Biomedical subjects

Toshiki Nagayama

Publications and source records attributed to Toshiki Nagayama.

8 recordsLinked to original sources

PubMed2005

Decrease in excitability of LG following habituation of the crayfish escape reaction.

Crayfish escapes from threatening stimuli to the abdomen by tailflipping upwards and forwards. This lateral giant (LG)-mediated escape reaction habituates readily upon repetitive sensory stimulation. Using an isolated abdominal nerve cord preparation, we have analyzed the change in LG activity by applying additional sensory stimulation after different periods following habituation to characterize the retention of LG habituation. Results show that the LG mediated response habituates more quickly, but the retention time is shorter, as repetitive sensory stimulation is applied at progressively shorter inter-stimulus time intervals. The spike response of LG recovers quickly, within several minutes after habituation, but they fail to spike when an additional stimulus is applied after specific long periods following habituation. The critical period of the delay for this decrease in excitability of LG is dependent on the inter-stimulus time interval of the initial repetitive stimulus. As the inter-stimulus interval became longer, the delay needed for decrease in excitability became shorter. Furthermore, the local injection of 10(-6) mol l-1 octopamine into the neuropil just following habituation promotes the achievement of decrease in excitability. No effects were observed when 10(-6) mol l-1 serotonin and tyramine were injected. These results suggested octopamine promotes decrease in excitability of LG following habituation.

Animals↗

PubMed2005

GABAergic and glutamatergic inhibition of nonspiking local interneurons in the terminal abdominal ganglion of the crayfish.

Nonspiking local interneurons in the terminal abdominal ganglion of the crayfish Procambarus clarkii receive inhibitory inputs from mainly glutamatergic spiking local interneurons and GABAergic nonspiking interneurons. In this study, the inhibitory responses of nonspiking interneurons to local application of glutamate and GABA into the neuropil were compared. Glutamate and GABA injection mediated the hyperpolarization of the nonspiking interneurons with an increase in membrane conductance. The glutamate-mediated membrane hyperpolarization was reversed by injection of 1 or 2 nA hyperpolarizing current. By contrast, more than 3 nA hyperpolarizing current was frequently necessary to reverse the GABA-mediated hyperpolarization. Bath application of a chloride channel blocker, 50 microM picrotoxin (PTX), reduced the glutamate-mediated hyperpolarization, but had no effect on the GABA-mediated hyperpolarization. The GABA-mediated hyperpolarization was not consistently affected by bath application of low chloride solution. These results suggest that the glutamate-mediated inhibition was related to the gating of a Cl(-) conductance, while the GABA-mediated inhibition was not. Electrical stimulation of sensory afferents innervating the exopodite elicited ipsps in uropod opener motor neurons. These sensory-evoked ipsps were also PTX-insensitive, suggesting GABAergic nonspiking interneurons could be the predominant premotor elements in organizing the uropod motor control system.

Animals↗

PubMed2005

Cyclic AMP mediates serotonin-induced synaptic enhancement of lateral giant interneuron of the crayfish.

The lateral giant (LG)-mediated escape behavior of the crayfish habituates readily on repetitive sensory stimulation. Recent studies suggested that the biogenic amines serotonin and octopamine modulate the time course of recovery and/or re-depression of the LG response after habituation. However, little is known of how serotonin and octopamine effect LG habituation and what second-messenger cascades they may activate. To investigate the effect of biogenic amines on LG habituation, serotonin and octopamine were superfused before presenting repetitive sensory stimulation. Serotonin and octopamine increased the number of stimuli needed to habituate the LG response. Their effects were mimicked by mixed application of a cAMP analogue [8-(4-chlorophenylthio)-cAMP (CPT-cAMP)] and a phosphodiesterase inhibitor [3-isobutyl-1-methylxanthine (IBMX)] but not by a cGMP analogue (8-bromoguanosine 3',5'-cyclic monophosphate). Perfusion of the adenylate cyclase inhibitor (SQ22536) abolished the effect of serotonin but not that of octopamine. To investigate the site of action of each biogenic amines in the neural circuit meditating LG escape, the effect of drugs on directly and indirectly elicited postsynaptic potentials in LG was investigated. Serotonin, octopamine, and a mixture of CPT-cAMP and IBMX increased both the direct and indirect synaptic inputs. Simultaneous application of SQ22536 abolished the effect of serotonin on both inputs but did not block the effect of octopamine. Direct injection of the cAMP analogue (Sp-isomer of adenosine-3',5'-cyclic monophosphorothioate) into LG increased both the direct and indirect inputs to LG. These results indicate that serotonin mediates an increase in cAMP levels in LG, but octopamine acts independently of cAMP and cGMP.

