Tyrosine Hydroxylase Protein Expression Research Articles

Estradiol increases catecholamine levels in the hypothalamus of ovariectomized rats during the dark-phase

Estrogens modulate critical homeostatic functions of the hypothalamus such as temperature regulation, sexual behavior and sleep with the most pronounced effects in rats occurring during the dark-phase. The neurochemical signals underlying estrogenic regulation of these hypothalamic functions have not been clearly identified, possibly due to the fact that previous studies have not explored the effects of estrogen treatments on neuronal signaling during the dark-phase. In the present study, ovariectomized rats received estradiol benzoate (5 μg/rat for 7 days, s.c.) and norepinephrine and dopamine levels were measured in the preoptic area of the hypothalamus across the light/dark cycle using in vivo microdialysis. Estradiol benzoate treatment increased extracellular norepinephrine and dopamine levels relative to vehicle treatment during the dark-phase. Increases in norepinephrine and dopamine were first detected by 30 min and 5.5 h after lights-off, respectively. Subsequent increases in norepinephrine and dopamine were also noted throughout the 9.5- h collection period. The effect of estradiol benzoate on catecholamine release did not correlate with increases in either tyrosine hydroxylase (TH) protein expression or activity levels in the anterior hypothalamus, although a marked decrease in TH activity correlated with a rise in extracellular norepinephrine at the beginning of the dark-phase. We conclude that subchronic estradiol benzoate treatment increases extracellular catecholamine levels in the preoptic area of the hypothalamus during the dark-phase without a concomitant increase in neurotransmitter biosynthesis. The estradiol benzoate-induced increases in norepinephrine and dopamine levels in the preoptic area during the dark-phase may play an important role in modulating critical hypothalamic functions.

Decreased expression of ErbB4 and tyrosine hydroxylase mRNA and protein in the ventral midbrain of aged rats

Decreased availability or efficacy of neurotrophic factors may underlie an increased susceptibility of mesencephalic dopaminergic cells to age-related degeneration. Neuregulins (NRGs) are pleotrophic growth factors for many cell types, including mesencephalic dopamine cells in culture and in vivo. The functional NRG receptor ErbB4 is expressed by virtually all midbrain dopamine neurons. To determine if levels of the NRG receptor are maintained during aging in the dopaminergic ventral mesencephalon, expression of ErbB4 mRNA and protein was examined in young (3 months), middle-aged (18 months), and old (24–25 months) Brown Norway/Fischer 344 F1 rats. ErbB4 mRNA levels in the substantia nigra pars compacta (SNpc), but not the adjacent ventral tegmental area (VTA) or subtantia nigra pars lateralis (SNl), were significantly reduced in the middle-aged and old animals when compared to young rats. Protein expression of ErbB4 in the ventral midbrain was significantly decreased in the old rats when compared to the young rats. Expression of tyrosine hydroxylase (TH) mRNA levels was significantly reduced in the old rats when compared to young animals in the SNpc, but not in the VTA or SNI. TH protein levels in the ventral midbrain were also decreased in the old animals when compared to the young animals. These data demonstrate a progressive decline of ErbB4 expression, coinciding with a loss of the dopamine-synthesizing enzyme TH, in the ventral midbrain of aged rats, particularly in the SNpc. These findings may implicate a role for diminished NRG/ErbB4 trophic support in dopamine-related neurodegenerative disorders of aging such as Parkinson's disease.

Rho Kinase Inhibitor Y‐27632 Down‐Regulates Norepinephrine Synthesis and Release in PC12 Cells

Rho kinase inhibition is beneficial for neurite outgrowth and nerve disorders, and the Rho kinase inhibitors have been regarded as promising agents to treat neural diseases. The main aim of the study was to elucidate how Rho kinase inhibitor Y-27632 regulates neurotransmitter norepinephrine synthesis and release in PC12 cells when neurite outgrowth was induced. PC12 cells were treated with Y-27632 for 6 days. The amount of norepinephrine synthesized in PC12 cells and the amount released evoked by acetylcholine or by KCl were determined by norepinephrine enzyme-linked immunosorbent assay kits. The results showed that the amount of norepinephrine both synthesized and released was down-regulated with a concentration-dependent relationship. Further results of Western blotting found that the protein expression of tyrosine hydroxylase and synapsin I (especially its active form, synapsin I phosphoSer603) was also down-regulated, which were directly related to synthesis and release of norepinephrine, respectively. All the results suggest that Y-27632 is able to down-regulate norepinephrine synthesis and release, the direct mechanism of which may be associated with down-regulation on expression of some proteins, including tyrosine hydroxylase and synapsin I.

