GABAA Receptor Subunits Research Articles

Transcriptomic and electrophysiological alterations underlying phenotypic variability in SCN1A-associated febrile seizures

Febrile seizures (FS) are a common childhood neurological condition triggered by fever in children without prior neurological disorders. While generally benign, some individuals, particularly those with complex FS or genetic predispositions, may develop epilepsy or other neurological comorbidities. The mechanisms underlying this transition remain unclear. Mutations in SCN1A, encoding the NaV1.1 sodium channel α-subunit, have been linked to several epilepsy syndromes associated with FS. This study examines phenotypic variability in individuals carrying the same SCN1A c.434T > C mutation, using induced pluripotent stem cell (iPSC)-derived neurons from two siblings with FS. Despite sharing the mutation, only the older sibling developed temporal lobe epilepsy (TLE). Transcriptomic analysis revealed downregulation of GABAergic pathway genes in both siblings’ neurons, aligning with SCN1A-associated epilepsy. However, neurons from the sibling with TLE exhibited additional abnormalities, including altered AMPA receptor subunit composition, changes in GABAA receptor subunits and chloride cotransporters expression, and reduced brain-derived neurotrophic factor (BDNF) levels, indicative of developmental immaturity. Voltage-clamp recordings confirmed impaired GABAergic and AMPA receptor-mediated synaptic activity. These findings suggest that combined GABAergic dysfunction, aberrant AMPA receptor composition, and reduced BDNF signaling contribute to the more severe phenotype and increased epilepsy susceptibility.

Effects of acamprosate on alprazolam-induced conditioned place preference in male rats: The role of GABA and NMDA receptor subunits.

Effects of acamprosate on alprazolam-induced conditioned place preference in male rats: The role of GABA and NMDA receptor subunits.

Increased GABAA receptor open probability: Adaptive mechanisms to cope with anoxia in the painted turtle.

Increased GABAA receptor open probability: Adaptive mechanisms to cope with anoxia in the painted turtle.

Bifenazate as compatible acaricide for biocontrol agent Transeius montdorensis (Acari: Phytoseiidae).

Integrated pest management strategies often include the use of pesticides that have minimal impact on biological control agents. The predatory mite Transeius montdorensis is a valuable agent for controlling whiteflies, thrips and spider mites, but its tolerance to commonly used pesticides is not well understood. This study aims to investigate the genetic basis of pesticide tolerance in Transeius montdorensis. Through genome sequencing, we examined 12 key pesticide target genes in Transeius montdorensis, including those related to acetylcholinesterase and gamma-aminobutyric acid (GABA) receptors. Our results show that Transeius montdorensis exhibits natural tolerance to bifenazate owing to the presence of the asparagine (N) residue at position 252 in cytochrome b, similar to resistance mechanisms observed in the two-spotted spider mite. In addition, we found multiple mutations in GABA receptor subunits, suggesting fipronil as a compatible pesticide for controlling thrips and whiteflies. This study provides critical genomic insights into pesticide resistance in Transeius montdorensis, advancing our understanding of its compatibility with various pesticides. These findings have significant implications for enhancing integrated pest management programmes by ensuring the effective use of Transeius montdorensis as a biological control agent without compromising its efficacy through pesticide exposure. © 2025 Society of Chemical Industry.

MicroRNA-502-3p Modulates the GABA A Subunits, Synaptic Proteins, and Mitochondrial Morphology in Hippocampal Neurons.

MicroRNA-502-3p (MiR-502-3p), a synapse-enriched miRNA, is significantly implicated in Alzheimer's disease (AD). Our previous study revealed a high expression level of miR-502-3p in AD synapses relative to controls. Additionally, miR-502-3p was found to modulate the GABAergic synapse function by modulating the GABA A receptor subunit α-1 (GABRA1) protein. The current study aims to investigate the impact of miR-502-3p on other GABA receptor subunit proteins, synaptic proteins, mitochondrial morphology, and other hippocampal neuron genes. Mouse hippocampal neuronal (HT22) cells were transfected with miR-502-3p overexpression (OE) vector, miR-502-3p suppression (sponge) vector, or scramble control vector. Transfection of miR-502-3p vectors was confirmed by fluorescence microscopy. MiR-502-3p and Gabra1 expressions were confirmed by qRT-PCR and RNAscope-based in situ hybridization analysis. GABA A subunits and synaptic protein levels were analyzed by immunoblotting, and mitochondrial morphology was examined by transmission electron microscopy. Additionally, Affymetrix gene array analysis was performed on miR-502-3p overexpressed and suppressed cells. Our results demonstrate that elevated levels of miR-502-3p negatively regulate the Gabra1 expression. The levels of GABA A subunit and synaptic proteins were reduced upon ectopic expression of miR-502-3p and increased upon miR-502-3p suppression. Mitochondrial morphology was improved in terms of mitochondrial number, length, and mitochondrial area in miR-502-3p suppressed cells. Furthermore, gene array analysis unveiled the modulatory effects of miR-502-3p on several specific genes, especially those that are associated with oxidative stress, immune response, and synaptic function. These findings provide a new insight into the molecular mechanism of miR-502-3p in regulating neuronal function and synaptic activity.

