ACS Chemical Neuroscience最新文献

This work describes the use of high resolution online microdialysis coupled with a wireless microfluidic electrochemical sensing platform for continuous monitoring of the effect of cardiac arrest and resuscitation methods on brain glucose and other key neurochemicals in a porcine model. The integrated portable device incorporates low-volume three-dimensional (3D) printed microfluidic flow cells containing enzyme-based biosensors for glucose, lactate and glutamate measurement and a complementary metal-oxide semiconductor (CMOS)-based ion-sensitive field effect transistor (ISFET) for potassium measurement. Both analysis systems incorporate wireless electronics forming a complete compact system that is ideal for use in a crowded clinical environment. Using this integrated system we were able to build a signature of the neurochemical impact of cardiac arrest and resuscitation. Our results demonstrate the almost complete depletion of brain glucose following cardiac arrest and the subsequent increase in lactate, highlighting the vulnerability of the brain while the blood flow is compromised. Following a return of spontaneous circulation, glucose levels increased again and remained higher than baseline levels. These trends were correlated with simultaneous blood measurements to provide further explanation of the metabolic changes occurring in the brain. In addition, the onset of cardiac arrest corresponded to a transient increase in potassium. In most cases glutamate levels remained unchanged after cardiac arrest, while in some cases a transient increase was detected. We were also able to validate the trends seen using online microdialysis with traditional discontinuous methods; the two methods showed good agreement although online microdialysis was able to capture dynamic changes that were not seen in the discontinuous data.

The in vivo protective mechanisms of two low-molecular-mass (∼1.4 kDa) novel custom peptides (CPs) against paraquat-induced neurodegenerative dysfunction in the Caenorhabditis elegans model were deciphered. CPs prevented the paraquat from binding to the nerve ring adjacent to the pharynx in C. elegans (wild-type) by stable and high-affinity binding to the tyrosine-protein kinase receptor CAM-1, resulting in significant inhibition of paraquat-induced toxicity by reducing the production of reactive oxygen species, mitochondrial membrane depolarization, and chemosensory dysfunction. The CPs inhibited paraquat-induced dopaminergic neuron degeneration and alpha-synuclein protein expression, the hallmarks of Parkinson's disease, in transgenic BZ555 and NL5901 strains of C. elegans. Transcriptomic, functional proteomics, and quantitative reverse transcription-polymerase chain reaction analyses show that CPs prevented the increased expression of the genes involved in the skn-1 downstream pathway, thereby restoring paraquat-mediated oxidative stress, apoptosis, and neuronal damage in C. elegans. The ability of CPs to repair paraquat-induced damage was demonstrated by a network of gene expression profiles, illustrating the molecular relationships between the regulatory proteins.

The post-mortem measurement of tissue neurotransmitters is an interesting technique to address the gross biochemical activity. Its primary limitation is a lack of temporal resolution, although this is mitigated by enhanced spatial resolution, compared to in vivo methods. This neurochemical data is quantitative and requires no complex transformation, making it ideal to analyze neurochemical connectivity via the correlation of the biochemical signals between brain regions. These correlative approaches to quantitative measurements are fundamentally based on the variability of the data, an underdeveloped area of analysis in neurochemistry. One of the main reasons, as discussed in this Viewpoint, is that neurochemists recognize that variability in quantitative data stems not only from the biological variability, such as interindividual differences, but also from factors such as analytical devices. There are several ways to reduce variability caused by analytical and experimental biases through well-designed, precise protocols, allowing for the study of meaningful biological variability, such as interindividual differences between subjects.

