The regionalization of [H]dihydrotetrabenazine binding sites in the mouse brain and its relationship to the distribution of monoamines and their metabolites

[³H]Dihydrotetrabenazine binding was measured in 8 areas of the mouse brain. In all areas, binding occurred on a homogeneous class of sites (K_d 2.6nM). The density of [³H]dihydrotetrabenazine binding sites strongly varied between the different brain structures; it was compared to endogenous levels of biogenic monoamines and their metabolites: the density is independent of the nature of the monoamine and of neuronal activity, but is highly correlated to the total amount of monoamines present in each structure.     

Age-related changes on monoamine turnover in hippocampus of rats

After pargyline treatment the turnover rates of dopamine (DA), noradrenaline (NA), 3,4-dihydroxyphenylacetic acid (DOPAC), serotonin (5-hydroxytryptamine (5-HT) and 5-hydroxy-3-indolacetic acid (5-HIAA) has been measured in control and aged hippocampus of the rats. In addition, the tyrosine hydroxylase (TH) activity and monoamine oxidase-A and monoamine oxidase-B activities have also been studied. The TH activity did not change in aged hippocampus as compared to controls. The monoamine oxidase-B:monoamine oxidase-A ratio increased in 26-month-old rats compared with controls. The turnover of DA, DOPAC and NA did not show significant changes while 5-HT synthesis, 5-HT accumulation rate and 5-HIAA turnover increased in aged rats. Serotonin fibers showed morphological dissimilarities between the hippocampus of young and aged rats using immunocytochemistry techniques. In aged rats aberrant serotoninergic fibers mainly appear in the molecular layer of the dentate gyrus and moleculare of the hippocampal CA1. It is suggested that the aberrant morphology of 5-HT fibers may reflect the local degeneration of serotoninergic hippocampal afferents during aging. Increase of 5-HT turnover in aged might be a signal of degeneration.     

Neurohormonal mechanisms of the antidepressant effect of ketanserin in ovariectomized rats

In the present study we investigated the effects of chronic administration of m-CPP (0.5 mg/kg, i.p.), a 5-HT_2B/2C receptor agonist, and ketanserin (0.1 mg/kg, i.p.), a 5-HT_2A/2C receptor antagonist, alone or in a combination with 17β-estradiol (0.5 μg per animal, i.m.) over 14 days on depressive behavior and the concentration of different monoamines in the hippocampus in adult ovariectomized (OVX) female rats. Depression-like behavior was assessed using the Porsolt’s test. The concentrations of monoamines and their metabolites were determined using HPLC. We found that chronic ketanserin administration has an antide-pressant effect in OVX rats. Chronic ketanserin administration in combination with 17β-estradiol in OVX females potentiated antidepressant action. The antidepressant effect of ketanserin in OVX rats correlated with the restoration of noradrenergic, serotonergic, and dopaminergic neurotransmission in the hippocampus. Our data indicate a close interaction between the ovary hormonal and serotonergic systems of the brain in mechanisms of depression.     

Impact of experimental hypothyroidism on monoamines level in discrete brain regions and other peripheral tissues of young and adult male rats

The levels of dopamine (DA), norepinephrine (NE) and serotonin (5-HT) in different brain regions as well as in blood plasma, cardiac muscle and adrenal gland of young and adult male albino rats were measured following experimentally induced hypothyroidism. Hypothyroidism induced by daily oral administration of propylthiouracil (PTU, 5 mg/kg body wt) caused a significant reduction in DA levels in most of the tissues examined of both young and adult rats after 21 and 28 days, in NE levels after all the time intervals studied in young rats, and after 21 and 28 days in adult rats. 5-HT exhibited a significant reduction in the selected brain regions and blood plasma after 21 and 28 days and in cardiac muscle after all the time intervals in the two age groups of animals. It may be suggested that the changes in monoamine levels induced by hypothyroidism may be due to disturbance in the synthesis and release of these amines through the neurons impairment or may be due to an alteration pattern of their synthesizing and/or degradative enzymes.     

