Brain Tyrosine Hydroxylase Activity Research Articles

Dopaminergic and serotonergic changes in rabbit fetal brain upon repeated gestational exposure to diesel engine exhaust.

Limited studies in humans and in animal models have investigated the neurotoxic risks related to a gestational exposure to diesel exhaust particles (DEP) on the embryonic brain, especially those regarding monoaminergic systems linked to neurocognitive disorders. We previously showed that exposure to DEP alters monoaminergic neurotransmission in fetal olfactory bulbs and modifies tissue morphology along with behavioral consequences at birth in a rabbit model. Given the anatomical and functional connections between olfactory and central brain structures, we further characterized their impacts in brain regions associated with monoaminergic neurotransmission. At gestational day 28 (GD28), fetal rabbit brains were collected from dams exposed by nose-only to either a clean air or filtered DEP for 2h/day, 5days/week, from GD3 to GD27. HPLC dosage and histochemical analyses of the main monoaminergic systems, i.e., dopamine (DA), noradrenaline (NA), and serotonin (5-HT) and their metabolites were conducted in microdissected fetal brain regions. DEP exposure increased the level of DA and decreased the dopaminergic metabolites ratios in the prefrontal cortex (PFC), together with sex-specific alterations in the hippocampus (Hp). In addition, HVA level was increased in the temporal cortex (TCx). Serotonin and 5-HIAA levels were decreased in the fetal Hp. However, DEP exposure did not significantly modify NA levels, tyrosine hydroxylase, tryptophan hydroxylase or AChE enzymatic activity in fetal brain. Exposure to DEP during fetal life results in dopaminergic and serotonergic changes in critical brain regions that might lead to detrimental potential short-term neural disturbances as precursors of long-term neurocognitive consequences.

Tetrahydrobiopterin treatment reduces brain L-Phe but only partially improves serotonin in hyperphenylalaninemic ENU1/2 mice.

Hyperphenylalaninemia (HPA) caused by hepatic phenylalanine hydroxylase (PAH) deficiency has severe consequences on brain monoamine neurotransmitter metabolism. We have studied monoamine neurotransmitter status and the effect of tetrahydrobiopterin (BH4) treatment in Pahenu1/enu2 (ENU1/2) mice, a model of partial PAH deficiency. These mice exhibit elevated blood L-phenylalanine (L-Phe) concentrations similar to that of mild hyperphenylalaninemia (HPA), but brain levels of L-Phe are still ~5-fold elevated compared to wild-type. We found that brain L-tyrosine, L-tryptophan, BH4 cofactor and catecholamine concentrations, and brain tyrosine hydroxylase (TH) activity were normal in these mice but that brain serotonin, 5-hydroxyindolacetic acid (5HIAA) and 3-methoxy-4-hydroxyphenylglycol (MHPG) content, and brain TH protein, as well as tryptophan hydroxylase type 2 (TPH2) protein levels and activity were reduced in comparison to wild-type mice. Parenteral L-Phe loading conditions did not lead to significant changes in brain neurometabolite concentrations. Remarkably, enteral BH4 treatment, which normalized brain L-Phe levels in ENU1/2 mice, lead to only partial recovery of brain serotonin and 5HIAA concentrations. Furthermore, indirect evidence indicated that the GTP cyclohydrolase I (GTPCH) feedback regulatory protein (GFRP) complex may be a sensor for brain L-Phe elevation to ameliorate the toxic effects of HPA. We conclude that BH4 treatment of HPA toward systemic L-Phe lowering reverses elevated brain L-Phe content but the recovery of TPH2 protein and activity as well as serotonin levels is suboptimal, indicating that patients with mild HPA and mood problems (depression or anxiety) treated with the current diet may benefit from supplementation with BH4 and 5-OH-tryptophan.

Brain tyrosine hydroxylase activity and systolic blood pressure in rats treated with either deoxycorticosterone and salt or angiotensin.

