Jan Kehr

Characterization and MRI study of surfactant-coated superparamagnetic nanoparticles administered into the rat brain

A pre-clinical studyhas been carried out for the utilization of magnetite (Fe O ) nanoparticles as a diagnostic tracer for MRI. Surfactant-coated Fe O nanoparticles have been synthesized by a chemical coprecipitation method with a narrow particle size of around 6 nm. Preliminaryexperiments demonstrated the feasibilityof using superparamagnetic Fe O nanoparticles as contrast agents in MR imaging. 2001 Elsevier Science B.V. All rights reserved.

Mephedrone, compared with MDMA (ecstasy) and amphetamine, rapidly increases both dopamine and 5-HT levels in nucleus accumbens of awake rats

BACKGROUND AND PURPOSE The designer drug 1‐(4‐methylphenyl)‐2‐methylaminopropan‐1‐one (4‐methylmethcathinone, mephedrone) is reported to possess psychostimulant, entactogenic and hallucinogenic effects. The purpose of this study was to examine the effects of acute administration of mephedrone on extracellular levels of dopamine (DA) and 5‐HT in the nucleus accumbens of awake rats and compare these effects with those induced by 3,4‐methylenedioxymethamphetamine (MDMA, ecstasy) and amphetamine.

The role of 5-HT(1A) receptors in learning and memory.

The ascending serotonin (5-HT) neurons innervate the cerebral cortex, hippocampus, septum and amygdala, all representing brain regions associated with various domains of cognition. The 5-HT innervation is diffuse and extensively arborized with few synaptic contacts, which indicates that 5-HT can affect a large number of neurons in a paracrine mode. Serotonin signaling is mediated by 14 receptor subtypes with different functional and transductional properties. The 5-HT(1A) subtype is of particular interest, since it is one of the main mediators of the action of 5-HT. Moreover, the 5-HT(1A) receptor regulates the activity of 5-HT neurons via autoreceptors, and it regulates the function of several neurotransmitter systems via postsynaptic receptors (heteroreceptors). This review assesses the pharmacological and genetic evidence that implicates the 5-HT(1A) receptor in learning and memory. The 5-HT(1A) receptors are in the position to influence the activity of glutamatergic, cholinergic and possibly GABAergic neurons in the cerebral cortex, hippocampus and in the septohippocampal projection, thereby affecting declarative and non-declarative memory functions. Moreover, the 5-HT(1A) receptor regulates several transduction mechanisms such as kinases and immediate early genes implicated in memory formation. Based on studies in rodents the stimulation of 5-HT(1A) receptors generally produces learning impairments by interfering with memory-encoding mechanisms. In contrast, antagonists of 5-HT(1A) receptors facilitate certain types of memory by enhancing hippocampal/cortical cholinergic and/or glutamatergic neurotransmission. Some data also support a potential role for the 5-HT(1A) receptor in memory consolidation. Available results also implicate the 5-HT(1A) receptor in the retrieval of aversive or emotional memories, supporting an involvement in reconsolidation. The contribution of 5-HT(1A) receptors in cognitive impairments in various psychiatric disorders is still unclear. However, there is evidence that 5-HT(1A) receptors may play differential roles in normal brain function and in psychopathological states. Taken together, the evidence indicates that the 5-HT(1A) receptor is a target for novel therapeutic advances in several neuropsychiatric disorders characterized by various cognitive deficits.

A survey on quantitative microdialysis: theoretical models and practical implications.

The existing methods of quantitative microdialysis are reviewed. The methods are divided into 8 groups, depending on the mathematical models and theoretical principles used to describe convective diffusion in the extracellular space of the brain. Special emphasis is made to describe each method from a historical perspective, showing its main contribution to recent knowledge, as well as its limitations and drawbacks. It is concluded that those methods based on explicitly derived equations for in vivo recovery are still too approximative and not suitable for routine application. Therefore, empirical models based on varying perfusion flow rates or concentrations of substances in the perfusion solution, found several practical implications. Methods using a reference substance as a marker of in vivo recovery are also discussed. The paper stresses the increasing importance of methods allowing the quantitative evaluation of microdialysis data whenever measuring neurotransmitter release, drug concentrations or pharmacokinetic variables.

