways by which ASH activity is regulated via its connectivity to other neurons and identifies a previously unknown role for ADL and RMG in the modulation of aversive behavior. [ABSTRACT FROM AUTHOR]
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- 2023
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18. Using a Robust and Sensitive GFP-Based cGMP Sensor for Real-Time Imaging in Intact Caenorhabditis elegans.
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Woldemariam, Sarah, Nagpal, Jatin, Hill, Tyler, Li, Joy, Schneider, Martin W., Shankar, Raakhee, Futey, Mary, Varshney, Aruna, Ali, Nebat, Mitchell, Jordan, Andersen, Kristine, Barsi-Rhyne, Benjamin, Tran, Alan, Costa, Wagner Steuer, Krzyzanowski, Michelle C., Yu, Yanxun V., Brueggemann, Chantal, Hamilton, Scott, Ferkey, Denise M., and VanHoven, Miri
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NUCLEOTIDE analysis , *PROTEIN analysis , *BIOSENSORS , *ENZYMES , *GENE expression , *HYDROCARBONS , *NEMATODES , *SALT , *SENSORY receptors - Abstract
cGMP plays a role in sensory signaling and plasticity by regulating ion channels, phosphodiesterases, and kinases. Studies that primarily used genetic and biochemical tools suggest that cGMP is spatiotemporally regulated in multiple sensory modalities. FRETand GFP-based cGMP sensors were developed to visualize cGMP in primary cell culture and Caenorhabditis elegans to corroborate these findings. While a FRET-based sensor has been used in an intact animal to visualize cGMP, the requirement of a multiple emission system limits its ability to be used on its own as well as with other fluorophores. Here, we demonstrate that a C. elegans codonoptimized version of the cpEGFP-based cGMP sensor FlincG3 can be used to visualize rapidly changing cGMP levels in living, behaving C. elegans. We coexpressed FlincG3 with the blue-light-activated guanylyl cyclases BeCyclOp and bPGC in body wall muscles, and found that the rate of change in FlincG3 fluorescence correlated with the rate of cGMP production by each cyclase. Furthermore, we show that FlincG3 responds to cultivation temperature, NaCl concentration changes, and sodium dodecyl sulfate in the sensory neurons AFD, ASEL/R, and PHB, respectively. Intriguingly, FlincG3 fluorescence in ASEL and ASER decreased in response to a NaCl concentration upstep and downstep, respectively, which is opposite in sign to the coexpressed calcium sensor jRGECO1a and previously published calcium recordings. These results illustrate that FlincG3 can be used to report rapidly changing cGMP levels in an intact animal, and that the reporter can potentially reveal unexpected spatiotemporal landscapes of cGMP in response to stimuli. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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19. Filamentation Regulatory Pathways Control Adhesion-Dependent Surface Responses in Yeast.
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Chow, Jacky, Starr, Izzy, Jamalzadeh, Sheida, Muniz, Omar, Kumar, Anuj, Gokcumen, Omer, Ferkey, Denise M., and Cullen, Paul J.
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CELL adhesion molecules , *CELL physiology , *CELLULAR signal transduction , *GENETIC mutation , *ONCOGENES , *OXYGEN , *TEMPERATURE , *TRANSFERASES , *MICROBIAL virulence , *YEAST , *PHENOTYPES , *GENE expression profiling - Abstract
Signaling pathways can regulate biological responses by the transcriptional regulation of target genes. In yeast, multiple signaling pathways control filamentous growth, a morphogenetic response that occurs in many species including fungal pathogens. Here, we examine the role of signaling pathways that control filamentous growth in regulating adhesion-dependent surface responses, including mat formation and colony patterning. Expression profiling and mutant phenotype analysis showed that the major pathways that regulate filamentous growth [filamentous growth MAPK (fMAPK), RAS, retrograde (RTG), RIM101, RPD3, ELP, SNF1, and PHO85] also regulated mat formation and colony patterning. The chromatin remodeling complex, SAGA, also regulated these responses. We also show that the RAS and RTG pathways coregulated a common set of target genes, and that SAGA regulated target genes known to be controlled by the fMAPK, RAS, and RTG pathways. Analysis of surface growth-specific targets identified genes that respond to low oxygen, high temperature, and desiccation stresses. We also explore the question of why cells make adhesive contacts in colonies. Cell adhesion contacts mediated by the coregulated target and adhesion molecule, Flo11p, deterred entry into colonies by macroscopic predators and impacted colony temperature regulation. The identification of new regulators (e.g., SAGA), and targets of surface growth in yeast may provide insights into fungal pathogenesis in settings where surface growth and adhesion contributes to virulence. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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20. The Protein Arginine Methyltransferase PRMT-5 Regulates SER-2 Tyramine Receptor-Mediated Behaviors in Caenorhabditis elegans.