The hypertension induced in adult male rats by doca/salt was found to be accompanied by a significant rise in whole brain tyrosine hydroxylase (TH) activity. A smaller hypertensive effect, produced by angiotensin (750 ng kg-1 daily) was also accompanied by a proportional rise in whole brain TH activity. The specific antagonists spironolactone and saralasin completely blocked both responses in the doca/salt- and angiotensin-treated animals respectively and spironolactone showed a partial inhibition of the effects of angiotensin. In all the animals treated there was a clear correlation between systolic blood pressure and whole brain TH activity. The significance of these changes is discussed in the light of the central mechanism of hypertension.

Whole brain tyrosine hydroxylase activity during the development of deoxycorticosterone acetate-1% sodium chloride-induced hypertension in rats.

Water was chosen as the optimal medium for the extraction of tyrosine hydroxylase (TH) from rat brain. Determination of TH activity in crude homogenates failed to exhibit a linear relationship between enzyme concentration and measured activity, however, when a supernatant was used, a linear relationship existed. At a time when TH activity is maximally increased in the locus coeruleus after reserpine treatment (Reis et al 1975; Zigmond.1979) (2.5 mg kg-1 day-1 for 3 days, kill 24 h after last dose) we could detect no alterations in whole brain TH, however if treatment continued for 4 days and animals were killed 72 h after the last dose it was possible to detect increases in TH activity in various brain regions. These results suggest that local changes in brain TH activity are not revealed in measurements made on whole brain. The early rise in blood pressure, following the administration of deoxycorticosterone acetate (doca) and 1% NaCl to male Wistar rats, was accompanied by bradycardia. Whole brain Th activity was determined in these hypertensive animals 6-21 days after the commencement of treatment and the results failed to confirm the reported elevation of TH activity (Rylett et al 1976). The results are discussed with reference to the TH assay employed.

Effect of Clonidine on Tyrosine Hydroxylase Activity in the Adrenal Medulla and Brain of Spontaneously Hypertensive Rats

In the present study, we evaluated the effect of the alpha(2)-adrenoceptor agonist clonidine on tyrosine hydroxylase activity in adrenal medulla and brain of spontaneously hypertensive rats and Wistar Kyoto rats. Six-week-old animals were treated with clonidine (100 microg/kg body weight, daily, i.p.) for 4 weeks. Treatment with clonidine significantly reduced mean arterial blood pressure in spontaneously hypertensive rats to values similar to normotensive Wistar Kyoto rats. In the adrenal medulla of spontaneously hypertensive rats, clonidine treatment produced a significant increase in tyrosine hydroxylase activity with higher V(max) values and no changes in K(M) values. This effect was accompanied by a significant increase in tyrosine hydroxylase protein expression and of noradrenaline and adrenaline levels. In the brain of spontaneously hypertensive rats, treatment with clonidine produced a significant decrease in tyrosine hydroxylase activity with lower V(max) values and no changes in K(M) values accompanied by a significant decrease in tyrosine hydroxylase protein expression and of dopamine and noradrenaline levels. In Wistar Kyoto rats, clonidine treatment had no effect on tyrosine hydroxylase activity and protein expression or catecholamine levels in adrenal medulla or brain. Clonidine treatment significantly reduced noradrenaline and adrenaline plasma levels in spontaneously hypertensive rats and Wistar Kyoto rats. In conclusion, treatment with the alpha(2)-adrenoceptor agonist clonidine prevented the increase in mean arterial blood pressure in young spontaneously hypertensive rats. This effect was accompanied by opposite effects on tyrosine hydroxylase activity in spontaneously hypertensive rat adrenal medulla and brain: an increase in adrenal medulla and a decrease in brain, bringing sympathetic function to a similar profile found in normotensive Wistar Kyoto rats.

