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4 Fig. 2. Duirnal variation of Dopamine contents in brain regions of young and aged rats. Horizontal bars represent standard errors of the mean. Statistical significance *: P<0.05, **: P<0.01, õ : P<0.005, õ õ: P<0.001, young vs aged at each time. _??_ : Dark period, _??_: Light period. Table 1. Mean contents of Dopamine in brain regions. Mean }SEM.ƒÊg/g wet tissue. Statistical significance *: <0.05, **: P<0.01, õ : P<0.005, õ õ: P>0.001, young vs aged at Pus, each time. Amy: amygdala, St.: corpus striatum, Cor.: cerebral cortex, Hip.: hippocampus, Die.: diencephalon, B. S.: brain stem D2 D6 D10: 2, 6, 10 hours after light off. L2 L6 L10: 2, 6, 10 hours after light on.
5 Fig. 3. Duirnal variation of Norepinephrine contents in brain regions of young and aged rats. Horizontal bars represent standard errors of the mean. Statistical significance *: P<0.05, **: P<0.01, time. _??_ : Dark period, _??_: Light period. Table 2. Mean contents of Norepinephrine in brain regions. Mean }SEMƒÊg/g wet tissue. young vs aged at each time. Amy.: amygdala, St.: corpus striatum, Cor.: cerebral cortex, Hip.: hippocampus Die: diencephalon, B. S.: brain stem D2 D6 D10: 2, 6, 10 hours after light off. L2 L6 L10: 2, 6, 10 hours after light on.,
6 young vs aged at each time. Amy.: amygdala, St.: corpus striatum, Cor.: cerebral cortex, Hip.: hippocampus, Die.: diencephalon, B. S.: brain stem, D2 D6 D10: 2, 6, 10 hours after light off, L2 L6 L10: 2, 6, 10 hours after light on. Table 4. Amplitude of monoamine diurnal variations (maximam mean/minimum mean)
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14 3. Bernard, B. K. and Finkelstein, E. R.: Alterations in Mouse Aggressive Behavior and Brain Mono amine Dynamics as a Function of Age. Physiol. & Behavior 15, , Black, I. B. and Geen S. C.: Postmortem Changes in Brain Catecholamine Enzymes. Arch NeuroL 32, 47-49, Bowers, M. B. and Gerbode, F. A.: Relationship of Monoamine Metabolites in Human Cerebrospinal Fluid to Age. Nature 219, , Dixit, B. N. and Buckley, J. P.: Circadian Changes in Brain 5-Hydroxytryptamine and Plasma Corti costerone in the Rat. Life Sci. 6, , Finch, C. E.: Catecholamine Metabolism in the Brains of Ageing Male Mice. Brain Research 52, , Friedman, A. H. and Walker, C. A.: Circadian Rhythms in Rat Mid-Brain and Caudate Nucleus Bio genic Amine Levels. J. Physiol. 197, 77-85, Glowinski, J. and Iversen, L. L.: Regional Studies of Catecholamines in the Rat Brain-I; The dispo sition of. [3H] Norepinephrine, [3H] Dopamine and [3H] Dopa in Various Regions of the Brain. J. Neurochem. 13, , Greengrass, P. M. and Tonge, S. R.: Changes in Brain Monoamine Concentrations During the Oestr ous Cycle in the Mouse; Possible Pharmacological Implications. J. Pharm. Pharmac. 23, , Grote, S. S.; Moses, S. G., Hudgens, R. W. and Croninger, A. B.: A Study of Selected Catecholamine Metabolizing Enzymes; A Comparison of Depressive Suicides and Alcoholic Suicides with Controls. J. Neurochem , Hillier, J. G., Martin, P. R. and Redfern, P. H.: A Possible Interaction Between the 24th Rhythms in
