The aggregation of α-synuclein plays a major role in Parkinson disease (PD) pathogenesis. Recent evidence suggests that defects in the autophagy-mediated clearance of α-synuclein contribute to the progressive loss of nigral dopamine neurons. Using an in vivo model of α-synuclein toxicity, we show that the PD-like neurodegenerative changes induced by excess cellular levels of α-synuclein in nigral dopamine neurons are closely linked to a progressive decline in markers of lysosome function, accompanied by cytoplasmic retention of transcription factor EB (TFEB), a major transcriptional regulator of the autophagy-lysosome pathway. The changes in lysosomal function, observed in the rat model as well as in human PD midbrain, were reversed by overexpression of TFEB, which afforded robust neuroprotection via the clearance of α-synuclein oligomers, and were aggravated by microRNA-128-mediated repression of TFEB in both A9 and A10 dopamine neurons. Delayed activation of TFEB function through inhibition of mammalian target of rapamycin blocked α-synuclein induced neurodegeneration and further disease progression. The results provide a mechanistic link between α-synuclein toxicity and impaired TFEB function, and highlight TFEB as a key player in the induction of α-synucleininduced toxicity and PD pathogenesis, thus identifying TFEB as a promising target for therapies aimed at neuroprotection and disease modification in PD.adeno-associated virus | Beclin | aggregates | synucleinopathy A major hallmark of Parkinson disease (PD) that contributes to the progressive loss of nigral dopamine (DA) neurons is α-synucleinopathy. Defects in clearance of oligomeric or misfolded proteins have been associated with aging and several neurodegenerative disorders (1-4). In human PD and related Lewy Body diseases, the presence of α-synuclein-positive (α-syn + ) aggregates is associated with accumulation of autophagosomes and reduction of lysosomal markers in affected nigral DA neurons, suggesting a defect in lysosome-mediated clearance of α-syn aggregates (5-7).How dysfunction of the autophagy-lysosome pathway (ALP) contributes to the pathogenesis of PD remains unclear. Under physiological conditions, α-syn is degraded by the ubiquitinproteasome system and the ALP, including macroautophagy and chaperone-mediated autophagy (6,(8)(9)(10)(11)(12). In cases of α-syn overload, however, misfolded or mutated α-syn fails to be processed and α-syn clearance by chaperone-mediated autophagy is blocked (12)(13)(14)(15). In this situation, processing of excess α-syn, or toxic α-syn species, will depend on the functional integrity of the macroautophagy pathway (14). In support, it has been shown that mice deficient in one of the autophagy-related (atg) proteins develop neurodegeneration, and that deficiency in atg7 or the PD-associated protein PARK9 (ATP13A2, a lysosomal ATPase) causes PD-like neurodegeneration, both in vitro and in vivo (7,(16)(17)(18)(19). In humans, PD has been genetically linked to the rare lysosomal storage diseases, Gaucher disease a...

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Highlights Extracellular levels of norepinephrine display dynamic changes during NREM and REM sleep  Phasic activity of locus coeruleus neurons during NREM underlies slow norepinephrine oscillations  Spindles occur at norepinephrine troughs and are abolished by norepinephrine increases  Increased spindles prior to REM reflect the beginning of a long-lasting norepinephrine decline  REM episodes are characterized by a sub-threshold continuous norepinephrine decline  The responsiveness of astrocytic Ca 2+ to norepinephrine is reduced during sleep

