Cannabinoid CB1 receptors activation and coactivation with D2 receptors modulate GABAergic neurotransmission in the globus pallidus and increase motor asymmetry.

The cannabinoid CB1 (CB1R) and dopaminergic D2 (D2R) receptors modify GABAergic transmission in the globus pallidus. Although dopaminergic denervation produces changes in the expression and supersensitization of these receptors, the consequences of these changes on GABAergic neurotransmission are unknown. The aim of this study was to show the effects of CB1R and D2R activation and coactivation on the uptake and release of [(3) H]GABA in the globus pallidus of hemiparkinsonian rats as well as their effects on motor behavior. The activation of CB1R blocked GABA uptake and decreased GABA release in the globus pallidus in the dopamine denervated side, whereas the co-activation of CB1R-D2R increased GABA release and had no effect on GABA uptake. A microinjection of the CB1R agonist ACEA into the globus pallidus ipsilaterally to a 6-OHDA lesion potentiated turning behavior that was induced by methamphetamine. However, a microinjection of the D2R agonist quinpirole did not modify this behavior, and a microinjection of a mixture of CB1R and D2R agonists significantly potentiated turning behavior. The behavioral effects produced after the activation of the CB1R and the co-activation of CB1R and D2R can be explained by increased GABAergic neurotransmission produced by a block of GABA uptake and an increase in the release of GABA in the globus pallidus, respectively.

Arachidonyl-2'-chloroethylamide (ACEA), a synthetic agonist of cannabinoid receptor, increases CB1R gene expression and reduces dyskinesias in a rat model of Parkinson's disease.

l-Dopa is the most effective drug used for Parkinson's disease (PD), but after long-term treatment, the vast majority of PD patients develop abnormal involuntary movements (AIMs) termed l-Dopa-induced dyskinesia (LID). Cannabinoid receptors in the basal ganglia can modulate motor functions, but their role in the treatment of LID is controversial. Therefore, the aim of this study is to evaluate the motor behavior and mRNA expression of the cannabinoid receptor-1 (CB1R), encoded by the Cnr1 gene, in the striatum and globus pallidus of a 6-hydroxydopamine rat model of PD. The evaluated rats had 6-hydroxydopamine-induced injury, LID, and LID treated with arachidonyl-2'-chloroethylamide (ACEA), a cannabinoid receptor agonist. Contralateral turns and AIMs were recorded to assess motor behavior. Gene expression was quantified by reverse transcription coupled with quantitative polymerase chain reaction using TaqMan probes. Behavioral evaluations demonstrated that dyskinetic rats treated with ACEA had a significant reduction in AIMs compared to the dyskinetic group. The expression of CB1R mRNA was significantly decreased in the 6-hydroxydopamine-injured and dyskinetic rats, compared to intact rats. The striata of dyskinetic rats treated with ACEA exhibited highly significant increases in CB1R mRNA expression. Contrary to results in the striatum, a lower CB1R expression was observed in globus pallidus from dyskinetic ACEA-treated group. In summary, significant differences in mRNA expression of CB1R were found between the evaluated groups of rats, suggesting the occurrence of compensatory mechanisms that may result in the ACEA-mediated reduction of dyskinesias in a rat model of PD.

Avenanthramide-C prevents amyloid formation of bovine serum albumin.

The misfolding of protein and its assembly into amyloid fibrils with a characteristic ß-sheet-rich secondary structure, cause a lot of illnesses. Polyphenols have been extensively studied as a class of amyloid inhibitors, whose effect depends on the position and number of hydroxyl groups around the flavone backbone. In this study, we used bovine serum albumin (BSA) as an amyloid model to test the anti-amyloid effects of Avenanthramide-C (Avn-C), a molecule with a long aliphatic linker between two aromatic rings. We used spectroscopy techniques like thioflavin T fluorescence and circular dichroism, to follow the ß-sheet-rich aggregates of BSA upon incubation at 68 °C. Our results demonstrated that Avn-C shows higher inhibitory effect on BSA oligomerization at micromolar concentrations, than Epigallocatechin gallate (EGCG) and Curcumin, proving for the first time, that Avn-C can serve as potential molecule in preventing protein aggregation.

Role of cannabinoid CB1 receptors on macronutrient selection and satiety in rats.