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Bowitch, Alexander, Michaels, Kerry L., Yu, Michael C., and Ferkey, Denise M.
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TYRAMINE , *G protein coupled receptors , *CAENORHABDITIS elegans genetics - Abstract
G protein-coupled receptors are 7-pass transmembrane receptors that couple to heterotrimeric G proteins to mediate cellular responses to a diverse array of stimuli. Understanding the mechanisms that regulate G protein-coupled receptors is crucial to manipulating their signaling for therapeutic benefit. One key regulatory mechanism that contributes to the functional diversity of many signaling proteins is post-translational modification. Whereas phosphorylation remains the best studied of such modifications, arginine methylation by protein arginine methyltransferases is emerging as a key regulator of protein function. We previously published the first functional evidence that arginine methylation of G protein-coupled receptors modulates their signaling. We report here a third receptor that is regulated by arginine methylation, the Caenorhabditis elegans SER-2 tyramine receptor. We show that arginines within a putative methylation motif in the third intracellular loop of SER-2 are methylated by PRMT5 in vitro. Our data also suggest that this modification enhances SER-2 signaling in vivo to modulate animal behavior. The identification of a third G protein-coupled receptor to be functionally regulated by arginine methylation suggests that this post-translational modification may be utilized to regulate signaling through a broad array of G protein-coupled receptors. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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21. Reprogramming the topology of the nociceptive circuit in C. elegans reshapes sexual behavior.
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Pechuk, Vladyslava, Goldman, Gal, Salzberg, Yehuda, Chaubey, Aditi H., Bola, R. Aaron, Hoffman, Jonathon R., Endreson, Morgan L., Miller, Renee M., Reger, Noah J., Portman, Douglas S., Ferkey, Denise M., Schneidman, Elad, and Oren-Suissa, Meital
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CAENORHABDITIS elegans , *HUMAN sexuality , *AVERSIVE stimuli , *TOPOLOGY , *DETECTOR circuits , *SENSORY neurons , *ELECTRIC network topology , *NEURAL circuitry - Abstract
The effect of the detailed connectivity of a neural circuit on its function and the resulting behavior of the organism is a key question in many neural systems. Here, we study the circuit for nociception in C. elegans , which is composed of the same neurons in the two sexes that are wired differently. We show that the nociceptive sensory neurons respond similarly in the two sexes, yet the animals display sexually dimorphic behaviors to the same aversive stimuli. To uncover the role of the downstream network topology in shaping behavior, we learn and simulate network models that replicate the observed dimorphic behaviors and use them to predict simple network rewirings that would switch behavior between the sexes. We then show experimentally that these subtle synaptic rewirings indeed flip behavior. Interestingly, when presented with aversive cues, rewired males were compromised in finding mating partners, suggesting that network topologies that enable efficient avoidance of noxious cues have a reproductive "cost." Our results present a deconstruction of the design of a neural circuit that controls sexual behavior and how to reprogram it. [Display omitted] • C. elegans exhibits sexually dimorphic responses to nociceptive stimuli • Computational models show that circuit topology accounts for the dimorphic behavior • Model-based rewiring of single neurons or synapses flips sex-specific behavior • Rewired males are compromised in finding mates when presented with aversive cues Pechuk et al. study the circuit for nociceptive sensing and processing in hermaphrodite and male C. elegans. Combining circuit models and experimental characterization, they demonstrate that network topology shapes sexually dimorphic behavior. They show that simple rewiring reprograms behavior and demonstrates its sexual implications. [ABSTRACT FROM AUTHOR]
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- 2022
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22. GBP, an inhibitor of GSK-3, is implicated in Xenopus...