Tyrosine hydroxylase expression and activity in the rat brain: differential regulation after long-term intermittent or sustained hypoxia

Tyrosine hydroxylase, a hypoxia-regulated gene, may be involved in tissue adaptation to hypoxia. Intermittent hypoxia, a characteristic feature of sleep apnea, leads to significant memory deficits, as well as to cortex and hippocampal apoptosis that are absent after sustained hypoxia. To examine the hypothesis that sustained and intermittent hypoxia induce different catecholaminergic responses, changes in tyrosine hydroxylase mRNA, protein expression, and activity were compared in various brain regions of male rats exposed for 6 h, 1 day, 3 days, and 7 days to sustained hypoxia (10% O(2)), intermittent hypoxia (alternating room air and 10% O(2)), or normoxia. Tyrosine hydroxylase activity, measured at 7 days, increased in the cortex as follows: sustained > intermittent > normoxia. Furthermore, activity decreased in the brain stem and was unchanged in other brain regions of sustained hypoxia-exposed rats, as well as in all regions from animals exposed to intermittent hypoxia, suggesting stimulus-specific and heterotopic catecholamine regulation. In the cortex, tyrosine hydroxylase mRNA expression was increased, whereas protein expression remained unchanged. In addition, significant differences in the time course of cortical Ser(40) tyrosine hydroxylase phosphorylation were present in the cortex, suggesting that intermittent and sustained hypoxia-induced enzymatic activity differences are related to different phosphorylation patterns. We conclude that long-term hypoxia induces site-specific changes in tyrosine hydroxylase activity and that intermittent hypoxia elicits reduced tyrosine hydroxylase recruitment and phosphorylation compared with sustained hypoxia. Such changes may not only account for differences in enzyme activity but also suggest that, with differential regional brain susceptibility to hypoxia, recruitment of different mechanisms in response to hypoxia will elicit region-specific modulation of catecholamine response.

Estrogen regulation of in vitro brain tyrosine hydroxylase activity in the catfish Heteropneustes fossilis: interactions with cAMP-protein kinase A and protein kinase C systems in enzyme activation

In the present in vitro study, interactions of both cAMP-protein kinase A (PKA) and protein kinase C (PKC) systems were investigated in the estradiol-17β (E 2) regulation of forebrain (hypothalamus and telencephalon) tyrosine hydroxylase (TH) activity in the female catfish Heteropneustes fossilis in vitellogenic phase. E 2 produced biphasic effects on TH activity: low concentrations (10 −12–10 −5M) stimulated, and high concentrations (10 −3–10 −4 M) inhibited enzyme activity (Tukey’s test, P < 0.05). Co-incubations of the enzyme preparations with cAMP (1.0 mM), IBMX (1.5 mM) or theophylline (1.5 mM) and a low concentration of E 2 (10 −9 M) increased TH activity significantly. However, the co-incubations with a high concentration of E 2 (10 −3 M) decreased it significantly. Pre-incubations of the enzyme preparations with cAMP (0.1 mM), followed by different concentrations of E 2 (10 −12, 10 −9, 10 −4, and 10 −3 M) produced concentration-dependent biphasic effects. The pre-incubations with a low concentration of E 2 (10 −9 M), followed by different concentrations of cAMP (0.05–1.0 mM) produced a significant concentration-dependent stimulation of TH activity and that with a high concentration of E 2 (10 −3 M) produced a significant decrease in TH activity. Co-incubations of high and low E 2, with or without cAMP, and PKA inhibitor (H-89) decreased TH activity significantly. The incubations with H-89 abolished the stimulatory effect of low E 2 or low E 2 + cAMP and intensified the inhibitory effect of high E 2 or high E 2 + cAMP combination. Co-incubations with PKC inhibitor (calphostin C) did not influence the stimulatory effect of low E 2 but lowered the stimulatory effect of low E 2 + cAMP treatment. Kinetic studies showed that the stimulatory effect of a low E 2 concentration was due to a decrease in apparent K m and an increase in apparent V max for both cofactor and substrate, and the inhibitory effect of a high E 2 concentration was due to reverse changes in the kinetics. The stimulatory effect of cAMP alone or in combination with low E 2 was related to decreased K m and increased V max for the cofactor. The inhibitory effect of PKA and PKC blockers, alone or in combination with E 2 and/or cAMP was due to increased K m and decreased V max of the enzyme for the cofactor. The present data suggest that E 2 modulates the short-term activation of brain TH activity differentially and may involve mainly the cAMP-PKA system.