15 Catecholamine and 5-Hydroxytryptamine Concentration in the Rat Brain. J. Pharm. Pharmacol. 27 (Suppl.), 40, Jonec, V. and Finch, C. E.: Ageing and Dopamine Uptake by Subcellular Fractions of the C57BL/ 6J Male Mouse Brain. Brain Research 91, , Karasawa, T., Furukawa, K., Yoshida, K. and Shimizu, M.: A Double Column Procedure for the Simultaneous Estimation of Norepinephrine, Normetanephrine, Dopamine, 3-Methoxytyramine and 5-Hydroxytryptamine in Brain Tissue. Japan. J. Pharrnacol , Kobayashi, K., Koide, Y., Doi, T., Kohsaka, M., Hosokawa, K., and Oku, Y.: Homovanillic Acid in Human Cerebrospinal Fluid-Its Concentration Gradient and Reduced Levels in Patient with Epil epsy-acta Medica Okayama 32, , Manshardt, J. and Wurtman, R. J.: Daily Rhythm in the Noradrenaline Content of Rat Hypothal amus. Nature 217, , Maura, G. and Vaccari, A.: Relationships between Age of Submission to Environmental Stress, and Monoamine Oxidase Activity in Rats. Experientia 31, , McGeer, E. G., McGeer, P. L. and Wada, J. A.: Distribution of Tyrosine Hydroxylase in Human and Animal Brain. J. Neurochem. 18, , Morgan, W. W., McFadin, L. S. and Harvey, C. Y.: A Daily Rhythm in Norepinephrine Content in Regions of the Hamster Brain. Comp. Gen. Pharmac. 4, 47-52, Morgan, W. W., Yndo, C. A. and McFadin, L. S.: Daily Rhythmic Changes in the Content of Seroto nin and 5-Hydroxyindoleacetic Acid in the Cerebral Cortex of Mice. Life Sci. 14, , Morgan, W. W., Pfeil, K. A., Reiter, R. J. and Gonzales, E.: Comparison of Changes in Tryptophan and Serotonin in Region of the Hamster and Rat Brain over a Twentyfour Hour Period. Brain Research 117, 77-84, Philo, R., Rudeen, P. K. and Reiter, R. J.: A Comparison of the Circadian Rhythms and Concentra tions of Serotonin and Norepinephrine in the Telencephalon of Four Rodent Species. Comp. Bioc hem. Physiol. 57, , Quay, W. B.: Differences in Circadian Rhythms in 5-Hydroxytryptamine according to Brain Region. Am. J. Physiol. 215 (6), , Reis D. J., Corelli A., and Conner J.: Circadian and ultradian rhythms of serotonin regionally in cat brain. J. Pharm. Fxperi. Therapeutics 167, , 1969, 30. Reis, D. J., Ross, R. A. and Job, T. H.: Changes in the Activity and Amounts of Enzymes Synthesiz ing Catecholamines and Acetylcholine in Brain, Adrenal Medulla, and Sympathetic Ganglia of Aged Rat and Mouse. Brain Research 136, , Robinson, D. S., Davis, J. M., Nies, A., Ravaris, C. L., and Sylwester, D.: Relation of Sex and Aging to Monoamine Oxidase Activity of Human Brain, Plasma, and Platelets. Arch Gen Psychiatry 24, , Robinson, D. S., Davis, J. M., Nies, A., Colburn, R. W. Davis, J. N., Bourne, H. R., Bunney, W. E., Shaw,
16 D. M. and Coppen, A. J.: Ageing, Monoamines, and Monoamine-Oxidase Levels. Lancet 5, , Robinson, D. S.: Changes in Monoamine Oxidase and Monoamines with Human Development and Aging. Fed. Proc. 34, , Robinson, D. S., Sourkes, T. L., Nies, A., Harris, L. S., Bartlett, D. L. and Kaye, I. S.: Monoamine Metabolism in Human Brain. Arch Gen Psychiatry 34, 89-92, Scheving, L. E., Harrison, W. H., Gordon, P. and Pauly, J. E.: Daily Fluctuation (Circadian and Ult radian) in Biogenic Amines of the Rat Brain. Am. J. Physiol. 214, , Schubert, J. and Sedvall, G.: Accumulation and Disappearance of 3H-5-Hydroxytryptamine Formed in Vivo from 3H-Tryptophan in Various Regions of the Rat Brain. Eur. J. Pharmacol. 17, 75-80, Snyder, S. H., Zweig, M., Axelrod, J. and Fischer, J. E.: Control of the Circadian Rhythm in Seroto nin Content of the Rat Pineal Gland. Physiology 53, , Takahashi, R. and Aprison, M. H.: Acetylcholine Content of Discrete Areas of the Brain Obtained by a Near-freezing Method. J. Neurochem. 11, , Wurtman, R.J. and Axelrod, J.: A 24-hour Rhythm in the Content of the Pineal and Salivary Glands of the Rat. Life Sci. 5, , Zolovick, A. J., Pearse, R., Boehlke, K. W. and Eleftheriou, B. E.: Monoamine Oxidase Activity in Various Parts of the Rat Brain during the Estrous Cycle. Science 154, 649, 1966.
