Kayo Akiyama

Regulation of brain function by exercise

The effect of excercise on brain function was investigated through animal experiments. Exercise leads to increased serum calcium levels, and the calcium is transported to the brain. This in turn enhances brain dopamine synthesis through a calmodulin-dependent system, and increased dopamine levels regulate various brain functions. There are abnormally low levels of dopamine in the neostriatum and nucleus accumbens of epileptic mice (El mice strain) and spontaneously hypertensive rats (SHR). The low dopamine levels in those animals were improved following intracerebroventricular administration of calcium chloride. Dopamine levels and blood pressure in SHR were also normalized by exercise. In epileptic El mice, convulsions normalized dopamine levels and physiologic function. These findings suggest that exercise or convulsions affect brain function through calcium/calmodulin-dependent dopamine synthesis. This leads to the possibility that some symptoms of Parkinsons disease or senile dementia might be improved by exercise.

Music improves dopaminergic neurotransmission: demonstration based on the effect of music on blood pressure regulation.

The mechanism by which music modifies brain function is not clear. Clinical findings indicate that music reduces blood pressure in various patients. We investigated the effect of music on blood pressure in spontaneously hypertensive rats (SHR). Previous studies indicated that calcium increases brain dopamine (DA) synthesis through a calmodulin (CaM)-dependent system. Increased DA levels reduce blood pressure in SHR. In this study, we examined the effects of music on this pathway. Systolic blood pressure in SHR was reduced by exposure to Mozarts music (K.205), and the effect vanished when this pathway was inhibited. Exposure to music also significantly increased serum calcium levels and neostriatal DA levels. These results suggest that music leads to increased calcium/CaM-dependent DA synthesis in the brain, thus causing a reduction in blood pressure. Music might regulate and/or affect various brain functions through dopaminergic neurotransmission, and might therefore be effective for rectification of symptoms in various diseases that involve DA dysfunction.

The mechanism by which exercise modifies brain function

The effect of exercise on central nervous system function was investigated in relation to the mechanism of calcium-calmodulin-dependent dopamine synthesis in the brain. It is shown here through animal experiments that exercise leads to an increase in the calcium level in the brain. This in turn enhances brain dopamine synthesis, and through this increased dopamine modifies and/or affects brain function, which might induce physiological, behavioral, and psychological changes.

Effect of calmodulin antagonists on calcium and ethanol-induced sleeping time in mice

This investigation was carried out to determine if calcium prolongation of ethanol-induced sleep is mediated by calmodulin and a calmodulin-dependent protein kinase. The duration of ethanol-induced sleeping time in ddY male mice was measured following the administration of CaCl2 (20, 40, 80 and 200 mumol/kg, intraperitoneally (IP) both with and without the calmodulin antagonists, W-7: [N-(6-Aminohexyl)-5-chloro-1-naphthalenesulfonamide] (4.2 micrograms/mouse, intraventricular (IVT) or trifluoperazine (TFP; 1.8 micrograms/mouse, IVT). When CaCl2 was administered in a dose dependent manner the duration of ethanol-induced sleep was prolonged. The prolongation was antagonized by W-7 and TFP. When mice were treated with W-7 or TFP together with serotonin (5-HT; 15 nmol/mouse, IVT), dopamine (DA; 30 nmol/mouse, IVT) or norepinephrine (NE; 30 nmol/mouse, IVT), the sleeping time induced by ethanol and calcium was enhanced. This finding suggests that W-7 and TFP selectively inhibit the synthesis of 5-HT, DA and NE, but they do not affect other neuronal functions of these biogenic amines. The results would suggest a probable mechanism in which Ca++ prolongs ethanol-induced sleeping time by activating tyrosine hydroxylase and tryptophan hydroxylase through intracerebral calmodulin and calmodulin-dependent protein kinase, which subsequently raise the levels of 5-HT, DA and NE.

A mechanism of cadmium poisoning: the cross effect of calcium and cadmium in the calmodulin-dependent system

The effects of the intraventricular (IVT) administration of cadmium on the amount of dopamine (DA) in various regions of the mice brain were analyzed immunohistochemically using a microphotometry system. DA levels in the neostriatum and nucleus accumbens were increased by approximately 30% (p<0.01) by the IVT administration of CdCl2 (1 μmol/kg). This effect was abolished by the calmodulin antagonist, W-7 (4.2 μg/mouse, IVT). The effects of cadmium on DA levels in the brain were very similar to those seen with calcium. Combining these results with our previous finding that calmodulin does not have the ability to distinguish between calcium and cadmium, a mechanism of cadmium poisoning is suggested in which cadmium activates catecholamine synthesizing enzyme and numerous other enzymes through calmodulin-dependent systems, thereby disturbing many functions in the organism.

Central dopamine-synthesis regulation by the calcium-calmodulin-dependent system

The effects of the intraventricular (IVT) administration of calcium on the amount of dopamine (DA) in various regions of the mouse brain were analyzed immunohistochemically by using a microphotometry system. The DA levels in the nucleus accumbens and the lateral part of the neostriatum were increased by approximately 45% (p less than 0.01) and 25-35% (p less than 0.01), respectively, by the IVT administration of CaCl2 (10 mumol/kg). It was also found that this effect was abolished by the calmodulin antagonist, W-7 (4.2 micrograms/mouse, IVT). The brain regions in which the amount of DA was increased by calcium were areas where high levels of calmodulin and tyrosine hydroxylase are distributed. These findings suggest that the synthesis of central DA is regulated by calcium through a calmodulin-dependent system.