From the Golgi–Cajal mapping to the transmitter-based characterization of the neuronal networks leading to two modes of brain communication: Wiring and volume transmission ☆

After Golgi-Cajal mapped neural circuits, the discovery and mapping of the central monoamine neurons opened up for a new understanding of interneuronal communication by indicating that another form of communication exists. For instance, it was found that dopamine may be released as a prolactin inhibitory factor from the median eminence, indicating an alternative mode of dopamine communication in the brain. Subsequently, the analysis of the locus coeruleus noradrenaline neurons demonstrated a novel type of lower brainstem neuron that monosynaptically and globally innervated the entire CNS. Furthermore, the ascending raphe serotonin neuron systems were found to globally innervate the forebrain with few synapses, and where deficits in serotonergic function appeared to play a major role in depression. We propose that serotonin reuptake inhibitors may produce antidepressant effects through increasing serotonergic neurotrophism in serotonin nerve cells and their targets by transactivation of receptor tyrosine kinases (RTK), involving direct or indirect receptor/RTK interactions. Early chemical neuroanatomical work on the monoamine neurons, involving primitive nervous systems and analysis of peptide neurons, indicated the existence of alternative modes of communication apart from synaptic transmission. In 1986, Agnati and Fuxe introduced the theory of two main types of intercellular communication in the brain: wiring and volume transmission (WT and VT). Synchronization of phasic activity in the monoamine cell clusters through electrotonic coupling and synaptic transmission (WT) enables optimal VT of monoamines in the target regions. Experimental work suggests an integration of WT and VT signals via receptor-receptor interactions, and a new theory of receptor-connexin interactions in electrical and mixed synapses is introduced. Consequently, a new model of brain function must be built, in which communication includes both WT and VT and receptor-receptor interactions in the integration of signals. This will lead to the unified execution of information handling and trophism for optimal brain function and survival.

Fast HPLC Estimation of γ‐Aminobutyric Acid in Microdialysis Perfusates: Effect of Nipecotic and 3‐Mercaptopropionic Acids

Abstract: A method for rapid, automated (<5 min), and sensitive (detection limit 50 fmol/10 μl) determination of γ‐aminobutyric acid (GABA) is described. The method is based on precolumn derivatiza‐tion with o‐phthaldialdehyde/t‐butylthiol reagent and separation by reverse‐phase HPLC with electrochemical detection under isocratic conditions. A 100 × 4 mm Nucleosil 3 C18 column was used; the mobile phase consisted of 0.15 M sodium acetate, 1 mM EDTA (pH 5.4), and 50% acetonitrile; the flow rate was 0.8 ml/min. The potential of the glassy carbon working electrode was +0.75 V. The method allows for the monitoring of GABA levels in the extracellular fluid sampled by microdialysis as documented in the present study when 0.5 mM nipecotic acid is infused via the probe, or 3‐mercaptopro‐pionic acid is injected at a dose of 100 mg/kg i.p. There was a 15‐fold increase of extracellular GABA after nipecotic acid, whereas in the second case the inhibition of GABA synthesis was followed by a 74% decrease of GABA as compared to basal levels.

Galanin Is a Potent In Vivo Modulator of Mesencephalic Serotonergic Neurotransmission

Neurochemical, molecular, immunohistochemical and behavioral methods were used to examine the in vivo effects of the neuropeptide galanin on central 5-HT neurotransmission and on 5-HT1A receptor-mediated responses. Intraventricularly infused galanin caused a long-lasting and dose-dependent reduction of basal extracellular 5-HT levels in the ventral hippocampus of awake rats as measured by microdialysis. Infusion of galanin into the dorsal raphe nucleus (DRN), but not intrahippocampally, reduced 5-HT release. The effect of i.c.v. galanin on 5-HT release was blocked by the galanin receptor antagonist M35, acting most likely via galanin receptors at the level of the DRN. Galanin also reduced the levels of tryptophanhydroxylase mRNA in the DRN. Therefore, the effects of galanin on 5-HT1A receptor-mediated responses were further investigated. Surprisingly, galanin significantly attenuated the reduction of hippocampal 5-HT release induced by systemic injection of the 5-HT1A receptor agonist 8-OH-DPAT. Galanin also attenuated 8-OH-DPAT-induced hypothermia and locomotor activity in rats. These results indicate that galanin has important inhibitory actions on central 5-HT neurotransmission and on 5-HT1A receptor-mediated events.

