Behavior, neurochemistry and histology after intranigral lipopolysaccharide injection.

Inflammation and neuronal degeneration of the substantia nigra (SN) occur in Parkinson's disease (PD). We studied the effects of intranigral lipopolysaccharide (LPS) injection on adult Sprague-Dawley rats. Locomotor activity measurement, neurotransmitter determination and perfusion fixation for immunohistochemistry were done on the 7th day. Bilateral LPS injection increased locomotor activity 2- to 3-fold. In the SN, dopamine (DA) and serotonin (5-HT) decreased but the ratios dihydroxyphenylacetic acid (DOPAC)/DA, homovanillic acid (HVA)/DA and 5-hydroxyindole-acetic acid (5-HIAA)/5-HT increased. In the striatum, DA, DOPAC, HVA, 3-methoxytyramine and epinephrine decreased but HVA/DA and 5-HIAA/5HT ratios increased. Unilateral LPS decreased dopamineric neurons ipsilaterally but increased contralaterally. This study provides the first evidence of behavioral hyperactivity, epinephrine suppression and neuronal plasticity in the LPS model of PD.

Calcium-induced proline accumulation contributes to amelioration of NaCl injury and expression of glutamine synthetase in greater duckweed (Spirodela polyrhiza L.).

The calcium-mediated proline accumulation is a critical response under NaCl stress and the function of the induced proline as a glutamine synthetase (GS) protectant in greater duckweed was investigated. The plants were treated with solutions containing 100mM NaCl, 200 mM NaCl, 200 mM NaCl plus 10mM CaCl2, or 10mM CaCl2 alone for 4 days. At the end of the experiment, the fronds of inoculum treated with 200 mM NaCl showed the chlorotic effect, higher glutamate dehydrogenase (NADH-GDH) activity and lower GS activity. At the lower salinity, the activities of GS and NADH-GDH were not altered markedly. A significant accumulation of proline was not found under either low or high salinity. The activity of Δ(1)-pyrroline-5-carboxylate reductase (P5CR) was enhanced only at 200 mM NaCl but remained unchanged at 100mM NaCl. The activity of Δ(1)-pyrroline-5-carboxylate synthetase (P5CS) did not change under salinity-stressed. Addition of CaCl2 to the salt stressed plants not only lowered NaCl injury but also showed an elevated level of proline contents in response to the salinity treatment. In addition, both GS activity and corresponding polypeptides were expressed close to the level of control. Exogenous proline protects GS2 and the 32 kDa protein in photosystem II reaction center (D1) from H2O2-induced redox degradation in the chloroplast lysates of duckweed. The results suggest that calcium-induced proline accumulation may play an important role as a GS protectant under NaCl exposure in S. polyrhiza.

Calcium-mediated responses and glutamine synthetase expression in greater duckweed (Spirodela polyrhiza L.) under diethyl phthalate-induced stress.

This study was carried out to assess the influence of diethyl phthalate (DEP) alone or associated with calcium chloride (CaCl2) on greater duckweed plants, emphasizing the implications of calcium in amelioration of DEP-induced stress on plant growth. Greater duckweed were treated with DEP in variable concentrations, as 0, 0.25, 0.5, 1.0 and 2.0mM for 7 days, or treated with the same concentration either 2mM DEP or 2mM DEP plus 10mM CaCl2·2H2O in different duration 0-7 days. Treatment with 2mM DEP resulted in increasing proline content, protease activity, and ammonia accumulation in duckweed tissues. NADH-glutamate dehydrogenase (NADH-GDH; EC 1.4.1.2) and Δ(1)-pyrroline-5-carboxylate reductase (P5CR; EC 1.5.1.2), two key enzymes in the glutamate pathway of proline synthesis, showed increase in activity with DEP treatment and positively correlated with proline accumulation. No further increase in proline accumulation was observed with addition of calcium chloride to the DEP-treated cultures. However, supplementation of Ca(2+) can mitigate the adverse effect of DEP, at least in part to decrease the DEP-induced superoxide accumulation and increase in GDH activity for ammonia assimilation in duckweed fronds. In addition, effects of calcium on mitigation of DEP injury were also observed in glutamine synthetase (GS; EC 6.3.1.2) expression. Both GS1 and GS2 polypeptide accumulation and the level of total GS activity were nearly equivalent to the control. Exogenous proline protects GS2 from DEP-modulated redox damage in the chloroplast lysates but there is no remarkable protection effects on D1 (the 32kDa protein in photosystem II reaction center) degradation. In conclusion, the glutamate pathway of proline synthesis might be involved in mitigation of DEP-induced injury, and calcium plays an important role in increasing GDH, P5CR, and GS expression.

Oxidative effects and metabolic changes following exposure of greater duckweed (Spirodela polyrhiza) to diethyl phthalate.

The toxicity and effects of diethyl phthalate (DEP), a potent allelochemical, on the growth of greater duckweed were studied. Biochemical analyses and physiological methods were combined to investigate oxidative stress, adverse effects and their mechanisms in greater duckweeds grown in 0-2 mM of diethyl phthalate (DEP) after cultivation for 7 days. The results showed that J-shaped concentration response curves were displayed in hydrogen peroxide (H2O2), ascorbic acid (ASA) and dehydroascorbate (DHA) levels, and ascorbate peroxidase (APX) and gualacol peroxidase (POD) activities, indicating reduced oxidative stress and toxic effect. The inverted U-shaped curves were exhibited in relative growth rate (RGR), fresh weight/dry weight (FW/DW) ratio, total chlorophyll content, total soluble thiols, and glutathione reductase (GR) activity, revealing beneficial effect in plant growth. The inverted U-shaped curves were also found in malondialdehyde (MAD) and superoxide radical (O2-) contents with the increasing concentration of DEP, indicative of enhanced oxidative stress. The results suggest that DEP is toxic to the greater duckweed by inducing oxidative stress and antioxidative enzymes may play important roles in the defense strategy against DEP toxicity.

Effects of the combined treatment of naloxone and indomethacin on catecholamines and behavior after intranigral lipopolysaccharide injection.

The present study examined effects of the combined administration of naloxone (NX) and indomethacin (IM) on nigrostriatal catecholamines and locomotor activity after intranigral lipopolysaccharide (LPS) injection in Sprague-Dawley rats. NX plus IM was given 3 days after LPS injection; it significantly (P < .05) reversed LPS inflammation on nigrostriatal dopamine (DA) and nigral serotonin (5-HT) and nigral homovanillic acid (HVA)/DA ratio and nigrostriatal 5-hydroxyindoleacetic acid (5-HIAA)/5-HT ratio. It also tended to ameliorate the locomotor hyperactivity. However, NX plus IM given 30 min before LPS could not satisfactorily protect against LPS's damage both biochemically and behaviorally. These results reveal that NX plus IM may protect against LPS on DA, 5-HT, and motor function after LPS injection but not before. Thus it suggests that the combined treatment of NX and IM gives a potent therapy, but not prevention, of LPS-induced inflammation and also protect nigrostriatal dopaminergic and serotoninergic systems against LPS in rats.