Effect of neonatal hypothyroidism on the serotonin system of the rat brain

The effects of neonatal thyroidectomy and thyroid hormone replacement therapy on the development of serotonin-contaning neurons in discrete rat brain nuclei were studied. Newborn male rats were rendered hypothyroid by the injection of 125 μCi¹³¹I, and, after 45 days, were compared with normal littermate controls and¹³¹I-injected animal subsequently maintainde by dialy T₄ injections. The serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) contents of discrete brain nuclei remove by punches of frozen brain slices were measured by HPLC with electrochemical detection. 5-HT and 5-HIAA contents were significantly increased in many nuclei of the hypothyroid rat brain. By blocking the biosynthesis ot 5-HT with p-chlorophynelalanine we found that the activity of tryptophan hydroxylase is an important step in the stimulatory effect of hypothyroidism on the 5HT and 5-HIAA contents. Futhermore, we demonstrated after blockage of monomine oxidase activity with pargyline, a less pronounced decline of 5-HIAA in neonatal hypothyroid animals, thus causing a relative accumulation of this metabolite. These results demonstrate theat there are important modifications of the 5-HT system in the brain of neonatal hypothyroid rats. This may have an important role in the development of hypothyroid-induced impairments of central nervous system function.     

Cerebrospinal fluid monoaminergic metabolites are elevated in adults with Down's syndrome

Under conditions of rest and a low monoamine diet, brain monoamine activity was examined in young (less than 35 years) and old (greater than 35 years) adults with Down's syndrome and in control subjects by measuring the cerebrospinal fluid (CSF) and plasma concentrations of the neurotransmitter norepinephrine, and of 5-hydroxyindoleacetic acid (5-HIAA), homovanillic acid (HVA), and 3-methoxy 4-hydroxyphenylglycol (MHPG), the respective metabolites of the neurotransmitters serotonin, dopamine, and norepinephrine. There were no age-related differences in metabolite concentrations in either the Down's syndrome or control subjects. CSF concentrations of 5-HIAA, HVA, and norepinephrine were significantly higher in young subjects with Down's syndrome as compared with young controls, and CSF concentrations of 5-HIAA and norepinephrine were significantly higher, by twofold or more, in old subjects with Down's syndrome as compared with older controls. The results suggest that monoamine turnover and brain functional activity involving monoamines is elevated in Down's syndrome, and that the early neuropathological changes in the disorder are not associated with a monoamine deficit.     

Effects of long-term hypoxia and hypoxic exercise on brain monoamine levels in rat

This study clarified the effects of long-term hypoxia and hypoxic exercise on monoamines in the whole brain, and in four specific regions of the rat brain. The male Wistar rat progenitors (P1 group) were randomly assigned to three groups: hypoxia (16.0% oxygen) and exercise (MHE-P1), hypoxia and sedentary (MHS-P1), normoxia and sedentary (MNS-P1). The male children of P1 (the first generation of hypoxic rats; F1) were randomly divided into two groups: hypoxia and exercise (MHE-F1) and hypoxic sedentary (MHS-F1). The monoamines of whole brain were measured in P1 males, and monoamines of cerebellum, frontal lobe, striatum and hippocampus were measured in F1 males. The monoamine levels of MHE-P1 were significantly lower than those of MHS-P1 and MNS-P1. No significant difference was found in monoamine levels between MHS-P1 and MNS-P1. Epinephrine, norepinephrine, and dopamine levels of the MHE-F1 group significantly decreased in the frontal lobe, cerebellum and striatum, compare with the other groups (hypoxic and sedentary; normoxic and sedentary, respectively). These monoamines in the hippocampus were not influenced by the hypoxia or hypoxic exercise conditions. This study suggests that long-term hypoxic exercise decreased monoamine levels in whole brain, and that sensitivity to hypoxia and hypoxic exercise differed according to brain region.     