Kam and colleagues discovered that FcγRIIb can specifically bind to Aβ42 oligomers (AβOs). The N-terminal residues F4 and D7 of Aβ42, as well as the W115 residue in domain D2 of FcγRIIb, are involved in this binding. However, the specificity of the FcγRIIb receptor's binding sites for AβOs and their dependence on different AβO species, including dimers (D/D_T), trimers (T/T_T), tetramers (Te/Te_T), and pentamers (P/P_T) during both the primary (P1) and secondary nucleation phases (P2), remains unknown. To address this, we employed molecular dynamics (MD) simulations to investigate the interactions between the extracellular domains D1 and D2 (FDD) of FcγRIIb and AβOs of varying sizes in the two different phases. We discovered that three specific fragments (f1, f2, and f3) of domain D2 in FDD are the primary binding sites for AβO species. Furthermore, among AβOs of the same molecular weight, those from the P2 phase exhibit a stronger binding affinity for FDD than those from the P1 phase. The distinction is ascribed to the stronger dependence on the hydrophobic residues in the β1 and β2 regions for the binding of AβOs in P2 (including T_T, Te_T, and P_T) than that (including D, Te, and P) in the P1 phase. In the P1 phase, these AβOs prefer to achieve binding to FDD through their N-terminal residues; however, by this, we identified that the species observed in Kam's experiment to bind FcγRIIb should probably be the tetrameric AβO (Te) in the P1 phase. Moreover, within both the P1 and P2 phases, we predicted that the trimeric AβO species in either the P1 or P2 phase is the strongest binding ligand for the FcγRIIb receptor. This study provides a comprehensive molecular perspective on the interaction between FcγRIIb and AβO in P2, which is of significant importance for the development of therapeutic strategies targeting Alzheimer's disease (AD) and autoimmune diseases.

Protein folding is crucial as it determines the three-dimensional structure and function of proteins, which are essential for biological processes, while misfolding can lead to the formation of aggregates and dysfunctional proteins, often associated with diseases. Histidine behaviors have been identified as a contributing factor to protein folding and misfolding due to changes in net charge and the diverse orientations of N/N-H groups on imidazole rings. In this viewpoint, we discuss misfolding diseases, the fundamental principles of histidine behaviors, and relevant studies in this field. Our current study helps elucidate histidine behaviors and their impact on secondary structure and aggregation characteristics, offering new insights into the mechanisms of histidine-related protein folding and misfolding.

The cholinergic deficits and deposition of β-amyloid (Aβ) species are regarded as the key events contributing to the progression of Alzheimer's disease (AD). Herein, a series of novel donor-acceptor architecture-type potential theranostic agents were designed, synthesized, and evaluated for their potential against cholinesterase (ChE) enzymes and detection of Aβ species, which are primary targets in the development of therapeutics for AD. The optimal compound/probe 18 containing a benzothiazolium fluorophore with a bifunctional electron-donating N-aryl piperazine scaffold exhibited potent inhibitory activities against acetylcholinesterase (AChE; IC₅₀ = 0.172 ± 0.011 μM) and butyrylcholinesterase (BuChE; IC₅₀ = 1.376 ± 0.141 μM). Measurement of fluorescence properties showed that probe 18 exhibited emission maxima (λ_em) of >610 nm in dimethyl sulfoxide (DMSO) and >590 nm in PBS, suitable for the fluorescence imaging. In vitro studies demonstrated a change in fluorescence characteristics and high binding affinities (18; K_d = 0.731 μM) upon binding with Aβ aggregates. The affinity of probe 18 toward Aβ aggregates was further observed in elavGAL4 > UAS Aβ, the Drosophila larval brain sections, using a fluorescence imaging technique. The in vivo acute oral toxicity evaluation indicated a safety profile of the lead probe 18. Moreover, in vivo behavioral studies including Y-maze and novel object recognition tests signified that the administration of compound 18 improved cognitive and spatial memory impairment at a dose of 10 and 20 mg/kg in the scopolamine-induced cognitive deficit model.

Traditional analgesic opioid compounds, which act through μ opioid receptors (MORs), engender a high risk for misuse and dependence. κ opioid receptor (KOR) activation, a potential target for pain treatment, produces antinociception without euphoric side effects but results in dysphoria and aversion. Triazole 1.1 is a KOR agonist biased toward G-protein coupled signaling, potentially promoting antinociception without dysphoria. We tested whether triazole 1.1 could provide antinociception and its effects in combination with morphine. We employed a lactic acid abdominal pain model, which induced acute pain behaviors, decreased basal dopamine levels in the nucleus accumbens (NAc), and increased KOR function. We administered several interventions including triazole 1.1 (30 mg/kg) and morphine (12 or 24 mg/kg), individually and in combination. Triazole 1.1 alone reduced the pain behavioral response and changes to KOR function but did not prevent the reduction in basal dopamine levels. Morphine not only dose-dependently prevented behavioral pain responses but also elevated NAc dopamine and did not prevent the pain-induced increase in KOR function. However, combining low-dose morphine with triazole 1.1 prevents behavioral pain responses, changes to NAc dopamine levels, and changes to KOR function. Therefore, we present triazole 1.1 as a dose-sparing pain treatment to be used in combination with a lower dose of morphine, thus reducing the potential for opioid misuse.