Involvement of cAMP in Basal and Estradiol-17β(E 2 ) Modulated Activity of Brain Tyrosine Hydroxylase in the Female Catfish Heteropneustes fossilis :An in vitro Study

In the present in vitro study, involvement of cAMP in basal and E 2 -modulated brain (hypothalamus and telencephalon) TH activity was demonstrated in female catfish Heteropneustes fossilis. Incubation of enzyme preparation (soluble active fraction) with cAMP (0.05-1 .OmM) and inhibitors of cAMP-specific phosphodiesterase (IBMX, 0.5-2.0mM) and theophylline, 0.5-2.0mM) stimulated enzyme activity in a concentration-dependent manner.

Thyroid hormone modulation of brain in vivo tyrosine hydroxylase activity and kinetics in the female catfish Heteropneustes fossilis.

In the female catfish Heteropneustes fossilis, administration of thyroxine (T(4))(,) 1 micro g/g body weight, i.p., in both gonadal resting and preparatory phases for 7, 14 and 21 days caused hyperthyroidism, as evidenced from a duration-dependent significant increase in serum triiodothyronine (T(3)), and of tyrosine hydroxylase (TH) activity in telencephalon, hypothalamus-pituitary and medulla oblongata (Newman-Keuls' test; P<0.05). Hypothyroidism induced by adding 0.03% thiourea to aquarium water holding the catfish for 7, 14 and 21 days decreased serum T(3) levels in a duration-dependent manner (Newman-Keuls' test; P<0.05) and inhibited TH activity in the brain regions. T(4) replacement in 21day thiourea-treated fish restored and even elevated significantly serum T(3) levels as well as brain TH activity in a duration-dependent manner. In general, the changes in enzyme activity were higher in the forebrain regions than medulla oblongata and in the resting phase than preparatory phase. Kinetic studies by Lineweaver-Burk plots showed that the stimulatory effect following T(4) administration and T(4) replacement on TH activity was due to increased affinity of the enzyme for its cofactor (6,7-dimethyl-2-amino-4-hydroxy-5,6,7,8-tetrahydropteridine), as evident from a significant decrease in apparent Michaelis-Menten constant (K(m)) and an increase in apparent velocity maximum (V(max)). The TH inhibition due to the thiourea treatment can be related to decreased affinity of the enzyme for its cofactor, as evident from a significant increase in apparent K(m) value and a significant decrease in V(max). These data clearly show that circulating levels of T(4)/T(3) modulate brain TH activity by altering the kinetic properties of the enzyme, which, in turn, influence catecholaminergic activity and dependent functions.

In vitro effects of catecholamines and catecholestrogens on brain tyrosine hydroxylase activity and kinetics in the female catfish Heteropneustes fossilis.

Effects of catecholamines and catecholestrogens on tyrosine hydroxylase (TH) activity and kinetics were investigated in the telencephalon and hypothalamus of female Heteropneustes fossilis in gonad quiescent (resting) and recrudescent (preparatory) phases. Dopamine, noradrenaline and adrenaline and the catecholestrogen, 2-hydroxyestradiol-17 beta inhibited TH activity in a concentration-dependent manner in both resting and preparatory phases, with a higher effect in the resting phase. Two- methoxyestradiol-17 beta did not alter TH activity in any season. The catecholamines inhibited TH in a competitive manner increasing apparent K(m) values significantly without altering the apparent V(max). Two-hydroxyestradiol-17 beta inhibited significantly the enzyme in a noncompetitive manner and decreased apparent V(max) without altering apparent K(m) values. The apparent K(i) is higher for dopamine than noradrenaline or adrenaline. The apparent K(i) for 2-hydroxyestradiol-17 beta is not significantly different from that of noradrenaline. The present results suggest an interaction between oestradiol-17beta (E2) and catecholamine metabolism at the level of tyrosine hydroxylation and E2 effects on catecholamines may be mediated through its 2-hydroxylation.