Up‐regulation of Adrenal Leptin Receptor Expression and Tyrosine Hydroxylase Activation in Age‐related Hypertension

Blood pressure (BP) and sympathetic activation have been shown to increase with age. The aim of the present study was to examine whether increased adrenal OB‐Rb expression and tyrosine hydroxylase (TH) activation mediated age‐related hypertension.Method4‐6 mth (young) and 20‐24 mth (old) male F344/BN rats (n=4‐6 rats/age) were implanted with telemetry for 24‐hrs arterial pressure (MAP) monitoring and were housed in individual metabolic cages. Adrenal leptin receptors (OB‐Ra & OB‐Rb) and TH protein expressions were determined by Western Blot.ResultsOld rats exhibited a significant increase in MAP (107±1 vs 98±1mmHg), body weight (556±11 vs 332±5g) and serum leptin (24±5 vs 5.1±0.7ng/ml) compared with young rats (p<0.05). The increased MAP was positively correlated with serum leptin (r2=0.7, p<0.0001). Western Blot analysis revealed a higher OB‐Rb expression (54%) than OB‐Ra in young adrenal. Although OB‐Ra expression did not change with age, OB‐Rb protein expression was markedly up‐regulated by 211±13% in old compared with young rats. TH activation (as measured by phosphorylated TH/total TH) in adrenal was also elevated by 185±10% in old compared with young rats.ConclusionResults of the present study suggest that the increased BP with age may, at least in part, be mediated by the elevated circulating leptin levels, up‐regulated adrenal OB‐Rb protein expression and TH activation.

Losartan affects Tyrosine Hydroxylase Protein Expression in the A2 Area and Hypoglossal Nucleus of Normal and Dystrophic Hamsters

Angiotensin AT1 receptors affect production of tyrosine hydroxylase (TH), the rate‐limiting enzyme for production of catecholamines. AT1 receptors are colocalized with TH in the A2 region of the nucleus tractus solitaries and hypoglossal nucleus. We hypothesized that blocking AT1 receptors with losartan (L), an AT1 receptor antagonist, would decrease TH expression in the A2 and hypoglossal nucleus in normal (N) and dystrophic (D) hamsters. Two month old male N and D hamsters received tap water or L in drinking water at a dose of 50 mg/kg/day for 2.5 months. At 4.5 months, animals were sacrificed, brains perfused, cryoprotected, and frozen at ‐70° C. Brains were sliced at 35‐40 micrometers. Immunohistochemistry was performed using a mouse monoclonal TH primary antibody (1:10,000) visualized with DAB. ImageJ was used to evaluate A2 and hypoglossal areas and number of TH positive neurons. Losartan had no effect on hypoglossal areas, but decreased the number and density of TH positive neurons and in D hamsters, but not in N hamsters. In N and D hamsters TH positive neurons were predominately located in ventral regions of the hypoglossal nuclei. Losartan treatment eliminated this localization in both groups. In the A2 region no significant differences were observed in areas or number of positive TH neurons, but in D hamsters L decreased TH positive neuron density.Funded by: APS Undergraduate Summer Fellowship & P20 RR‐015567