Central monoamine metabolism in the male Brown-Norway rat in relation to aging and testosterone

Concentrations of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), noradrenaline (NA), free 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG), serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were determined in brain regions of 5-, 20-, and 32-month-old male Brown-Norway rats using high pressure liquid chromatography. In view of the activating effects of sex steroids on peptide and monoamine transmitter systems and the declining plasma testosterone levels with aging, the effects of testosterone supplementation on age-related changes in central monoamine metabolism were also studied. Age-related decreases in monoamine metabolism were observed in nigrostriatal, mesocortical and coeruleohippocampal systems. Marked reductions in DOPAC (35%) and HVA (50%) occurred in the ventral tegmental area between 20 and 32 months of age. 5-HT and 5-HIAA levels showed reductions and increases depending on the brain region. Testosterone administration resulted in elevations of HVA in the substantia nigra and MHPG in the locus coeruleus and hippocampus, which were most pronounced in young animals. It is concluded that there are marked differences in age-related changes between nigrostriatal, mesocortical and coeruleohippocampal systems and that testosterone exerts a stimulatory influence on some aspects of monoamine metabolism in young but not in aged animals.     

Relationship of Gonadal Hormone Administration, Sex, Reproductive Status and Age to Monoamine Oxidase Activity Within the Hypothalamus

Activity of Type A monoamine oxidase (MAO) was measured in microdissected samples from preoptic-hypothalamic and hindbrain areas of young male and female rats, and aged female rats. Administration of estradiol and progesterone, in doses sufficient to facilitate lordosis behavior and induce a luteinizing hormone surge in ovariectomized, but not castrated rats, was associated with sexually dimorphic changes of MAO activity within the hypothalamus. Forty-two h following estradiol benzoate administration, increased MAO activity was measured in the ventromedial nucleus (VML) and midbrain central gray of females, while decreased MAO activity was measured in the VML and arcuate-median eminence (ArME) of males. Progesterone administration to estradiol benzoate-primed rats was associated with decreased MAO activity in the VML and medial preoptic nucleus (mPOA) of females and decreased activity in the dorsal raphe nucleus of males. Activity of MAO on diestrus, proestrus and estrus was assessed in ten preoptic-hypothalamic and hindbrain sites. Differences between days of the cycle were limited to the mPOA, ArME and VML. While activities were generally lowest at estrus, these areas exhibited different patterns of activity across the cycle. Activity was highest at proestrus in the mPOA and highest at diestrus in the VML and ArME. Activity of MAO in some areas of 25-month old, diestrus rats was altered as compared to young, cycling rats; however, ageing was not associated with widespread changes in MAO activity. In the suprachiasmatic nucleus, aged rats showed approximately 30% less activity than young rats. In the mPOA, VML and ArME, activity in aged females was different from some, but not all, days of the estrous cycle. These results show that MAO activity changes within specific hypothalamic sites when the neuroendocrine axis is altered. Since the changes are present in areas where activity of rnonoaminergic systems is critical for initiating gonadotrophin surges and inducing lordosis behavior, these results provide initial evidence that catabolism of monoamines by MAO may contribute to rnonoaminergic regulation of reproductive function.     

Changes in the activity and amounts of enzymes synthesizing catecholamines and acetylcholine in brain, adrenal medulla, and sympathetic ganglia of aged rat and mouse.

The effects of aging on the activities of the catecholamine synthesizing enzymes, tyrosine hydroxylase (TH), aromatic L-amino acid decar?ylase (DDC), dopamine-β-hydroxylase (DBH), and phenylethanolamine-N-methyltransferase (PNMT), and on choline acetyltransferase (CAT), the enzyme synthesizing acetylcholine, were investigated regionally in brain, in adrenal medulla, and in sympathetic ganglia of young (4 month) and aged (24–26 month) rats and mice. In brain, significant changes in enzyme activity were variable, regional, present only in rats and were small, usually less than 20%. In both species, however, the activities of CAT, TH and DDC, but not DBH, were increased 1.5–2.5-fold in the adrenal of aged animals when compared with 4 month old animals. The increase in adrenal TH and DDC activities with age occurred without a change in the K_m for substrate or cofactor and could be demonstrated by immunotitration with specific antibodies to be due to accumulation of enzyme protein. A comparable pattern of changes in enzyme activities occurred in rat superior cervical ganglion. We conclude that significant elevations in the activities and amounts of several enzymes synthesizing catecholamines and acetylcholine occur in the adrenals of aging rats and mice and are not paralleled by comparable changes in these enzymes in brain. The pattern of change in adrenal enzymes appears unique for aging and suggests an increased capacity for biosynthesis of circulating catecholamines. The absence of changes in brain TH and DBH activity, found in this study and contradicting observations made by others in man and another strain of rat, suggests that some changes in brain catecholamine enzymes in aging may be a species- and even strain-specific event and not a universal concomitant of aging.     