Parkinson's disease (PD) is one of the most common progressive neurodegenerative pathologies that leads to dopaminergic deficiency and motor manifestations. Alpha-synuclein aggregation is a characteristic hallmark of PD pathogenesis. These aggregates facilitate the formation of Lewy bodies and degeneration. The epidemiological evidence demonstrates a definitive association of diabetes with PD risk. Considering this, many antidiabetic agents such as GLP-1 agonists and DPP-4 inhibitors are being explored as alternative PD therapeutics. This study evaluated the neuroprotective effect of the DPP-4 inhibitor sitagliptin mediated by the PI3K/AKT and Nrf2 pathways in PD models. In silico studies were conducted to determine the binding affinity, stability, and ADMET properties of DPP-4 inhibitors with target proteins. Sitagliptin (15 mg/kg p.o.) was administered in rotenone (30 mg/kg p.o. for 28 days)-induced and MPTP/P (25 mg/kg i.p. MPTP and 100 mg/kg probenecid i.p. twice a week for 5 weeks)-induced PD mouse (C57/BL6) models. Neurobehavioral assessments were carried out throughout the study. Biochemical (GSH, MDA), molecular estimations (AKT, Nrf2, PI3K, GSK-3β, GLP1, CREB, BDNF, NF-κB, alpha-synuclein), histopathological studies, and immunohistochemistry were carried out at the end of the study. The in silico studies demonstrate better binding, stability, and ADMET profile of sitagliptin with both target proteins. Sitagliptin restored cognitive and motor deficits in both rotenone- and MPTP/P-induced mouse models. There was upregulation of PI3K, AKT, Nrf2, CREB, and BDNF levels and downregulation of GSK-3β, NF-κB, and alpha-synuclein levels in both models after treatment with sitagliptin. However, GLP1 levels were not significantly restored, indicating a GLP1-independent mechanism. It also restored histopathological alterations and TH+ neuronal loss induced by rotenone and MPTP/P. These findings demonstrate that sitagliptin exhibits neuroprotective action mediated by upregulation of the PI3K/AKT and Nrf2 pathways in rotenone and MPTP/P mouse models of PD.

Intrinsically disordered proteins (IDPs) are highly flexible molecules often linked to the onset of incurable diseases. Despite their great therapeutic potential, IDPs are often considered as undruggable because they lack defined binding pockets, which constitute the basis of drug discovery approaches. However, small molecules that interact with the intrinsically disordered state of α-synuclein, the protein linked to Parkinson's disease (PD), were recently identified and shown to act as chemical chaperones. Glucocerebrosidase (GCase) is an enzyme crucially involved in PD, since mutations that code for GCase are among the most frequent genetic risk factors for PD. Following the "dual-target" approach, stating that one carefully designed molecule can, in principle, interfere with more than one target, we identified a pharmacological chaperone for GCase that interacts with the intrinsically disordered monomeric form of α-synuclein. This result opens novel avenues to be explored in the search for molecules that act on dual targets, in particular, with challenging targets such as IDPs.

Cerebral dysfunctions give rise to a wide range of neurological diseases due to the structural and functional complexity of the human brain stemming from the interactive cellular metabolism of its specific cells, including neurons and glial cells. In parallel with advances in isolation and measurement technologies, genome-scale metabolic models (GEMs) have become a powerful tool in the studies of systems biology to provide critical insights into the understanding of sophisticated eukaryotic systems. In this study, brain cell-specific GEMs were reconstructed for neurons, astrocytes, microglia, oligodendrocytes, and oligodendrocyte precursor cells by integrating single-cell RNA-seq data and global Human1 via a task-driven integrative network inference for tissues (tINIT) algorithm. Then, intercellular reactions among neurons, astrocytes, microglia, and oligodendrocytes were added to generate a combined brain model, iHumanBrain2690. This brain network was used in the prediction of metabolic alterations in glucose, ketone bodies, oxygen change, and reporter metabolites. Glucose supplementation increased the subsystems' activities in glycolysis, and ketone bodies elevated those in the TCA cycle and oxidative phosphorylation. Reporter metabolite analysis identified L-carnitine and arachidonate as the top reporter metabolites in gray and white matter microglia in multiple sclerosis (MS), respectively. Carbamoyl-phosphate was found to be the top reporter metabolite in primary progressive MS. Taken together, single and integrated iHumanBrain2690 metabolic networks help us elucidate complex metabolism in brain physiology and homeostasis in health and disease.