Neuroprotective effects of enriched environment in MPTP-treated SAMP8 mice

Neuroprotective effects of enriched environment (EE) have been well established. Recent study suggests that exposure to EE can protect dopaminergic neurons against MPTP-induced Parkinsonism. After 64 female SAMP8 mice were reared in EE and standard environment (SE) for 3 months, the effects of EE and SE were compared on behavioural change, tyrosine hydroxylase (TH) immunoreaction positive neuron and dopaminetransporter (DAT) expression in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-treated SAMP8. EE mice showed decreased spontaneous activity compared with SE mice. But EE + MPTP mice showed less decreased spontaneous activity compared with SE + MPTP mice. Otherwise, EE mice showed increased percentage of entries into the open arms and percentage of time spent in the open arms. Furthermore, EE mice demonstrated reduced neurotoxicity, with less decreased TH mRNA and protein expression in Substantia Nigra (SN) after MPTP administration compared with SE mice. SE mice showed a 53.77% loss of TH-positive neurons, whereas EE mice only showed a 42.28% loss. Moreover, EE mice showed decreased DAT mRNA and protein expression compared with SE mice. These data demonstrate that EE can protect dopaminergic neurons against MPTP-induced neuronal damage, which suggest that the probability of developing Parkinson's disease (PD) may be related to life environment.

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.

Absence of age-related changes in nigral dopaminergic neurons of Asian Indians: Relevance to lower incidence of Parkinson's disease

Age-related loss of melanized nigral neurons reported in the British Caucasians is not observed in Asian Indian, American and French adults. In the Americans, loss of dopaminergic phenotype occurs from midlife, without frank neurodegeneration. Here, we investigated whether nigral dopaminergic neurons in Asian Indians are lost with age or undergo morphological or biochemical dysfunction. Using unbiased stereology we estimated volume, number of melanized, borderline/non-melanized (n=34, 28 gestational weeks to 80 years) and tyrosine hydroxylase (TH)–Nurr1 co-labeled neurons (n=32, 28 gestational weeks to 80 years) in substantia nigra pars compacta. We quantified Nurr1 and TH proteins by immunoblotting (n=18, 28 gestational weeks to 69 years) and apoptotic neurons by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) staining. Nuclear and soma size was estimated by morphometry. There was no age-related decline in volume, neuronal density, neuronal numbers and TH-Nurr1 co-labeled neurons. TH and Nurr1 protein expression remained stable. Lack of TUNEL-TH co-labeled cells confirmed absence of neuronal apoptosis. The neuronal size remained unaltered. Our findings of preserved nigral dopaminergic neurons suggest no age-related loss of nigral function in Asian Indians, unlike the Americans. This may explain the lower incidence of Parkinson's disease in Asian Indians.

Changes in NGF and NT-3 protein species in the superior cervical ganglion following axotomy of postganglionic axons

Mature sympathetic neurons in the superior cervical ganglion (SCG) are regulated by target-derived neurotrophins such as nerve growth factor (NGF) and neurotrophin-3 (NT-3). High molecular weight NGF species and mature NT-3 are the predominant NGF and NT-3 protein isoforms in the SCG, yet it is unknown whether the presence of these species is dependent on intact connection with the target tissues. In an attempt to determine the role of peripheral targets in regulating the neurotrophin species found in the SCG, we investigated the NGF and NT-3 protein species present in the SCG following axotomy (transection) or injury of the post-ganglionic axons. Following a 7 day axotomy, the 22–24 kDa NGF species and the mature 14 kDa NT-3 species in the SCG were significantly reduced by 99% and 66% respectively, suggesting that intact connection with the target is necessary for the expression of these protein species. As expected, tyrosine hydroxylase (TH) protein in the SCG was significantly reduced by 80% at 7 days following axotomy. In order to distinguish between the effects of injury and loss of target connectivity, the SCG was examined following compression injury to the post-ganglionic nerves. Following injury, no reduction in the 22–24 kDa NGF or 14 kDa mature NT-3 species was observed in the SCG. TH protein was slightly, yet significantly, decreased in the SCG following injury. The findings of this study suggest that the presence of the 22–24 kDa NGF and mature 14 kDa NT-3 species in the SCG is dependent on connection with peripheral targets and may influence, at least in part, TH protein expression in adult sympathetic neurons.

