Shyh Yuh Liou

Influence of excitatory amino acid receptor antagonists and of baclofen on synaptic transmission in the optic nerve to the suprachiasmatic nucleus in slices of rat hypothalamus.

Electrical stimulation of the optic nerve evoked two positive waves with short latency, followed by a large negative wave in the suprachiasmatic nucleus of slices of hypothalamus of the rat. The latency to peak of the two positive waves and the large negative wave were 2.7 +/- 0.1, 6.1 +/- 0.1 and 10.3 +/- 0.5 msec, respectively. Only the large negative wave disappeared in low calcium Ca2+-high magnesium (Mg2+) Krebs solution and with the addition of tetrodotoxin (1 microM) all the waves disappeared. Baclofen inhibited the large negative wave in a dose-dependent manner but not the two positive waves. Excitatory amino acid antagonists also inhibited only the large negative wave, i.e. it was reduced to about 70% by 1 mM glutamic acid diethyl ester and to about 50% by both 1 mM 2-amino-4-phosphonobutyric acid and 1 mM DL-2-amino adipic acid. All waves were unaffected by 0.1 mM atropine, hexamethonium and curare. These results indicate that two positive waves, induced by stimulation of the optic nerve are attributed to nerve conduction and the large negative wave to the neurons of the suprachiasmatic nucleus, and that the neuronal pathway from the optic nerve to the suprachiasmatic nucleus may include aspartate and/or glutamate as an excitatory neurotransmitter.

Different effects of amino acids, acetylcholine and monoamines on neuronal activity of suprachiasmatic nucleus in rat pups and adults

A study was undertaken of the effects of iontophoretically applied amino acids, acetylcholine and monoamines on suprachiasmatic nucleus (SCN) neurons of rat pups, at the 11th postnatal day, using a hypothalamic slice preparation. GABA and taurine inhibited 97 and 42% of SCN neurons, respectively, while glycine had no effect. Glutamate excited 68% of them. These effects were similar to those in adult rats. Serotonin and noradrenaline inhibited 6 and 10% of SCN neurons, respectively, in rat pups, whereas 32 and 26% of them were inhibited in adult rats. Acetylcholine excited 7% but inhibited 26% of SCN neurons in rat pups and these values were almost the same as those of adult rats. These results reveal that acetylcholine and amino acids regulate SCN neurons even in rat pups, whereas monoamines play an important regulatory role in SCN activity in adult rats only.

Inhibitory action of insulin on suprachiasmatic nucleus neurons in rat hypothalamic slice preparations

In order to elucidate a role of insulin on neurons in the suprachiasmatic nucleus (SCN) involved in the generation of circadian feeding behavior, we examined the effect of insulin on SCN neuronal activity at in vitro condition. Bath application of insulin (1-100 microU) mainly inhibited the SCN neuronal activity, and this inhibitory effect was still observed in a Ca2+-free Krebs solution. The present result strongly suggests that insulin directly inhibits SCN neurons and may explain an increase in food intake by insulin infusion into the SCN during the light period.

Effect of monoamines on field potentials in the suprachiasmatic nucleus of slices of hypothalamus of the rat evoked by stimulation of the optic nerve

The effects of the application of serotonin, histamine, noradrenaline and dopamine to the bath on field potentials in the suprachiasmatic nucleus evoked by stimulation of the optic nerve were studied using a hypothalamic slice. Stimulation of the contralateral optic nerve evoked fast positive and late large negative waves in the suprachiasmatic nucleus. The monoamines produced a dose dependent suppression of the amplitude of the negative wave but did not affect that of the positive waves, and the order of potency was serotonin greater than noradrenaline greater than dopamine greater than or equal to histamine. The negative wave was suppressed by phenylephrine (0.1-10 microM) in a dose-dependent manner, whereas it was unaffected by isoproterenol (0.1-10 microM). The suppression of the negative wave produced by the application of histamine and noradrenaline was antagonized by the H1-receptor antagonist, diphenhydramine and the alpha 1-receptor antagonists, phenoxybenzamine and phentolamine. Therefore, the suppression of the negative wave by histamine and noradrenaline was mediated by the H1-receptor and alpha 1-receptor, respectively. The present study indicates that monoamines may play an inhibitory role in the regulation of neurotransmission in the retinohypothalamic pathway to the suprachiasmatic nucleus.