Effects of ovariectomy and oestradiol-17beta replacement on brain tyrosine hydroxylase in the catfish Heteropneustes fossilis: changes in in vivo activity and kinetic parameters.

In Heteropneustes fossilis, ovariectomy inhibited in vivo brain (hypothalamus-pituitary, telencephalon and medulla oblongata) tyrosine hydroxylase (TH) activity with significant effects in weeks 2, 3, 4 and 5 of the gonadal resting phase and in weeks 3, 4 and 5 of the prespawning phase (P<0.05, Tukey's test). Oestradiol-17beta (OE(2)) replacement in 3-week ovariectomised fish produced biphasic responses in both seasons; the low dosages of 0.05 and 0.5 micro g/g body weight (BW) elevated TH activity, whereas the high dosages of 1.0 and 2.0 micro g/g BW decreased it. The magnitude of the inhibition was higher in the resting phase than in the prespawning phase. The inhibitory effect of ovariectomy may be produced by elevating the apparent K(m) values (decreased affinity) of the enzyme for both L-tyrosine (substrate) and dimethyltetrahydropteridine (cofactor) and consequently decreasing the V(max). Significant changes (P<0.05) in both these parameters were noticed but showed minor differences with regard to the length of ovariectomy, season or brain regions. The biphasic effects of OE(2) replacement on TH activity seemed to be produced by differential effects on apparent K(m) and V(max). The stimulatory effect of the low dosages of OE(2) coincides with a decrease in the apparent K(m) values (increased affinity) for both substrate and cofactor and an increase in the V(max) of the enzyme. The inhibitory effect of the high dosages of OE(2) correlated with an increase in the apparent K(m) values (decreased affinity) for both substrate and cofactor, and a decrease in the V(max) compared with the lower dosage groups. The results strongly suggested that OE(2) can modulate brain catecholaminergic activity at the level of tyrosine hydroxylation which, in turn, may alter gonadotrophin secretion. OE(2) may elicit biphasic effects by differentially altering the enzyme affinity towards the substrate and cofactor.

Effects of altered photoperiod and temperature, serotonin-affecting drugs, and melatonin on brain tyrosine hydroxylase activity in female catfish, Heteropneustes fossilis: a study correlating ovarian activity changes.

Exposure of female catfish, Heteropneustes fossilis to 30-day regimes of long photoperiod (16L), elevated temperature (28 +/- 2 degrees C), or a combination of both stimulated brain tyrosine hydroxylase (TH) activity significantly over that of control fish held in natural conditions in gonad resting (10.5L:13.5D, 10 +/- 2 degrees C) and preparatory (12.5L:11.5D,18 +/- 2 degrees C) phases. The response was high in the combination group in both phases. The increase in TH activity was higher in forebrain regions (telencephalon and hypothalamus) than medulla oblongata. Exposure of the fish to short photoperiod (8L:16D) and total darkness decreased the enzyme activity significantly in both resting and preparatory phases regardless of the temperature. The inhibition was high in fish held under total darkness. Gonadosomatic index (GSI) was significantly elevated in long photoperiod and high temperature groups, alone or in combination, and decreased significantly in short photoperiod (only in preparatory phase) and total darkness groups. Administration of the serotonin precursor 5-hydroxytryptophan (5-HTP; 5mg/100g body weight [BW], three daily intraperitoneal [i.p.] injections prior to sacrificing) stimulated TH activity in fish held under long and normal photoperiods in both phases. Three daily injections of the serotonin blocker parachlorophenylalanine (p-CPA; 10mg/100g BW) and melatonin (75 microg/100g BW) prior to sacrificing inhibited brain TH activity significantly in both phases. GSI was significantly stimulated by 5-HTP, and inhibited by both p-CPA and melatonin injections. Changes in TH activity and GSI can be correlated and explained on the basis of previous reports on changes in catecholamine activity that modulates gonadotropin secretion in the catfish. Further, the photoperiod and temperature-induced changes in TH activity may be modulated by alterations in serotonergic activity.

