Age and injury severity influence patient outcomes and recovery trajectories after traumatic brain injury (TBI). However, there is a paucity of understanding the effects of age and biomechanical load on molecular changes...Age and injury severity influence patient outcomes and recovery trajectories after traumatic brain injury (TBI). However, there is a paucity of understanding the effects of age and biomechanical load on molecular changes to the brain after pediatric TBI. We examined frontal lobe transcriptional changes and axonal injury 1 day after sagittal rapid non-impact head rotation (RNR) in newborn 3-5 day old piglets (N = 26) exposed to either Low (43.9 ± 6.3 rad/s) or High (150 ± 2.6 rad/s) loads compared to juvenile 4 week old piglets (N = 40) exposed to scaled High loads (124.3 ± 1.7 rad/s) and to age-matched Shams. In newborns, we identified 780 differentially expressed genes (DEGs) and 333 DEGs in the Low and High groups, respectively, with 91 of these overlapping. While overlapping DEGs and gene sets were consistent with stress, inflammation, BBB disruption, mitochondrial maintenance, extracellular matrix (ECM) and collagen degradation and formation, DEGs specific to the High group signaled damage and exacerbated immune response. Axonal injury volume was significantly larger in the High group than the Low and Sham newborn groups, and increased axon growth and neuronal dysfunction DEGs were also correlated with increased rotational loads. Importantly, age had a profound influence, despite mechanically equivalent rotational loads. Upregulated DEGs from newborn piglets were associated with endothelial cells, while juvenile piglet DEGs were associated with neurons. Additionally, sham newborns and juveniles displayed distinct microglia morphologies, and TBI-induced changes were observed only in the juveniles. In summary, we conclude that biological differences associated with developmental stage, rather than biomechanical load, dominate the tissues level response. In total, our data reveal the influence of rotational magnitude and emphasize distinct changes in gene expression, cell response, and tissue pathology in newborn versus juvenile brains after TBI.
The hippocampus is a principal brain region of adult neurogenesis and a key model for studying synaptic plasticity underlying learning and memory. Previously, our work suggested a role for the adaptor protein, PLEKHA2, i...The hippocampus is a principal brain region of adult neurogenesis and a key model for studying synaptic plasticity underlying learning and memory. Previously, our work suggested a role for the adaptor protein, PLEKHA2, in adult hippocampal neurogenesis; however, Plekha2 has been studied primarily in immune regulation, leaving its neural functions largely unexplored. To understand its functions in the brain, we generated a brain-specific Plekha2 conditional knockout (cKO) mouse model using CRISPR/Cas9 and Cre/loxP systems. Our findings showed that Plekha2 deletion resulted in reduced adult hippocampal neurogenesis, particularly impairing neuronal lineage entry and subsequent neuronal output. Electrophysiological recordings further revealed decreased excitability of dorsal, but not ventral, dentate granule cells, a region difference consistent with the higher endogenous Plekha2 expression in the dorsal hippocampus. Impaired synaptic plasticity, as indicated by a reduced paired-pulse ratio and attenuated long-term potentiation, was also observed. Behaviorally, Plekha2 cKO mice exhibited increased anxiety and impaired avoidance learning. To explore potential molecular mechanisms, RNA-sequencing identified down-regulated genes enriched for functions related to neuronal development, in line with the observed deficits in neurogenesis and synaptic functions, although the precise pathways remain unclear. Collectively, these findings establish Plekha2 as a critical regulator of adult hippocampal neurogenesis, synaptic plasticity, neuronal excitability, and behavior.
Brain protease-resistant misfolded proteins have been described in Alzheimer (AD), Parkinson (PD), Lewy Body (LBD), Amyotrophic Lateral Sclerosis (ALS), Progressive Supranuclear Palsy (PSP) and Creutzfeldt Jakob (CJD) di...Brain protease-resistant misfolded proteins have been described in Alzheimer (AD), Parkinson (PD), Lewy Body (LBD), Amyotrophic Lateral Sclerosis (ALS), Progressive Supranuclear Palsy (PSP) and Creutzfeldt Jakob (CJD) diseases. The role of free radicals in generating these protease resistant structures has been experimentally demonstrated in prion bovine spongiform encephalopathy (BSE), when manganese is substituted for copper (Cu), in bovine brain homogenates in reductive medium, while Cu protective effect against free radicals can be restored by Cu supplementation in oxidative medium. These facts can suggest a free radical-induced epimerization process in neuroprotein misfolding leading to the transformation of physiological L-amino acid brain proteins into abnormal D-structures which will be deposited in the brain as observed in neurodegenerative diseased brains. A blood Cu increase, not ceruloplasmin (CP) bound correlated with a Cu increase in the cerebrospinal fluid (CSF) and a Cu decrease in the brain have been described in AD, PD, ALS, or CJD. This indicates that following neuronal death, Cu might be expelled from brain proteins and subsequent to redistribution between brain, CSF and blood, it will result a brain Cu deficiency and a decrease in Cu brain protection against free radicals. In the aim of repairing this deficiency and slow down the neurodegenerative disease process, a brain Cu complexes vectorization through the blood-brain barrier might restore brain Cu homeostasis.
