Objective To investigate the real-time biological effects of 2650 MHz radiofrequency radiation(RFR)on learning and memory as well as calcium activity patterns of hippocampal CA1 pyramidal neurons of mice.Methods Adult male C57BL/6N mice were randomly assigned to control(CON)and RFR groups before being exposed to 2650 MHz RFR for 3 hours in an electromagnetic reverberation chamber.Within 1 hour after exposure,changes in learning and memory were assessed using novel object recognition(NOR),object location recognition(OLR),and temporal order recognition(TOR)tests.Calcium activity of CA1 pyramidal neurons was monitored before,during,and after exposure using genetically encoded calcium imaging combined with fiber photometry.Neuronal activation was evaluated by c-Fos immunofluorescence staining.Results Compared with the CON group,RFR-exposed mice showed significantly reduced preference indices for target objects in the NOR(P<0.001),OLR(P<0.01),and TOR(P<0.01)tests within 1 hour post-exposure.During RFR exposure,neuronal calcium signals exhibited an abnormal high-frequency but low-amplitude pattern,characterized by increased event frequency(P<0.05)and decreased single-event amplitude(P<0.05).Furthermore,the immunofluorescence intensity of c-Fos in the hippocampal CA1 region was significantly reduced(P<0.05).Conclusion 2650 MHz radiofrequency radiation exposure can induce immediate effects,such as abnormalities in calcium signaling patterns and suppressed excitability of pyramidal neurons in the hippocampal CA1 region,ultimately leading to impaired learning and memory functions.

BACKGROUND:Electromagnetic field (EMF) exposure is increasingly common and has been implicated in a range of effects on human health. Conditioned fear memory plays a critical role in enabling organisms to respond appropriately to previously encountered threats. Despite growing interest in the neurobiological consequences of EMF exposure, its impact on the neural circuits underlying conditioned fear responses has not been clearly defined. METHODS:Using a mouse model exposed to combined microwave and static magnetic fields, we examined the involvement of the primary auditory cortex-basolateral amygdala (Au1-BLA) circuit in EMF-associated alterations in conditioned fear retrieval. A multifaceted experimental approach was employed, including behavioral assays, viral tracing, genetically encoded calcium imaging, chemogenetic modulation, histopathological analysis, and immunofluorescence. RESULTS:Exposure was associated with reduced conditioned fear memory retrieval, pathological changes in Au1 and BLA tissue ultra-structures, and decreased Nissl bodies in Au1 neurons and Au1-BLA neuronal fiber projections. The attenuation of conditioned fear memory retrieval coincided with decreased calcium activity in Au1 and BLA neurons. Consistently, chemogenetic activation of Au1 calcium-dependent protein kinase II (CaMKII)-expressing neurons enhanced calcium activity in BLA neurons during fear retrieval and was accompanied by changes in cholinergic signaling in the BLA. These findings suggest that cholinergic neuronal populations downstream of the Au1-BLA circuit are sensitive to EMF exposure and may participate in EMF-related modulation of fear retrieval. CONCLUSIONS:Our findings support an association between EMF exposure and altered conditioned fear expression involving functional changes within the Au1-BLA circuit, especially for the changes in calcium activity and chemogenetic modulation of Au1 CaMKII-expressing neurons. This study provides direct experimental evidence linking EMF exposure to circuit-level functional interactions underlying fear memory retrieval.