It has been shown that endogenous and exogenous cannabinoids substantially increase feeding. Despite evidence for a role of endocannabinoids in mediating food ingestion, the mechanisms by which CB1 receptor agonists and antagonists have an effect on motivational processes (hunger, satiety) as well as on specific food preference are not entirely understood. The purpose of this study was to investigate the effects of systemic injection of the CB1 receptor agonist, ACEA, on protein, carbohydrates and fat intake as well as on the behavioural satiety sequence (BSS) in pre-satiated rats. Following a 120-min access to a three pure nutrient diet (protein, carbohydrates and fat) at dark onset, male Wistar rats were injected intraperitoneally with ACEA (0.1, 0.25, 0.5 and 1.0 mg/kg). Immediately after the injection, animals were placed into separate experimental cages with free access to food and a single 60-min period was video recorded to evaluate the BSS; protein, carbohydrates and fat intake (g) was measured at the same period of time. Intake of carbohydrates was significantly increased and this effect was prevented by the pre-treatment with AM 251. Analysis of BSS showed that administration of 0.5 mg/kg of ACEA reversed the satiation induced by food ingestion by increasing the time spent eating and decreasing the time resting without altering the overall activity. The present results suggest that the stimulation of food intake induced by activation of CB1 receptors involves a specific dietary component and behavioural selective mechanisms (stimulating hunger and inhibiting satiety).

Maternal Flavonoids Intake Reverts Depression-Like Behaviour in Rat Female Offspring.

Maternal hypercaloric exposure during pregnancy and lactation is a risk factor for developing diseases associated with inflammation such as obesity, diabetes and, neurological diseases in the offspring. Neuroinflammation might modulate neuronal activation and flavonoids are dietary compounds that have been proven to exert anti-inflammatory properties. Thus, the aim of the present study is to evaluate the effect of maternal supplementation with flavonoids (kaempferol-3-O-glucoside and narirutin) on the prevention of depression-like behaviour in the female offspring of dams fed with an obesogenic diet during the perinatal period. Maternal programming was induced by high fat (HFD), high sugar (HSD), or cafeteria diets exposure and depressive like-behaviour, referred to as swimming, climbing, and immobility events, was evaluated around postnatal day 56⁻60 before and after 30 mg/kg i.p. imipramine administration in the female offspring groups. Central inflammation was analyzed by measuring the TANK binding kinase 1 (TBK1) expression. We found that the offspring of mothers exposed to HSD programming failed to show the expected antidepressant effect of imipramine. Also, imipramine injection, to the offspring of mothers exposed to cafeteria diet, displayed a pro-depressive like-behaviour phenotype. However, dietary supplementation with flavonoids reverted the depression-like behaviour in the female offspring. Finally, we found that HSD programming increases the TBK1 inflammatory protein marker in the hippocampus. Our data suggest that maternal HSD programming disrupts the antidepressant effect of imipramine whereas cafeteria diet exposure leads to depressive-like behaviour in female offspring, which is reverted by maternal flavonoid supplementation.

The effects of 5-HT1A and 5-HT2C receptor agonists on behavioral satiety sequence in rats.

The present study examined the effects of 5-HT1A and 5-HT2C receptor agonists on behavioral satiety sequence (BSS) in rats. The 5-HT1A receptor agonist, 8-OH-DPAT (0.5 microg), and the 5-HT2C receptor agonist, Ro-60-0175 (3.0 microg), were injected into the paraventricular nucleus (PVN) of rats. The animals were maintained on an ad libitum feeding paradigm with access to water and individual sources of protein, carbohydrate, and fat. Intra-PVN administration of each agonist was associated with decreased carbohydrate consumption. The effect was enhanced by the administration of both agonists together. Behavioral analysis indicated that co-administration of 8-OH-DPAT and Ro-60-0175 interrupted the natural BSS with an increase in non-feeding behavior, whereas the 8-OH-DPAT alone promoted early development of the natural BSS. In conclusion, the 5-HT receptor agonists affected serotonergic modulation of feeding behavior in a functionally selective way.

Dopamine D4 receptor stimulation in GABAergic projections of the globus pallidus to the reticular thalamic nucleus and the substantia nigra reticulata of the rat decreases locomotor activity.

