Frontiers in Cellular Neuroscience最新文献

Neurodegenerative diseases such as Alzheimer's, Parkinson's, Huntington's, ALS, and spinocerebellar ataxia are becoming more prevalent as populations age, posing major global health challenges. Despite decades of research, effective treatments that halt or reverse these conditions remain elusive. Aging is the most significant risk factor in the development of these diseases, intertwining with molecular processes like DNA damage, mitochondrial dysfunction, and protein aggregation. Recent advances in gene-editing technologies, particularly CRISPR-Cas9, are beginning to shift the therapeutic landscape. This revolutionary tool allows for precise correction of genetic mutations associated with neurodegeneration, offering the potential for disease modification rather than symptom management alone. In this review, we explore how CRISPR-Cas9 is being leveraged to target key genes implicated in various neurodegenerative conditions and how it may overcome barriers posed by aging biology. We also examine the delivery systems and safety challenges that must be addressed before clinical application. With continued progress, CRISPR-Cas9 could mark a turning point in our ability to treat or even prevent age-related neurological decline.

Sustained synaptic transmission requires the continuous replenishment of released synaptic vesicles (SVs). This process is particularly critical in neuronal circuits that operate at high rates and with high temporal precision, such as those in the auditory brainstem. Here, we investigated the effect of SV (re-)filling on inhibitory synapses between the medial nucleus of the trapezoid body (MNTB) and the lateral superior olive (LSO). These synapses transmit information with high speed and fidelity, properties essential for auditory computations such as sound localization. We specifically examined the role of the vacuolar ATPase (V-ATPase), a proton pump that acidifies the SV lumen to enable neurotransmitter loading. Using patch-clamp recordings in acute mouse slices, we assessed synaptic function under control conditions and during continuous V-ATPase inhibition with bafilomycin or folimycin. Contrary to our initial hypothesis, pharmacological inhibition caused only moderate impairment of sustained transmission. Even under high drug concentrations and intense stimulation (e.g., 100 Hz for 4 min), steady-state responses declined only to ~33% of control. Similar reductions were observed in the replenishment rate, the size of the readily releasable pool, and the cumulative eIPSC amplitude. Quantal size decreased gradually, reaching ~70% of control. Recovery from synaptic depression persisted in the presence of V-ATPase blockade, although it was less efficient. Together, these findings indicate that MNTB-LSO synapses are relatively resistant to V-ATPase inhibition, suggesting that SV replenishment does not rely solely on V-ATPase activity. Alternative acidification mechanisms may contribute, and among potential candidates, the Na⁺/H⁺ exchanger isoform NHE6 showed strong immunoreactivity in glycinergic MNTB axon terminals contacting LSO somata. This identifies NHE6 as a promising target for future investigation.

Occupational noise-induced hearing loss (NIHL) is linked to the overproduction of mitochondrial reactive oxygen species after noise exposure. This cross-sectional study investigated the relationship between mitochondrial DNA (mtDNA) D-loop region methylation and oxidative stress in 150 participants divided into three age and sex matched groups: a control group (n = 50, workers without noise exposure and with normal hearing), an exposed group (n = 50, workers with significant noise exposure but normal hearing), and a case group (n = 50, workers diagnosed with NIHL). The subjects among groups were matched for sex and age to control confounding factors. Methylation levels of the mtDNA D-loop region were determined by the quantitative PCR following bisulfite conversion, while mitochondrial DNA copy number (mtDNA-CN) was assessed using the real-time PCR. Oxidative stress markers-including superoxide dismutase (SOD), glutathione peroxidase (GPX), total antioxidant status (TAS), and malondialdehyde (MDA)-were quantified via substrate-specific assays, ultraviolet enzymatic methods, and colorimetric techniques. Results showed the case group (141.6 ± 46.80 U/mL) showed lower SOD than the control (159.5 ± 18.68 U/mL, p < 0.05) and exposed groups (164.0 ± 15.44 U/mL, p < 0.01), MDA was higher in the case group (232.8 ± 134.5 nmol/mL) than in the control (193.5 ± 84.13 nmol/mL) and exposed groups (187.3 ± 60.76 nmol/mL), with a significant overall difference (F = 3.162, p < 0.05). The case group showed lower methylation [1.205 (0.595, 2.748) %] than both the control [1.710 (0.912, 3.225) %] and exposed groups [1.850 (0.987, 4.093) %] (H = 7.492, p < 0.05). The case group exhibited higher mtDNA-CN levels [397.7 (205.9, 532.1)] compared to both the blank control group [317.4 (234.6, 549.6)] and the exposed group [225.1 (125.3, 445.0)] (H = 9.213, p < 0.05). Methylation levels of the D-loop region were positively correlated with SOD and negatively correlated with MDA. Mediation analysis indicated that SOD may mediate the relationship between D-loop methylation and bilateral high-frequency hearing thresholds, suggesting an indirect epigenetic regulatory mechanism. These findings imply that noise-induced oxidative imbalance, reflected by reduced SOD, may lead to D-loop hypomethylation, contributing to the development of NIHL. These methylation sites may serve as preliminary biomarkers for further research on preventive strategies.

