Tohru Kodama

State-dependent activity of neurons in the perifornical hypothalamic area during sleep and waking

Neurons containing orexins are located in the perifornical hypothalamic area and are considered to have a role in sleep-wake regulation. To examine how this area is involved in the regulation of sleep and wakefulness, we recorded neuronal activity in undrugged, head-restrained rats across sleep-waking cycles. Recordings were made in the perifornical hypothalamic area where orexin-immunoreactive neurons are distributed (PFH), and in the area dorsal to the PFH, including the zona incerta and subincertal nucleus (collectively referred to as ZI). The 40 neurons recorded from in the PFH were divided into five groups: (1) neurons most active during paradoxical sleep (PS, n=14, 35%), (2) neurons active during both waking (W) and PS (n=12, 30%), (3) neurons most active during W (n=7, 18%), (4) neurons most active during slow-wave sleep (SWS, n=3, 7.5%), and (5) neurons whose activity had no correlation with sleep-waking states (n=4, 10%). Of 30 neurons recorded from in the ZI, the corresponding numbers were 13 (43%), seven (23%), six (20%), three (10%), and one (3.3%). In both areas, neuronal activity fluctuated more during PS than during W. Waking-specific neurons (group 3) in the PFH generated action potentials with longer durations than those produced by other types of neurons. About half of the neurons in the PFH that were classified in groups 1, 2, and 3 increased their firing rate after the transition from one state to another, while higher percentages of neurons of groups 1 and 2 in the ZI than those in the PFH increased their firing rate prior to the state shift from SWS to PS. In these ZI neurons, however, the firing rate varied considerably at the state shift. These results suggest that the PFH and ZI are involved in the regulation of PS or W, especially the regulation of phasic events during PS or the maintenance of W. The ZI appears to be more closely involved than the PFH in the induction of PS or some phasic phenomena associated with PS.

Arousal effects of orexin-A correlate with GLU release from the locus coeruleus in rats

Although orexin was found to promote food intake, recent reports proposed its involvement in the regulation of vigilance. To study the mechanism of how orexin affects arousal, we analyzed glutamate (GLU) release from the locus coeruleus (LC) in rats after systemic injection of orexin-A. Baseline levels of orexin-A in the LC were significantly higher during the dark period than the light period. Intravenous administration of orexin-A increased GLU levels as well as orexin in the LC, simultaneously promoting wakefulness. These results suggest that increases in GLU release may reflect the arousal-inducing effects of orexin.

Intravenously administered hypocretin‐1 alters brain amino acid release: an in vivo microdialysis study in rats

We have reported that intravenous administration of hypocretin (Hcrt or orexin) reverses the symptoms of narcolepsy in genetically narcoleptic dogs. We have also reported that the onset of symptoms in canine genetic narcolepsy is accompanied by degenerative changes in forebrain regions, particularly the septal nucleus and amygdala. In the present in vivo microdialysis study we have investigated the effect of intravenous administration of Hcrt‐1 (orexin‐A) to anaesthetized rats on glutamate and GABA release in the amygdala, a region with moderate Hcrt innervation, and in the cerebellar cortex, a region with sparse or no Hcrt innervation. We found that intravenous Hcrt administration caused a marked (> 60 %) and sustained (> 50 min) increase in glutamate release within the amygdala, but no change in release in the cerebellar cortex. We did not detect a significant change in GABA release. When calcium‐free artificial cerebrospinal fluid was used as the microdialysis perfusate, Hcrt‐1 no longer produced an increase in glutamate release. Hcrt may act via the calcium‐dependent regulation of glutamate release in certain nuclei of the central nervous system.

Differential changes in glutamate concentration in the primate prefrontal cortex during spatial delayed alternation and sensory-guided tasks