BACKGROUND: Postoperative delirium (POD) is a serious complication in elderly patients. Dexmedetomidine (DEX) can reduce POD incidence, but its neuroprotective mechanisms are unclear. This study investigated whether DEX...BACKGROUND: Postoperative delirium (POD) is a serious complication in elderly patients. Dexmedetomidine (DEX) can reduce POD incidence, but its neuroprotective mechanisms are unclear. This study investigated whether DEX alleviates POD-like phenotypes and whether hippocampal transcription factor EB (TFEB) contributes to this effect, with a focus on microglial lysosomal function. METHODS: An aged mouse POD model was established via laparotomy with visceral manipulation. Mice were assigned to Sham, POD, and POD+DEX groups. Behavioral tests assessed cognitive and affective functions. Hippocampal tissues were analyzed for neuroinflammation and lysosomal markers. Primary microglia were isolated for in vitro studies involving TFEB knockdown. RESULTS: DEX treatment significantly ameliorated POD-like behaviors and reduced hippocampal microglial activation and pro-inflammatory cytokine levels. DEX treatment was associated with increased lysosomal protein expression and promoted TFEB nuclear translocation in microglia, correlating with enhanced lysosomal gene transcription. In vitro, DEX increased lysosomal acidity and improved functional readouts in LPS-stimulated microglia in a TFEB-dependent manner, suggesting partial recovery of lysosome-dependent clearance. DEX also inhibited the NLRP3 inflammasome pathway, an effect substantially attenuated by TFEB knockdown. Mechanistically, DEX-mediated TFEB activation was associated with reduced mTOR activity. CONCLUSION: DEX mitigates POD-like phenotypes in aged mice, with evidence that TFEB activation in the hippocampus contributes to restoration of lysosomal function and attenuation of neuroinflammation. Microglia appear to be a key cellular compartment in this process. These findings suggest that enhancing lysosomal function may represent a potential target for POD.
m6A methylation is the most abundant modification in eukaryotic mRNA and has been implicated in epitranscriptomic regulation of various cellular functions. Recent studies have demonstrated its significance in brain devel...m6A methylation is the most abundant modification in eukaryotic mRNA and has been implicated in epitranscriptomic regulation of various cellular functions. Recent studies have demonstrated its significance in brain development, neuronal signalling and memory formation; however, the precise mechanisms by which m6A RNA methylation affects synaptic transmission and plasticity in memory-related neuronal circuits remain unclear. Here, we have studied the effects of newly developed pharmacological compounds that target m6A methylation on excitatory synaptic transmission and plasticity in the hippocampus, using a combination of electrophysiological and immunohistological techniques in infant and adult rats. We demonstrate that STM2457, a highly potent catalytic inhibitor of the m6A methyltransferase METTL3, blocks long-term potentiation (LTP) without affecting basal synaptic transmission in area CA1. Moreover, our findings support that LTP in vivo is associated with elevated m6A immunostaining, suggesting that LTP induction triggers METTL3 activation and a subsequent rise in m6A methylation. Interestingly, pharmacological activation of METTL3/14 or inhibition of the m6A demethylase FTO increased synaptic m6A levels in vivo, yet attenuated LTP in adult hippocampal slices. METTL3/14 activation also diminished long-term depression (LTD). These findings align with a model where elevated m6A methylation acts as a stabilizing signal, limiting excessive activity-dependent plasticity of synaptic transmission across development. Furthermore, they add to the growing body of evidence supporting that dysregulation of m6A RNA methylation perturbs synaptic plasticity - the neurobiological foundation of memory - and demonstrate that these processes can be pharmacologically targeted.