Effects of aging on agmatine levels in memory-associated brain structures

Agmatine is a metabolite of L-arginine by arginine decarboxylase. Recent evidence suggests that it exists in mammalian brain and is a novel neurotransmitter. The present study measured agmatine levels in several memory-associated brain structures in aged (24-month-old), middle-aged (12-month-old), and young (4-month-old) male Sprague Dawley rats using liquid chromatography/mass spectrometry. Agmatine levels were significantly decreased in the CA1, but increased in the CA2/3 and dentate gyrus, subregions of the hippocampus in aged and middle-aged rats relative to the young adults. In the prefrontal cortex, a dramatic decrease in agmatine level was found in aged rats as compared with middle-aged and young rats. There were significantly increased levels of agmatine in the entorhinal and perirhinal cortices in aged relative to middle-aged and young rats. In the postrhinal and temporal cortices, agmatine levels were significantly increased in aged and middle-aged rats as compared with young adults. The present findings, for the first time, demonstrate age-related changes in agmatine levels in memory-associated brain structures and raise a novel issue of the potential involvement of agmatine in the aging process.     

Effects of 8-OH-DPAT and NAN-190 on anxiety behavior,monoamine metabolism,and the level of sex hormones in female rats during the estrus cycle

In this study, we performed a comparative analysis of the effects of chronic administration of an agonist of 5-HT_1A receptors, 8-OH-DPAT, (0.05 mg/kg, subcutaneously) and an antagonist of 5-HT_1A receptors, NAN-190, (0.1 mg/kg, intraperitoneally) for 14 days on the anxiety behavior, monoamine metabolism in the hippocampus and amygdala, and the level of sex hormones in the blood of adult female rats during the estrus cycle. Anxiety behavior was tested in an elevated X-maze. The level of monoamines and their metabolites was evaluated using HPLC with an electrochemical detector; the content of sex hormones in the blood was measured using an enzyme immunoassay. We found that chronic administration of 8-OH-DPAT has an anxiolytic effect in rats in estrus and proestrus. After treatment with NAN-190, we found clear modulation of anxiety behavior that depended on the phase of the estrus cycle, an anxiolytic effect at a high level of endogenous estrogens and an anxiogenic effect at a low level of estrogens. The anxiolytic effect of 8-OH-DPAT in female rats was accompanied by its suppressive influence on the pituitary-ovarian and noradrenergic systems and by its activating effect on the dopaminergic system of the brain. In contrast, the behavioral effects of NAN-190 in female rats during key phases of the estrus cycle corresponded to its stimulating effect of the pituitary-ovarian system and the absence of any effect on the monoaminergic systems of the brain. These data suggest that the mechanisms of anxiety include tight interaction between the ovarian hormonal system and the serotonergic brain system.     

Thyroid hormone and conditioned medium effects on astroglial cells from hypothyroid and normal rat brain: Factor secretion,cell differentiation,and proliferation

The effects of triiodothyronine (T₃)on cell morphology were examined in cerebral hemisphere and cerebellar astrocyte cultures obtained from normal and hypothyroid neonatal rats. T₃-treatment induced morphological changes in astrocytes from cerebral hemispheres. This morphological effect was produced earlier if astrocytes were treated with conditioned medium obtained from cerebral hemisphere astrocyte cultures previously exposed to 50 nM T₃. T₃ or conditioned medium-treatment produced faster morphological changes in hypothyroid rat cerebral hemisphere astrocyte monolayers. Cerebellar astrocytes from normal brain did not respond to thyroid hormone with morphological changes, but proliferated after T₃-treatment. However, hypothyroid cerebellar astrocyte cultures exhibited morphological changes, differently than normal cells. We verified that T₃ may induce astrocyte secretion of factor(s) that promotes morphological differentiation in cerebral hemisphere astroglial cultures and stimulates the proliferation of cerebellar astrocytes. Astrocytes obtained from hypothyroid animals were more sensitive to secreted factors than normal cells. These results emphasize the heterogeneity and the importance of glial cells to normal brain development and open new questions about thyroid hormone therapy in hypothyroidism. © 1995 Wiley-Liss, Inc.     