ObjectiveTo investigate the effect of in situ simulation (ISS) versus off-site simulation (OSS) on knowledge, patient safety attitude, stress, motivation, perceptions of simulation, team performance and organisational impact.DesignInvestigator-initiated single-centre randomised superiority educational trial.SettingObstetrics and anaesthesiology departments, Rigshospitalet, University of Copenhagen, Denmark.Participants100 participants in teams of 10, comprising midwives, specialised midwives, auxiliary nurses, nurse anaesthetists, operating theatre nurses, and consultant doctors and trainees in obstetrics and anaesthesiology.InterventionsTwo multiprofessional simulations (clinical management of an emergency caesarean section and a postpartum haemorrhage scenario) were conducted in teams of 10 in the ISS versus the OSS setting.Primary outcomeKnowledge assessed by a multiple choice question test.Exploratory outcomesIndividual outcomes: scores on the Safety Attitudes Questionnaire, stress measurements (State-Trait Anxiety Inventory, cognitive appraisal and salivary cortisol), Intrinsic Motivation Inventory and perceptions of simulations. Team outcome: video assessment of team performance. Organisational impact: suggestions for organisational changes.ResultsThe trial was conducted from April to June 2013. No differences between the two groups were found for the multiple choice question test, patient safety attitude, stress measurements, motivation or the evaluation of the simulations. The participants in the ISS group scored the authenticity of the simulation significantly higher than did the participants in the OSS group. Expert video assessment of team performance showed no differences between the ISS versus the OSS group. The ISS group provided more ideas and suggestions for changes at the organisational level.ConclusionsIn this randomised trial, no significant differences were found regarding knowledge, patient safety attitude, motivation or stress measurements when comparing ISS versus OSS. Although participant perception of the authenticity of ISS versus OSS differed significantly, there were no differences in other outcomes between the groups except that the ISS group generated more suggestions for organisational changes.Trial registration numberNCT01792674.

Glucagon-like peptide 1 (GLP-1) analogues are used for the treatment of type 2 diabetes. The ability of the GLP-1 system to decrease food intake in rodents has been well described and parallels results from clinical trials. GLP-1 receptors are expressed in the brain, including within the ventral tegmental area (VTA) and the nucleus accumbens (NAc). Dopaminergic neurons in the VTA project to the NAc, and these neurons play a pivotal role in the rewarding effects of drugs of abuse. Based on the anatomical distribution of GLP-1 receptors in the brain and the well-established effects of GLP-1 on food reward, we decided to investigate the effect of the GLP-1 analogue Exendin-4 on cocaine- and dopamine D1-receptor agonist-induced hyperlocomotion, on acute and chronic cocaine self-administration, on cocaine-induced striatal dopamine release in mice and on cocaine-induced c-fos activation. Here, we report that GLP-1 receptor stimulation reduces acute and chronic cocaine self-administration and attenuates cocaine-induced hyperlocomotion. In addition, we show that peripheral administration of Exendin-4 reduces cocaine-induced elevation of striatal dopamine levels and striatal c-fos expression implicating central GLP-1 receptors in these responses. The present results demonstrate that the GLP-1 system modulates cocaine's effects on behavior and dopamine homeostasis, indicating that the GLP-1 receptor may be a novel target for the pharmacological treatment of drug addiction.

