The Dunce cAMP Phosphodiesterase PDE-4 Negatively Regulates Gαs-Dependent and Gαs-Independent cAMP Pools in the <i>Caenorhabditis elegans</i> Synaptic Signaling Network

Charlie, Nicole K.; Thomure, Angela M.; Schade, Michael A.; Miller, Kenneth G.

doi:10.1534/genetics.105.054007

Cited by 51 publications

(66 citation statements)

References 60 publications

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“…By RNAi and/or mutant analyses of the six PDE homologs in C. elegans (Charlie et al 2006), we found that the pde-2 gene is required for normal body size regulation (Figure 9 and data not shown). pde-2 encodes the closest C. elegans homolog of human cGMPdependent 39,59-cyclic phosphodiesterase 2A (PDE2A).…”

Section: Role Of the Extracellular Domain Of Gcy-12mentioning

confidence: 94%

The Importance of cGMP Signaling in Sensory Cilia for Body Size Regulation in Caenorhabditis elegans

et al. 2015

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The body size of Caenorhabditis elegans is thought to be controlled by sensory inputs because many mutants with sensory cilium structure defects exhibit small body size. The EGL-4 cGMP-dependent protein kinase acts in sensory neurons to reduce body size when animals fail to perceive sensory signals. In addition to body size control, EGL-4 regulates various other behavioral and developmental pathways, including those involved in the regulation of egg laying and chemotaxis behavior. Here we have identified gcy-12, which encodes a receptor-type guanylyl cyclase, as a gene involved in the sensory regulation of body size. Analyses with GFP fusion constructs showed that gcy-12 is expressed in several sensory neurons and localizes to sensory cilia. Genetic analyses indicated that GCY-12 acts upstream of EGL-4 in body size control but does not affect other EGL-4 functions. Our studies indicate that the function of the GCY-12 guanylyl cyclase is to provide cGMP to the EGL-4 cGMP-dependent kinase only for limited tasks including body size regulation. We also found that the PDE-2 cyclic nucleotide phosphodiesterase negatively regulates EGL-4 in controlling body size. Thus, the cGMP level is precisely controlled by GCY-12 and PDE-2 to determine body size through EGL-4, and the defects in the sensory cilium structure may disturb the balanced control of the cGMP level. The large number of guanylyl cyclases encoded in the C. elegans genome suggests that EGL-4 exerts pleiotropic effects by partnering with different guanylyl cyclases for different downstream functions.

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Section: Role Of the Extracellular Domain Of Gcy-12mentioning

confidence: 94%

The Importance of cGMP Signaling in Sensory Cilia for Body Size Regulation in Caenorhabditis elegans

et al. 2015

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“…The antibody was affinity purified using purified GST-UNC-29 fusion protein coupled to Aminolink Plus (Pierce) after presorbing 10 ml of serum to 10 mg of pure GST to remove GST-specific antibodies. GST and GST-UNC-29 fusions were produced using methods similar to those previously described (Charlie et al 2006b). To confirm the specificity of the UNC-29 antibody, we tested it on unc-29(x29) null mutants.…”

Section: Methodsmentioning

confidence: 99%

HID-1, a New Component of the Peptidergic Signaling Pathway

et al. 2011

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hid-1 was originally identified as a Caenorhabditis elegans gene encoding a novel conserved protein that regulates the decision to enter into the enduring dauer larval stage. We isolated a novel allele of hid-1 in a forward genetic screen for mutants mislocalizing RBF-1 rabphilin, a RAB-27 effector. Here we demonstrate that HID-1 functions in the nervous system to regulate neuromuscular signaling and in the intestine to regulate the defecation motor program. We further show that a conserved N-terminal myristoylated motif of both invertebrate and vertebrate HID-1 is essential for its association with intracellular membranes in nematodes and PC12 cells. C. elegans neuronal HID-1 resides on intracellular membranes in neuronal cell somas; however, the kinesin UNC-104 also transports HID-1 to synaptic regions. HID-1 accumulates in the axons of unc-13 and unc-31 mutants, suggesting it is associated with neurosecretory vesicles. Consistent with this, genetic studies place HID-1 in a peptidergic signaling pathway. Finally, a hid-1 null mutation reduces the levels of endogenous neuropeptides and alters the secretion of fluorescent-tagged cargos derived from neuronal and intestinal dense core vesicles (DCVs). Taken together, our findings indicate that HID-1 is a novel component of a DCV-based neurosecretory pathway and that it regulates one or more aspects of the biogenesis, maturation, or trafficking of DCVs.