Visualization of spatiotemporal differentiation of dopaminergic interneurons in adult mouse olfactory bulb using transgenic mice

Olfactory bulb (OB) interneurons, predominantly periglomerular (PG) and granule (GR) cells, are derived from neural stem cells in the subventricular zone (SVZ) throughout life and migrate to the OB in the rostral migratory stream (RMS). In the adult superficial GR layer transgene expression was found, either enhanced green fluorescent protein or LacZ reporter driven by a 9 kb tyrosine hydroxylase (TH) promoter, that marked the dopamine (DA) phenotype. To demonstrate that the reporters and endogenous TH were similarly regulated expression of both parameters was shown to decline in the OB PG cells ipsilateral to odor deprivation produced by adult unilateral naris closure. The present findings suggested that DAergic differentiation might begin prior to progenitors reaching a PG position despite evidence that TH protein expression occurred only after PG cells received olfactory afferent stimulation. Of many genes previously hypothesized to regulate OB DA expression, regulated expression of the orphan receptor, Nurr1, but not the homeobox-containing genes, Dlx-1 and -2, was consistent with a role in regulation of the DA phenotype in adult mice OB. The studies show that transgenic lines are useful for analyzing spatiotemporal regulation by both intrinsic (programmed) and environmental factors in the neurogenesis of adult mouse OB DAergic interneurons.

Potassium channels are involved in zinc‐induced apoptosis in MES23.5 cells

There is evidence that zinc may be involved in the pathogenesis of Parkinson's disease by an apoptotic pathway. However, the mechanisms underlying zinc-induced apoptosis are unknown. Previous studies showed that 6-hydroxydopamine (6-OHDA)-enhanced potassium channels are involved in apoptosis of dopaminergic neurons. Our study was designed to test whether zinc-induced apoptosis was mediated by potassium channels. First we demonstrated cell apoptosis with zinc treatment by Hoechst staining assay. The results showed that 13.38% +/- 0.6% of MES23.5 cells were apoptotic after 24 hr of incubation with 60 microM zinc sulfate. Then we observed that the tyrosine hydroxylase (TH) protein expression and the dopamine content decreased, as detected by Western blots and high-performance liquid chromatography-electrochemical detection (HPLC-ECD). We further elucidated the mechanism of cell apoptosis by using whole-cell patch clamp recording. The data demonstrated that MES23.5 cells exhibited a tetraethylammonium (TEA)-sensitive outward K(+) current with delayed rectifier characteristics. Increases of K(+) current density were recorded following the treatment with 60 microM zinc for 4-8 hr. After incubation with 20 mM TEA, the zinc-induced enhancement of K(+) currents was fully blocked. Furthermore, incubation with TEA blocked zinc-mediated caspase-3 activation and cell apoptosis. These data suggest that zinc-induced apoptosis of MES23.5 dopaminergic cells may due to the enhancement of TEA-sensitive K(+) channel activity.

The Effects of Piroxicam in the Attenuation of MPP+/MPTP Toxicity In Vitro and In Vivo

Several lines of evidence support the neuroprotective action of cyclooxygenase-2 (COX-2) inhibitors in various models of Parkinson's disease (PD). In the current study, we investigated the neuroprotective properties of several COX inhibitors against 1-methyl-4-phenylpyridinium (MPP+) in neuroblastoma Neuro 2A (N-2A) cells in vitro and the protection against degeneration of substantia nigra pars compacta (SNc) dopaminergic (DA) neurons after the administration of 1-methyl 4-phenyl 1,2,3,6-tetrahydropyridine (MPTP) in C57/BL6 male mice. The data obtained demonstrate a lack of protective effects observed by COX 1-2 inhibitors ibuprofen and acetylsalicylic acid against MPP+ toxicity in N-2A, where piroxicam was protective in a dose dependent manner (MPP+ control: 15 +/- 2% MPP+ piroxicam: 5 mM 89 +/- 4%). The data also indicate a drop in mitochondrial oxygen (O(2)) consumption and ATP during MPP+ toxicity with no restoration of mitochondrial function concurrent to a heightened concentration of somatic ATP during piroxicam rescue. These findings indicate that the neuroprotective effects of COX inhibitors against MPP+ are not consistent, but that piroxicam may work through an unique mechanism to propel anaerobic energy metabolism. On the other hand, using mice, piroxicam (20 mg/kg) was effective against MPTP-induced dopaminergic degeneration in the (SNc) and loss of locomotive function in mice. Administering a 3 day pre-treatment of piroxicam (20 mg/kg) was effective in antagonizing the losses in SNc tyrosine hydroxylase protein expression, SNc DA concentration and associated anomaly in ambulatory locomotor activity. It was concluded from these findings that piroxicam is unique among COX inhibitors in providing very significant neuroprotection against MPP+ in vitro and in vivo.