Parvalbumin (PV) interneurons in the dorsal striatum (DS) are fast-spiking GABAergic cells critical for feedforward inhibition and synaptic integration within basal ganglia circuits. Despite their well-characterized electrophysiological roles, their molecular identity remains incompletely defined. Using the Ribotag approach in Pvalb-Cre mice, we profiled the translatome of DS PV interneurons and identified over 2,700 transcripts significantly enriched (fold-change > 1.5) in this population. Our data validate established PV markers and reveal a distinct molecular signature of DS PV neurons compared to PV interneurons from the nucleus accumbens. Gene ontology analyses highlight prominent expression of genes related to extracellular matrix components, cell adhesion molecules, synaptic organization, ion channels, and neurotransmitter receptors, particularly those mediating glutamatergic and GABAergic signaling. Notably, perineuronal net markers were robustly expressed in DS PV interneurons and confirmed by immunofluorescence. Transcriptomic analysis of DS PV neurons following repeated d-amphetamine exposure identified Gm20683 as the only differentially expressed transcript between treated groups. Furthermore, RNAseq analysis of mice subjected to an operant behavior paradigm with two types of food reward (high-palatable diet or standard chow) identified over 1,000 and 100 genes enriched in DS PV neurons from standard and high-palatable masters, respectively. These findings provide a comprehensive molecular profile of DS PV interneurons, distinguishing them from other striatal PV populations, and reveal specific gene expression changes associated with psychostimulant exposure and reward-driven behaviors. Our findings deepen insight into the molecular mechanisms of PV interneuron activity in striatal circuits and their potential roles in neuropsychiatric, motor and reward-related disorders.

Spreading depolarization (SD) is a wave of profound cellular depolarization that propagates primarily across gray matter of central nervous system tissue and causes a near-complete collapse of ionic gradients. Implicated in neuropathologies including seizures, migraine with aura, traumatic brain injury, and stroke, SD is experimentally induced in animals by electrical stimulation, mechanical injury, hypoxia, elevated extracellular potassium, and various other techniques. Despite extensive research, the mechanisms underlying SD initiation remain unclear. Prior research in rodents found that simultaneously blocking sodium, calcium, and glutamatergic (AMPA and NMDA) channels prevents SD induction whereas inhibiting any two of these three currents is insufficient. This suggests that SD induction could be a product of overstimulation of any single known inward cationic current. However, some researchers propose that SD induction occurs via an unknown "SD channel." To further explore the role of known inward cationic currents in SD induction, we applied high potassium to two biological models, namely zebrafish and mice. First, we developed a novel ex vivo zebrafish model to assess SD induction in the optic tectum. Using KCl microinjection and DC recordings, we found that inhibition of sodium, calcium, and glutamatergic channels significantly decreased SD amplitude but never blocked SD induction in the zebrafish optic tectum. Similar pharmacological experiments in hippocampal mouse slices (CA1 subregion) also confirmed that SDs persist despite the same pharmacological cocktail. These findings suggest that additional mechanisms beyond sodium, calcium, and glutamatergic signaling contribute to SD induction, supporting the hypothesis that an unknown channel is critical in SD physiology.