Regulation of blood pressure with calcium-dependent dopamine synthesizing system in the brain and its related phenomena

The effects of calcium on blood pressure regulation remain controversial. Although the mechanism by which calcium increases blood pressure when it is given intravenously and acutely has been elucidated, that by which calcium reduces blood pressure when it is supplemented chronically and slightly through daily diet is unclear. From a number of animal experiments concerning the effects of calcium on blood pressure, we believe that calcium ions have two separate roles in the regulation of blood pressure through both central and peripheral systems: (1) calcium ions reduce blood pressure through a central, calcium/calmodulin-dependent dopamine-synthesizing system and (2) calcium ions increase blood pressure through an intracellular, calcium-dependent mechanism in the peripheral vasculature. These concepts were applied to elucidate the mechanisms underlying hypertension in spontaneously hypertensive rats (SHR) and changes in blood pressure in other experimental animals, and the following conclusions were reached. The decrease of the serum calcium level in spontaneously hypertensive rats (SHR) causes a decrease in calcium/calmodulin-dependent dopamine synthesis in the brain. The subsequent low level of brain dopamine induces hypertension. The increase in susceptibility to epileptic convulsions and the occurrence of hypertension in epileptic mice (El mice) may be linked through a lowering of calcium-dependent dopamine synthesis in the brain, and epilepsy and hypertension may be associated. Exercise leads to increases in calcium-dependent dopamine synthesis in the brain, and the increased dopamine levels induce physiological changes, including a decrease in blood pressure. Cadmium which is not distinguished from calcium by calmodulin, activates calmodulin-dependent functions in the brain, and increased dopamine levels may decrease blood pressure. In this report, our studies are considered in light of reports from many other laboratories.

FORMATION OF GUANIDINOSUCCINIC ACID, A STABLE NITRIC OXIDE MIMIC, FROM ARGININOSUCCINIC ACID AND NITRIC OXIDE-DERIVED FREE RADICALS

Guanidinosuccinic acid (GSA) is noted for its nitric oxide (NO) mimicking actions such as vasodilatation and activation of the N-methyl-D-aspartate (NMDA) receptor. We have reported that GSA is the product of argininosuccinate (ASA) and some reactive oxygen species, mainly the hydroxyl radical. We tested for GSA synthesis in the presence of NO donors. ASA (1 mM) was incubated with NOR-2, NOC-7 or 3-morpholinosydomine hydrochloride (SIN-1) at 37 degrees C. GSA was determined by HPLC using a cationic resin for separation and phenanthrenequinone as an indicator. Neither NOR-2 or NOC-7 formed GSA. SIN-1, on the other hand, generates NO and the superoxide anion which, in turn, generated peroxynitrite which was then converted to the hydroxyl radical. Incubation of ASA with SIN-1 leads, via this route, to GSA. When ASA was incubated with 1 mM SIN-1, the amount of GSA produced depended on the incubation time and the concentration of ASA. Among the tested SIN-1 concentrations, from 0.5 to 5 mM, GSA synthesis was maximum at 0.5 mM and decreased with increasing concentrations of SIN-1. Carboxy-PTIO, a NO scavenger, completely inhibited GSA synthesis. SOD, a superoxide scavenger, decreased GSA synthesis by 20%, and catalase inhibited GSA synthesis only by 12%; DMSO, a hydroxyl radical scavenger completely inhibited GSA synthesis in the presence of SIN-1. These data suggest that the hydroxyl radical derived from a combination of NO and the superoxide anion generates GSA, a stable NO mimic. Meanwhile, synthesis of GSA by NO produces reactive oxygen and activates the NMDA receptor that generates NO from GSA, suggesting a positive feed back mechanism.

Quantitative analysis of immunohistochemical distributions of cholinergic and catecholaminergic systems in the human brain

The distributions of the cholinergic system and catecholaminergic system in the normal human brain were analysed quantitatively by a microphotometry system. Consecutive coronal sections were obtained from the anterior area of the left hemisphere and were stained alternately with fluorescent immunohistochemical staining for choline acetyltransferase or tyrosine hydroxylase. Each stained section was divided into approximately 120,000 areas and the fluorescence intensity in each area was measured by a fluorescence microphotometry system which is a measuring microscope for distribution of fluorescence intensity in the tissue slice. Nonspecific autofluorescence was distributed in myelinated nerve fiber throughout the entire area, which was subtracted from the fluorescence intensity value in each measuring area. The obtained immunohistochemical fluorescence intensities of choline acetyltransferase and tyrosine hydroxylase were classified into eight ranks and were indicated by color graphics. Also, the intensity values of actual immunohistochemical fluorescence in the various brain regions were presented. The choline acetyltransferase and tyrosine hydroxylase concentrations varied greatly depending on the brain region. Relatively high levels of choline acetyltransferase and tyrosine hydroxylase were distributed in the putamen, caudate nucleus, claustrum, insula and some cortical regions. The immunohistochemical level of tyrosine hydroxylase was lower than that of choline acetyltransferase in a few brain regions such as the globus pallidus and amygdala. High levels of choline acetyltransferase and tyrosine hydroxylase were localized in the one area of the basal ganglia which developed from the telencephalic area, whereas middle levels of these were distributed in another, part of which developed from the diencephalic area.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of intraventricular administration of calcium and calmodulin antagonist on the blood pressure in the rat.

The effect of intraventricular administration of calcium on the mean arterial pressure in the conscious rat was investigated. Biphasic changes of blood pressure, fugitive rapid increment during the first 3 min and gradual lengthy decrement during the next 60 min were observed by the administration of CaCl2 (30 mumol/kg). Both these changes were significant as compared with the control level. On the other hand, the biphasic response of blood pressure by CaCl2 was abolished by the administration of calmodulin antagonist, W-7, or catecholamine synthesizing enzyme inhibitor, alpha-methyltyrosine. These results are discussed on the basis of our calcium-calmodulin-dependent biogenic amine-synthesizing mechanism.