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Yost, Cynthia, Farr III, Gist H., Pierce, Sarah B., Ferkey, Denise M., Chen, Michelle Mingzi, and Kimelman, David
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GLUTAMINE , *XENOPUS , *GENETICS , *STRUCTURE-activity relationships - Abstract
Provides information on a study which identifies the role of glutamine binding proteins (GBP), a maternal Xgsk-3-binding proteins, on the establishment of the dorsal-ventral axis in Xenopus. Cloning and expression of a novel Xgsk-3-binding proteins; Effect of GBP on axis duplication; Role of GBP on the regulation of stable beta-catenin.
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- 1998
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23. The C. elegans OCTR-1 and Human Alpha-2A Adrenergic Receptors are Methylated within the Third Intracellular Loop by Human PRMT5 in vitro .
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Bowitch A, Chinsky TM, Yu MC, and Ferkey DM
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- 2022
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24. Corrigendum to: Chemosensory signal transduction in Caenorhabditis elegans.
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Ferkey DM, Sengupta P, and L'Etoile ND
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- 2022
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25. Methylation of the D2 dopamine receptor affects binding with the human regulatory proteins Par-4 and Calmodulin.
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Bowitch A, Sahoo A, Clark AM, Ntangka C, Raut KK, Gollnick P, Yu MC, Pascal SM, Walker SE, and Ferkey DM
- Published
- 2021
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26. The C. elegans TRPV channel proteins OSM-9 and OCR-2 contribute to aversive chemical sensitivity.
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Mehle EA, Sojka SE, K C M, Zel RM, Reese SJ, and Ferkey DM
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- 2020
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27. The protein arginine methyltransferase PRMT5 promotes D2-like dopamine receptor signaling.
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Likhite N, Jackson CA, Liang MS, Krzyzanowski MC, Lei P, Wood JF, Birkaya B, Michaels KL, Andreadis ST, Clark SD, Yu MC, and Ferkey DM
- Subjects
- Amino Acid Motifs, Amino Acid Sequence, Animals, Animals, Genetically Modified, Arginine chemistry, Caenorhabditis elegans drug effects, Caenorhabditis elegans genetics, Caenorhabditis elegans metabolism, Caenorhabditis elegans Proteins chemistry, Caenorhabditis elegans Proteins genetics, Caenorhabditis elegans Proteins metabolism, Computational Biology, Conserved Sequence, Dopamine metabolism, Dopamine pharmacology, HEK293 Cells, Humans, Locomotion drug effects, Locomotion genetics, Locomotion physiology, Methylation, Molecular Sequence Data, Octanols pharmacology, Odorants, Protein-Arginine N-Methyltransferases deficiency, Protein-Arginine N-Methyltransferases genetics, Receptors, Dopamine D2 chemistry, Receptors, Dopamine D2 genetics, Receptors, G-Protein-Coupled chemistry, Receptors, G-Protein-Coupled genetics, Receptors, G-Protein-Coupled metabolism, Sequence Homology, Amino Acid, Signal Transduction, Protein-Arginine N-Methyltransferases metabolism, Receptors, Dopamine D2 metabolism
- Abstract
Protein arginine methylation regulates diverse functions of eukaryotic cells, including gene expression, the DNA damage response, and circadian rhythms. We showed that arginine residues within the third intracellular loop of the human D2 dopamine receptor, which are conserved in the DOP-3 receptor in the nematode Caenorhabditis elegans, were methylated by protein arginine methyltransferase 5 (PRMT5). By mutating these arginine residues, we further showed that their methylation enhanced the D2 receptor-mediated inhibition of cyclic adenosine monophosphate (cAMP) signaling in cultured human embryonic kidney (HEK) 293T cells. Analysis of prmt-5-deficient worms indicated that methylation promoted the dopamine-mediated modulation of chemosensory and locomotory behaviors in C. elegans through the DOP-3 receptor. In addition to delineating a previously uncharacterized means of regulating GPCR (heterotrimeric guanine nucleotide-binding protein-coupled receptor) signaling, these findings may lead to the development of a new class of pharmacological therapies that modulate GPCR signaling by changing the methylation status of these key proteins., (Copyright © 2015, American Association for the Advancement of Science.)