1-Methyl-3-isobutylxanthine↗

PubMed2005

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.

Animals↗

PubMed2004

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. gamma-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.

Abdomen↗

PubMed2004

Nitric oxide modulates local reflexes of the tailfan of the crayfish.

Electrical stimulation of sensory neurons that innervate receptors on the tailfan of crayfish evokes a reflex response of motor neurons that produce movements of the blades of the tailfan, the uropods. We analyzed the modulatory effects of nitric oxide (NO) on the spike frequency of the reflex response. Bath application of L-arginine and SNAP, which elevate endogenous and exogenous NO levels, increased the frequency of the evoked response, whereas the application of L-NAME and PTIO, which reduce NO levels, decreased the frequency of the response. To determine through what pathway and target NO exerted these effects we bath applied ODQ, an inhibitor of soluble guanylyl cyclase (sGC), which decreased the frequency of response, and 8-br-cGMP, which increased the spike frequency of response. To provide further evidence that NO acts via sGC, we elevated NO levels with L-arginine while simultaneously inhibiting sGC with ODQ. This application reduced the response to control levels, indicating that NO in the terminal ganglion of crayfish acts via sGC to modulate cGMP levels, which in turn regulate the responses of the uropod motor neurons.

Analysis of Variance↗

PubMed2004

Effects of nitric oxide on proprioceptive signaling.

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.

Animals↗

PubMed2002

Serotonergic modulation of nonspiking local interneurones in the terminal abdominal ganglion of the crayfish.

The modulatory effect of serotonin on local circuit neurones forming the uropod motor control system of the crayfish Procambarus clarkii Girard was analysed electrophysiologically. Bath application of 10 micro mol l(-1) serotonin caused a decrease in the tonic spike activity of the exopodite reductor motor neurone. The inhibitory effect of serotonin on the motor neurone was dose-dependent and its spike discharge was completely suppressed for long periods by 1 mmol l(-1) serotonin perfusion. Nonspiking local interneurones in the terminal abdominal ganglion showed either a membrane depolarization (N=6) or hyperpolarization (N=9) of 10-30 mV in amplitude when 100 micro mol l(-1) serotonin was perfused for 3-5 min. By contrast, spiking local interneurones and intersegmental ascending interneurones showed no observable excitatory responses to the perfusion of serotonin but instead some showed a small membrane hyperpolarization of 2-5 mV. These results indicate that the nonspiking interneurones could contribute substantially to the level of tonic excitation of the uropod motor neurones. Sensory stimulation elicited depolarizing or hyperpolarizing potentials in the nonspiking interneurones and excitatory postsynaptic potentials (EPSPs) and spikes in the spiking interneurones. The sensory responses of spiking interneurones increased during bath application of serotonin and were reduced after 20-30 min of washing with normal saline. In the nonspiking interneurones, the amplitude of both depolarizing and hyperpolarizing potentials increased without any direct correlation with the serotonin-mediated potential change. This effect of serotonin was long-lasting and continued to enhance the responses of the nonspiking interneurones after washing. This postserotonin enhancement persisted for over 1 h.

Abdomen↗