Induction of Human Mesenchymal Stem Cells into Dopamine-Producing Cells with Different Differentiation Protocols

Several reports have shown that human mesenchymal stem cells (MSCs) are capable of differentiating outside the mesenchymal lineage. We sought to induce MSCs to differentiate into dopamine-producing cells for potential use in autologous transplantation in patients with Parkinson's disease (PD). Following cell culture with various combinations of differentiation agents under serum-free defined conditions, different levels of up-regulation were observed in the protein expression of tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis. Further analysis of selected differentiation protocols revealed that the induced cells displayed a neuron-like morphology and expressed markers suggesting neuronal differentiation. In addition, there was an increase in Nurr 1, the dopaminergic transcription factor gene, concomitant with a decrease gamma-aminobutyric acid (GABA)ergic marker expression, suggesting a specific dopaminergic direction. Moreover, the induced cells secreted dopamine in response to depolarization. These results demonstrate the great therapeutic potential of human MSCs in PD.

Alteration of dopaminergic markers in gastrointestinal tract of different rodent models of Parkinson's disease

Parkinson's disease (PD) is a progressive neurological disorder that is often associated with various gastrointestinal (GI) symptoms. The link between the alteration of dopaminergic system and the symptoms of the GI tract in PD is complicated. To determine the changes in the dopaminergic system in the GI tract in PD, two kinds of rodent PD models were used in the present study. One was 6-hydroxydopamine (6-OHDA) -treated rats in which 6-OHDA was microinjected in the bilateral substantia nigra (SN). The other was 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) -treated mice in which MPTP was injected intraperitoneally. Immunofluorescence, reverse transcription (RT)-real time polymerase chain reaction (PCR) and Western blot were used to evaluate and compare the levels of mRNA and protein expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the GI tract between normal and rodent PD models, as well as between 6-OHDA-treated rats and MPTP-treated mice. The results indicated that TH- and DAT-positive cells were widely distributed in the GI tract. There were significant differences in TH and DAT expression in the GI tract between normal and PD models, as well as between 6-OHDA-treated rats and MPTP-treated mice. The protein levels of TH and DAT in the GI tract were significantly increased in 6-OHDA-treated rats, but the protein level of TH was significantly decreased in MPTP-treated mice. In addition, there was visible atrophy of gastric epithelial parietal cells in MPTP-treated mice, although the protein level of DAT was not significantly changed. The different alterations of dopaminergic system in the GI tract of the two kinds of PD models might underline the differences in GI symptoms in PD patients and might be correlated with the disease severity and disease process affecting the GI tract.

Intermittent hypoxia and sleep restriction: Motor, cognitive and neurochemical alterations in rats

The present study evaluated the effects of intermittent hypoxia (IH) and sleep restriction (SR) upon motor and cognitive function in rats. Also evaluated were catecholamine concentrations and tyrosine hydroxylase (TH) protein expression in different regions of the forebrain. Wistar Hannover rats were submitted to IH for 4 days or 21 days (2min room air to 2min 10% O2 for 10:00–16:00h), followed by SR for 18h (16:00–10:00h). Rats were randomly assigned into four experimental groups: (1) control (2) IH (3) SR and (4) IH–SR. In the inhibitory avoidance task, an additional group of rats was submitted to paradoxical sleep deprivation (PSD) for 96 consecutive hours. Results showed that SR induced an increase in motor activity without modifying catecholaminergic turnover in the frontal cortex and striatum. The increase in exploratory activity in SR rats could be the result of impaired habituation. Neither SR periods induced cognitive deficits in the inhibitory avoidance task after 5 or 21 days. However, 96h of PSD impaired acquisition/retention in rats. Exposure to IH did not affect motor and cognitive function but IH was associated with SR in increased motor activity. After 21 days, IH and IH–SR reduced striatal norepinephrine concentration although neither SR nor IH affected TH protein expression. The results presented here suggest that hypoxia and sleep loss exert distinct deleterious effects upon the central nervous system.