- Published
- 2015
- Full Text
- View/download PDF
28. Epitope-guided engineering of monobody binders for in vivo inhibition of Erk-2 signaling.
- Author
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Mann JK, Wood JF, Stephan AF, Tzanakakis ES, Ferkey DM, and Park S
- Subjects
- Animals, Caenorhabditis elegans metabolism, Fibronectins chemistry, Fibronectins genetics, Fibronectins metabolism, HEK293 Cells, Humans, Mitogen-Activated Protein Kinase 1 chemistry, Models, Molecular, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, Structure-Activity Relationship, Epitopes genetics, Mitogen-Activated Protein Kinase 1 antagonists & inhibitors, Mitogen-Activated Protein Kinase 1 metabolism, Protein Engineering, Protein Kinase Inhibitors chemistry, Protein Kinase Inhibitors pharmacology, Signal Transduction drug effects
- Abstract
Although the affinity optimization of protein binders is straightforward, engineering epitope specificity is more challenging. Targeting a specific surface patch is important because the biological relevance of protein binders depends on how they interact with the target. They are particularly useful to test hypotheses motivated by biochemical and structural studies. We used yeast display to engineer monobodies that bind a defined surface patch on the mitogen activated protein kinase (MAPK) Erk-2. The targeted area ("CD" domain) is known to control the specificity and catalytic efficiency of phosphorylation by the kinase by binding a linear peptide ("D" peptide) on substrates and regulators. An inhibitor of the interaction should thus be useful for regulating Erk-2 signaling in vivo. Although the CD domain constitutes only a small percentage of the surface area of the enzyme (~5%), sorting a yeast displayed monobody library with wild type (wt) Erk-2 and a rationally designed mutant led to isolation of high affinity clones with desired epitope specificity. The engineered binders inhibited the activity of Erk-2 in vitro and in mammalian cells. Furthermore, they specifically inhibited the activity of Erk-2 orthologs in yeast and suppressed a mutant phenotype in round worms caused by overactive MAPK signaling. The study therefore shows that positive and negative screening can be used to bias the evolution of epitope specificity and predictably design inhibitors of biologically relevant protein-protein interaction.
- Published
- 2013
- Full Text
- View/download PDF
29. The C. elegans cGMP-dependent protein kinase EGL-4 regulates nociceptive behavioral sensitivity.
- Author
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Krzyzanowski MC, Brueggemann C, Ezak MJ, Wood JF, Michaels KL, Jackson CA, Juang BT, Collins KD, Yu MC, L'etoile ND, and Ferkey DM
- Subjects
- Animals, Caenorhabditis elegans physiology, Caenorhabditis elegans Proteins metabolism, Cyclic GMP-Dependent Protein Kinases metabolism, GTP-Binding Protein alpha Subunits, Gi-Go genetics, GTP-Binding Protein alpha Subunits, Gi-Go metabolism, Phosphorylation, RGS Proteins genetics, RGS Proteins metabolism, Sensory Receptor Cells metabolism, Sensory Receptor Cells physiology, Signal Transduction genetics, Behavior, Animal physiology, Caenorhabditis elegans genetics, Caenorhabditis elegans Proteins genetics, Cyclic GMP metabolism, Cyclic GMP-Dependent Protein Kinases genetics
- Abstract
Signaling levels within sensory neurons must be tightly regulated to allow cells to integrate information from multiple signaling inputs and to respond to new stimuli. Herein we report a new role for the cGMP-dependent protein kinase EGL-4 in the negative regulation of G protein-coupled nociceptive chemosensory signaling. C. elegans lacking EGL-4 function are hypersensitive in their behavioral response to low concentrations of the bitter tastant quinine and exhibit an elevated calcium flux in the ASH sensory neurons in response to quinine. We provide the first direct evidence for cGMP/PKG function in ASH and propose that ODR-1, GCY-27, GCY-33 and GCY-34 act in a non-cell-autonomous manner to provide cGMP for EGL-4 function in ASH. Our data suggest that activated EGL-4 dampens quinine sensitivity via phosphorylation and activation of the regulator of G protein signaling (RGS) proteins RGS-2 and RGS-3, which in turn downregulate Gα signaling and behavioral sensitivity., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2013
