Mitochondrial biogenesis in fast skeletal muscle of CK deficient mice.

Creatine kinase (CK) is a phosphotransfer kinase that catalyzes the reversible transfer of a phosphate moiety between ADP and creatine and that is highly expressed in skeletal muscle. In fast glycolytic skeletal muscle, deletion of the cytosolic M isoform of CK in mice (M-CK-/-) leads to a massive increase in the oxidative capacity and of mitochondrial volume. This study was aimed at investigating the transcriptional pathways leading to mitochondrial biogenesis in response to CK deficiency. Wild type and M-CK-/- mice of eleven months of age were used for this study. Gastrocnemius muscles of M-CK-/- mice exhibited a dramatic increase in citrate synthase (+120%) and cytochrome oxidase (COX, +250%) activity, and in mitochondrial DNA (+60%), showing a clear activation of mitochondrial biogenesis. Similarly, mRNA expression of the COXI (mitochondria-encoded) and COXIV (nuclear-encoded) subunits were increased by +103 and +94% respectively. This was accompanied by an increase in the expression of the nuclear respiratory factor (NRF2alpha) and the mitochondrial transcription factor (mtTFA). Expression of the co-activator PGC-1alpha, a master gene in mitochondrial biogenesis was not significantly increased while that of PGC-1beta and PRC, two members of the same family, was moderately increased (+45% and +55% respectively). While the expression of the modulatory calcineurin-interacting protein 1 (MCIP1) was dramatically decreased (-68%) suggesting inactivation of the calcineurin pathway, the metabolic sensor AMPK was activated (+86%) in M-CK-/- mice. These results evidence that mitochondrial biogenesis in response to a metabolic challenge exhibits a unique pattern of regulation, involving activation of the AMPK pathway.

Cocaine and amphetamine-regulated transcript peptide (CART(62-76))-induced changes in regional monoamine levels in ratbrain.

Cocaine- and amphetamine-regulated transcript (CART) is a novel neuropeptide with neurotransmitter-like effects. In the present study we examined the influence of CART peptide fragment, CART(62-76), on the levels of catecholamines (dopamine and norepinephrine), serotonin and their metabolites in five regions of the rat brain. CART(62-76) was administered at 0.5 or 5.0 microg dose intracerebroventricularly. A high-pressure liquid chromatograph coupled to electrochemical array detector was used to analyse the tissue homogenates. In the frontal cortex, CART(62-76) increased the levels of 5-hydroxyindoleacetic acid (5-HIAA), a metabolite of serotonin (5-HT). In the striatum, CART(62-76) decreased the levels of noradrenaline and 5-HT. In the hypothalamus, CART(62-76) increased the levels of 5-HIAA. CART(62-76) had no significant effect in the hippocampus and cerebellum. Our data suggest that CART(62-76) peptide has no major effect on dopaminergic pathways, but it modulates the activity of striatal noradrenergic and corticostriatal and hypothalamic serotoninergic system in the rat brain. These regionally selective neurochemical changes may explain the effects of CART peptides on appetitive, emotional and locomotor behaviour.

Apomorphine-induced aggressive behaviour and post-mortem monoamine content in male Wistar rats.

The aim of this study was to investigate the monoamine content in post-mortem brain samples of control, apomorphine-aggressive, and apomorphine-non-aggressive adult male Wistar rats. The repeated apomorphine (1.0 mg/kg, (s.c.) once daily during 2 weeks) gradually induced aggressive behaviour in 18 animals out of 24. No unidirectional changes in the brain monoamine contents in four regions (frontal cortex, striatum, hippocampus, and hypothalamus) were detected as measured by high pressure liquid chromatography-electrochemical detection. In conclusion, our present experiment demonstrates that the development and intensity of apomorphine-induced aggressive behaviour do not correlate with the brain post-mortem monoamine content.

1-(1-naphthyl)-piperazine, a mixed 5-HT1A and 5-HT2A/2C receptor ligand, elicits an anxiolytic-like effect in the open-field test without changes in 5-HT metabolism.

The effects of acute 1-(1-naphthyl)-piperazine (1-NP, 2 mg/kg i.p.) on rat open-field and social interaction behaviors were studied followed by postmortem and in vivo microdialysis measurement of serotonin (5-HT) and 5-hydroxyidolacetic acid (5-HIAA) content. 1-NP treatment elicited an anxiolytic-like effect in the open-field test, which was not modified by citalopram (5 mg/kg i.p.) challenge. A statistically insignificant tendency toward prolongation of the social interaction time was also found. The only significant change in the postmortem experiment was found in the striatum: 5-HIAA content was reduced after combined 1-NP plus citalopram treatment. Using the in vivo microdialysis technique, no difference in the 5-HT or 5-HIAA output between the treatment groups was found in the frontal cortex of anaesthetized rats. Our present study demonstrates the anxiolytic-like properties of 1-NP in the open-field test, but this effect is irrelevant to changes in 5-HT metabolism.

