Basal Forebrain Modulation of Olfactory Coding In Vivo.

Sensory perception is one of the most fundamental brain functions, allowing individuals to properly interact and adapt to a constantly changing environment. This process requires the integration of bottom-up and topdown neuronal activity, which is centrally mediated by the basal forebrain, a brain region that has been linked to a series of cognitive processes such as attention and alertness. Here, we review the latest research using optogenetic approaches in rodents and in vivo electrophysiological recordings that are shedding light on the role of this region, in regulating olfactory processing and decisionmaking. Moreover, we summarize evidence highlighting the anatomical and physiological differences in the basal forebrain of individuals with autism spectrum disorder, which could underpin the sensory perception abnormalities they exhibit, and propose this research line as a potential opportunity to understand the neurobiological basis of this disorder.

La percepción sensorial es una de las funciones cerebrales más fundamentales, permitiendo a los individuos interactuar de manera apropiada con el entorno y adaptarse a un ambiente en constante cambio. Este proceso requiere la integración de la actividad neuronal ascendente y descendente, que es mediada por el cerebro basal (BF), una región cerebral que ha sido asociada a una serie de procesos cognitivos, como estados de atención y alerta.En este trabajo revisamos las últimas investigaciones que han utilizado optogenética y registros electrofisiológicos in vivo que han iluminado el rol del BF en el procesamiento olfatorio y la toma de decisiones. Además, resumimos la literatura que destaca las alteraciones fisiológicas y anatómicas del BF de individuos con trastornos del espectro autista, que podrían subyacer las anormalidades en la percepción que presentan, y proponemos esta línea de investigación como una posible oportunidad para entender las bases neurobiológicas de este trastorno.

OPTO-BLUE: An Integrated Bidirectional Optogenetic Lentiviral Platform for Controlled Light-Induced Gene Expression.

Regulated systems for transgene expression are useful tools in basic research and a promising platform in biomedicine due to their regulated transgene expression by an inducer. The emergence of optogenetics expression systems enabled the construction of light-switchable systems, enhancing the spatial and temporal resolution of a transgene. The LightOn system is an optogenetic tool that regulates the expression of a gene of interest using blue light as an inducer. This system is based on a photosensitive protein (GAVPO), which dimerizes and binds to the UASG sequence in response to blue light, triggering the expression of a downstream transgene. Previously, we adapted the LightOn system to a dual lentiviral vector system for neurons. Here, we continue the optimization and assemble all components of the LightOn system into a single lentiviral plasmid, the OPTO-BLUE system. For functional validation, we used enhanced green fluorescent protein (EGFP) as an expression reporter (OPTO-BLUE-EGFP) and evaluated the efficiency of EGFP expression by transfection and transduction in HEK293-T cells exposed to continuous blue-light illumination. Altogether, these results prove that the optimized OPTO-BLUE system allows the light-controlled expression of a reporter protein according to a specific time and light intensity. Likewise, this system should provide an important molecular tool to modulate gene expression of any protein by blue light.

Parvalbumin Role in Epilepsy and Psychiatric Comorbidities: From Mechanism to Intervention.

Parvalbumin is a calcium-binding protein present in inhibitory interneurons that play an essential role in regulating many physiological processes, such as intracellular signaling and synaptic transmission. Changes in parvalbumin expression are deeply related to epilepsy, which is considered one of the most disabling neuropathologies. Epilepsy is a complex multi-factor group of disorders characterized by periods of hypersynchronous activity and hyperexcitability within brain networks. In this scenario, inhibitory neurotransmission dysfunction in modulating excitatory transmission related to the loss of subsets of parvalbumin-expressing inhibitory interneuron may have a prominent role in disrupted excitability. Some studies also reported that parvalbumin-positive interneurons altered function might contribute to psychiatric comorbidities associated with epilepsy, such as depression, anxiety, and psychosis. Understanding the epileptogenic process and comorbidities associated with epilepsy have significantly advanced through preclinical and clinical investigation. In this review, evidence from parvalbumin altered function in epilepsy and associated psychiatric comorbidities were explored with a translational perspective. Some advances in potential therapeutic interventions are highlighted, from current antiepileptic and neuroprotective drugs to cutting edge modulation of parvalbumin subpopulations using optogenetics, designer receptors exclusively activated by designer drugs (DREADD) techniques, transcranial magnetic stimulation, genome engineering, and cell grafting. Creating new perspectives on mechanisms and therapeutic strategies is valuable for understanding the pathophysiology of epilepsy and its psychiatric comorbidities and improving efficiency in clinical intervention.

