Search

The release of [3H]dopamine ([3H]DA) and [3H]noradrenaline ([3H]NA) in acutely perfused rat striatal and cortical slice preparations was measured at 37 °C and 17 °C under ischemic conditions. The ischemia was simulated by the removal of oxygen and glucose from the Krebs solution. At 37 °C, resting release rates in response to ischemia were increased; in contrast, at 17 °C, resting release rates were significantly reduced, or resting release was completely prevented. The removal of extracellular Ca2+ further increased the release rates of [3H]DA and [3H]NA induced by ischemic conditions. This finding indicated that the Na+/Ca2+ exchanger (NCX), working in reverse in the absence of extracellular Ca2+, fails to trigger the influx of Ca2+ in exchange for Na+ and fails to counteract ischemia by further increasing the intracellular Na+ concentration ([Na+]i). KB-R7943, an inhibitor of NCX, significantly reduced the cytoplasmic resting release rate of catecholamines under ischemic conditions and under conditions where Ca2+ was removed. Hypothermia inhibited the excessive release of [3H]DA in response to ischemia, even in the absence of Ca2+. These findings further indicate that the NCX plays an important role in maintaining a high [Na+]i, a condition that may lead to the reversal of monoamine transporter functions; this effect consequently leads to the excessive cytoplasmic tonic release of monoamines and the reversal of the NCX. Using HPLC combined with scintillation spectrometry, hypothermia, which enhances the stimulation-evoked release of DA, was found to inhibit the efflux of toxic DA metabolites, such as 3,4-dihydroxyphenylacetaldehyde (DOPAL). In slices prepared from human cortical brain tissue removed during elective neurosurgery, the uptake and release values for [3H]NA did not differ from those measured at 37 °C in slices that were previously maintained under hypoxic conditions at 8 °C for 20 h. This result indicates that hypothermia preserves the functions of the transport and release mechanisms, even under hypoxic conditions. Oxidative stress (H2O2), a mediator of ischemic brain injury enhanced the striatal resting release of [3H]DA and its toxic metabolites (DOPAL, quinone). The study supports our earlier findings that during ischemia transmitters are released from the cytoplasm. In addition, the major findings of this study that hypothermia of brain slice preparations prevents the extracellular calcium concentration ([Ca2+]o)-independent non-vesicular transmitter release induced by ischemic insults, inhibiting Na+/Cl−-dependent membrane transport of monoamines and their toxic metabolites into the extracellular space, where they can exert toxic effects.

The concentrations of amino acids in the cerebrospinal fluid (CSF) (n = 20) and serum (n = 20) taken from patients with essential tremor were measured by HPLC and compared with those of controls (n = 10). Reduced concentrations of some amino acids (asparagine, glutamine, glycine, threonine, isoleucine, leucine) were observed in serum taken from patients with tremor. Significant increases were detected in the concentrations of glutamate (p < 0.001) and aspartate (p < 0.01). The general tendency of the changes in CSF and serum was similar; although the highest differences were observed in amino acid concentrations in the serum of patients with essential tremor. Opposite shifts of some amino acids were detected, in the concentrations of aspartate, serine, tyrosine, leucine, and isoleucine, which may indicate the independence of the changes in the serum from those in the CSF. This study raises the possibility that a genetically determined metabolic disorder is involved in the etiology of essential tremor that appears peripherally and, partly, centrally. The slight increase in the concentration of glutamate together with the reduced levels of GABA, glycine, and serine in CSF may form the neurochemical basis of the central oscillation observed in essential tremor.

Methods PC12 cells and primary mesencephalic neurons were used in culture, and the striatum and the substantia nigra were prepared from wild-type and P2X7 receptor knockout mice. Rotenone and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treatments were applied in vitro and in vivo to reproduce neurochemical hallmarks of PD. Receptor expression, cell survival indicators, and endogenous biogenic amine, amino acid, adenine nucleotide and endocannabinoid contents were analyzed.

Parkinson’s disease (PD) is a neurodegenerative syndrome associated with selective loss of dopaminergic neurons in the striatum. It is believed that both mitochondrial dysfunction and oxidative stress (OS) play important role in the pathogenesis of PD. Development of an effective causal therapy should be focused on preventing or at least retarding the neurodegenerative process underlying the disease. In chronic in fusion PD model studied the effect of rotenone on the content, and release of striatal neurotransmitters of rat upon OS by H2O 2 perfusion. The effects of anti-Parkinsonian drugs were examined in vitro rotenone PD, OS slices model. The role of P2X7 receptor (P2X7r) formed by 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine (MPTP) or in vitro rotenone treatment using C57/BI6 and P2X7r knockout mice. Neurochemical response was determined using microvolume perfusion method, HPLC separation of dansylated samples. Dopamine o-quinone (DAQ) oxidized metabolite of DA was detected electrochemically at -100 mV potential in reduction working mode. The dopamine content in the rat striatum is decreased in response to chronic intravenous rotenone infusion. However, surviving dopaminergic neurons take up and release only a slightly lower amount of DA in response to electrical stimulation. Striatal dopaminergic neurons showed increased susceptibility to OS, responding with enhanced release of DA and with formation of toxic metabolite DAQ. The loss of DA and ATP and formation of DAQ induced by OS confirmed the impairment of striatum upon in vitro rotenone. Formation of DAQ at the end effect of OS confirmed the impairment of dopaminergic function by rotenone. However, when applied rotenone with l-DOPA treatment the endogenous DAQ was detected in perfusion samples. However, when applied rotenone with l-DOPA treatment the endogenous DAQ was detected in the micro-volume perfusion samples. Conversely, when rasagiline and trolox were in use these DA-quinones metabolites were not detectable. Rotenone treatment elicited a similar reduction in ATP content in the substantia nigra of both genotypes, whereas reduction of ATP was more pronounced after rotenone treatment in striatal slices of P2X7 deficient mice. Although the level of the endocannabinoid, 2-AG, was elevated by rotenone in the striatum of wild-type mice, an effect that was absent in mice deficient in P2X7 receptors. Genetic deletion of P2X7 receptors did not change depletion of striatal endogenous DA content after in vivo MPTP or in vitro rotenone treatment. Although the endogenous amino acids content and the level of the endocannabinoid, AEA was elevated by MPTP in the striatum of mice, but 2-AG remained unchanged in mice deficient in P2X7 receptors. In conclusion, oxidative stress induced, pathological DA release and the formation of toxic metabolites of DA are distinctly modified by the tested antiparkinsonian drugs and P2X7 receptor deficiency does not promote the survival of dopaminergic neurons.