Dopamine protects neurons against glutamate-induced excitotoxicity.

Glutamate excitotoxicity is responsible for neuronal death in acute neurological disorders including stroke, trauma and neurodegenerative disease. Loss of calcium homeostasis is a key mediator of glutamate-induced cell death. The neurotransmitter dopamine (DA) is known to modulate calcium signalling, and here we show that it can do so in response to physiological concentrations of glutamate. Furthermore, DA is able to protect neurons from glutamate-induced cell death at pathological concentrations of glutamate. We demonstrate that DA has a novel role in preventing delayed calcium deregulation in cortical, hippocampal and midbrain neurons. The effect of DA in abolishing glutamate excitotoxicity can be induced by DA receptor agonists, and is abolished by DA receptor antagonists. Our data indicate that the modulation of glutamate excitotoxicity by DA is receptor-mediated. We postulate that DA has a major physiological function as a safety catch to restrict the glutamate-induced calcium signal, and thereby prevent glutamate-induced cell death in the brain.

Alpha-synuclein A30P point-mutation generates age-dependent nigrostriatal deficiency in mice.

Lewy bodies are mainly composed of alpha-synuclein (SNCA) and specific mutations in SNCA gene are related to familial forms of Parkinson's disease (PD). The purpose of our study was to generate a mouse line with A30P knock-in point mutation in SNCA gene and to test if a single point-mutation is able to turn otherwise normal SNCA into a toxic form. The behavioral profile of SNCA A30P mice was followed for 16 months. Generally, these mice are healthy and viable without any obvious abnormalities. Starting from the age of 13 months mice developed a significant deficit in motor performance tests related to nigrostriatal function (ink-test and beam walk). In other tests (motility boxes, rotarod) mice continuously performed normally. Moreover, SNCA A30P mice expressed the altered sensitivity to VMAT2 inhibitor reserpine, possibly reflecting a functional deficiency of dopamine. Indeed, mice at 15 months of age had significantly reduced levels of dopamine and its major metabolite DOPAC in the striatum, and reduced levels of dopamine in the mesolimbic system. The present study confirms that SNCA plays an important role in the development of PD and an insertion of a single point mutation is sufficient to generate age-related decline in specific motor performance. The generated mouse line has a potential to become a model for PD with comparable time course and phenotype.

Apomorphine-induced aggressive behaviour in para-chlorophenylalanine-treated male rats: implications to brain.

The aim of this study was to investigate the role of serotoninergic neurotransmission and the effect of acute para-chlorophenylalanine (350 mg/kg, intraperitoneally) treatment on apomorphine-induced aggressive behaviour in adult male Wistar rats. In addition, [3H]ketanserin binding and monoamine content were studied. Repeated administration of apomorphine (1.0 mg/kg, subcutaneously, once daily for two weeks) gradually induced aggressive behaviour. Acute p-chlorophenylalanine treatment down-regulated the [3H]ketanserin binding and reduced over 90 per cent the content of serotonin and 5-hydroxyindolacetic acid. In a half of the p-chlorophenylalanine-treated animals, the aggressive behaviour was suppressed, while there was no difference in [3H]ketanserin binding or monoamine content between the p-chlorophenylalanine treated aggressive and non-aggressive animals. In conclusion, the acute p-chlorophenylalanine treatment attenuates the aggressiveness only in half of the animals, while the latter phenomenon is independent on the CNS monoamine content or [3H]ketanserin binding.

Acute trazodone and quipazine treatment attenuates apomorphine-induced aggressive behaviour in male rats without major impact on emotional behaviour or monoamine content post mortem.

We have studied the effect of acute trazodone (3--20 mg kg(-1)) and quipazine (1--3 mg kg(-1)) treatment on the apomorphine-induced (1 mg kg(-1), once daily over 2 weeks) aggressive behaviour in male Wistar rats. All doses of trazodone and quipazine tested attenuated the aggressiveness as evidenced by the abolished intensity of aggressive behaviour and increased time of latency before the first attack. The acute trazodone (3--10 mg kg(-1)) or quipazine (1--3 mg kg(-1)) treatment had no or only a minor effect on rat behaviour in the elevated plus-maze, open field, and forced swimming test. Concomitant apomorphine (1 mg kg(-1)) plus trazodone (3 mg kg(-1)), but not apomorphine (1 mg kg(-1)) plus quipazine (1 mg kg(-1)), treatment slowed the development of aggressive behaviour. Repeated apomorphine treatment moderately reduced the dopamine post mortem and increased the DOPAC and HVA contents in striatum. Other monoamines or their metabolites were unchanged. Neither trazodone and quipazine treatment nor forced swimming stress induced any changes in the monoamine contents. In conclusion, our results indicate that acute trazodone and quipazine treatment attenuates the apomorphine-induced aggressive behaviour in male rats, but this phenomenon cannot be implicated in the changes in emotional and motivational behaviour, or in changes of monoamine content post mortem.