The potential health hazards caused by microwave exposure have attracted increasing attention. Microwave radiation has been reported to induce oxidative stress in neural tissues, which is considered one of the primary mechanisms underlying its adverse effects on central nervous system function. The hippocampus is sensitive to microwave radiation, whereas underlying cellular and molecular mechanisms remain incompletely understood. In this study, microwave-exposed mice exhibited significantly impaired performance in the Go/No-go, Y-maze, and novel object recognition tests at 6 h and 7 days post-exposure, indicating deficits in hippocampus-dependent working memory. Single-cell RNA sequencing of hippocampal tissues from control and microwave-exposed mice yielded 94,088 high-quality cells across eight major cell types. Astrocyte sub-clustering identified five transcriptionally distinct subpopulations, with Astrocyte_S100a6 and Astrocyte_Son proportions increased and Astrocyte_Serpinf1 decreased in the radiation group. Analysis of astrocyte transcriptional state transitions showed microwave-exposed astrocytes were preferentially distributed toward terminal reactive states with depletion at early homeostatic nodes. Cell-cell communication analysis revealed increased total interactions and interaction strength following radiation. Astrocyte outgoing signaling was increased for pathways associated with vascular remodeling, phagocytic regulation, and neuroinflammation, while pathways related to trophic support were decreased. Incoming signaling showed increased activity in pathways linked to phagocytic recruitment and inflammatory mediation. Taken together, these findings indicate that microwave exposure is associated with hippocampus-dependent working memory deficits accompanied by transcriptional remodeling of astrocyte subpopulation composition, directional astrocyte state transitions toward reactive phenotypes, and broad alterations in astrocyte-centered intercellular communication, providing a cellular and molecular framework for understanding astrocyte involvement in microwave radiation-associated hippocampal dysfunction.

Background: Acute lung injury (ALI) is characterized by excessive inflammation and oxidative stress, with macrophages playing pivotal roles in disease progression. The differential survival mechanisms of macrophage subsets during ALI remain poorly understood. Methods: We performed integrated analysis of ALI datasets (GSE11434, GSE237260, GSE264032) and isolated CD14 + macrophages from bronchoalveolar lavage fluid of ALI patients. Murine alveolar macrophages (MH-S) were polarized to M1/M2 phenotypes and subjected to ferroptosis induction. HIF-1 A knockdown and lactate supplementation experiments were performed to investigate metabolic regulation. Myeloid-specific HIF-1 A knockout mice were generated and subjected to LPS-induced ALI. Results: Transcriptomic analysis revealed ferroptosis as a key pathway in ALI, with macrophages showing the highest expression of ferroptosis-related genes. ALI-derived macrophages exhibited enhanced ferroptosis-defense gene expression and elevated lactate production. M1 macrophages demonstrated superior ferroptosis resistance compared to M2 macrophages, associated with higher baseline expression of antioxidant genes and enhanced HIF-1 A signaling. Mechanistically, HIF-1 A in M1 macrophages promoted lactate metabolism, which activated ferroptosis-defense genes through histone lactylation. Myeloid-specific HIF-1 A knockout prevented M1 macrophage survival advantage, reduced inflammatory tissue damage, and alleviated ALI severity. Conclusions: HIF-1 A-mediated lactate metabolism confers ferroptosis resistance in M1 macrophages through epigenetic activation of antioxidant defense genes. This metabolic reprogramming represents a novel therapeutic target for modulating macrophage survival and inflammation in ALI.