Hyperbaric oxygen (HBO) therapy can enhance motor recovery when initiated shortly after spinal cord injury (SCI). HBO can also induce a biphasic ventilatory response that includes hypo- and hyperventilation, but this has...Hyperbaric oxygen (HBO) therapy can enhance motor recovery when initiated shortly after spinal cord injury (SCI). HBO can also induce a biphasic ventilatory response that includes hypo- and hyperventilation, but this has not been evaluated after SCI. We studied adult Sprague-Dawley rats with cervical SCI to determine how an acute bout of HBO impacts diaphragm activation and to test the hypothesis that daily HBO treatment can accelerate recovery of diaphragm activation. Indwelling electromyogram (EMG) electrodes were placed in the mid-costal diaphragm prior to lateral mid-cervical contusion injury. Rats were treated with HBO (10 days, 1 h/day, 100% O, 3 atm, n = 9) or normobaric normoxia (NORM, n = 9) beginning the day of SCI. EMG was recorded daily in freely behaving rats before and during HBO. The diaphragm EMG inspiratory burst rate and peak amplitude (ipsi- and contralateral to SCI) decreased during HBO (p < 0.05) on days 3-10. Daily HBO increased baseline (recorded prior to HBO on days 3-10) ipsilateral "minute EMG" output (area under curve x rate; p = 0.038). RNAseq evaluation of ipsilateral hemidiaphragm tissues harvested, from a separate cohort (n=4 per group) at day 11 revealed SCI-induced changes in gene expression related to muscle structure and function, some of which were normalized by HBO treatment. We conclude that acute exposure to HBO reduces diaphragm activity after cervical SCI, and daily bouts of HBO may increase the rate of diaphragm recovery.
BACKGROUND: Postoperative delirium (POD) is a common and serious complication of surgery, driven in part by neuroinflammation mediated by microglial activation. However, the molecular mechanisms underlying this process r...BACKGROUND: Postoperative delirium (POD) is a common and serious complication of surgery, driven in part by neuroinflammation mediated by microglial activation. However, the molecular mechanisms underlying this process remain poorly defined. This study investigates the role of glucose transporter 1 (Glut1) in microglial activation and the pathogenesis of POD. METHODS: A mouse model of POD-like behaviour was established via partial hepatectomy. Microglia were depleted using PLX3397 and isolated to confirm their role, and that of Glut1, in POD pathogenesis. Glut1 function was inhibited pharmacologically with BAY-876, and its expression in microglia was modulated using microglia-specific adeno-associated viruses (AAV-mir-Glut1 and AAV-mir-Glut1). In vitro, BV2 microglial cells and BV2-HT22 neuron co-cultures were stimulated with lipopolysaccharide (LPS) to assess how inflammatory activation alters glucose metabolism and affects neuronal glucose uptake. Regulatory effects of microglial Glut1 were evaluated using PET-CT imaging, biodistribution analysis, histology, and biochemical assays. RESULTS: An increase in brain glucose metabolism was observed during POD-like behaviour, corresponding with microglial activation following anesthesia and surgery. Inhibition of Glut1 with BAY-876 reduced cerebral glucose uptake, suppressed microglial activation, and improved cognitive performance. Critically, microglia-specific modulation of Glut1 expression attenuated neuroinflammation, corrected metabolic abnormalities, and mitigated POD-like behaviour. CONCLUSIONS: Glut1-mediated glucose hypermetabolism in microglia contributes to POD through a metabolic-inflammatory cascade. These findings reveal a key role for microglial Glut1 in linking energy metabolism to neuroinflammation and suggest that targeting this pathway may offer a novel strategy for the prevention and treatment of POD and related perioperative neurocognitive disorders.