Effects of Repetitive Hypoglycemia on Neuroendocrine Response and Brain Tyrosine Hydroxylase Activity in the Rat

Hypoglycemia-associated autonomic failure (HAAF) is a syndrome of acute adaptation to a metabolic stressor, in which neuroendocrine responses to repetitive hypoglycemic bouts are blunted. The CNS mechanisms that contribute to HAAF are unknown. In the present study, we modeled HAAF in the rat and measured the activity of tyrosine hydroxylase (TH) as an index of acute noradrenergic activation, to test the hypothesis that noradrenergic activation of the hypothalamus might be impaired. In association with a significant counter-regulatory response to a single bout of hypoglycemia (elevated corticosterone, catecholamines, and glucagon), TH activity was elevated overall in brainstem NE cell body areas and hypothalamus. With multiple hypoglycemic episodes in a 24 h period, the counter-regulatory response was blunted, and hypothalamic TH activity was comparable to that of saline-infused controls. In a similar paradigm, multiple bouts of CNS neuroglucopenia did not blunt the hyperglycemic or corticosterone responses, and were required for elevation of TH activity. This alternate response pattern suggests that insulin-induced hypoglycemia and cerebral neuroglucopenia represent somewhat different metabolic stressors at the CNS.

Effects of imidazoline drugs on tyrosine and tryptophan hydroxylase activity in rat brain in vivo.

Effects of imidazoline drugs on tyrosine and tryptophan hydroxylase activity in rat brain in vivo.

Regional distribution and control of tyrosine hydroxylase activity in the quail brain

Tyrosine hydroxylase (TH) activity, the rate-limiting step in the synthesis of catecholamines, was quantified in the preoptic area-hypothalamus of adult male Japanese quail by a new assay measuring the tritiated water production from 3,5-[ 3H]-L-tyrosine. Maximal levels of activity were observed at a 20–25 μM concentration of substrate, with more than 50% inhibition of the activity being recorded at a 100 μM concentration. TH activity was linear as a function of the incubation time during the first 20 min and maximal at a pH of 6.0. TH was heterogeneously distributed in the quail brain with highest levels of activity being found (in decreasing order) in the mesencephalon, diencephalon, and telencephalon. Given the large size of the telencephalon, this is the brain area that contains, as a whole, the highest level of enzyme activity. TH inhibitors that have been well-characterized in mammals, such as 3-iodo- l-tyrosine and l-α-methyl-p-tyrosine (AMPT) completely inhibited the enzyme activity at a 100 μM concentration. In mammals, the accumulation of catecholamines exerts a negative feedback control on TH activity. Similar controls were observed in the quail brain. Two inhibitors of the DOPA decarboxylase that should lead to accumulation of DOPA depressed TH activity by 60% or more, and the inhibitor of the dopamine β-hydroxylase, fusaric acid that should cause an accumulation of dopamine, suppressed 90% of the TH activity. The addition of exogenous DOPA, dopamine, or norepinephrine to the brain homogenates also strongly inhibited TH activity, independently confirming the feedback effects of the enzyme products on the enzyme activity. These data demonstrate that TH activity in the quail brain is heterogeneously distributed and acutely regulated, as it is in mammals, by the accumulation of its products and of the derived catecholamines.

Biological research on schizophrenia.