Dopamine D4 receptors are localized in the GABAergic projections that globus pallidus (GP) neurons send to the reticular nucleus of the thalamus (RTN), the substantia nigra reticulata (SNr) and the subthalamic nucleus (STN). Deficient D4 function in this network could lead to hyperactivity and thus be important in generating some of the symptoms of ADHD (attention deficit hyperactivity disorder), a condition associated with polymorphisms of dopamine D4 receptors. It is then, unexpected that systemic injections of D4 ligands have no significant effects on the motor activity of normal rats. We further examined this issue by microinjecting D4 ligands and psychostimulant drugs in relevant structures. Interstitial dopamine overflow in the RTN was increased by reverse microdialysis of both methylphenidate and methamphetamine. Intranuclear injections in the RTN of methylphenidate, methamphetamine and the selective D4 agonist PD 168,077 reduced motor activity. Intraperitoneal injection of the D4 antagonist L 745,870 blocked the effects of these intranuclear injections. Similarly, intranuclear injections of PD 168,077 in the SNr inhibited motor activity, an effect that was also blocked by intraperitoneal L 745,870. In rats with 6-OHDA induced hemiparkinsonism, intraperitoneal PD 168,077 produced ipsilateral turning behavior that was blocked by L 745,870. Our results suggest that diminished D4 signaling in GP projections could lead to increased traffic through the relay nuclei of the thalamus and hyperactivity. Hence this basal-ganglia-thalamus network may be one of the targets of the beneficial effects that psychostimulant drugs have in disorders associated with D4 receptor abnormalities. This article is part of a Special Issue entitled 'Post-Traumatic Stress Disorder'.

Análisis funcional de oligómeros de alfa-sinucleína en la permeabilidad de membranas / Functional analysis of oligomeric alpha-synuclein in membrane permeabilization / Análise funcional dos olig¶meros de alfa-sinucleína na permeabilidade das membranas

La enfermedad de Parkinson es un desorden neurodegenerativo común originado por la muerte celular dentro de la substancia nigra pars compacta. Se caracteriza por la presencia de agregados intracelulares proteicos compuestos principalmente por la alfa-sinucleína. Aunque los mecanismos moleculares por los cuales esta proteína contribuye a la toxicidad neuronal todavía se desconocen, se ha sugerido que los intermediarios oligoméricos son citotóxicos y permeabilizan las membranas celulares posiblemente a través de complejos que forman un poro en la bicapa; sin embargo, este mecanismo es altamente controversial. Así pues es necesario identificar el mecanismo por el cual ocurre la permeabilización de membranas para entender la interacción entre oligómeros y lípidos y poder estimar la relevancia biológica de este proceso. En este trabajo se evaluó por espectroscopía de fluorescencia la liberación de contenidos acuosos originados por oligómeros de alfa-sinucleína desde vesículas fosfolipídicas de distinta composición mediante el método ANTS/ DPX. Los resultados muestran que la disrupción de membranas solo ocurre en presencia de lípidos aniónicos y también por parámetros de empaquetamiento lipídico, lo que sugiere que la accesibilidad a la región hidrofóbica de las vesículas modula la interacción lípido-oligómero.(AU)

Parkinsons disease is a common neurodegenerative disorder marked by increased cell death within the substantia nigra pars compacta. It is characterized by the presence of intracellular aggregates composed primarily of the protein alpha-synuclein. How the aggregation of a-synuclein is related to neuronal degeneration is an important unresolved question. Oligomeric intermediates have been found to be more toxic to cells than monomeric or fibrillar forms of the protein. A possible mechanism by which oligomers could be toxic is through the disruption and permeabilization of cellular membranes. The proposed disruption mechanism is the formation of pore-like structures within the lipid bilayer although this mechanism is still highly controversial. To identify the mechanism through which membrane permeabilization is facilitated and to estimate the biological relevance of this process, it is crucial to have a greater knowledge of the lipid-oligomer interaction. The membrane disruptive effect of Alpha-synuclein oligomers on lipid vesicles of different headgroup composition using the ANTS/DPX assay was evaluated in this work. It was shown that membrane permeabilization is mainly determined by the presence of negatively charged lipids and also by lipid packing parameters, suggesting that the accessibility to the bilayer hydrocarbon core modulates oligomer-membrane interaction.(AU)

A doenþa de Parkinson é uma condiþÒo neuro-degenerativa comum que se origina na morte celular dentro da substÔncia nigra pars compacta. é caracterizada pela presenþa de agregados intracelulares protéicos compostos especialmente pela alfa-sinucleína.Mesmo que se desconheþa os mecanismos moleculares por onde essa proteína contribui para toxicidade neuronal, existe uma possibilidade de que os intermediários oligoméricos sejam citotóxicos, e permeabilizam as membranas celulares possivelmente através de complexos que formam um poro na capa dupla, entretanto, este mecanismo gera muita controvérsia. Sendo assim, é necessário identificar o mecanismo responsável pela permeabilidade das membranas para entender a interaþÒo entre os olig¶meros e os lipídios, e para ter uma estimativa da relevÔncia biológica deste processo. Neste trabalho, analisamos através de espectroscopia de fluorescÛncia, a liberaþÒo dos conteúdos aquoso originados por olig¶meros de alfa-sinucleína a partir das vesículas fosfolídicas de composiþÒo diferente através do método ANTS/DPX. Os resultados mostraram que a ruptura de membranas ocorrerá somente diante da presenþa de lipídios ani¶nicos, e também pelos parÔmetros de empacotamento lipídico, e isso sugere que a acessibilidade O regiÒo hidrofóbica das vesículas faz a modulaþÒo da interaþÒo lípidio-olig¶mero.(AU)