GABRA2-related encephalopathy: Identification of two phenotypes with distinctive electroclinical features.

Pathogenic variants in γ-aminobutyric acid type A (GABAA) receptor subunit genes are increasingly associated with epilepsy and neurodevelopmental disorders. Pathogenic variants in GABRA2, encoding the α-2 subunit of GABAA receptors, have been recently reported. This study aims to better delineate the phenotypic spectrum of GABRA2 pathogenic variants. We conducted a retrospective multicenter study, analyzing six new patients with GABRA2 pathogenic variants identified through a French national collaboration. Clinical, electroencephalographic (EEG), and genetic data were reviewed alongside a literature analysis of eight previously reported cases. Two distinct electroclinical phenotypes were identified. The most severe, in four of six patients, featured early infantile developmental and epileptic encephalopathy with an EEG pattern of rapid rhythms suggestive of GABAergic hyperactivity. The milder phenotype, in two of six patients, included later onset, drug-responsive epilepsy with moderate developmental delay. A literature review confirmed these phenotypes and supported genotype-phenotype correlations, with transmembrane domain variants more frequently associated with severe phenotypes. This study refines the phenotypic spectrum of GABRA2-related disorders, highlighting two distinct electroclinical phenotypes. The identification of a recognizable EEG pattern of unusual rapid rhythms for age may be a biomarker for early diagnosis of a severe phenotype and suggests a potential underlying gain-of-function mechanism, to be confirmed by functional studies.

Identification and Classification of Position-specific GABAA Receptor Subunit Missense Variants for Their Role In Hippocampal Pyramidal Neurons.

Understanding the effects of functionally unknown variants in epilepsy associated genes is crucial for elucidating disease pathophysiology and developing personalized therapeutics. With a multiscale framework, spanning from DNA sequence to protein function and neural behavior, we describe a novel approach for predicting and investigating pathogenic mutations, hypothesizing that epileptogenic mutations in the GABAA receptor subunit and nearby predicted mutations may produce similar effects on the CA1 pyramidal neuron model. By exploring the characteristic relationships between predicted pathogenic mutations and proximal epileptogenic mutations, the study aims to estimate the effects of predicted mutations based on the effects of epileptogenic mutations on hippocampal pyramidal neuron simulations. The methodology begins with the collection of GABAA receptor γ2 subunit genetic data, followed by data cleaning and formatting performed in R using a custom script. Next, ensemble predictors will be applied to identify and prioritize the pathogenic missense variants of the γ2 subunit. Mapping a specific pathogenic variant (predicted) to the subunit structural domains shared by epileptogenic mutations will be illustrated, accompanied by molecular modeling of their effects and consideration of evolutionary conservation. Then, variant-specific meta-analysis and parameter normalization will be performed, followed by correlation analysis to identify any significant relationships between predicted mutations and proximal epileptogenic mutations. Using a Python-based neural simulator, multi-compartmental conductance-based neuron model, reflecting the effect of wild-type and epileptogenic mutants will be described. Simulation of neural responses generated by epileptogenic GABAA receptor subtype will be considered for the rough estimation of the predicted pathogenic variants' effect on neural response. To our knowledge, this is the first protocol exploring a multiscale framework to estimate the effects of GABAA receptor variants on neuronal behavior, crucial for epilepsy research. This protocol can serve as a foundation for enhancing predictions of cellular phenotypes caused by potentially pathogenic variants of GABAA receptors associated with epilepsy.

The genetic and phenotypic spectrum of GABRB1-related disorders.

Pathogenic variants in GABAA receptor subunits genes (GABR*) are important contributors to rare and common genetic epilepsies. Here, we present a comprehensive analysis of variants in GABRB1, which encodes the GABAA receptor β1 subunit, by revealing their functional implications, establishing genotype-phenotype correlations, and evaluating treatment response. Clinical information on individuals carrying a GABRB1 variant was obtained through an international collaboration and literature review. Our cohort included 19 individuals (7 males, 12 females) from 15 families harboring 13 different GABRB1 variants (11 missense, 1 indel, 1 stop). Functional analysis was performed using two-electrode voltage-clamp recordings in Xenopus laevis oocytes. For all eleven missense variants, α1β1γ2 GABAA receptors with a single mutant β1 subunit were used. Four missense variants were selected for further functional analysis using α5β1γ2 GABAA receptors with two mutant β1 subunits. Gain-of-function (GoF) effects, characterised by increased GABA-sensitivity, were observed for eight missense variants. Loss-of-function (LoF) effects were observed for one, and no functional effects for two variants. Clinically, GoF variants were only observed in individuals with severe early-onset disease including profound intellectual disability, hypotonia, and early mortality. Additionally, cortical visual impairment, dysmorphisms and cortical atrophy were exclusive to this cohort. By integrating previously reported clinical data for variants in other GABR* genes we validated that these features were associated with GoF variants more broadly. The only LoF variant was identified in a nuclear family with the relatively milder syndrome of genetic epilepsy with febrile seizures plus. Seizures were therapy-resistant in all individuals with GoF variants, and in a single individual with a LoF variant. The GABAergic anti-seizure medication (ASM) vigabatrin caused life-threatening side-effects in two individuals with GoF variants, while the sodium-channel blocker (SCB) lamotrigine exacerbated seizures in a single individual carrying a LoF variant. By integrating data from literature on all GABR* variants, we observed a potential dichotomy in treatment responses: GABAergic and broad-spectrum ASMs, such as valproate and levetiracetam, were more effective for individuals with LoF variants in GABR* genes, while SCBs showed greater benefit for GoF variants. Additionally, there is an increased risk of adverse effects of SCBs in LoF and vigabatrin in GoF variants. Our results highlight the importance of functional characterisation of variants and clinical predictors in guiding treatment strategies for individuals with GABRB1 and other GABR* variants, though larger prospective studies are needed to confirm these observations.