Homo-and heteromeric complexes of KCNQ channel subunits are the molecular correlate of the M-current, a neuron-specific voltage-dependent K ϩ current with a well established role in control of neural excitability. We investigated the effect of KCNQ channel modulators on the activity of dopaminergic neurons in vitro and in vivo in the rat ventral mesencephalon. The firing of dopaminergic neurons recorded in mesencephalic slices was robustly inhibited in a concentration-dependent manner by the KCNQ channel opener N-(2-amino-4-(4-fluorobenzylamino)-phenyl) carbamic acid ethyl ester (retigabine). The effect of retigabine persisted in the presence of tetrodotoxin and simultaneous blockade of GABA A receptors, smallconductance calcium-activated K ϩ (SK) channels, and hyperpolarization-activated (I h ) channels, and it was potently reversed by the KCNQ channel blocker 4-pyridinylmethyl-9(10H)-anthracenone (XE991), indicating a direct effect on KCNQ channels. Likewise, in vivo single unit recordings from dopaminergic neurons revealed a prominent reduction in spike activity after systemic administration of retigabine. Furthermore, retigabine inhibited dopamine synthesis and c-Fos expression in the striatum under basal conditions. Retigabine completely blocked the excitatory effect of dopamine D 2 autoreceptor antagonists. Again, the in vitro and in vivo effects of retigabine were completely reversed by preadministration of XE991. Dual immunocytochemistry revealed that KCNQ4 is the major KCNQ channel subunit expressed in all dopaminergic neurons in the mesolimbic and nigrostriatal pathways. Collectively, these observations indicate that retigabine negatively modulates dopaminergic neurotransmission, likely originating from stimulation of mesencephalic KCNQ4 channels.KCNQ (also termed Kv7) channels are voltage-dependent potassium channels composed of homo-and heteromeric complexes of five different KCNQ subunits (KCNQ1-5, Kv7.1-Kv7.5). Unlike KCNQ1, all other KCNQ subunits (KCNQ2-5) are expressed in the CNS (Jentsch, 2000). Opening of KCNQ channels leads to neuronal hyperpolarization, thereby stabilizing the membrane potential and decreasing excitability. This makes them particularly interesting as targets in CNS diseases linked to hyperexcitability, including epilepsy, anxiety, pain, and migraine (Blackburn-Munro et al., 2005). The attractiveness of neuronal KCNQ channels in the treatment of such disease states is strongly supported by the identification of mutations within the human KCNQ genes. Thus, mutations in the KCNQ2 and KCNQ3 genes are associated with benign familial neonatal convulsions (Biervert et al., 1998), and certain mutations in the KCNQ4 gene result in progressive hearing loss (Kubisch et al., 1999). Several attempts have been made to find pharmacological KCNQ modulators. N-(2-Amino-4-(4-fluorobenzylamino)-This work was supported by the European Union 6th Framework Program (LSHM-CT-2004-503038) (to H.E.H., C.E., C.M., P.W., and L.C.R.) and by a grant from the Fonds National de la Recherche Scienti...

Agonism of the glucagon-like peptide 1 (GLP-1) receptor (GLP-1R) has been effective at treating aspects of addictive behavior for a number of abused substances, including cocaine. However, the molecular mechanisms and brain circuits underlying the therapeutic effects of GLP-1R signaling on cocaine actions remain elusive. Recent evidence has revealed that endogenous signaling at the GLP-1R within the forebrain lateral septum (LS) acts to reduce cocaine-induced locomotion and cocaine conditioned place preference, both considered dopamine (DA)-associated behaviors. DA terminals project from the ventral tegmental area to the LS and express the DA transporter (DAT). Cocaine acts by altering DA bioavailability by targeting the DAT. Therefore, GLP-1R signaling might exert effects on DAT to account for its regulation of cocaine-induced behaviors. We show that the GLP-1R is highly expressed within the LS. GLP-1, in LS slices, significantly enhances DAT surface expression and DAT function. Exenatide (Ex-4), a long-lasting synthetic analog of GLP-1 abolished cocaine-induced elevation of DA. Interestingly, acute administration of Ex-4 reduces septal expression of the retrograde messenger 2-arachidonylglycerol (2-AG), as well as a product of its presynaptic degradation, arachidonic acid (AA). Notably, AA reduces septal DAT function pointing to AA as a novel regulator of central DA homeostasis. We further show that AA oxidation product γ-ketoaldehyde (γ-KA) forms adducts with the DAT and reduces DAT plasma membrane expression and function. These results support a mechanism in which postsynaptic septal GLP-1R activation regulates 2-AG levels to alter presynaptic DA homeostasis and cocaine actions through AA.

SUMMARY The emerging interest in brain fluid transport has prompted a need for techniques that provide an understanding of what factors regulate cerebrospinal fluid (CSF) production. Here, we describe a methodology for direct quantification of CSF production in awake mice. We measure CSF production by placing a catheter in a lateral ventricle, while physically blocking outflow from the 4 th ventricle. Using this methodology, we show that CSF production increases during isoflurane anesthesia, and to a lesser extent with ketamine/xylazine anesthesia, relative to the awake state. Aged mice have reduced CSF production, which is even lower in aged mice overexpressing amyloid-β. Unexpectedly, CSF production in young female mice is 30% higher than in age-matched males. Altogether, the present observations imply that a reduction in CSF production might contribute to the age-related risk of proteinopathies but that the rate of CSF production and glymphatic fluid transport are not directly linked.