[Background]Co-exposure to noise and microwave radiation occurs frequently.The central ner-vous system has been identified as a sensitive target organ for both noise and microwave exposure individually,and the underlying mechanisms remain poorly understood.The specific biological effects resulting from co-exposure to these two factors have yet to be fully elucidated. [Objective]To clarify the effects of co-exposure to noise and microwave on neurobehavior and hippocampal tissue structure,and to explore the underlying mechanism through the assessment of serum cytokines. [Methods]C57BL/6N mice were selected and randomly assigned to a blank control group,a noise group,a microwave group,and a com-bined noise&microwave exposure group.To establish the exposure models,the noise group was subjected to broadband noise at 100 dB for 2 h,while the microwave group received radiation at a central frequency of 9.375 GHz with an average power density of 12 mW·cm-2 and a specific absorption rate of 2.58 W·kg-1 for 15 min.Open field and tail suspension tests assessed anxiety-like emotional behaviour;novel object recognition and Y-maze tests evaluated cognitive function.Histological changes in hippocampal tissue were ex-amined using haematoxylin and eosin(HE)staining,and Nissl staining under light microscopy.Serum cytokine levels were measured using radioimmunoassay and enzyme-linked immunosorbent assay(ELISA). [Results]After 3 d of exposure,the noise,microwave,and combined exposure groups showed significant reductions in exploration fre-quency,duration,and distance within the central zone of the open field test compared to the control group(P<0.01);the combined ex-posure group exhibited increased ratios of peripheral-to-central exploration time and distance(P<0.05).After 7 d of exposure,compared with the control group,the noise group maintained a decrease in central zone exploration time(P<0.01),while the combined exposure group showed persistent decline across all central zone metrics(P<0.05)and elevated peripheral-to-central ratios(P<0.05);compared to the microwave group,the combined exposure group showed significant less time in the central zone(P<0.05)and higher peripheral-to-central ratios(P<0.05).Regarding behaviour and cognition,compared with the control group,the combined exposure group showed increased immobility time in the tail suspension test after 3 d of exposure(P<0.01).At this interval,all exposure groups demonstrated reduced frequency and duration of novel object recognition(P<0.05),with the combined exposure group showing a marked decrease in novel arm exploration time(P<0.01).After 7 d of exposure,compared with the control group,the noise group showed reduced novel object recognition frequency(P<0.05),and both the noise and microwave groups exhibited decreased novel arm exploration time(P<0.05).Pathological alterations including an increased number of hyperchromatic nuclei and depleted Nissl bodies were observed in the CA3 and DG regions across all exposure groups with the most severe lesions observed in the combined exposure group.Serum levels of central nervous system-specific protein β(S-100β),glial fibrillary acidic protein(GFAP),and corticosterone(CORT)were significantly el-evated in all exposure groups compared with the control group(P<0.05).Aquaporin-4(AQP4)levels increased in the combined exposure group(P<0.05),while CXC chemokine ligand 10(CXCL10)levels rose in both the noise and combined groups compared with the control group(P<0.05).Specifically,S-100β and CXCL10 levels in the combined exposure group were higher than those in the microwave group(P<0.05);moreover,levels of S-100β,GFAP,CORT,AQP 4,and CXCL10 in the combined exposure group were significantly higher than those in the noise group(P<0.05). [Conclusion]Combined exposure to noise and microwave radiation induces pathological changes in the hippocampus of mice,increases levels of serum stress hormones and neuro-specific biomarkers.These impairments are more severe than those observed following single-factor exposure.The underlaying mechanism may be related to systemic stress response,neuronal damage,astrocyte activation,and changes in blood-brain barrier permeability,leading to emotional behavioral abnormalities and cognitive decline.

Endotoxemia represents a life-threatening clinical disorder driven by an aberrant host immune response to pathogenic infection, often resulting in severe multiple organ dysfunction. Among its most devastating complications are acute lung injury (ALI) and endotoxemia-associated encephalopathy (EAE), both of which are associated with elevated mortality and currently lack effective targeted interventions. This study evaluated the therapeutic efficacy and underlying molecular mechanisms of recombinant human thymosin β4 (rhTβ4) in a murine model of lipopolysaccharide (LPS)-induced endotoxemia. Our results showed that treatment with rhTβ4 markedly enhanced survival rates and diminished the systemic overproduction of diverse proinflammatory cytokines and chemokines in endotoxemic mice. These systemic protective actions were achieved through the inhibition of the TLR4/NF-κB signaling cascade, the reduction in M1 macrophage polarization, and the simultaneous alleviation of mitochondrial impairment and oxidative stress. Moreover, rhTβ4 treatment significantly rescued EAE-related cognitive deficits and attenuated neuronal damage, primarily through the suppression of neuroinflammation and microglial overactivation. Integrative transcriptomic profiling and functional assays identified lysophosphatidic acid receptor 3 (LPAR3) as an important contributor, suggesting that rhTβ4 suppresses microglial-mediated neurotoxicity at least in part through LPAR3 downregulation. In conclusion, rhTβ4 confers robust multi-organ protection against endotoxemic injury by orchestrating the inhibition of systemic and central neuroinflammatory cascades, positioning it as a promising candidate for the treatment of endotoxemia-induced ALI and EAE.

