Strongly alkaline pH avoidance mediated by ASH sensory neurons in C. elegans.

High pH is a noxious stimulus to animals, and their ability to avoid dangerously alkaline pH is critical for survival. However, the means by which they sense high pH has not been determined. The nematode Caenorhabditis elegans (C. elegans) avoids environmental pH above 10.5. In contrast, C. elegans mutants with structurally, developmentally, and/or functionally abnormal sensory cilia fail to avoid high pH, suggesting that sensory neurons in the cilia participate in sensing. Genetic rescue of the mutants indicates that ASH polymodal sensory neurons play a vital role in the process. Consistently, specific laser ablation of ASH neurons made animals insensitive to high pH. Furthermore, avoidance assays of other mutants also indicated that transient receptor potential vanilloid type (TRPV) ion channels encoded by osm-9 and ocr-2 are involved in sensing. Indeed, genetic rescue of osm-9 mutants by specifically expressing OSM-9 in ASH showed that TRPV channels play an essential role in sensing of high pH. Ca(2+) imaging in vivo also revealed that ASH neurons were activated by high pH stimulation, but ASH of osm-9 or ocr-2 mutants were not. These results demonstrate that in C. elegans, high pH is sensed by ASH nociceptors through opening of OSM-9/OCR-2 TRPV channels.

Decision making in C. elegans chemotaxis to alkaline pH: Competition between two sensory neurons, ASEL and ASH.

Monitoring of environmental and tissue pH is critical for animal survival. The nematode, Caenorhabditis elegans (C. elegans), is attracted to mildly alkaline pH, but avoids strongly alkaline pH. However, little is known about how the behavioral switching or decision making occurs. Genetic dissection and Ca(2+) imaging have previously demonstrated that ASEL and ASH are the major sensory neurons responsible for attraction and repulsion, respectively. Here we report that unlike C. elegans wild type, mutants deficient in ASEL or ASH were repelled by mildly alkaline pH, or were attracted to strongly alkaline pH, respectively. These results suggest that signals through ASEL and ASH compete to determine the animal's alkaline-pH chemotaxis. Furthermore, mutants with 2 ASEL neurons were more efficiently attracted to mildly alkaline pH than the wild type with a single ASEL neuron, indicating that higher activity of ASEL induces stronger attraction to mildly alkaline pH. This stronger attraction was overridden by normal activity of ASH, suggesting that ASH-mediated avoidance dominates ASEL-mediated attraction. Thus, C. elegans chemotactic behaviors to alkaline pH seems to be determined by signal strengths from the sensory neurons ASEL and ASH, and the behavior decision making seems to be the result of competition between the 2 sensory neurons.

A G-protein α subunit, GOA-1, plays a role in C. elegans avoidance behavior of strongly alkaline pH.

The ability of animals to avoid strongly alkaline pH is critical for survival. However, the means by which they sense high pH has not been determined. We have previously found that the nematode Caenorhabditis elegans (C. elegans) avoids environmental pH above 10.5. Detection involves ASH nociceptive neurons as the major sensors. Upon stimulation, transient receptor potential vanilloid-type (TRPV) ion channels encoded by osm-9 and ocr-2 play an essential role in Ca(2+) entry into ASH. Here we report that C. elegans mutants deficient in a G-protein α subunit, GOA-1, failed to avoid strongly alkaline pH with normal Ca(2+) influx into ASH. These results suggest that GOA-1 regulates signal transmission downstream of Ca(2+) influx through OSM-9/OCR-2 TRPV channels in ASH.

A ubiquitin E2 variant protein acts in axon termination and synaptogenesis in Caenorhabditis elegans.

In the developing nervous system, cohorts of events regulate the precise patterning of axons and formation of synapses between presynaptic neurons and their targets. The conserved PHR proteins play important roles in many aspects of axon and synapse development from C. elegans to mammals. The PHR proteins act as E3 ubiquitin ligases for the dual-leucine-zipper-bearing MAP kinase kinase kinase (DLK MAPKKK) to regulate the signal transduction cascade. In C. elegans, loss-of-function of the PHR protein RPM-1 (Regulator of Presynaptic Morphology-1) results in fewer synapses, disorganized presynaptic architecture, and axon overextension. Inactivation of the DLK-1 pathway suppresses these defects. By characterizing additional genetic suppressors of rpm-1, we present here a new member of the DLK-1 pathway, UEV-3, an E2 ubiquitin-conjugating enzyme variant. We show that uev-3 acts cell autonomously in neurons, despite its ubiquitous expression. Our genetic epistasis analysis supports a conclusion that uev-3 acts downstream of the MAPKK mkk-4 and upstream of the MAPKAPK mak-2. UEV-3 can interact with the p38 MAPK PMK-3. We postulate that UEV-3 may provide additional specificity in the DLK-1 pathway by contributing to activation of PMK-3 or limiting the substrates accessible to PMK-3.

All EGF(ErbB) receptors have preformed homo- and heterodimeric structures in living cells.

The epidermal growth factor receptor (EGFR) family of receptor tyrosine kinases, also known as ErbB or HER, plays crucial roles in the development of multicellular organisms. Mutations and over-expression of the ErbB receptors have been implicated in a variety of human cancers. It is widely thought that the ErbB receptors are located in the plasma membrane, and that ligand binding to the monomeric form of the receptors induces its dimeric form for activation. However, it still remains controversial whether prior to ligand binding the receptors exist as monomers or dimers on the cell surface. Using bimolecular fluorescence complementation (BiFC) assays in the present study, we demonstrate that in the absence of bound ligand, all the ErbB family members have preformed, yet inactive, homo- and heterodimers on the cell surface, except for ErbB3 homodimers and heterodimers with cleavable ErbB4, which exist primarily in the nucleus. BiFC assays of the dimerization have also suggested that the ligand-independent dimerization of the ErbB receptors occurs in the endoplasmic reticulum (ER) before newly synthesized receptor molecules reach the cell surface. Based on BiFC and mammalian two-hybrid assays, it is apparent that the intracellular domains of the receptors are responsible for the spontaneous dimer formation. These provide new insights into an understanding of transmembrane signal transduction mediated by the ErbB family members, and are relevant to the development of anti-cancer drugs.