17 Diurnal variations in motor activity and regional brain monoamine content in young adult and aged rats Tohru DOI Institute for Neurobiology, Okayama University Medical School, Okayama, Japan Diurnal variations in motor activity and regional monoamine levels of the brain in both young adult and aged wista; male rats were compared. The young adult group (Y group) con sisted of 16 week old rats. The aged group (A group) comprised 19 month old rats. All the animals were housed in a room with an alternating 12 hour darklight cycle. The temperature was kept at 24 Ž and the moisture at 55%. Motor activity was measured in selected rats by Animex. Both groups were then subdivided into six smaller groups. The animals of these groups were sacrificed by decapitation every 4 hours of the 24 hour period (2, 6 and 10 hours in the dark cycle, and 2, 6 and 10 hours in the light cycle; D2, D6, D10, L2, L6 and L10). Immedi ately after decapitation, brains were collected, dissected into six different regions (amygdala [Amy], corpus striatum [St], cerebral cortex [Cor], hippocampus [Hip], diencephalon [Die] and brain stem [B. S.]), and the levels of dopamin [DA], norepinephrine [NE] and serotonin [5-HT] in each determined fluorometrically. The results were as follows. 1. Diurnal variation of motor activity. Both Y and A groups showed a marked difference between the dark and light cycles. Activity increased during the first 4 hours of the dark cycle, then decreased gradually during the rest of the dark cycle. Motor activity decreased markedly immediately after begin ning of the light cycle, and remained decreased throughout the light period. The motor activity of the both groups increased transiently a little before the light cycle. Y group activity increas ed directly before the dark time. A group activity was significantly less than that of Y group, except for 2 time points during the light cycle. A group motor activity was about 50% lower than that of the Y group. 2. Diurnal variation of DA. Amy from both groups showed a higher levels in the dark time than in the light. St showed two peaks in both groups but the range of variation was wider in A group than in Y group. Cor from both groups showed a higher level in the light time than in the dark. The levels of Hip, Die and B. S. in both groups showed little variation, the range being much smaller in A group. There was no significant difference in the levels of Amy, Cor and Hip in either group, but St, Die and B. S. were significantly lower in A group throughout almost the entire 24 hour period. 3. Diurnal variation of NE levels. Mark ed variations in Amy, Cor, Hip and Die occurred in both groups. The NE level in the 4 brain regions tended to be lower during the dark time than during the light. The diurnal variation of amine levels in St was smaller in the groups than in the other five regions. B. S. in the Y group showed lower levels during the dark time; however, B. S. from A group showed almost constant levels during the 24 hour period. The NE levels of each groups showed little difference in brain regions other than B. S.. In A group, B. S. showed 1.8 to 9 times higher levels than Y group at all examined time points. 4. Diurnal variation in 5-HT levels. Both groups showed higher amine levels in Amy, Hip and Die during the dark period than in the light. St in Y group showed two peaks at D10 and L6 but the same region in A group showed almost constant amine levels during the diurnal period except for D6. The level in Cor from both groups showed little varia tion, the range being much smaller in A group than in Y group. The level in B.S. of Y group showed a marked dark-light difference with a high peak at L2 but the same region from A group showed a gradual variation in the amine level with a peak between D10 and L2. Com parison of the 5-HT levels in Y and A groups showed that there was no significant difference in Amy, Hip and Die. The level in St remained high throughout the diurnal period in Y group.
18 The level in Cor showed a higher value during the 24h period in A group. The amine level in B. S. was higher in the A group at two time points of the dark period than in Y group. The data indicate a characteristic influence of aging on diurnal variations of motor activity and three monoamine contents of the brain. Some regions of the brain showed no difference in DA levels of Y and A groups, whereas other regions showed significantly lower levels in A group. Some regions showed no difference in NE levels of both groups and the other showed significantly higher levels in A group. These results suggest that the turnover rate of DA in some brain regions is accelerated in A group compared to Y group. A group tended to show less diurnal variation in the three monoamine levels of the brain than Y group when related to the reduced motor activity of A group, this suggests that metabolism of these monoamines in related to behavior.
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