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INTRODUCTION: Dopaminergic (DA) neurons in the ventral tegmental area (VTA) are key players in regulating motivation and learning. Such control is mediated by DA innervation of other brain regions, such as the nucleus accumbens (NAc) and the prefrontal cortex (PFC). AIM(S): With use of optogenetics we aimed to delineate if both electrophysiological activity of DA neurons and DA release in target brain structures follow the optogenetic light stimulation protocols. Additionally, our goal was to compare results obtained with these two approaches. METHOD(S): To address these questions we used genetically modified rats expressing Cre recombinase gene under control of tyrosine hydroxylase gene – a marker of catecholaminergic neurons. Rats were stereotaxically injected into the VTA with adenoviral vectors carrying the Cre-dependent genes for channelrhodopsin-2 (ChR2) and yellow fluorescent protein. After proper expression of ChR2 in DA neurons in vivo electrophysiological or electrochemical experiments (single-unit recordings or fast-scan cyclic voltammetry, respectively) combined with optogenetic stimulation of the VTA were conducted. RESULTS: We demonstrated that laser blue light (473 nm, 5–60 Hz) stimulation alters both the activity of ChR2-expressing TH-positive VTA neurons and DA release in target brain regions. We showed that both DA neuronal firing and DA release elevates not linearly with increasing frequencies of light stimulation. High stimulation frequencies (>20 Hz) decreases both fidelity and amplitude of action potentials, preventing further increase in DA release. Finally, we demonstrated differentiation in DA release within the mesocorticolimbic brain subregions, with higher light-evoked DA concentration in the NAc than in the medial PFC. CONCLUSIONS: ChR2 enables selective control of DA neurons’ activity and subsequent DA release in the target brain regions with high spatial but limited temporal resolution. We demonstrated that light-evoked DA release differ in mesolimbic brain regions. FINANCIAL SUPPORT: This work was supported by the Polish National Science Center (Research grants UMO-2013/11/D/NZ4/02371 and UMO-2014/13/B/ NZ4/00146).
The increased inclination to natural rewards like food and sensation seeking have been associated with elevated drug intake in humans. We aimed to compare four inbred mouse strains with known different susceptibility to drug addiction in respect to response to natural rewards. In this study, we investigated four inbred strains C57Bl/6J, DBA/2J, SWR/J and 129P3/J in the selfadministration of food (SAF) and operant sensation seeking (OSS) models in three paradigms: (1) the fi xed ratio operant task (FR), (2) the progressive ratio operant task and (3) the concurrent choice operant task. No difference in response to reward in SAF FR paradigm was found among the studied strains. In contrast C57Bl/6J mice performed better then DBA/2J and 129P3/J strains in OSS paradigm while SWR mice did not react at all. In turn, the highest score of correct choice in SAF was observed in 129P3/J mice. The systemic administration of dopamine D1 receptor antagonist SCH 23390 or opioid receptor antagonists (naloxone or naltrexone) reduced the response to reward in the C57Bl/6J and DBA/2J mice. Our data suggest that genotype background is responsible for reward-seeking in SAF and OSS models with C57Bl/6J mice being the most responsive to the reward. We observed also that food as a reward has different motivational value than sensation seeking. Also our study indicates that opioid and dopaminergic systems may be involved in the response to natural rewards.
The noradrenergic system of the brain has been postulated to mediate arousal, selective attention, as well as anxiety, depression and learning and memory. Here we report the behavioral phenotype of transgenic mouse (NR1DbhCre) with selective ablation of the NMDA receptor subunit NR1 in noradrenergic cells. We focused on behaviors modulated by the noradrenergic system, in particular anxiety-like behaviors in the light/dark box, and open fi eld as well as depressive-like behaviors in forced swim test. Furthermore, memory performance was measured using elevated plus maze spatial learning (EPMSL), Y maze, right/left discrimination, habituation to open fi led and novel object recognition and motivation-dependent behaviors was studied using food pellet self-administration paradigm. While no alterations were found in anxiety- and depressive-like behaviors in NR1DbhCre mice, the transgenic animals displayed impaired object recognition memory, but increased performance in right/left discrimination test. In addition, the NR1DbhCre mice displayed higher food reward sensitivity as evidenced by faster acquisition of food self-administration and higher breaking point in progressive ratio schedule of reinforcement. In conclusion, the ablation of NMDA receptors in noradrenergic neurons in mice produce impairment of recognition memory and alteration of motivational systems.