Regulation of neurofilament gene expression by thyroid hormone in the developing rat brain.

The role of thyroid hormone (TH) in the expression of neurofilament (NF) genes during the first 3-4 postnatal weeks of rat brain development has been examined. I.p. administration of TH to 2-day-old hypothyroid rats resulted in a 2-fold increase in cerebral NF-M and NF-L mRNAs, while administration to 15-day-old hypothyroid rats led to a 1.5- to 3-fold increase in NF-H mRNA within 2-4 h of hormone injection. Comparison of the level of these mRNAs in cerebra from 5, 10, 15 and 20-day-old normal and hypothyroid rats by Northern blot analysis revealed that hypothyroidism declined the expression of all three mRNAs by 50-70% at all ages examined. Western blot analysis of total protein and cytoskeletal proteins isolated from cerebras of 5, 10, 15, 20 and 25-day-old normal and hypothyroid rats demonstrated an even greater reduction (60-90%) in the expression of NF proteins in the hypothyroid cerebra during the period examined. The overall results show that TH plays an important role in regulating the expression of all three NF genes during rat brain development.     

Thyroid hormone-induced GLUT-1 expression in rat cerebral tissue: effect of age

To determine if thyroid hormone (TH) modulates the expression of cerebral glucose transporter one (GLUT-1) and whether there are age-related differences in TH effect, young male Fischer 344 rats (6 months old) and aged rats (26 months old) were studied at euthyroid, hyperthyroid and hypothyroid conditions. Immunoblot analysis indicated that 55 kDa GLUT-1 mass was decreased in hypothyroid young rats (174±15 arbitrary units) and hyperthyroid rats (144±22) compared to euthyroid young rats (395±57) P<0.01. In aged rats, the 55 kDa GLUT-1 mass was significantly increased in hyperthyroidism (392±49) compared to euthyroid aged rats (237±27) P<0.05. The 45 kDa isoform of GLUT-1 in rat cerebral tissue did not significantly change with age or thyroidal state. The changes in 55 kDa GLUT-1 mass did not correlate with the changes in GLUT-1 mRNA content. It is concluded that alterations in cerebral GLUT-1 content in response to altered thyroid state are age-specific.     

The regionalization of [3H]dihydrotetrabenazine binding sites in the mouse brain and its relationship to the distribution of monoamines and their metabolites

[3H]Dihydrotetrabenazine binding was measured in 8 areas of the mouse brain. In all areas, binding occurred on a homogeneous class of sites (Kd approximately equal to 2.6 nM). The density of [3H]dihydrotetrabenazine binding sites strongly varied between the different brain structures; it was compared to endogenous levels of biogenic monoamines and their metabolites: the density is independent of the nature of the monoamine and of neuronal activity, but is highly correlated to the total amount of monoamines present in each structure.     

Semax,synthetic ACTH(4–10) analogue,attenuates behavioural and neurochemical alterations following early-life fluvoxamine exposure in white rats