IntroductionChronic stress is known to exacerbate the malignant progression of colorectal cancer (CRC), a leading cause of cancer-related mortality worldwide. While outer membrane vesicles (OMVs) derived from Akkermansia muciniphila (A. muciniphila) exhibit anti-tumor potential, their underlying mechanisms in the context of stress remain unclear. This study investigated whether A. muciniphila OMVs could mitigate chronic stress-induced CRC progression by regulating the pro-tumorigenic protein Fetuin-A.Methods and resultsIn vitro, the β-adrenergic agonist isoproterenol (ISO), used to simulate chronic stress, significantly promoted CRC cell proliferation, migration, and invasion while inhibiting cellular uptake of OMVs. Notably, A. muciniphila OMVs effectively rescued the ISO-driven malignant progression in CRC cells by counteracting the upregulation of Fetuin-A. In vivo, using a CRC mouse model combined with chronic unpredictable mild stress, we found that OMV administration markedly suppressed tumor growth. This therapeutic effect was associated with a significant downregulation of Fetuin-A expression in tumor tissues.ConclusionCollectively, our findings demonstrate that A. muciniphila OMVs inhibit chronic stress-driven CRC progression by downregulating Fetuin-A. This study uncovers a novel regulatory mechanism within the “stress–Fetuin-A–CRC” axis and highlights A. muciniphila OMVs as a promising therapeutic strategy for managing stress-associated CRC.

Objective To investigate the transcription of neutrophils(Neu)expressing high levels of Csf3rhiCD14hiNeu in C3H/HeN mouse lung tissues before and after irradiation.Methods Twelve C3H/HeN mice were randomly divided into an irradiation group and a control group before being exposed to a single dose of 20 Gy chest irradiation.Twenty-four weeks after irradiation,two Neu subsets,Csf3rhiCD14hiNeu and Csf3rlowCD14hiNeu,were selected from lung tissues of irradiated and non-irradiated mice by flow cytometry.Transcriptomic sequencing(RNA-Seq)was performed on the two types of cells.Differentially expressed genes(DEGs)were identified using the DESeq2 package.Functional enrichment analysis and gene set enrichment analysis(GSEA)were performed using the gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)databases.Results At 24 weeks post-irradiation,the alveolar space was significantly reduced,collagen deposition increased,and fibrosis localized in C3H/HeN mice.RNA-seq analysis suggested that there were 1499 DEGs in Csf3rhiCD14hi Neu compared with Csf3rlowCD14hi Neu in the irradiated group,179 of which were upregulated and 1320 downregulated.In the control group,there were 701 significantly upregulated and 431 downregulated genes in Csf3rhiCD14hi Neu compared with Csf3rlowCD14hi Neu.Compared with non-irradiated Csf3rlowCD14hi Neu,there were 1319 upregulated and 219 downregulated genes in the irradiated group.Compared with non-irradiated Csf3rhiCD14hi Neu,the irradiated group had 202 upregulated and 540 downregulated genes.GO enrichment analysis indicated that these DEGs were involved in immune response,Wnt signaling pathway,and cytokine production.KEGG pathway analysis pointed to significant enrichment in the TGF-β signaling pathway,cytokine-cytokine receptor interactions,and Jak-STAT signaling pathway.GSEA revealed significant alterations in non-canonical NF-κB signaling,PD-1 signaling,and laminin interaction-related pathways.Conclusion Thoracic γ-ray irradiation can cause pulmonary fibrosis in C3H/HeN mice,with marked transcriptomic differences between Csf3rhiCD14ʰⁱand Csf3rlowCD14hi Neu subsets in lung tissue post-irradiation.