Análisis funcional de oligómeros de alfa-sinucleína en la permeabilidad de membranas / Functional analysis of oligomeric alpha-synuclein in membrane permeabilization / Análise funcional dos oligômeros de alfa-sinucleína na permeabilidade das membranas

La enfermedad de Parkinson es un desorden neurodegenerativo común originado por la muerte celular dentro de la substancia nigra pars compacta. Se caracteriza por la presencia de agregados intracelulares proteicos compuestos principalmente por la alfa-sinucleína. Aunque los mecanismos moleculares por los cuales esta proteína contribuye a la toxicidad neuronal todavía se desconocen, se ha sugerido que los intermediarios oligoméricos son citotóxicos y permeabilizan las membranas celulares posiblemente a través de complejos que forman un poro en la bicapa; sin embargo, este mecanismo es altamente controversial. Así pues es necesario identificar el mecanismo por el cual ocurre la permeabilización de membranas para entender la interacción entre oligómeros y lípidos y poder estimar la relevancia biológica de este proceso.En este trabajo se evaluó por espectroscopía de fluorescencia la liberación de contenidos acuosos originados por oligómeros de alfa-sinucleína desde vesículas fosfolipídicas de distinta composición mediante el método ANTS/DPX. Los resultados muestran que la disrupción de membranas solo ocurre en presencia de lípidos aniónicos y también por parámetros de empaquetamiento lipídico, lo que sugiere que la accesibilidad a la región hidrofóbica de las vesículas modula la interacción lípido-oligómero.

Parkinson's disease is a common neurodegenerative disorder marked by increased cell death within the substantia nigra pars compacta. It is characterized by the presence of intracellular aggregates composed primarily of the protein alpha-synuclein. How the aggregation of a-synuclein is related to neuronal degeneration is an important unresolved question. Oligomeric intermediates have been found to be more toxic to cells than monomeric or fibrillar forms of the protein. A possible mechanism by which oligomers could be toxic is through the disruption and permeabilization of cellular membranes. The proposed disruption mechanism is the formation of pore-like structures within the lipid bilayer although this mechanism is still highly controversial. To identify the mechanism through which membrane permeabilization is facilitated and to estimate the biological relevance of this process, it is crucial to have a greater knowledge of the lipid-oligomer interaction. The membrane disruptive effect of Alpha-synuclein oligomers on lipid vesicles of different headgroup composition using the ANTS/DPX assay was evaluated in this work. It was shown that membrane permeabilization is mainly determined by the presence of negatively charged lipids and also by lipid packing parameters, suggesting that the accessibility to the bilayer hydrocarbon core modulates oligomer-membrane interaction.

pars compacta. é caracterizada pela presença de agregados intracelulares protéicos compostos especialmente pela alfa-sinucleína.Mesmo que se desconheça os mecanismos moleculares por onde essa proteína contribui para toxicidade neuronal, existe uma possibilidade de que os intermediários oligoméricos sejam citotóxicos, e permeabilizam as membranas celulares possivelmente através de complexos que formam um poro na capa dupla, entretanto, este mecanismo gera muita controvérsia. Sendo assim, é necessário identificar o mecanismo responsável pela permeabilidade das membranas para entender a interação entre os oligômeros e os lipídios, e para ter uma estimativa da relevância biológica deste processo. Neste trabalho, analisamos através de espectroscopia de fluorescência, a liberação dos conteúdos aquoso originados por oligômeros de alfa-sinucleína a partir das vesículas fosfolídicas de composição diferente através do método ANTS/DPX. Os resultados mostraram que a ruptura de membranas ocorrerá somente diante da presença de lipídios aniônicos, e também pelos parâmetros de empacotamento lipídico, e isso sugere que a acessibilidade à região hidrofóbica das vesículas faz a modulação da interação lípidio-oligômero.