RESUMO
Behavioral persistency reflects internal brain states, which are the foundations of multiple brain functions. However, experimental paradigms enabling genetic analyses of behavioral persistency and its associated brain functions have been limited. Here, we report novel persistent behavioral responses caused by electric stimuli in the nematode Caenorhabditis elegans. When the animals on bacterial food are stimulated by alternating current, their movement speed suddenly increases 2- to 3-fold, persisting for more than 1 minute even after a 5-second stimulation. Genetic analyses reveal that voltage-gated channels in the neurons are required for the response, possibly as the sensors, and neuropeptide signaling regulates the duration of the persistent response. Additional behavioral analyses implicate that the animal's response to electric shock is scalable and has a negative valence. These properties, along with persistence, have been recently regarded as essential features of emotion, suggesting that C. elegans response to electric shock may reflect a form of emotion, akin to fear.
Assuntos
Proteínas de Caenorhabditis elegans , Caenorhabditis elegans , Animais , Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/genética , Neurônios , Movimento , Transdução de Sinais/fisiologiaRESUMO
The precipice response in Caenorhabditis elegans is a little-understood phenomenon in which worms move rapidly away from edges. We hypothesized that mechanosensation underlies the precipice response and that mechanosensory mutants would exhibit the precipice response less often than N2 wild type worms. We found that mec-3 mutants, with severe loss of mechanosensation, exhibited the precipice response at a lower rate than N2, but mec-10 and trp-4 mutants, with partial loss of response to mechanical stimuli, responded at a similar rate to N2. These results provide a characterization of the precipice response and implicate a role for mechanosensation in this behavior.
RESUMO
Leaf stomata close in response to high carbon dioxide levels and open at low CO(2). CO(2) concentrations in leaves are altered by daily dark/light cycles, as well as the continuing rise in atmospheric CO(2). Relative to abscisic acid and blue light signaling, little is known about the molecular, cellular, and genetic mechanisms of CO(2) signaling in guard cells. Interestingly, we report that repetitive Ca(2+) transients were observed during the stomatal opening stimulus, low [CO(2)]. Furthermore, low/high [CO(2)] transitions modulated the cytosolic Ca(2+) transient pattern in Arabidopsis guard cells (Landsberg erecta). Inhibition of cytosolic Ca(2+) transients, achieved by loading guard cells with the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid and not adding external Ca(2+), attenuated both high CO(2)-induced stomatal closing and low CO(2)-induced stomatal opening, and also revealed a Ca(2+)-independent phase of the CO(2) response. Furthermore, the mutant, growth controlled by abscisic acid (gca2) shows impairment in [CO(2)] modulation of the cytosolic Ca(2+) transient rate and strong impairment in high CO(2)-induced stomatal closing. Our findings provide insights into guard cell CO(2) signaling mechanisms, reveal Ca(2+)-independent events, and demonstrate that calcium elevations can participate in opposed signaling events during stomatal opening and closing. A model is proposed in which CO(2) concentrations prime Ca(2+) sensors, which could mediate specificity in Ca(2+) signaling.