The role of calcium ions in circadian rhythm of suprachiasmatic nucleus neuron activity in rat hypothalamic slices

The role of calcium ions in maintaining the circadian rhythm of suprachiasmatic nucleus (SCN) neuron activity was investigated using rat hypothalamic slice preparations. In normal Krebs solution, the firing rate of SCN neurons was higher in the light period than in the dark period. In Ca2+-free Krebs solution, SCN neuron activity was low during all periods and did not show diurnal rhythm. These results suggest that the disappearance of circadian rhythmic change of SCN neuron activity in Ca2+-free Krebs solution may be due to the disappearance of synaptic transmission in the SCN.

Inhibitory and excitatory effects of histamine on suprachiasmatic neurons in rat hypothalamic slice preparation

The effects of histamine (HA) and serotonin (5-HT) on single neurons of the suprachiasmatic nucleus (SCN) in hypothalamic slices were investigated by iontophoretic application. Sixty-two of 181 cells were inhibited by HA, whereas 30 were excited. Forty-nine of 163 cells were inhibited by 5-HT, whereas 9 were excited. Pyrilamine, an H1-antagonist, slightly (2/7), and cimetidine, an H2-antagonist, strongly (10/13) antagonized the inhibitory effect of HA. The excitatory effect of HA was not antagonized by cimetidine (0/4), but was perfectly antagonized by pyrilamine (5/5). The present results suggest that HA and 5-HT play mainly an inhibitory role in the SCN neuronal activity, and that inhibition and excitation induced by HA may be mediated by H2- and H1-receptors, respectively.

Effects of adrenergic blockers on the inhibition of muricide by desipramine and noradrenaline injected into the amygdala in olfactory bulbectomized rats

Muricide in olfactory bulbectomized rats (OB rats) is readily inhibited by systemic administration of desipramine (DMI) or microinjection of DMI and noradrenaline (NA) into the medial amygdaloid nucleus. The present experiment investigated whether the muricide inhibition produced by these forms of drug treatment was mediated by alpha- or beta-noradrenergic receptors in the central nervous system. Muricide inhibition produced by systemic administration of DMI was antagonized by an alpha-blocker phenoxybenzamine but unaffected by a beta-blocker sotalol, although administration of these adrenergic blockers alone had no effect on muricide. Muricide inhibition induced by the microinjection of DMI and NA into the medial amygdaloid nucleus was similarly antagonized by pretreatment of phenoxybenzamine injected into the same site, but sotalol had no effect. Injection of phenoxybenzamine or sotalol alone into the medial amygdaloid nucleus did not elicit any changes in muricide. These findings suggest that mechanisms mediated by brain noradrenergic alpha-receptor play an important role in muricide inhibition by tricyclic antidepressants in rats and that the medial amygdaloid nucleus is an important site of action of these drugs.

Electrophysiological studies of the development of suprachiasmatic neuronal activity in hypothalamic slice preparations

The rate and pattern of neuronal discharge in the suprachiasmatic nucleus (SCN) during developmental stages were studied and compared with those of the ventromedial (VMH) and the lateral hypothalamus (LHA) using rat hypothalamic slices. The firing rate of the SCN neurons was low on the 7th and 11th days; however, it dramatically increased by the 14th day to reach the adult rate, while firing rates of VMH and LHA neurons increased gradually with age. The discharge rate of neurons in the ventrolateral part of the SCN (VL-SCN) was higher than that of the dorsomedial SCN (DM-SCN) neurons in 14-, 21- and 70-100-day-old rats. Activity of the DM-SCN neurons on day 21 were unaffected by bilateral enucleation on the third day, while activity in the VL-SCN decreased; that of both parts was significantly decreased by a constant light schedule.

Field potentials in the suprachiasmatic nucleus of rat hypothalamic slice produced by optic nerve stimulation

Spatial profile of a projection from retina to the suprachiasmatic nucleus (SCN) was studied by electrophysiological method. Stimulation of contralateral optic nerve evoked the fast positive and late large negative waves in only the ventrolateral but not the dorsolateral parts of the posterior SCN. This result, in agreement with anatomical evidence suggests that the late negative wave is due to excitatory synaptic potential in the SCN and has a role for elevating the neuronal activity in the ventrolateral part of the posterior SCN daytime.

Effects of restricted feeding on single neuron activity of suprachiasmatic neurons in rat hypothalamic slice preparation

Influence of restricted feeding on single neuron activity in the rat suprachiasmatic nucleus (SCN) was examined using hypothalamic slice preparations. Feeding restriction for one month did not change SCN neuron activity in either normal light-dark schedule (LD) or constant light schedule (LL). Locomotor activity in vivo was increased for a few hours before and during feeding time by food restriction of both LD and LL rats. The present experiment suggested that the memory of feeding time produced by food restriction is independent of SCN neuron activity.