INTRODUCTION: Intraventricular hemorrhage (IVH) is a major contributor to acute brain injury and post-hemorrhagic hydrocephalus, especially in older adults. CD47 on erythrocytes delivers a "don't-eat-me" signal that inhi...INTRODUCTION: Intraventricular hemorrhage (IVH) is a major contributor to acute brain injury and post-hemorrhagic hydrocephalus, especially in older adults. CD47 on erythrocytes delivers a "don't-eat-me" signal that inhibits macrophage/microglia (M/MΦ) mediated phagocytosis, slowing hematoma resolution. While CD47 blockade enhances hematoma clearance after IVH in young animals and in intracerebral hemorrhage models, its therapeutic potential in aged IVH has not been examined. METHODS: Eighteen-month-old male Fischer 344 rats received intraventricular injections of autologous arterial blood mixed with either anti-CD47 antibody or isotype IgG. Saline-injected rats served as additional controls. MRI was performed post-IVH at 4 h, days 1 and 3 to quantify ventricular volume, T2* intraventricular hematoma volume, and T2* non-hypointense lesion volume (early hemolysis). Histological analyses assessed M/MΦ activation, choroid plexus and periventricular immune cell accumulation, ventricular wall injury, and hippocampal neuronal survival. RESULTS: CD47-blocking antibody markedly alleviated hemorrhage-induced hydrocephalus in aged rats by day 3 after IVH. Treatment with CD47 inhibition significantly reduced intraventricular T2* hematoma volume on both days 1 and 3, and decreased T2* non-hypointense lesion size (an indicator of early hematoma lysis) at both time points. On post-hemorrhage day 3, CD47 blockade robustly increased hematomal Iba1, CD68, and HO-1 M/MΦ infiltration. Enhanced Iba1 cell accumulation was also observed in the choroid plexus, white matter, and hippocampus, alongside increased CD68 cells in white matter. CD47 inhibition further attenuated ventricular wall damage on day 3 and reduced neuronal loss in the hippocampal CA1 region. CONCLUSION: Blocking erythrocyte CD47 facilitates hematoma resolution, reduces hemolysis and hydrocephalus, enhances M/MΦ recruitment across multiple anatomical routes, and mitigates ependymal damage and neuronal injury after IVH in aged rats. These findings support CD47 as a promising therapeutic target for IVH, particularly in the aging population.
The cranio-cervical junction (CCJ) represents a critical transitional zone between the cranium and cervical spine. It is speculated that changes of tissue compliance within the CCJ may disrupt normal cerebrospinal fluid...The cranio-cervical junction (CCJ) represents a critical transitional zone between the cranium and cervical spine. It is speculated that changes of tissue compliance within the CCJ may disrupt normal cerebrospinal fluid (CSF) flow, but this hypothesis has not been sufficiently validated. Therefore, this research study investigated the influence of tissue compliance changes at the CCJ on CSF dynamics and its potential role in related neurological disorders. A collagen fibrous hyperplasia model was established by injecting bleomycin (BLM) into the posterior atlanto-occipital interspace (PAOiS) in mice. After four weeks, the soft tissues of the PAOiS were harvested for histological and molecular analyses to verify the successful establishment of the animal model. A biomechanical investigation was performed to assess tissue compliance in the PAOiS. CSF pressure in the lateral ventricle was monitored to evaluate the influence of the tissue compliance changes on CSF dynamics. After BLM treatment, the deposition of collagen fibers and the expression levels of Col Iα1 and α-SMA in the PAOiS were significantly increased, which indicated that collagen fibrous hyperplasia model in mice was successfully established. Uniaxial tensile testing results revealed a significant increase of the elastic modulus in the PAOiS, indicating enhanced stiffness and reduced compliance of the PAOiS tissue. Furthermore, CSF pressure in the lateral ventricle was elevated following these changes in the PAOiS tissue. This study demonstrated that reduced tissue compliance caused by hyperplasia can disrupt normal CSF dynamics, offering new insights into the pathogenesis of neurological disorders associated with CSF dynamics.
Ischaemic stroke is a leading global cause of death and long-term disability. Current acute interventions, notably thrombolysis and mechanical thrombectomy, are limited by narrow therapeutic windows and primarily focus o...Ischaemic stroke is a leading global cause of death and long-term disability. Current acute interventions, notably thrombolysis and mechanical thrombectomy, are limited by narrow therapeutic windows and primarily focus on vascular recanalization. These approaches often fail to address the sustained neurovascular injury, persistent inflammation, and insufficient endogenous repair that underlie long-term functional impairment. In this context, mesenchymal stem cells (MSCs) have emerged as a highly promising regenerative strategy for ischemic stroke, targeting the multifaceted pathophysiology beyond acute revascularization. Sourced predominantly from bone marrow, umbilical cord, and adipose tissue, MSCs exert effects primarily through their multimodal paracrine actions. They home to the injury site and orchestrate the local immune microenvironment by dampening excessive inflammation and promoting a shift in macrophages toward a pro-repair phenotype. Furthermore, MSCs enhance neurovascular plasticity and tissue regeneration through the release of bioactive factors and extracellular vesicles. This review provides a comprehensive analysis of MSCs sources and their multimodal mechanisms of action, synthesizes preclinical evidence and clinical trial advancements in stroke, discusses persisting translational challenges, and outlines future directions in the field. Collectively, MSCs-based therapies represent a transformative approach with significant potential to bridge a critical gap in stroke care, advancing the goal of long-term tissue regeneration and meaningful functional recovery.