Based on the biochemical analysis of postmortem brains from chronic schizophrenic patients, we found abnormalities of glutamatergic neurons as well as dopaminergic neurons. Glutamate receptors, such as the kainate receptor labeled by 3H-kainate, the N-methyl-D-aspartate (NMDA) receptor by 3H MK801, and the strychnine-insensitive glycine sites in the NMDA receptor by 3H-glycine, increased significantly in various cortical areas of schizophrenic brains. According to the animal experiments and a significant negative correlation between kainate binding values and glutamate concentrations, it is suggested that glutamate receptors increased due to hypoglutamatergic function in the brain of chronic schizophrenia. Hyperdopamine hippothesis of schizophrenia is supported by the correlation between affinity to dopamine receptor and clinical potency of antipsychotic drugs. Measurement of tyrosine hydroxylase activity and dopamine D2 receptor in the schizophrenic brain provided evidence of hyperdopaminergia. Association study of a missense variant in the dopamine D2 receptor gene (Cys311) revealed that the allele frequency of the variant was significantly higher in the schizophrenic patients than the controls. The patients carrying this variant had less severe negative symptoms and better response to antipsychotic drug treatment. Dopamine-induced sequestration of dopamine D2S receptor with Cys variant expressed in CHO cells was shown to a lesser extent than wild-type receptor. This experimental result may be consistent with better responsiveness of the patients with Cys311 to antipsychotic drugs.

Pb-induced alterations in tyrosine hydroxylase activity in rat brain.

Our previous studies have shown that exposure to low levels of Pb results in significant reductions in dopamine (DA) and its metabolites (3,4-dihyroxyphenylacetic acid, DOPAC and homovanillic acid, HVA) in nucleus acumbens (NA). This area of brain receives dopaminergic projections from the ventral tegmentum and is considered vital in manifestation of many behavioral responses. Similarly, basal and K(+)-induced release of DA was found significantly reduced in the Pb-exposed rats as compared to the controls in this brain region. Additional studies indicated that acute infusion of Pb in nucleus acumbens caused significant release of DA. Based on these observations it was postulated that the reductions in DA contents and in the basal and stimulus-induced release of DA in NA were manifestations of attenuated dopaminergic activity in this brain region. However, the mechanism of this attenuation is not yet clear. Studies reported here were designed to evaluate the role of a key regulatory enzyme in biosynthesis of DA, i.e. tyrosine hydroxylase (TH) in Pb-induced reductions in dopaminergic activity. The results of these studies indicated that 50 and 500 ppm Pb produced 22.8 and 56% inhibition of TH activity in vitro respectively, and that the enzyme activity was reduced to 43% in rats exposed to 50 ppm lead for 30 days as compared to the controls. The alterations in TH activity in Pb-exposed animals were further confirmed by Western blot analysis. Collectively, these results suggest that Pb-induced inhibition of TH activity in rat brain may contribute to the reductions in dopaminergic activity observed in Pb-exposed animals.

Biochemical and physiological evidence that carnosine is an endogenous neuroprotector against free radicals.

1. Carnosine, anserine, and homocarnosine are endogenous dipeptides concentrated in brain and muscle whose biological functions remain in doubt. 2. We have tested the hypothesis that these compounds function as endogenous protective substances against molecular and cellular damage from free radicals, using two isolated enzyme systems and two models of ischemic brain injury. Carnosine and homocarnosine are both effective in activating brain Na, K-ATPase measured under optimal conditions and in reducing the loss of its activity caused by incubation with hydrogen peroxide. 3. In contrast, all three endogenous dipeptides cause a reduction in the activity of brain tyrosine hydroxylase, an enzyme activated by free radicals. In hippocampal brain slices subjected to ischemia, carnosine increased the time to loss of excitability. 4. In in vivo experiments on rats under experimental hypobaric hypoxia, carnosine increased the time to loss of ability to stand and breath and decreased the time to recovery. 5. These actions are explicable by effects of carnosine and related compounds which neutralize free radicals, particularly hydroxyl radicals. In all experiments the effective concentration of carnosine was comparable to or lower than those found in brain. These observations provide further support for the conclusion that protection against free radical damage is a major role of carnosine, anserine, and homocarnosine.