Glutamate is a major neurotransmitter in the mammalian brain and glutamatergic neurotransmission in the frontal cortex is indicated to play important roles in cognitive operations. We previously examined changes in extracellular dopamine in the primate frontal cortex in cognitive tasks, and in this paper we extend this to glutamate. We employed, as cognitive tasks, a delayed alternation task where the animal must retain information in working memory, and a sensory-guided task in which there is no working memory requirement but there may be more sensory processing requirements. Using the in vivo microdialysis method, we examined changes in extracellular glutamate concentration in the dorsolateral, arcuate, orbitofrontal, and premotor areas of the primate frontal cortex. Compared to basal rest levels, we observed significant increases in glutamate concentration in dorsolateral and arcuate areas of the prefrontal cortex during the sensory-guided task, but did not find significant changes in any of the frontal areas examined during the delayed alternation task. When glutamate concentration was compared between the delayed alternation and sensory-guided tasks, difference was observed only in the dorsolateral prefrontal cortex, especially in the ventral lip area of the principal sulcus. The results indicate the importance of glutamate in processing sensory information but not in retaining information in working memory in the primate dorsolateral and arcuate prefrontal cortex. We also compared the concentration of glutamate and dopamine in the tasks. We found a double dissociation in the concentration of glutamate and dopamine in the dorsolateral area: there was an increase in glutamate but no change in dopamine during the sensory-guided task, whereas there was an increase in dopamine but no change in glutamate during the delayed alternation task. It is thus suggested that in the primate dorsolateral prefrontal cortex, increased glutamate tone without dopamine increase facilitates sensory-guided task performance, while increased dopamine tone without glutamate increase is beneficial for working memory task performance.

Nitric oxide from the laterodorsal tegmental neurons: its possible retrograde modulation on norepinephrine release from the axon terminal of the locus coeruleus neurons.

Nitric oxide released from the cholinergic neurons in the pons may play important roles in sleep-wake regulation. However, there are few reports demonstrating the mechanisms of nitric oxide release in the cholinergic neurons in the pons. The present study investigated the effects of drug delivery of N-methyl-D-aspartic acid on nitric oxide and the neurotransmitters released in the laterodorsal tegmental nucleus (LDT), one of the major cholinergic cell groups in the pons, in rats by in vivo microdialysis with a view to clarifying nitric oxide functions in the cholinergic system. The application of N-methyl-D-aspartic acid (1 mM) into the LDT induced a significant increase in NO(2)and NO(3) for 40 min (P<0.001). Furthermore the same dose of N-methyl-D-aspartic acid induced a significant increase in cyclic GMP for 30 min (P<0.05), as well as in acetylcholine (P<0.001) and norepinephrine for 15 min (P<0.001). 3-(4-Morpholinyl)-sydonone imine hydrochloride (a nitric oxide donor, 5 mM) also induced significant increase in norepinephrine (P<0.05). Pretreatment with 1 mM 2-amino-5-phosphonopentanoic acid (an antagonist of N-methyl-D-aspartic acid receptor) prevented the N-methyl-D-aspartic acid-induced increase in cyclic GMP (P<0.01), acetylcholine and norepinephrine (P<0.01), while that with 1 mM N(G)-nitro-L-arginine (an inhibitor of nitric oxide synthase) prevented the increase in cyclic GMP (P<0.01) and norepinephrine (P<0.01) but not in acetylcholine. These results suggested that nitric oxide release in the LDT induced by activation of the N-methyl-D-aspartic acid receptor on the cholinergic neurons of the LDT, then through the cyclic GMP system, facilitates norepinephrine release from the terminals of noradrenergic neurons in the locus coeruleus. Based on these findings, we propose a possible role of nitric oxide in the LDT is as a retrograde regulator of norepinephrine release from the locus coeruleus.

High Fos expression during the active phase in orexin neurons of a diurnal rodent, Tamias sibiricus barberi

To investigate whether a diurnal animal possesses the orexinergic system implicating vigilance and behavior, we examined Fos immunoreactivity (IR) in orexinergic neurons of Korean chipmunks raised under 12h light-dark cycles. Brain tissue, collected at four different zeitgeber times (ZT), was double-labeled with Fos and orexin-A antibodies. There was no difference in the number of orexin-IR neurons in the hypothalamus across all ZTs. However, more orexin-IR neurons expressing Fos-IR were found at ZTs 3 and 9 than ZTs 15 and 21. The results demonstrate circadian variations in the activation of orexin neurons corresponding with locomotor cycles, similarly seen in nocturnal rodents.

Release of neurotransmitters in the monkey frontal cortex is related to level of attention

Abstract Attention is reported to be maintained by monoamines, acetylcholine and amino acids systems. Changes in the releases of these neurotransmitters during the three stages comprising quiet wake (QW) and two arousal states (AW), which are activated from different sources, were investigated. Norepinephrine releases during AW were significantly higher than that during QW. Conversely, the levels of acetylcholine and serotonin that were released did not change significantly among these three stages. The interesting observation was the dissociation of the increase between glutamate and dopamine releases in the two AW states. These results indicate that attention level is related to the amount of norepinephrine release, and that attention quality is related to the interaction between dopamine and glutamate releases.