BACKGROUND: Emerging evidence highlights a bidirectional link between Alzheimer's disease (AD) and sleep disturbances. This study investigates whether early chronic sleep deprivation (CSD) exacerbates AD progression by i...BACKGROUND: Emerging evidence highlights a bidirectional link between Alzheimer's disease (AD) and sleep disturbances. This study investigates whether early chronic sleep deprivation (CSD) exacerbates AD progression by impairing the antioxidant transcription factor Nrf2 and promoting hippocampal ferroptosis. METHODS: 5xFAD transgenic mice underwent early CSD. Behavioral tests (Morris water maze, novel object recognition, open field) assessed cognition and anxiety. Histological and molecular analyses evaluated neuronal loss, phosphorylated tau (p-tau), oxidative stress markers (Fe, ROS, MDA, GSH), and expression/localization of Nrf2, Keap1, and ferroptosis-related proteins (GPX4, HO-1, ACSL4, SLC7A11). Interventions included Nrf2 knockout, AAV-mediated Nrf2 overexpression, melatonin, vitamin E (Vit.E), their combination, and the ferroptosis inhibitor liproxstatin1(Lip-1). Data were analyzed with t-tests and ANOVA (p < 0.05). RESULTS: Early CSD accelerated cognitive decline, hippocampal neuronal loss, and p-tau pathology in 5xFAD mice. CSD triggered Nrf2 depletion, suppressed its nuclear translocation, and downregulated GPX4 and HO-1, leading to oxidative stress and ferroptosis. Nrf2 knockout worsened these deficits. While melatonin or Lip-1 attenuated damage, combined melatonin and Vit.E most effectively reactivated Nrf2, inhibited ferroptosis, reduced p-tau, and restored cognitive function. CONCLUSIONS: Early CSD promotes AD pathogenesis via Nrf2 dysfunction, driving oxidative stress and ferroptosis in the hippocampus. Preemptive intervention targeting sleep and Nrf2 activation-particularly through combined melatonin and vitamin E-represents a promising strategy to delay neurodegeneration in at-risk individuals.
Deferoxamine (DFX), a classic iron-chelating agent, has exhibited neuroprotective effects in preclinical studies of hemorrhagic stroke. However, its clinical translation remains limited, and its precise mechanisms of act...Deferoxamine (DFX), a classic iron-chelating agent, has exhibited neuroprotective effects in preclinical studies of hemorrhagic stroke. However, its clinical translation remains limited, and its precise mechanisms of action in subarachnoid hemorrhage (SAH) have not been fully elucidated. This study aimed to investigate the therapeutic efficacy and underlying molecular mechanisms of DFX in a mouse model of SAH, with a particular focus on ferroptosis and astrocyte polarization. Our results demonstrated that DFX alleviated early brain injury (EBI) by reducing iron overload, oxidative stress, and ferroptosis induced by heme degradation. DFX treatment also inhibited the pathological activation of both pro-inflammatory A1 and anti-inflammatory A2 reactive astrocytes, but failed to fully suppress the overall neuroinflammatory response. Notably, biliverdin reductase A (BLVRA) was found to modulate inflammatory cytokine production via inducible nitric oxide synthase (NOS2) and TLR4 signaling, independent of astrocyte polarization. Consequently, combined treatment with DFX and siBLVRA exerted a synergistic therapeutic effect, resulting in significantly improved neurological outcomes relative to DFX monotherapy. In conclusion, DFX effectively mitigates iron-dependent ferroptosis and pathological astrocyte activation after SAH, but shows limited efficacy in controlling neuroinflammation. The enhanced anti-inflammatory effect achieved by BLVRA inhibition highlights a novel and promising therapeutic strategy that simultaneously targets iron overload and the BLVRA pathway for the treatment of SAH.