Effects of Polychlorinated Biphenyls (PCBs) on Brain Tyrosine Hydroxylase Activity and Dopamine Synthesis in Rats

Literature reports suggest that polychlorinated biphenyls (PCBs) may alter dopaminergic neurotransmission in mammalian forebrain. In vitro, PCBs can decrease dopamine levels in PC 12 cells and studies of the structure-activity relationship (SAR) indicate that ortho-substituted (non-coplanar) PCB congeners are more active than para-substituted (coplanar) congeners. This report tested the hypothesis that ortho-substituted PCBs can selectively (vs para-substituted congeners) decrease dopamine synthesis in mammalian forebrain by inhibiting the activity of tyrosine hydroxylase, the rate-limiting enzyme in dopamine biosynthesis. In vitro effects of individual PCB congeners on activity of striatal tyrosine hydroxylase from two different rat strains were assessed. It was found that certain ortho-substituted PCB congeners (e.g., 2,2'-DCB) can inhibit tyrosine hydroxylase activity and dopamine synthesis by nearly 40% in minces of corpus striatum prepared from Sprague-Dawley and Long-Evans hooded rats. Comparatively, the ortho, meta-substituted PCB congener 2,2',5,5'-TeCB inhibited tyrosine hydroxylase activity only in striatal minces obtained from Sprague-Dawley rats, suggesting that genetic factors may influence the susceptibility of mammals to effects of PCBs that compromise brain dopamine synthesis. The PCB-induced inhibition of tyrosine hydroxylase activity in mammalian forebrain observed here appears to occur through indirect and as yet unknown mechanisms.

Effects of zonisamide on dopaminergic system.

Effects of zonisamide (ZNS) on extracellular dopamine (DA), its precursor 3,4-dihydroxyphenylalanine (DOPA), its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) levels in the striatum as well as hippocampus of freely moving rats were studied. Intracellular DA, DOPA, DOPAC and HVA levels, as well as DOPA accumulation as an index of tyrosine hydroxylase activity in the rat brain in vivo, DA re-uptake in the striatum and hippocampus, and monoamine oxidase (MAO) activities were also determined. Acute administrations of therapeutic ZNS doses (20 and 50 mg/kg) increased striatal extracellular DOPA levels, intracellular striatal and hippocampal DOPA levels, and stimulated DOPA accumulation in both brain regions. ZNS also increased striatal and hippocampal intracellular as well as extracellular DA and HVA levels, but decreased those of DOPAC levels. Chronic (3 weeks) administrations of therapeutic ZNS doses (20 and 50 mg/kg/day) increased intracellular DA, DOPA, DOPAC and HVA levels in striatum and hippocampus. ZNS-induced changes were greater in intracellular levels than in extracellular levels. Acute and chronic supratherapeutic ZNS dose (100 mg/kg) administration decreased intracellular levels of all substances detectable in both brain regions, and inhibited DOPA accumulation. Both subtypes of MAO (type A and type B) activities were weakly inhibited by ZNS. ZNS showed no effect on DA re-uptake in striatum nor in hippocampus. These results suggest that therapeutic ZNS doses increase DOPA accumulation as well as both intracellular and extracellular DA, DOPA and HVA levels. However, such doses also decrease extracellular and intracellular DOPAC levels by enhancing DA synthesis and/or by selectively inhibiting MAO-B activities. In addition, chronic therapeutic ZNS dose administration enhances DA synthesis, which results in increased intracellular DA, its precursor and its metabolites levels. On the other hand, both acute and chronic supratherapeutic ZNS dose administrations inhibit DA turnover. These ZNS effects on DA metabolism are at least partly involved in the mechanisms of action of ZNS.