Effect of the Co-Administration of Vitamin C and Vitamin E on Tyrosine Hydroxylase and Nurr1 Expression in the Prenatal Ray Ventral Mesencephalon

Tyrosine hydroxylase (TH) is the rate-limiting enzyme of dopamine (DA) biosynthesis, which is up-regulated by vitamin C administration. Nurr1 gene is highly expressed in brain and important for midbrain DAergic cell development and survival. But, the role of vitamin C and/or vitamin E during Nurr1 expression is yet to be known. Further, the synergistic effect of vitamins C and E on TH expression has not yet been explored clinically. Therefore, we studied the effects of single or co- administration of vitamin C (0.5 mM) and vitamin E (1 mM) for 72 hr, on both TH and Nurr1 expression in in vitro primary cultured gestational days (GD) 13.5 rat ventral mesencephalon (VM) by Western blot and immunocytochemistry. Our study revealed highest expressions of both TH and Nurr1 in the vitamin C + vitamin E treated group. TH expression was also increased in the vitamin C treated group than that of the control group, but the vitamin E treated group did not show any statistically significant effect. However, both the vitamin C and the vitamin E treated groups revealed increased expression of Nurr1 protein as compared with the control group. The present experimental data suggest that vitamin C can up-regulate the protein expression of TH, but Nurr1 level was elevated either by vitamin C or by vitamin E administration. Further, vitamin E acts synergistically with vitamin C to elevate TH and Nurr1 expression, which is the most novel finding of our study and for the first time; we reported this result, since there have been no published data on the synergistic effect of both the antioxidant vitamins on TH and Nurr1 expression in VM. As the motor function defect due to the progressive loss of DAergic neuron is the major reason of Parkinsonism, therefore, the results of our study finally suggest the effective role of vitamin C and vitamin E during early treatment of Parkinsonism.

Effect of pravastatin on sympathetic reinnervation in postinfarcted rats

We assessed whether pravastatin attenuates cardiac sympathetic reinnervation after myocardial infarction through the activation of ATP-sensitive K(+) (K(ATP)) channels. Epidemiological studies have shown that men treated with statins appear to have a lower incidence of sudden death than men without statins. However, the specific factor for this has remained disappointingly elusive. Twenty-four hours after ligation of the anterior descending artery, male Wistar rats were randomized to groups treated with either vehicle, nicorandil (a specific mitochondrial K(ATP) channel agonist), pinacidil (a nonspecific K(ATP) channel agonist), pravastatin, glibenclamide (a K(ATP) channel blocker), or a combination of nicorandil and glibenclamide, pinacidil and glibenclamide, or pravastatin and glibenclamide for 4 wk. Myocardial norepinephrine levels revealed a significant elevation in vehicle-treated rats at the remote zone compared with sham-operated rats (2.54 +/- 0.17 vs. 1.26 +/- 0.36 mug/g protein, P < 0.0001), consistent with excessive sympathetic reinnervation after infarction. Immunohistochemical analysis for tyrosine hydroxylase, growth-associated factor 43, and neurofilament also confirmed the change of myocardial norepinephrine. This was paralleled by a significant upregulation of tyrosine hydroxylase protein expression and mRNA in vehicle-treated rats, which was reduced after the administration of either nicorandil, pinacidil, or pravastatin. Arrhythmic scores during programmed stimulation in vehicle-treated rats were significantly higher than those treated with pravastatin. In contrast, the beneficial effects of pravastatin were reversed by the addition of glibenclamide, implicating K(ATP) channels as the relevant target. The sympathetic reinnervation after infarction is modulated by the activation of K(ATP) channels. Chronic use of pravastatin after infarction, resulting in attenuated sympathetic reinnervation by the activation of K(ATP) channels, may modify the arrhythomogenic response to programmed electrical stimulation.