Regional specific effects of clozapine and haloperidol on GABA and dopamine release in rat basal ganglia

gamma-Aminobutyric acid (GABA) and dopamine release were measured concomitantly in rat dorsolateral striatum, fundus striati (a ventral region of striatum) and globus pallidus following s.c. administration of haloperidol or clozapine. Release was measured by microdialysis in halothane-anesthetized rats. Clozapine (5.0 mg/kg) increased GABA release in the fundus striati and haloperidol (0.5 mg/kg) increased GABA release in the globus pallidus. In contrast, clozapine (2.5-40 mg/kg) failed to increase GABA release in the globus pallidus and haloperidol (0.1-2.0 mg/kg) failed to increase GABA release in the fundus. Thus, haloperidol and clozapine are clearly distinguished by their effects on GABA release in the fundus striati and globus pallidus (both drugs increased GABA release in the dorsolateral striatum). Dopamine release was increased by haloperidol and clozapine in the two regions of the striatum. However, except in the fundus striati where clozapine-induced increases in dopamine and GABA occurred in parallel, both drugs were more potent in releasing dopamine than GABA. Drug-induced increases in GABA and dopamine release were reversed by addition of 1 microM tetrodotoxin to the perfusion medium. These data suggest that (1) regional differences in the effects of haloperidol and clozapine on GABA release in the basal ganglia may parallel the unique clinical profiles of these drugs; and (2) increases in dopamine release may occur independently of a GABAergic component in the dorsolateral striatum following low doses of haloperidol and clozapine and in the fundus striati following all effective doses of haloperidol.

Intraseptal muscarinic ligands and galanin: influence on hippocampal acetylcholine and cognition

The cholinergic neurons in the septohippocampal projection are implicated in hippocampal functions such as spatial learning and memory. The aim of this study was to examine how septohippocampal cholinergic transmission is modulated by muscarinic inputs and by the neuropeptide galanin, co-localized with acetylcholine (ACh) in septohippocampal cholinergic neurons, and how spatial learning assessed by the Morris water maze test is affected. Muscarinic inputs to the septal area are assumed to be excitatory, whereas galanin is hypothesized to inhibit septohippocampal cholinergic function. To test these hypotheses, compounds were microinjected into the medial septum and hippocampal ACh release was assessed by microdialysis probes in the ventral hippocampus of the rat. Blockade of septal muscarinic transmission by intraseptal scopolamine increased hippocampal ACh release suggesting that septal cholinergic neurons are under tonic inhibition. Stimulation of septal muscarinic receptors by carbachol also increased hippocampal ACh release. Despite this increase, both scopolamine and carbachol tended to impair hippocampus-dependent spatial learning. This finding also suggests a revision of the simplistic notion that an increase in hippocampal ACh may be facilitatory for learning and memory. Galanin infused into the medial septum enhanced hippocampal ACh release and facilitated spatial learning, suggesting that septal galanin, contrary to earlier claims, does not inhibit but excites septohippocampal cholinergic neurons. Galanin receptor stimulation combined with muscarinic blockade in the septal area resulted in an excessive increase of hippocampal ACh release combined with an impairment of spatial learning. This finding suggests that the level of muscarinic activity within the septal area may determine the effects of galanin on hippocampal cognitive functions. In summary, a limited range of cholinergic muscarinic transmission may contribute to optimal hippocampal function, a finding that has important implications for therapeutic approaches in the treatment of disorders of memory function.

Characterization of gamma-aminobutyric acid and dopamine overflow following acute implantation of a microdialysis probe

The present study characterized the voltage and calcium dependence of gamma-aminobutyric acid and dopamine overflow after the acute implantation of a microdialysis probe. Probes were implanted in dorsolateral striatum and globus pallidus. Experiments were performed under light halothane anesthesia. Basal, extracellular levels of GABA were not affected by tetrodotoxin (TTX) and were increased to 140 percent of basal values by calcium free Ringer. Basal, extracellular levels of dopamine were reduced to 14 percent of basal values by the addition of TTX and to 30 percent of basal values by the removal of calcium from the Ringer solution. The results suggest that in this in vivo preparation basal extracellular dopamine is largely of vesicular origin while GABA is not.