BACKGROUND:Radiation-induced colorectal fibrosis (RICF) is a chronic condition that can develop after pelvic radiation therapy for colorectal cancer. Adipose-derived mesenchymal stem cells (ADSCs) have emerged as promising candidate for fibrosis treatment, yet the mode of action of ADSC upon RICF remains obscure. This study aimed to investigate the optimal delivery route, treatment timing, anti-fibrotic effects, and underlying mechanisms of ADSCs upon RICF. METHODS:The RICF rat model was constructed by single dose of 20 Gy irradiation, and ADSCs were delivered via diverse ways (e.g., tail vein injection, abdominal aorta injection, peritoneal injection, or perianal tissue injection) at different frequencies (once, twice, or thrice a week) for 10 weeks. TMT-labelled proteomic and phosphoproteomic analysis was conducted for dissecting the underlying mechanisms of ADSCs upon RICF. ADSCs were co-cultured with primary human intestinal fibroblasts to verify the anti-fibrotic effects upon radiation-induced fibroblasts. Additionally, 4D label-free proteomic analysis and 4D-parallel reaction monitoring were carried out to explore their molecular mechanisms. RESULTS:RICF rats revealed better outcomes after intraperitoneal injection of ADSCs rather than the relative ways, and in particular, those with thrice-weekly injections showed effective prevention and improvement in RICF. Proteomic and phosphoproteomic analyses, together with multifaceted analyses (e.g., co-culture, 4D-PRM analysis), indicated Cytochrome b-245 alpha chain (Cyba) as a candidate target in mediating the efficacy of ADSCs upon RICF. CONCLUSIONS:This comprehensive multilevel proteomic study provides valuable insights into the molecular mechanisms underlying RICF and enhances understanding of the potential of ADSCs-based cytotherapy.

Excessive use of artificial light sources has led to a significant increase in light pollution, which has raised serious concerns due to its serious adverse effects on lipid metabolism. Although moderate static magnetic fields (SMFs) have shown potential in health intervention and treatment as non-invasive and highly permeable physical field, the influence of SMFs on lipid metabolic disturbance induced by lights remains largely unknown. In this study, we explored the lipid metabolism of Caenorhabditis elegans (C. elegans) under varying wavelengths of light ranging from 395 nm to 635 nm, both in the presence and absence of a 0.5 T SMF, and elucidated their underlying mechanisms. Exposure of C. elegans to artificial light at 200 lux resulted in a shortened lifespan while significantly increasing fat accumulation in a wavelength-dependent manner. The presence of 0.5 T SMF significantly extended the lifespan and reduced the size of fat droplets, as well as the content of triglyceride in light exposed worms. These effects were achieved by upregulating the expression of genes related to lipolysis and downregulating the expression of genes related to lipid synthesis. Moreover, the 0.5 T SMF alleviated abnormalities in lipid metabolism caused by light through the regulation of iron ions. Our findings provided clear evidence that moderate SMFs have significant protective effects on lipid metabolism abnormalities induced by artificial light via mediating iron homeostasis, which might contribute to a better understanding of the combined photomagnetic effects in living organisms.