Degeneration or damage of neuronal circuits in the central nervous system can lead to an irreversible loss of neurons and function in the affected brain region. Neuronal transplantation is a promising therapeutic approach consisting of introducing healthy cells into the damaged or diseased regions to restore lost circuits. To achieve successful neuronal transplantation, proper integration of the graft in the host circuitry is necessary. This includes the restoration of connectivity as well as the recapitulation of the physiological characteristics of the lost endogenous neurons. An often-overlooked aspect to assess the integration of transplanted neurons is the acquisition of cell-extrinsic features, such as myelination. This review explores the interaction between transplanted cells and endogenous oligodendroglia, the evidence of myelination in different neuronal transplantation models, and the checkpoints that can influence graft myelination in the injured or diseased brain. Additionally, it discusses how appropriate myelin ensheathment could help overcome some challenges faced in the field of neuronal replacement.

Introduction: Neuroinflammation is a critical factor contributing to secondary brain injury following traumatic brain injury (TBI). This process engages diverse cell types within the central nervous system (CNS), including significant infiltration of myeloid lineage cells-primarily neutrophils and macrophages-during the acute and subacute phases of TBI. These myeloid-derived cells represent a major population that critically influences the development and progression of neuroinflammation. Microglia and peripherally infiltrating macrophages exhibit polarization phenotypes that play a pivotal role in modulating inflammatory changes. Due to their functional and phenotypic similarities, their distinct contributions to the inflammatory response in TBI remain a subject of considerable debate. Lysozyme 2 (Lyz2) is a well-established marker for myeloid lineage cells (including monocytes, macrophages, and neutrophils) in mice, allowing specific targeting and depletion of these cells to dissect their functional roles in TBI.

Methods: In the present study, we investigated the trend of inflammatory factors during the early stage of TBI using Lyz2-IRES-DTREGFP transgenic mice, which specifically target and deplete Lyz2-positive myeloid cells. Tissue samples for RT-qPCR and flow cytometry were harvested from the perilesional cortex (within a 2-mm radius of the impact site) and the underlying hippocampus.

Results and discussion: Our findings revealed a considerable reduction in the expression of pro-inflammatory factors (e.g., IL-1β, iNOS, IL-6, IFN-γ) and an increase in the expression of anti-inflammatory factors (e.g., IL-4, IL-10, IL-13, Arg-1). Furthermore, we observed a shift in polarization phenotypes, characterized by a decreased proportion of M1 macrophages and an increased proportion of M2 macrophages. However, during the chronic phase, behavioral and histological analyses revealed worse outcomes. These findings demonstrate that targeted depletion of Lyz2-positive myeloid cells during acute TBI attenuates neuroinflammation. However, this early immunomodulatory shift correlates paradoxically with exacerbated chronic neurological deficits, suggesting that transient suppression of myeloid-driven inflammation may disrupt long-term reparative processes critical for functional recovery after TBI.

Prominent histopathological features of Parkinson's disease (PD) include the presence of Lewy bodies, intra-neural protein aggregates mainly composed of α-synuclein (α-syn), and cell death of dopaminergic neurons. Epidemiological studies have revealed a correlation between exposure to environmental neurotoxins, such as rotenone, and an increased risk of developing PD. In this study, we evaluated the role of rotenone in α-syn spreading and accumulation, with the aim of developing a mouse model of accelerated PD. Human α-synuclein pre-formed fibrils (PFF) were injected into the mouse striatum by stereotactic surgery. Rotenone (2.5 mg/kg-body-weight) was administered intraperitoneally once daily for four consecutive weeks one day or three weeks after the PFF injection. Brains were collected twenty-four hours after the last injection for immunohistochemical analysis. In this study, rotenone significantly synergized PFF induced α-syn spreading, neuroinflammation, in addition to augmented loss of dopaminergic neurons along the nigrostriatal pathway.

Microglia, essential for brain development, homeostasis, and neuroinflammation, originate from the yolk sac during embryogenesis and migrate into the developing brain. Because of this developmental origin, many brain organoid models naturally lack microglia and require co-culture. To address this issue, we developed a microglia-integrated brain organoid model (immune-competent brain microphysiological system, μbMPS) by aggregating hiPSC-derived neural and microglia progenitors in U-bottom 96-well plates, allowing controlled and reproducible incorporation of microglia progenitors. We demonstrated that microglia integrated, matured, and survived long-term in the neural environment without the need for costly exogenous microglia-specific growth factors or cytokines. We maintained microglia-containing organoids for over 9 weeks, demonstrating functional activity, phagocytosis, and neuroinflammatory responses. The μbMPS also exhibited enhanced neuronal activity and maturity, providing a scalable, reproducible model for neurodevelopment, disease modeling, and neurotoxicology research.