Progress in Neurobiology, Journal Year: 2024, Volume and Issue: 244, P. 102707 - 102707
Published: Dec. 25, 2024
Language: Английский
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown
Published: Jan. 16, 2025
Abstract Cis -acting regulatory enhancer elements are valuable tools for gaining cell type-specific genetic access. Leveraging large chromatin accessibility atlases, putative sequences can be identified and deployed in adeno-associated virus (AAV) delivery platforms. However, a significant bottleneck AAV discovery is charting their detailed expression patterns vivo , process that currently requires gold-standard one-by-one testing. Here we present barcoded multiplex strategy screening AAVs at type resolution using single RNA sequencing taxonomy mapping. We executed proof-of-concept study small pool of validated expressing variety neuronal non-neuronal types across the mouse brain. Unexpectedly, encountered substantial technical biological noise including chimeric packaging products, necessitating development novel techniques to accurately deconvolve patterns. These results underscore need improved methods mitigate highlight complexity biology .
Language: Английский
Citations
0
Cells, Journal Year: 2025, Volume and Issue: 14(3), P. 236 - 236
Published: Feb. 6, 2025
The problem of drug resistance in epilepsy means that many cases, a surgical treatment may be advised. But this is only possible if there an epileptic focus, and resective brain surgery have adverse side effects. One the promising alternatives gene therapy, which allows targeted expression therapeutic genes different regions, even specific cell types. In review, we provide detailed explanations some key terms related to genetic engineering, describe various regulatory elements already been used development approaches treating using viral vectors. We compare few universal promoters for their strength duration transgene expression, our description cell-specific promoters, focus on driving glutamatergic neurons, GABAergic neurons astrocytes. also explore enhancers other cis-regulatory currently vectors consider future perspectives state-of-the-art technologies designing new, stronger more elements. Gene therapy has multiple advantages should become common future, but still lot study invent field.
Language: Английский
Citations
0
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown
Published: Feb. 26, 2025
Cortical GABAergic interneurons (INs) are comprised of distinct types that provide tailored inhibition to pyramidal cells (PCs) and other INs, thereby enabling precise control cortical circuit activity. INs expressing the neuropeptide vasoactive-intestinal peptide (VIP) have attracted attention recently following discovery they predominantly function by inhibiting dendritic-targeting somatostatin (SST) disinhibiting PCs. This VIP-SST disinhibitory motif is observed throughout neocortex from mice humans, serves as a key mechanism for top-down (feedback) context-dependent information processing. Thus, VIP IN-mediated disinhibition has been found play an important role in sensory processing, executive functions, attention, sensorimotor integration cortico-cortical thalamocortical feedback interactions. Furthermore, implicated mediating effects reinforcement signals, both reward aversive, via their responsiveness neuromodulators such acetylcholine (ACh), facilitating synaptic plasticity learning. While it evident transcriptomic analyses molecularly heterogeneous group, physiological significance this diversity unclear at present. Here, we characterized functional primary somatosensory cortex leveraging intersectional genetic approaches study IN subtypes. We can be divided into four different populations: group expresses Ca 2+ -binding protein calretinin (CR), two groups express cholecystokinin (CCK), does not either CR or CCK (non-CCK non-CR; nCCK nCR). neurons each exhibit laminar distributions, axonal dendritic arbors, intrinsic electrophysiological properties, efferent connectivity, VIP/CR target almost exclusively SST VIP/nCCK nCR also mainly but connections parvalbumin (PV) INs. These essentially no connectivity (PCs). On hand, VIP/CCK PCs, differ degree which release type modulated endocannabinoids. long-range inputs differentially recruit groups. Intriguingly, find populations subtypes turn, indicating presence specialized subcircuits. Activation subpopulations vivo results differential on network, thus providing evidence modularity actions during
Language: Английский
Citations
0
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown
Published: March 3, 2025
ABSTRACT Gene therapy for neurodegenerative diseases faces significant challenges due to the blood-brain barrier (BBB), which limits drug delivery central nervous system (CNS). While clinical trials Parkinson’s disease (PD) have progressed, administration of vectors expressing enzymatic or neurotrophic factor transgenes required extensive optimization method achieve potentially therapeutic levels transgene expression. Focused ultrasound (FUS) combined with microbubbles has emerged as a promising non-invasive strategy transiently open BBB targeted gene via viral nanocarriers including recombinant adeno-associated viruses (AAVs). However, key factors influencing FUS-mediated AAV delivery, dose distribution and efficacy, remain underexplored in non-human primates (NHPs). Here, we evaluated feasibility AAV9-CAG-GFP using two portable modalities: ultrasound-guided, spherically-focused FUS (USgFUS) novel low-frequency linear array configuration imaging called theranostic (ThUS). In mice, FUS-sonicated regions exhibited 25-fold increase AAV9 biodistribution compared systemic injection alone. Extending this approach NHPs, observed up 200-fold DNA treated brain regions, PD-relevant structures. assessing translational potential technique, ThUS-mediated AAV9-hSyn-hNTRN (human neurturin) toxin mouse model PD facilitated rescue 80% 75% degenerated dopaminergic neurons substantia nigra striatum, respectively. These findings demonstrate that technologies can non-invasively enhance both mice NHPs relative what be achieved intravenous (IV) same capsid With further development, these approaches may offer clinically viable, alternative diseases. One sentence summary opening increased after vector rhesus macaques.