Noradrenergic Function in Suicide

Although abnormalities in serotonergic function have been the major focus of studies on suicidal behavior, several studies indicate that abnormalities of noradrenergic function may also be involved in the pathophysiology of suicide. In this paper, we have reviewed some of the noradrenergic studies in sucide, including studies of the biosynthetic enzyme for norepinephrine, tyrosine hydroxylase (TH), the receptors for norepinephrine, α- and β-adrenergic receptors, as well as the signaling cascades linked to β-adrenergic receptors. In general, these studies indicate that the protein expression of TH, as well as α2- and β2-adrenergic receptors, is increased in the postmortem brain of suicide victims. More studies are needed in order to examine extensively the role of noradrenergic function in suicidal behavior.

Neurotoxicity in murine striatal dopaminergic pathways following long-term application of low doses of permethrin and MPTP

The long-term effects of permethrin (PM) and its interaction with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on striatal dopaminergic pathways were investigated in C57BL/6 mice. In a 3-month exposure, technical PM (1.5 mg/kg) was administered once per week, with MPTP (20 mg/kg) given once on either the 7th week or 11th week. In a 6-month exposure, PM (0.8 mg/kg or 1.5 mg/kg) was administered once per week for 26 weeks, with MPTP (20 mg/kg) given once, on week 24. Alterations in the expression of tyrosine hydroxylase (TH), dopamine transporter (DAT), and α-synuclein proteins were analyzed 1 day after the last PM treatment using western blot assay. PM had no significant effect on striatal dopaminergic pathways by itself, whereas MPTP significantly reduced the expression of TH and DAT proteins. In both exposure paradigms, weekly 1.5 mg/kg PM treatments antagonized the toxic effect of MPTP on TH and DAT expression ( p < 0.05). There was no significant alteration of α-synuclein expression following any exposure to PM and/or MPTP. [ 3H]Tetrabenazine (TBZ) binding assay for expression of striatal vesicular monoamine transporter (VMAT) showed no effect of PM, but the reduction in this protein caused by MPTP was attenuated by PM, consistent with effects on other dopaminergic biomarkers. The overall findings demonstrate that long-term, low-dose exposure to PM alone did not cause signs of neurotoxicity to striatal dopaminergic neural terminals, or enhance the effects of MPTP. We conclude that under typical use conditions, PM poses little Parkinsonian hazard to humans, including when impregnated into clothing for control of biting flies.

Nuclear factor kappa‐B p50 and p65 subunits expression in dementia with Lewy bodies

Dementia with Lewy bodies (DLB) is the second most common cause of neurodegenerative dementia after Alzheimer's disease (AD). Parkinsonism in DLB is mainly caused by neuronal loss with Lewy bodies (LBs) in the substantia nigra, thereby inducing degeneration of the nigrostriatal dopaminergic pathway similar to that in Parkinson's disease (PD). To clarify the pathogenesis of DLB, it is important to investigate the mechanisms involved in the degenerative process of LB-bearing neurones. Several reports suggest a role for nuclear factor kappa-B (NFkappaB) in the manifestation of neurodegenerative conditions such as AD and PD. The aim of the present study was to investigate whether NFkappaB subunits are involved in the pathogenesis of neurodegeneration in DLB by measuring tyrosine hydroxylase (TH), NFkappaB p65 and p50 protein expression in frontal cortex and substantia nigra pars compacta of DLB and control human brains. An increase, although not statistically significant, in nigral TH expression in DLB cases was observed. There were no differences in the cortical and nigral expression levels of NFkappaB p65 subunit between control and DLB cases. Western blots of the frontal cortex showed no differences in the expression levels of NFkappaB p50 subunit. However, NFkappaB p50 levels were significantly decreased (P < 0.05) in the pars compacta of the substantia nigra in the DLB cases in comparison with controls. The decrease in the expression of the p50 subunit in the substantia nigra of DLB cases achieved in the present study may increase the vulnerability of the dopaminergic neurones to a possible neurotoxic effect of p65 subunit. Thus, normal levels of NFkappaB p65 might be toxic in neurones with a low expression of the NFkappaB p50 subunit.