Optimization of the Light-On system in a lentiviral platform to a light-controlled expression of genes in neurons

BACKGROUND: Molecular brain therapies require the development of molecular switches to control gene expression in a limited and regulated manner in time and space. Light-switchable gene systems allow precise control of gene expression with an enhanced spatio-temporal resolution compared to chemical inducers. In this work, we adapted the existing light-switchable Light-On system into a lentiviral platform, which consists of two modules: (i) one for the expression of the blue light-switchable transactivator GAVPO and (ii) a second module containing an inducible-UAS promoter (UAS) modulated by a light-activated GAVPO. RESULTS: In the HEK293-T cell line transfected with this lentiviral plasmids system, the expression of the reporter mCherry increased between 4 to 5 fold after light induction. A time expression analysis after light induction during 24 h revealed that mRNA levels continuously increased up to 9 h, while protein levels increased throughout the experiment. Finally, transduction of cultured rat hippocampal neurons with this dual Light-On lentiviral system showed that CDNF, a potential therapeutic trophic factor, was induced only in cells exposed to blue light. CONCLUSIONS: In conclusion, the optimized lentiviral platform of the Light-On system provides an efficient way to control gene expression in neurons, suggesting that this platform could potentially be used in biomedical and neuroscience research, and eventually in brain therapies for neurodegenerative diseases.

Perturbations in the Activity of Cholinergic Interneurons in the Dorsomedial Striatum Impairs the Update of Actions to an Instrumental Contingency Change.

The striatal cholinergic system is key in detecting changes in instrumental contingencies. While recent evidence supports this vision, cell type-specific online control on the activity of the cholinergic striatal neurons is necessary to empirically test it. In this study, we performed optogenetic manipulations of the activity of striatal cholinergic interneurons (CINs) to evaluate their contribution to the updating of a previously learned instrumental contingency. By modulating the activity of CINs, we identified that the inhibition of CINs impairs the update of actions to a contingency change. Remarkably, a manipulation that perturbs the activity of CINs, rather than inhibiting them also impaired the encoding of the change in contingency. These results emphasize that beyond an increase in the activity of CINs, the intact activity of these cells is required for the identification of an instrumental contingency change.

Optogenetics: the new molecular approach to control functions of neural cells in epilepsy, depression and tumors of the central nervous system.

The optogenetic tools have been described as valuable techniques to study neural activity through light stimulation, as well as potential neuromodulator approaches in the management of several central nervous system (CNS) diseases. Since the first bacteriorhodopsin protein described as a single-component light-activated regulator of transmembrane ion flow description, in 1980's, the focus has been on channel proteins for neurobiology; however, the advances in engineering techniques showed involvement changes in cellular biological behavior in several types of proteins involved in cell cytoskeleton regulation, motility and gene expression. Although the use of this technology has been published in many papers, a question still remains regarding real results and potential clinical applicability in CNS diseases, as well as the publications scarcity that systematically analyses the published results. Lastly, the aim of this review is to discuss the experimental results, molecular mechanisms and potential clinical applications of optogenetic tools in epilepsy and depression treatment, as well as its applicability in the treatment of CNS tumors.

Input-Specific NMDAR-Dependent Potentiation of Dendritic GABAergic Inhibition.

Preservation of a balance between synaptic excitation and inhibition is critical for normal brain function. A number of homeostatic cellular mechanisms have been suggested to play a role in maintaining this balance, including long-term plasticity of GABAergic inhibitory synapses. Many previous studies have demonstrated a coupling of postsynaptic spiking with modification of perisomatic inhibition. Here, we demonstrate that activation of NMDA-type glutamate receptors leads to input-specific long-term potentiation of dendritic inhibition mediated by somatostatin-expressing interneurons. This form of plasticity is expressed postsynaptically and requires both CaMKIIα and the ß2 subunit of the GABA-A receptor. Importantly, this process may function to preserve dendritic inhibition, as genetic deletion of NMDAR signaling results in a selective weakening of dendritic inhibition. Overall, our results reveal a new mechanism for linking excitatory and inhibitory input in neuronal dendrites and provide novel insight into the homeostatic regulation of synaptic transmission in cortical circuits.