Post-traumatic stress disorder (PTSD) is a chronic anxiety condition that develops as a result of a terrifying event. Clinical studies show that only about 10% of trauma-exposed people suffer from PTSD. Our research was focused on endophenotypes and molecular biomarkers of PTSD in an animal model. Differences in response to stress among inbred mouse strains (C57BL/6J, DBA/2J, SWR/J and 129P3/J) were compared: a single intense footshock was applied and ultrasonic vocalization during/after the stress was measured. Long-lasting effects were assessed 4-6 weeks after the traumatic event: conditioned and sensitized fear, social withdrawal, depressive-like behavior and susceptibility to drug addiction. SWR/J strain displayed the lowest conditioned fear, whereas sensitized fear was increased over time in C57BL/6J mice. Moreover, C57BL/6J strain exhibited increase in depressive-like behavior, while DBA/2J strain displayed increased social withdrawal. In addition, it was observed that exposition to traumatic stress increased sensitivity to rewarding properties of morphine in 129P3/J mice. Diverse long-lasting behavioral consequences of exposition to stress were associated with changes in basal and stress-induced profile of glucocorticoid receptor-dependent (GR) genes (e.g., Fkbp5 and Tsc22d3) in amygdala. Furthermore, our research showed that administration of GR antagonist disrupted reconsolidation of the traumatic event memory. Our research supports a model in which genetic factors are important for phenotypic variation in responsivity to stress. These genes may provide novel insight into mechanisms of stress-related disorders. This work was supported by Polish MSHE grants NN405 274137, N405 143238, IUVENTUS Plus and POIG De-Me-Ter 3.1
Single morphine administration to mice leads to changes in the expression of numerous genes in mouse striatum. Among them are genes regulated by the glucocorticoid receptor activation, like Sgk1, Gilz and Plzf [Korostynski et al. (2007) Genome Biol]. Analysis of their expression patterns with the use of in situ hybridization revealed their different regional distribution throughout the brain. To verify the cellular specifi city of expression, the double immunohistochemical stainings were performed with antibodies directed against investigated proteins and cellular markers (NeuN, GFAP, CC1, OX42). The results show that certain morphine- and GR-induced proteins are expressed in neurons, whereas others in glial cells. Hence, their regulation depends on the cell-type specifi c context.
The brain’s noradrenergic system system provides essential modulation to neuronal activity, with well established roles in control of arousal and selective attention, as well as memory, learning and response to stress. We have generated a transgenic mouse, NR1DbhCre, with ablation of the essential NMDA receptor subunit NR1 (Grin1) in noradrenergic cells. Since no functional NMDA receptors may be formed in the absence of NR1, and their expression is restricted to the central nervous system, the NR1DbhCre mice have impaired glutamate-dependent plasticity in the central noradrenergic neurons, without observable alterations in the sympathetic system or the hypothalamus-pituitary-adrenal axis. Transgenic animals were born at expected ratios and developed normally, displaying no obvious impairments. The general anatomy of the noradrenergic system in the mutant mice was normal, no loss of cells was observed and noradrenaline content in the prefrontal cortex was not altered. Interestingly, preliminary electrophysiological analysis indicates that loss of functional NMDA receptors attenuates the spontaneous activity in current-clamped locus coeruleus noradrenergic neurons manually held at −50 mV potential. In summary the NR1DbhCre transgenic mice are a novel model for the study of the roles of the noradrenergic system in the central nervous system.
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