Assuntos
Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/citologia , Arabidopsis/metabolismo , Cálcio/metabolismo , Dióxido de Carbono/metabolismo , Transdução de Sinais , Ácido Abscísico/farmacologia , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Dióxido de Carbono/farmacologia , Citosol/metabolismo , Mutação/genéticaRESUMO
Abscisic acid (ABA) regulates developmental processes and abiotic stress responses in plants. We recently characterized a new Arabidopsis mutant, abh1, which shows ABA-hypersensitive regulation of seed germination, stomatal closing, and cytosolic calcium increases in guard cells (V. Hugouvieux, J.M. Kwak, J.I. Schroeder [2001] Cell 106: 477-487). ABH1 encodes the large subunit of a dimeric Arabidopsis mRNA cap-binding complex and in expression profiling experiments was shown to affect mRNA levels of a subset of genes. Here, we show that the dimeric ABH1 and AtCBP20 subunits are ubiquitously expressed. Whole-plant growth phenotypes of abh1 are described and properties of ABH1 in guard cells are further analyzed. Complemented abh1 lines expressing a green fluorescent protein-ABH1 fusion protein demonstrate that ABH1 mainly localizes in guard cell nuclei. Stomatal apertures were smaller in abh1 compared with wild type (WT) when plants were grown at 40% humidity, and similar at 95% humidity. Correlated with stomatal apertures from plants grown at 40% humidity, slow anion channel currents were enhanced and inward potassium channel currents were decreased in abh1 guard cells compared with WT. Gas exchange measurements showed similar primary humidity responses in abh1 and WT, which together with results from abh1/abi1-1 double-mutant analyses suggest that abh1 shows enhanced sensitivity to endogenous ABA. Double-mutant analyses of the ABA-hypersensitive signaling mutants, era1-2 and abh1, showed complex genetic interactions, suggesting that ABH1 and ERA1 do not modulate the same negative regulator in ABA signaling. Mutations in the RNA-binding protein sad1 showed hypersensitive ABA-induced stomatal closing, whereas hyl1 did not affect this response. These data provide evidence for the model that the mRNA-processing proteins ABH1 and SAD1 function as negative regulators in guard cell ABA signaling.
Assuntos
Ácido Abscísico/farmacologia , Proteínas de Arabidopsis/genética , Canais Iônicos/genética , Proteínas de Ligação ao Cap de RNA/genética , Transdução de Sinais/genética , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Canais Iônicos/metabolismo , Potenciais da Membrana/fisiologia , Mutação , Fosfoproteínas Fosfatases/efeitos dos fármacos , Fosfoproteínas Fosfatases/genética , Fosfoproteínas Fosfatases/fisiologia , Reguladores de Crescimento de Plantas/farmacologia , Canais de Potássio/genética , Canais de Potássio/metabolismo , Proteínas de Ligação ao Cap de RNA/efeitos dos fármacos , Proteínas de Ligação ao Cap de RNA/metabolismo , Proteínas de Ligação ao Cap de RNA/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Nuclear/genética , RNA Nuclear/metabolismo , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologiaRESUMO
A variety of stimuli, such as abscisic acid (ABA), reactive oxygen species (ROS), and elicitors of plant defense reactions, have been shown to induce stomatal closure. Our study addresses commonalities in the signaling pathways that these stimuli trigger. A recent report showed that both ABA and ROS stimulate an NADPH-dependent, hyperpolarization-activated Ca(2+) influx current in Arabidopsis guard cells termed "I(Ca)" (Z.M. Pei, Y. Murata, G. Benning, S. Thomine, B. Klüsener, G.J. Allen, E. Grill, J.I. Schroeder, Nature [2002] 406: 731-734). We found that yeast (Saccharomyces cerevisiae) elicitor and chitosan, both elicitors of plant defense responses, also activate this current and activation requires cytosolic NAD(P)H. These elicitors also induced elevations in the concentration of free cytosolic calcium ([Ca(2+)](cyt)) and stomatal closure in guard cells. ABA and ROS elicited [Ca(2+)](cyt) oscillations in guard cells only when extracellular Ca(2+) was present. In a 5 mM KCl extracellular buffer, 45% of guard cells exhibited spontaneous [Ca(2+)](cyt) oscillations that differed in their kinetic properties from ABA-induced Ca(2+) increases. These spontaneous [Ca(2+)](cyt) oscillations also required the availability of extracellular Ca(2+) and depended on the extracellular potassium concentration. Interestingly, when ABA was applied to spontaneously oscillating cells, ABA caused cessation of [Ca(2+)](cyt) elevations in 62 of 101 cells, revealing a new mode of ABA signaling. These data show that fungal elicitors activate a shared branch with ABA in the stress signal transduction pathway in guard cells that activates plasma membrane I(Ca) channels and support a requirement for extracellular Ca(2+) for elicitor and ABA signaling, as well as for cellular [Ca(2+)](cyt) oscillation maintenance.