Retrieval-extinction training based on the theory of memory reconsolidation has promising intervention effects for addiction. However, the conventional conditioned stimuli used in retrieval-extinction training have limitations in lack of contextual and selective activation of memories, which limits intervention efficacy and clinical translation. Therefore, we developed a novel imagery-based retrieval-extinction training (I-RE) and examined its effects on nicotine addiction. This study included 57 nicotine-dependent individuals randomly assigned to either the experimental (n = 29) or control (n = 28) group. Participants were exposed to a 5-min imagery script cue, followed by a 10-min rest period and 60-min extinction training session. Short- and long-term (1 week, 1 month, 3 months, 6 months, 12 months) intervention effects were assessed via the smoking imagery vividness score, smoking craving, and daily cigarette consumption. Electroencephalogram (EEG) data were collected pre- and post-intervention. Regarding short-term effects, smoking imagery vividness score [pre- vs. post-intervention: p < 0.001; pre- vs. 1-day follow-up (FU): p = 0.003] and craving significantly decreased (pre- vs. post-intervention: p < 0.001; pre- vs. 1-day FU: p < 0.001). Decreased imagery vividness score mediated decreased smoking craving induced by smoking-related I-RE. Moreover, the significant correlation observed between these variables at pre-intervention disappeared at post-intervention. For effects on EEG microstate, a significant decrease was observed in microstate C duration induced by the smoking-related imagery script cue reactivity task post-intervention (p < 0.001). This mediated a decreased smoking craving induced by smoking-related I-RE. Degree of decrease in duration was positively correlated with addict imagery ability (p = 0.035). Consistently, the microstate C occurrence rate significantly decreased during the memory reconsolidation phase (p < 0.001). Regarding long-term effects, the smoking imagery vividness score (1-week FU: p = 0.004; 1-month FU: p < 0.001), smoking craving (1-week FU: p < 0.001; 1-month FU: p < 0.001), and daily cigarette consumption (1-week FU: p < 0.001; 1-month FU: p < 0.001) significantly decreased at 1-week and 1-month FU. Furthermore, decreased smoking craving mediated decreased Daily cigarette consumption in the experimental group. The significant correlation observed between the imagery vividness score and craving at pre-intervention disappeared at the 1-week and 1-month FU. This novel I-RE demonstrated significant effects on nicotine addiction for 1 month after a single intervention session, suggesting that it is a promising treatment tool. Chinese Clinical Trial Registry identifier: ChiCTR2200064469.

Magnetic fields are widely used in medical diagnostics because of their superior non-invasive properties. In addition, with the widespread use of magnetic fields in transportation and other areas, their potential hazards to human health and the assessment of their safety have attracted considerable attention. The effects of magnetic fields on living organisms have a long history. The biological effects of magnetic field exposure in mice and rats depend on the magnetic field strength, exposure time, and direction; depending on these and potentially other factors, magnetic fields can cause a series of neurobiological effects. We reviewed global research on the neurobiological effects of magnetic fields from recent years to provide an overview and insights into the underlying mechanisms. This review focuses on the biological effects of static and dynamic magnetic fields of different frequencies and intensities on animals and nerve cells and their mechanisms of action.

Progress in understanding early human development has been impeded by the scarcity of reference datasets from natural embryos, particularly those with spatial information during crucial stages like gastrulation. We conducted high-resolution spatial transcriptomics profiling on 38,562 spots from 62 transverse sections of an intact Carnegie stage (CS) 8 human embryo. From this spatial transcriptomic dataset, we constructed a 3D model of the CS8 embryo, in which a range of cell subtypes are identified, based on gene expression patterns and positional register, along the anterior-posterior, medial-lateral, and dorsal-ventral axis in the embryo. We further characterized the lineage trajectories of embryonic and extra-embryonic tissues and associated regulons and the regionalization of signaling centers and signaling activities that underpin lineage progression and tissue patterning during gastrulation. Collectively, the findings of this study provide insights into gastrulation and post-gastrulation development of the human embryo.

In the original publication [...].