- Full Text
- View/download PDF
30. Remote control of ion channels and neurons through magnetic-field heating of nanoparticles.
- Author
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Huang H, Delikanli S, Zeng H, Ferkey DM, and Pralle A
- Subjects
- Animals, Behavior, Animal physiology, Caenorhabditis elegans, Cells, Cultured, Drosophila, Electromagnetic Fields, Equipment Design, Equipment Failure Analysis, Heating methods, Ion Channels physiology, Nanoparticles chemistry, Neurons radiation effects, Behavior, Animal radiation effects, Heating instrumentation, Ion Channels radiation effects, Nanoparticles radiation effects, Nanotechnology instrumentation, Neurons physiology, Telemetry instrumentation
- Abstract
Recently, optical stimulation has begun to unravel the neuronal processing that controls certain animal behaviours. However, optical approaches are limited by the inability of visible light to penetrate deep into tissues. Here, we show an approach based on radio-frequency magnetic-field heating of nanoparticles to remotely activate temperature-sensitive cation channels in cells. Superparamagnetic ferrite nanoparticles were targeted to specific proteins on the plasma membrane of cells expressing TRPV1, and heated by a radio-frequency magnetic field. Using fluorophores as molecular thermometers, we show that the induced temperature increase is highly localized. Thermal activation of the channels triggers action potentials in cultured neurons without observable toxic effects. This approach can be adapted to stimulate other cell types and, moreover, may be used to remotely manipulate other cellular machinery for novel therapeutics.
- Published
- 2010
- Full Text
- View/download PDF
31. Glycogen synthase kinase-3 beta mutagenesis identifies a common binding domain for GBP and Axin.
- Author
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Ferkey DM and Kimelman D
- Subjects
- Amino Acid Substitution, Animals, Axin Protein, Binding Sites, Blastomeres physiology, Calcium-Calmodulin-Dependent Protein Kinases chemistry, Glycogen Synthase Kinase 3, Glycogen Synthase Kinases, Mice, Models, Molecular, Mutagenesis, Site-Directed, Protein Binding, Protein Conformation, RNA, Messenger genetics, Xenopus embryology, Xenopus Proteins, Calcium-Calmodulin-Dependent Protein Kinases genetics, Calcium-Calmodulin-Dependent Protein Kinases metabolism, Carrier Proteins metabolism, Phosphoproteins metabolism, Plant Proteins, Proteins metabolism, Repressor Proteins
- Abstract
Glycogen synthase kinase-3 beta (GSK-3) is a key downstream target of Wnt signaling and is regulated by its interactions with activating and inhibitory proteins. We and others have shown that GSK-3 activity toward non-primed substrates is regulated in part through a competition between its activating (Axin) and inhibitory (GBP/FRAT) binding partners. Here we use a reverse two-hybrid screen to identify mutations in GSK-3 that alter binding to GBP and Axin. We find that these mutations overlap and propose that GBP and Axin compete for binding to the same region of GSK-3. We use these mutations to examine the ability of GSK-3 to block eye development in Xenopus embryos and suggest that GSK-3 regulates eye development through a non-Wnt pathway.
- Published
- 2002
- Full Text
- View/download PDF
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