Language: Английский
Citations
0
Neuron, Journal Year: 2025, Volume and Issue: unknown
Published: March 1, 2025
Language: Английский
Citations
0
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown
Published: Sept. 29, 2024
We present an enhancer AAV toolbox for accessing and perturbing striatal cell types circuits. Best-in-class vectors were curated major neuron populations including medium spiny neurons (MSNs), direct indirect pathway MSNs, as well Sst-Chodl, Pvalb-Pthlh, cholinergic interneurons. Specificity was evaluated by multiple modes of molecular validation, three different routes virus delivery, with diverse transgene cargos. Importantly, we provide detailed information necessary to achieve reliable type specific labeling under experimental contexts. demonstrate circuit-selective optogenetic perturbation behavior multiplex interneuron targeted analysis cellular features. Lastly, show conserved
Language: Английский
Citations
3
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown
Published: Jan. 21, 2024
SUMMARY Within the adult rodent hippocampus, opioids suppress inhibitory parvalbumin-expressing interneurons (PV-INs), thus disinhibiting local micro-circuits. However, it is unknown if this disinhibitory motif conserved in other cortical regions, species, or across development. We observed that PV-IN mediated inhibition robustly suppressed by hippocampus proper but not primary neocortex mice and nonhuman primates, with spontaneous tone resected human tissue also following a consistent dichotomy. This hippocampal was established early development when PV-INs were found to regulate population activity. Acute opioid-mediated modulation partially occluded morphine pretreatment, implications for effects of on network activity important learning memory. Together, these findings demonstrate exhibit divergence opioid sensitivity brain regions remarkably evolution highlights underappreciated role acting through immature shaping
Language: Английский
Citations
2
Research Square (Research Square), Journal Year: 2024, Volume and Issue: unknown
Published: Aug. 2, 2024
Cholinergic projection neurons of the nucleus basalis and substantia innominata (NBM/SI) densely innervate basolateral amygdala (BLA) have been shown to contribute encoding fundamental life-threatening experiences. Given vital importance these circuits in acquisition retention memories that are essential for survival a changing environment, it is not surprising basic anatomical organization NBM/SI well conserved across animal classes as diverse teleost mammal. What known extent which physiology morphology also conserved. To address this issue, we made patch-clamp recordings from
Language: Английский
Citations
0
Cellular and Molecular Life Sciences, Journal Year: 2024, Volume and Issue: 81(1)
Published: Oct. 25, 2024
Thermogenetics is a promising neuromodulation technique based on the use of heat-sensitive ion channels. However, way to its clinical application, number questions have be addressed. First, avoid immune response in future human applications, channels should studied as thermogenetic actuators. Second, heating levels necessary activate these vivo brain tissue and cytotoxicity temperatures Third, possibility safety chronic has demonstrated. In this study, we present comprehensive framework for using thermosensitive channel hTRPV1. By targeting hTRPV1 expression excitatory neurons mouse activating them within non-harmful temperature range with fiber-coupled infrared laser, not only induced neuronal firing stimulated locomotion mice, but also demonstrated that thermogenetics can employed repeated without causing evident injury. Our results lay foundation research therapy neuropathologies.
Language: Английский
Citations
0
bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown
Published: Nov. 1, 2024
The interaction of mammals with a novel environment (NE) results in structural and functional changes multiple brain areas, including the hippocampus. This experience-dependent circuit reorganization is driven part by gene expression however, dynamic sensory experience-driven chromatin states diverse cell type specific programs that are regulated experiences not well described. We employed single- nucleus multiomics (snRNA- ATAC-seq) bulk RNA-seq hippocampal DG, CA3, CA1 regions to characterize temporal evolution cell-type-specific accessibility occur 14 different types hippocampus upon exposure mice environment. observe strong regional specificity excitatory neuron as great diversity inhibitory non-neuronal transcriptional responses. environment-regulated genes each were enriched for encode secreted factors, their cognate receptors identified promising candidates modulation learning memory processes. Our characterization effect experience on revealed thousands accessibility. Coordinated analysis within individual Fos/AP-1 key driver experience-induced expression. Together, these data provide rich resource profiles across response experience, physiological stimulus affects memory.
Language: Английский
Citations
0