Alzheimer's disease (AD) is a major neurodegenerative disorder characterized by β-amyloid (Aβ) deposition and pathological tau phosphorylation and aggregation, frequently accompanied by chronic pain. Pain sensitization i...Alzheimer's disease (AD) is a major neurodegenerative disorder characterized by β-amyloid (Aβ) deposition and pathological tau phosphorylation and aggregation, frequently accompanied by chronic pain. Pain sensitization is closely linked to AD progression. Transient receptor potential vanilloid 1 (TRPV1), a key cation channel in pain transduction, is expressed not only in peripheral sensory neurons but also widely in central neurons and glial cells, where it contributes to pain sensitization and neuroinflammation. Emerging evidence indicates a bidirectional regulatory interplay between TRPV1 and tau, forming a "TRPV1-tau axis." This axis acts as a core molecular bridge connecting pain sensitization and AD pathology via calcium dyshomeostasis, mTOR/AMPK, PI3K/Akt/GSK3β, and neuroinflammatory pathways. TRPV1 overactivation promotes tau hyperphosphorylation and aggregation through calcium-dependent kinases, metabolic dysregulation, and inflammatory signaling. Conversely, pathological tau modulates TRPV1 expression and function via transcriptional regulation, protein interactions, and impaired axonal transport, establishing a deleterious feedback cycle. Therapeutic interventions targeting this axis, including TRPV1 modulators, tau-directed agents, anti-inflammatory drugs, and natural compounds, demonstrate potential in alleviating both pain sensitivity and cognitive deficits through multi-target mechanisms. However, clinical translation remains challenging due to issues including blood-brain barrier penetration, target selectivity, and a lack of reliable biomarkers. This review systematically outlines the bidirectional mechanisms of the TRPV1-tau axis, current intervention strategies, and translational challenges, with the goal of informing future development of disease-modifying therapies that concurrently address cognitive decline and chronic pain in AD.
Hydrocephalus is a common neurological disorder characterized by pathological dilation of the ventricular system. Its pathogenesis involves multiple factors, including cerebrospinal fluid (CSF) dynamic imbalance (encompa...Hydrocephalus is a common neurological disorder characterized by pathological dilation of the ventricular system. Its pathogenesis involves multiple factors, including cerebrospinal fluid (CSF) dynamic imbalance (encompassing overproduction, circulation obstruction, and impaired absorption) and neuroinflammatory responses. Astrocytes, as core components of the neurovascular unit, mediate CSF transport via their specialized end-foot structures and the glymphatic system formed by perivascular spaces (PVS). Simultaneously, they serve as key effector cells in neuroinflammatory regulation, participating in various disease processes. This article systematically reviews the role of astrocytes in the pathogenesis and progression of hydrocephalus, with a focus on their molecular mechanisms in CSF circulation disorders and neuroinflammatory responses. The aim is to provide novel insights for targeted therapies against hydrocephalus.
Lanthionine synthetase C-like protein (Lancl) gene family members 1-3 play roles in responses to oxidative stress and cellular metabolic processes. Lancl1 in particular functions as a neuroprotective factor in the centra...Lanthionine synthetase C-like protein (Lancl) gene family members 1-3 play roles in responses to oxidative stress and cellular metabolic processes. Lancl1 in particular functions as a neuroprotective factor in the central nervous system (CNS), but it is unknown whether experimentally upregulating Lancl1 in neurons could also repair injured circuits by regenerating damaged axons. We recently identified Lancl1 as a gene upregulated in response to an axon-regenerating treatment, which prompted us to ask here if Lancl1 could directly repair injured axons. First, we characterized expression of Lancl1-3 family genes in single-cell RNA-seq (scRNA-seq)-profiled retinal ganglion cell (RGC) CNS projection neurons, then we investigated through gain-of-function experiments whether Lancl1 promotes axon regeneration after CNS injury in vivo, and finally we tested if Lancl1 transgene activates the mTOR pathway's pS6 marker, which is associated with Pten knockout-promoted axon regeneration. We show that within the retina, Lancl1 and Lancl2 are enriched in the RGCs, whereas Lancl3 is not considerably expressed in retinal cell types. Within the RGCs, there is moderate subtype-to-subtype variability in Lancl1 and Lancl2 expression, whereas Lancl3 is enriched in αRGCs but expressed at modest levels. We then found that after optic nerve injury, Lancl1 transgene targeted to RGCs through intravitreal AAV2 promotes neuroprotection and axon regeneration, with axons extending through the full-length of the optic nerve when co-treated with a fibronectin-based recombinant small protein. However, we did not find that Lancl1 transgene activates the mTOR pathway's pS6 marker in injured RGCs. Thus, Lancl1 is a novel axon regeneration-promoting factor with a therapeutic potential for treating CNS injury and disease, and future studies need to investigate downstream mechanisms of its neurotherapeutic efficacy.