Selective serotonin reuptake inhibitors (SSRI) are commonly used to treat depression during pregnancy. SSRIs cross the placenta and may influence the maturation of the foetal brain. Clinical and preclinical findings suggest long-term consequences of SSRI perinatal exposure for the offspring. The mechanisms of SSRI effects on developing brain remain largely unknown and there are no directional approaches for prevention of the consequences of maternal SSRI treatment during pregnancy. The heptapeptide Semax (MEHFPGP) is a synthetic analogue of ACTH(4–10) which exerts marked nootropic and neuroprotective activities. The aim of the present study was to investigate the long-term effects of neonatal exposure to the SSRI fluvoxamine (FA) in white rats. Additionally, the study examined the potential for Semax to prevent the negative consequences of neonatal FA exposure. Rat pups received FA or vehicle injections on postnatal days 1–14, a time period equivalent to 27–40 weeks of human foetal age. After FA treatment, rats were administered with Semax or vehicle on postnatal days 15–28. During the 2nd month of life, the rats underwent behavioural testing, and monoamine levels in brain structures were measured. It was shown that neonatal FA exposure leads to the impaired emotional response to stress and novelty and delayed acquisition of food-motivated maze task in adolescent and young adult rats. Furthermore, FA exposure induced alterations in the monoamine levels in brains of 1- and 2- month-old rats. Semax administration reduced the anxiety-like behaviour, improved learning abilities and normalized the levels of brain biogenic amines impaired by the FA exposure. The results demonstrate that early-life FA exposure in rat pups produces long-term disturbances in their anxiety-related behaviour, learning abilities, and brain monoamines content. Semax exerts a favourable effect on behaviour and biogenic amine system of rats exposed to the antidepressant. Thus, peptide Semax can prevent behavioural deficits caused by altered 5-HT levels during development.     

Kainic acid-induced seizures in aged rats: neurochemical correlates.

This study was conducted to assess the functional integrity of the kainate receptor-mediated seizure response in aged rats. Kainic acid was administered systemically to aged female Long-Evans (LE) rats and aged male F344 rats and the proconvulsant actions of kainic acid was compared to adult controls. The effects of kainic acid on brain regional content of monoamines and amino acids was also determined in the aged female LE and adult control rats. The latency to full clonic-tonic seizures was significantly reduced in aged female LE rats, and the number of seizures was significantly increased above that of the controls. There was increased mortality and a reduction in the latency to exhibit wet dog shakes in the aged F344 rats. Studies were also conducted to evaluate the role of ovarian hormones, route of administration, and dose of kainic acid in mediating the enhanced proconvulsant actions of kainic acid in aged rats. The neurochemical studies suggested that kainic acid significantly enhanced the release of ASP, GLU, and norepinephrine (NE) in the aged rats exhibiting clonic-tonic seizures. The adult rats given the same dose of kainic acid (15 mg/kg, IP) did not exhibit any significant change in brain content of monoamines or amino acids except for a reduction in mediobasal hypothalamic NE. An in vitro study was also conducted using brain slices from adult and aged F344 and it was found that aged rats released significantly more ASP than adults in response to kainic acid. These neurochemical findings were discussed in relation to previous studies of age-related alterations in excitatory amino acids (EAAs) and the role of EAA and NE in modulating limbic seizures. This study has clearly demonstrated that aged rats may be more susceptible to the excitotoxic action of EEAs acting through kainetic receptors.     

Regional differences in brain monoamine oxidase subtypes in an animal model of geriatric depression: effects of olfactory bulbectomy in young versus aged rats

Geriatric depression is often associated dysregulation of the hypothalamus-pituitary-adrenal (HPA) axis, and with poor responsiveness to antidepressants that work through inhibition of monoamine reuptake; accordingly, it has been suggested that MAO inhibitors may represent a therapeutic alternative in this group. In the current study, we evaluated expression of MAO subtypes in brain regions of young and aged rats subjected to olfactory bulbectomy (OBX), a procedure that reproduces many of the biochemical and functional changes associated with human depression. Activities of both MAO A and B were elevated in aged rats as compared to young rats in most regions, but not in the midbrain, and the OBX lesion failed to produce any change in this pattern. These results stand in contrast to the differential effects of glucocorticoids, which reduce brain MAO in young animals but induce activity in aged rats. Our results support the view that the aged brain possesses biochemical characteristics that distinguish its monoamine biochemistry from that of young brain, and that these distinctions may work in conjunction with HPA axis dysregulation to influence the etiology and therapy of geriatric depression. The use of appropriate animal models for depression and for disruption of HPA axis function can allow for the testing of potential human biomarkers (such as platelet MAO) that may serve to predict treatment outcome.