Introduction Ultra-high static magnetic fields (SMFs) have unique advantages in improving medical and academic research. However, the research on the early embryo exposure of ultra-high SMFs is minimal, extensive exploration is indispensable in living organisms. Objectives The present study was aimed to study the effects of ultra-high SMFs on the early embryonic division and development of Caenorhabditis elegans (C. elegans). Methods Early adult parents containing fertilized eggs in vivo were exposed to SMFs at intensities ranging from 4 T to 27 T. The number of mitotic cells in the reproductive glands of the P0 worms, early embryonic cell spindle localization, embryo hatching and the reproductive as well as developmental indicators of F1 and F2 nematodes were examined as endpoints. Results Our results indicated that ultra-high SMFs has no obvious effect on the germ cell cycle, while 14 T and 27 T SMFs significantly increased the proportion of multi-polar spindle formation in early embryonic cells, and reduced the developmental rate and lifespan of C. elegans exposed at the embryonic stage. Spindle abnormalities of early embryonic cells, as well as the down-regulation of genes related to asymmetric embryonic division and the abnormal expression of the non-muscle myosin NMY-2 in the division grooves played a critical role in the slowing down of embryonic development induced by ultra-high SMFs. Conclusions This study provided novel information and a new sight for evaluating the biosafety assessment by exposure to ultra-high SMFs at the early embryonic stage in vivo.

Macroautophagy/autophagy is a homeostatic process in response to multiple signaling, such as the lysosome-dependent recycling process of cellular components. Starvation-induced MTOR inactivation and PPP3/calcineurin activation were shown to promote the nuclear translocation of TFEB. However, the mechanisms via which signals from endomembrane damage are transmitted to activate PPP3/calcineurin and orchestrate autophagic responses remain unknown. This study aimed to show that autophagy regulator SMURF1 controlled TFEB nuclear import for transcriptional activation of the lysosomal biogenesis. We showed that blocking SMURF1 affected lysosomal biogenesis in response to lysosomal damage by preventing TFEB nuclear translocation. It revealed galectins recognized endolysosomal damage, and led to recruitment of SMURF1 and the PPP3/calcineurin apparatus on lysosomes. SMURF1 interacts with both LGALS3 and PPP3CB to form the LGALS3-SMURF1-PPP3/calcineurin complex. Importantly, this complex further stabilizes TFEB, thereby activating TFEB for lysosomal biogenesis. We determined that LLOMe-mediated TFEB nuclear import is dependent on SMURF1 under the condition of MTORC1 inhibition. In addition, SMURF1 is required for PPP3/calcineurin activity as a positive regulator of TFEB. SMURF1 controlled the phosphatase activity of the PPP3CB by promoting the dissociation of its autoinhibitory domain (AID) from its catalytic domain (CD). Overexpression of SMURF1 showed similar effects as the constitutive activation of PPP3CB. Thus, SMURF1, which bridges environmental stress with the core autophagosomal and autolysosomal machinery, interacted with endomembrane sensor LGALS3 and phosphatase PPP3CB to control TFEB activation.Abbreviations: ATG: autophagy-related; LLOMe: L-Leucyl-L-Leucine methyl ester; ML-SA1: mucolipin synthetic agonist 1; MTOR: mechanistic target of rapamycin kinase; PPP3CB: protein phosphatase 3 catalytic subunit beta; RPS6KB1/p70S6K: ribosomal protein S6 kinase B1; SMURF1: SMAD specific E3 ubiquitin protein ligase 1; TFEB: transcription factor EB.

Whether the nonthermal effects of radiofrequency radiation (RFR) exist and how nonthermal RFR acts on the nervous system are unknown. An animal model of spatial memory impairment is established by exposing mice to 2856-MHz RFR in the range of thermal noise (≤1 °C). Glutamate release in the dorsal hippocampus (dHPC) CA1 region is not significantly changed after radiofrequency exposure, whereas dopamine release is reduced. Importantly, RFR enhances glutamatergic CA1 pyramidal neuron calcium activity by nonthermal mechanisms, which recover to the basal level with RFR termination. Furthermore, suppressed dHPC dopamine release induced by radiofrequency exposure is due to decreased density of dopaminergic projections from the locus coeruleus to dHPC, and artificial activation of dopamine axon terminals or D1 receptors in dHPC CA1 improve memory damage in mice exposed to RFR. These findings indicate that nonthermal radiofrequency stimulation modulates ongoing neuronal activity and affects nervous system function at the neural circuit level.