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“…The density of synapses in the sublateral processes is low (Charlie et al 2006b), thus permitting resolution of what are likely to be individual DCVs.…”

Section: Figure S2mentioning

confidence: 99%

“…We prepared freeze-cracked 4% paraformaldehyde fixed worms for immunostaining as described (Charlie et al 2006a;Charlie et al 2006b), with the exception that we fixed worms for anti-FMRFamide staining for 16 h at 6° C . For immunostaining of FMRFamide neuropeptides, we co-immunostained N2 and unc-43(ce685) fixed animals using Rabbit antiFMRFamide antiserum (1/2000) and Goat anti-UNC-47 antiserum (1/12,000) ), using Donkey anti-Rabbit Dylight 550 and Donkey anti-Goat Dylight 488 secondary antibodies (Pierce).…”

Section: Immunostainingmentioning

confidence: 99%

“…For immunostaining of UNC-31 (CAPS), we triple-stained wild type and unc-43(ce685) fixed animals using Goat anti-UNC-31 (1/200; KM16C-3.1; Charlie et al, 2006a), Mouse anti-UNC-17 (1/5000 from monoclonal ascites fluid; , and Rabbit anti-UNC-10 (1/240; RIM5431; (Koushika et al 2001). We used previously described methods for immunostaining of fixed animals (Charlie et al 2006a;Charlie et al 2006b), with the exception that we only spread 750 animals per slide at the final mounting step.…”

Section: Immunostainingmentioning

confidence: 99%

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A Novel CaM Kinase II Pathway Controls the Location of Neuropeptide Release fromCaenorhabditis elegansMotor Neurons

Hoover

Edwards

et al. 2014

Genetics

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Neurons release neuropeptides via the regulated exocytosis of dense core vesicles (DCVs) to evoke or modulate behaviors. We found that Caenorhabditis elegans motor neurons send most of their DCVs to axons, leaving very few in the cell somas. How neurons maintain this skewed distribution and the extent to which it can be altered to control DCV numbers in axons or to drive release from somas for different behavioral impacts is unknown. Using a forward genetic screen, we identified loss-of-function mutations in UNC-43 (CaM kinase II) that reduce axonal DCV levels by 90% and cell soma/dendrite DCV levels by 80%, leaving small synaptic vesicles largely unaffected. Blocking regulated secretion in unc-43 mutants restored near wild-type axonal levels of DCVs. Time-lapse video microscopy showed no role for CaM kinase II in the transport of DCVs from cell somas to axons. In vivo secretion assays revealed that much of the missing neuropeptide in unc-43 mutants is secreted via a regulated secretory pathway requiring UNC-31 (CAPS) and . DCV cargo levels in unc-43 mutants are similarly low in cell somas and the axon initial segment, indicating that the secretion occurs prior to axonal transport. Genetic pathway analysis suggests that abnormal neuropeptide function contributes to the sluggish basal locomotion rate of unc-43 mutants. These results reveal a novel pathway controlling the location of DCV exocytosis and describe a major new function for CaM kinase II. BOTH neurons and neuroendocrine cells rely on the controlled release of neuropeptides via dense core vesicle (DCV) exocytosis to evoke or modulate behaviors (Scheller and Axel 1984;Kupfermann 1991;T. Liu et al. 2007;Li and Kim 2008). The DCVs in neuroendocrine and PC12 cells are much more abundant and accessible to biochemical and physiological experiments than those in neurons. For example, in chromaffin and pancreatic b-cells (both neuroendocrine cells), DCVs can number in the tens of thousands per cell and can occupy 31 and 12% of the cell volume, respectively (Dean 1973;Plattner et al. 1997). Exploiting these advantages, studies in PC12 and neuroendocrine cells have revealed that DCVs arise from a regulated secretory pathway. The pathway begins in the trans Golgi, where various sorting mechanisms cause regulated secretory proteins, such as neuropeptides and their processing enzymes, to coalesce into vesicles that bud from the trans Golgi to form immature DCVs. Additional sorting of non-DCV cargos away from DCV cargos occurs as DCVs mature through this pathway (Arvan and Castle 1998;Tooze et al. 2001;Borgonovo et al. 2006).DCVs must selectively retain and protect several distinct cargos that have different physical states as they mature. These include the neuropeptide core, which is thought to be in an aggregated state, the neuropeptide-processing enzymes PC-2 convertase and carboxypeptidase E, possibly soluble cargos, and transmembrane cargos.While neurons and neuroendocrine cells share this core pathway for DCV production, neurons have evolved additional...

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The Dunce cAMP Phosphodiesterase PDE-4 Negatively Regulates Gαs-Dependent and Gαs-Independent cAMP Pools in the Caenorhabditis elegans Synaptic Signaling Network

Cited by 51 publications

References 60 publications

The Importance of cGMP Signaling in Sensory Cilia for Body Size Regulation in Caenorhabditis elegans

The Importance of cGMP Signaling in Sensory Cilia for Body Size Regulation in Caenorhabditis elegans

HID-1, a New Component of the Peptidergic Signaling Pathway

A Novel CaM Kinase II Pathway Controls the Location of Neuropeptide Release fromCaenorhabditis elegansMotor Neurons

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