Optogenetic switches for light-controlled gene expression in yeast.

Light is increasingly recognized as an efficient means of controlling diverse biological processes with high spatiotemporal resolution. Optogenetic switches are molecular devices for regulating light-controlled gene expression, protein localization, signal transduction and protein-protein interactions. Such molecular components have been mainly developed through the use of photoreceptors, which upon light stimulation undergo conformational changes passing to an active state. The current repertoires of optogenetic switches include red, blue and UV-B light photoreceptors and have been implemented in a broad spectrum of biological platforms. In this review, we revisit different optogenetic switches that have been used in diverse biological platforms, with emphasis on those used for light-controlled gene expression in the budding yeast Saccharomyces cerevisiae. The implementation of these switches overcomes the use of traditional chemical inducers, allowing precise control of gene expression at lower costs, without leaving chemical traces, and positively impacting the production of high-value metabolites and heterologous proteins. Additionally, we highlight the potential of utilizing this technology beyond laboratory strains, by optimizing it for use in yeasts tamed for industrial processes. Finally, we discuss how fungal photoreceptors could serve as a source of biological parts for the development of novel optogenetic switches with improved characteristics. Although optogenetic tools have had a strong impact on basic research, their use in applied sciences is still undervalued. Therefore, the invitation for the future is to utilize this technology in biotechnological and industrial settings.

Optogenética e estimulação óptica neural: estado atual e perspectivas / Optogenetics and neural optical stimulation: current state and perspectives

Ao longo dos últimos 50 anos, o uso da luz, em especial o laser, vem promovendo grandes avanços em diversas áreas da ciência e da tecnologia. Na última década o uso de estímulos ópticos no campo da biomédica tem despertado grande interesse no meio acadêmico e na indústria. Dois ramos que se destacam pelo seu crescimento são: a estimulação óptica direta e a optogenética. A primeira utiliza diferentes parâmetros da luz para adequar o efeito desejado na interação com o tecido biológico. A segunda faz uso de engenharia genética para tornar os tecidos biológicos sensíveis à luz. A estimulação neural por infravermelho (estimulação óptica direta) não necessita de contato direto com o tecido e apresenta maior seletividade especial se comparada à estimulação elétrica, mas tem a capacidade restrita de ativar (despolarizar) os neurônios. A optogenética, entretanto, pode ser utilizada para manipular o tecido neural tornando-o sensível à luz; sendo, então, possível despolarizar ou hiperpolarizar os neurônios codificados, assim como monitorar as ativações por meio de codificação de proteínas fluorescentes sensíveis à tensão elétrica. Tanto a técnica de estimulação óptica por infravermelho ou a técnica de optogenética, vêm sendo aplicadas apenas à modelos animais. Os resultados mostram, entretanto, que há grande viabilidade de aplicação da estimulação óptica em seres humanos. Futuramente, tais técnicas poderão substituir o atual padrão ouro para a ativação neural, a estimulação elétrica, em aplicações envolvendo doenças neurológicas específicas.

Within the last 50 years the light and specially the laser has fomented great advances in several areas of science and technology. During the past decade the use of optical stimuli in the biomedical research field have been of great interest for both academy and industry. Two research branches that can be highlighted due to its growth are: direct optical stimulation and optogenetic. The first one uses different parameters of light to optimize the desired effect on the tissue interaction. The other branch works with genetic engineering technics to make cells sensitive to light. The neural stimulation by infrared (direct optical stimulation) does not require direct contact with the tissue and has higher spatial selectivity when compared to electrical stimulation, but it has restricted ability to activate (depolarize) neurons. The optogenetic, however, can be used to manipulate the neural tissue depolarizing or hyperpolarizing encoded neurons, as well as monitor activations by encoding fluorescent proteins sensitive to voltage. The stimulation by infrared optical or optogenetic, has been applied only to animal models although there is a great possibility for human applications. In the future, it may even replace existing techniques such as electrical brain stimulation to treat specific neurological diseases.