Makoto Araki

Behavioral and Electrophysiological Experiments Suggest That the Antennular Outer Flagellum Is the Site of Pheromone Reception in the Male Helmet Crab Telmessus cheiragonus

Sexually competent females of Telmessus cheiragonus (helmet crab) release two pheromones that elicit grasping and copulation behaviors in males (Kamio et al., 2000, 2002, 2003). Our study aimed to use behavioral and electrophysiological techniques to identify the site of reception of these sex pheromones. In behavioral experiments, either the inner or the outer flagella of the antennules were ablated bilaterally from male crabs, and responses of male crabs to female odor were examined. When the inner flagella were surgically ablated, the sexual response (i.e., grasping and copulation behavior) of male crabs was not significantly changed relative to control animals that had their second antennae ablated. In contrast, the sexual response was significantly reduced when the outer flagella of the antennules were ablated, suggesting that the outer flagellum is the receptor organ that detects the sex pheromones. In electrophysiological experiments, urine, which in females contains the pheromone that elicits grasping behavior by males but does not contain the pheromone eliciting copulation, whose release site is not known, was tested. Female and male urine as well as shrimp extract evoked phasic responses of chemosensory afferents innervating aesthetasc sensilla on the outer flagellum of male crabs. The response of the afferents had significantly higher magnitude and lower threshold when female urine was applied. Thus, behavioral and electrophysiological observations suggest that in male helmet crabs, the outer flagellum of the antennule is the chemosensory organ that detects female sex pheromone.

Social status-dependent modulation of LG-flip habituation in the crayfish.

SUMMARY Strong stimuli applied to the tailfan of the crayfish Procambarus clarkii evoked lateral giant interneurone (LG)-mediated tailflips. When the sensory stimulus was applied repeatedly, the response of the LG habituated until it failed to give rise to a spike. We found that this LG-flip habituation was dependent on social status. With a short interstimulus interval of 5 s, the rate of habituation of the LG in both socially dominant and subordinate crayfish was lower than that in socially isolated animals. By contrast, with a long interstimulus interval of 60 s, the rate of habituation of subordinate animals was lower than that of both socially isolated and dominant animals. The excitability of the LGs following habituation was also dependent on social status. Following habituation, the spike response of LGs recovered within several minutes; however, they showed significant depression with a decrease in excitability. With a 5 or 60 s interstimulus interval, subordinate animals showed longer delays of depression compared with dominant animals. A decrease in the rate of habituation and a delay of depression in subordinate crayfish would be advantageous for maintaining an active escape response to evade repeated attacks of dominant animals and a reduced learning ability to adapt to social status.

IP3-mediated octopamine-induced synaptic enhancement of crayfish LG neurons

The biogenic amines, octopamine and serotonin, modulate the synaptic activity of the lateral giant interneuron (LG) circuitry of the crayfish escape behavior. Bath application of both octopamine and serotonin enhances the synaptic responses of LG to sensory stimulation. We have shown previously (Araki et al. J Neurophysiol 94:2644–2652, 2005) that a serotonin-induced enhancement of the LG response was mediated by an increase in cAMP levels following activation of adenylate cyclase; however, octopamine acts independently. Here, we clarify how octopamine enhances the LG response during sensory stimulation using physiological and pharmacological analyses. When phospholipase C inhibitor U-73122 was directly injected into the LG before biogenic amine application, it abolished the enhancing effect of octopamine on direct sensory input to the LG, but did not block indirect input via sensory interneurons or the effect of serotonin. Direct injection of IP3, and its analogue adenophostin A, into the LG increased the synaptic response of the LG to sensory stimulation. Thus, IP3 mediates octopamine-induced synaptic enhancement of the LG, but serotonin acts independently. These results indicate that both octopamine and serotonin enhance the synaptic responses of the LG to sensory stimulation, but that they activate two different signaling cascades in the LG.

Distribution of glutamatergic immunoreactive neurons in the terminal abdominal ganglion of the crayfish

Using an antiserum directed against glutamate, we have analyzed the distribution of glutamate‐like immunoreactive neurons in the terminal abdominal ganglion of the crayfish Procambarus clarkii. Approximately 160 central neurons (157 ± 8; mean ± SEM, n = 8) showed positive glutamate‐like immunoreactivity, which represents approximately 25% of the total number of neurons in the terminal ganglion. Using a combination of intracellular staining with the marker Lucifer yellow and immunocytochemical staining has shown that most excitatory motor neurons are glutamatergic and that glutamate acts as an excitatory transmitter at peripheral neuromuscular junctions. Seven of 10 identified spiking local interneurons and only 2 of 19 identified ascending interneurons, showed positive immunoreactivity. Our observation that inhibitory spiking interneurons were immunopositive, whereas excitatory ascending interneurons were immunonegative, indicates that glutamate is likely to act as an inhibitory neurotransmitter within the central nervous system. Local pressure injection of L‐glutamate into the neuropil of the ganglion caused a hyperpolarization of the membrane potentials of many interneurons. γ‐Aminobutyric acid (GABA)ergic posterolateral nonspiking interneurons and the bilateral nonspiking interneuron LDS showed no glutamate‐like immunoreactivity, whereas non‐GABAergic anterolateral III nonspiking interneurons showed glutamate‐like immunoreactivity. Thus, not only GABA but also glutamate are used in parallel as inhibitory neurotransmitters at central synapses. J. Comp. Neurol. 474:123–135, 2004.

Lateral giant fibre activation of exopodite motor neurones in the crayfish tailfan

Abstract. The uropods of decapod crustaceans play a major role in the production of thrust during escape swimming. Here we analyse the output connections of a pair of giant interneurones, that mediate and co-ordinate swimming tail flips, on motor neurones that control the exopodite muscles of the uropods. The lateral giants make short latency output connections with phasic uropod motor neurones, including the productor, the lateral abductor and adductor exopodite motor neurones that we have identified both physiologically and anatomically. On the other hand, tonic motor neurones, including the ventral abductor and reductor exopodite motor neurones, receive no input from the lateral giants. We show that there is no simple reciprocal activation of the phasic opener (lateral abductor) and closer (adductor) motor neurones of the exopodite, but instead both phasic motor neurones are activated in parallel with the productor motor neurone during a tail flip. Our results show that the neuronal pathways activating the tonic and phasic motor neurones of the exopodite are apparently independent, with phasic motor neurones being activated during escape movements and tonic motor neurones being activated during slow postural movements.

Effects of nitric oxide on proprioceptive signaling

Abstract We have analysed the effects of the neuromodulator nitric oxide (NO) on proprioceptive information processing by ascending intersegmental interneurons that form part of the local circuits within the terminal abdominal ganglion of the crayfish. NO modulates the synaptic inputs to ascending interneurons, enhancing the amplitude of class I interneurons and reducing the amplitude of class II interneurons. Repetitive proprioceptive stimulation leads to rapid depression in a specific set of identified interneurons but not in others. Bath application of a nitric oxide scavenger, PTIO, causes a significant decrease in the rate of depression of the interneurons showing a rapid depression, independent of interneuron class, but has no effect on the dynamic responses of the interneurons that show little initial depression. These results indicate that NO exerts multiple effects at the very first stage of synaptic integration in local circuits.