Zolpidem for Catatonia Refractory to Benzodiazepines in Resource-Limited Settings

Aims: Catatonia is a neuropsychiatric syndrome characterized by a paucity of movement and speech. Benzodiazepines are the mainstay of treatment for catatonia, but a subset of patients do not respond. While electroconvulsive therapy is another treatment option for catatonia, access can be limited. This case report discusses the use of zolpidem, a hypnotic non-benzodiazepine GABA-A receptor modulator, in a patient with benzodiazepine-resistant catatonia.Methods: A 19-year-old male presented to a hospital in the United States from jail custody with altered mental status. His medical history was notable for psychosis with paranoid delusions and severe catatonia.The patient reportedly had severely reduced oral intake for the past month, with almost no intake at all in the prior eight days. His presentation, which included psychomotor retardation, mutism, and posturing, raised a high clinical suspicion for catatonia.Initial treatment with 1 mg of lorazepam every 6 hours was ineffective, and the patient developed tachycardia, raising concern for malignant catatonia. At this time, his differential diagnosis included schizophrenia. His treatment regimen was adjusted to include olanzapine for suspected schizophrenia, memantine for catatonia treatment augmentation, and metoprolol for tachycardia. Despite this regimen, along with escalating lorazepam doses up to 7 mg three times daily, the patient remained severely catatonic. A trial of clozapine also failed to yield significant improvement.Given the patient’s limited response to benzodiazepines, zolpidem was introduced. He showed rapid improvement in his catatonic symptoms, including markedly improved speech, oral intake, and overall participation. Zolpidem was dosed throughout the day to limit drowsiness. By day 30, the patient demonstrated substantial recovery, with minimal catatonic symptoms and improved engagement in daily activities. He was discharged home on a regimen of zolpidem, clozapine, and memantine.Results: A subset of patients with catatonia fail to respond to benzodiazepines, necessitating alternative treatments. Prompt intervention is crucial due to the life-threatening nature of catatonia and its numerous complications. While electroconvulsive therapy is often effective, its availability can be limited. Zolpidem, acting through a distinct mechanism of GABA-A receptor subunit binding, may offer an effective alternative for benzodiazepine-resistant cases.Conclusion: Further research into zolpidem and other alternative therapies for catatonia is warranted, especially in settings where electroconvulsive therapy is not accessible. Zolpidem’s potential as a treatment for benzodiazepine-refractory catatonia deserves further investigation.

A CRISPR/Cas9-induced point mutation on the GABA receptor subunit RDL confers high resistance to phenylpyrazole insecticides in the rice planthopper Laodelphax striatellus.

A CRISPR/Cas9-induced point mutation on the GABA receptor subunit RDL confers high resistance to phenylpyrazole insecticides in the rice planthopper Laodelphax striatellus.

Update on inherited disorders of GABA metabolism.

Update on inherited disorders of GABA metabolism.

Regulation of neutrophil migration in acute pulmonary inflammation by extraneuronal α1 gamma-aminobutyric acidA receptors

Acute pulmonary inflammation is a common disease on intensive care. Due to the lack of specific treatments, lethality is still very high. The ionotropic GABAA-receptors are known from the central nervous system (CNS) and have recently been detected in the lung. These receptors have been shown to influence inflammatory processes. Opposing data has been reported, where both, GABA site agonists and antagonists achieved anti-inflammatory effects. The distribution of the 19 known GABAA-receptor subunits (α1-6, β1-3, γ1-3, δ, ε, π, θ and ρ1-3) and their role in inflammation remain unclear. In murine models of LPS- and bacteria-induced inflammation, Muscimol (GABAA-receptor agonist) and Bicuculline (antagonist) were administered before the onset of inflammation. Transcription of GABAA-receptor subunits was evaluated by real-time polymerase chain reaction. Neutrophil counts and adhesion molecule expression in wild type and GABAα1 knock-in mice were determined by flow-cytometry. Myeloperoxidase, neutrophil extracellular traps and cytokines were determined. In a model of ventilator-induced lung injury, blood gas analysis was performed using arterial blood. A multiplex western blot was done to assess signaling proteins. Muscimol and Bicuculline inhibited neutrophil influx in the bronchoalveolar lavage but did not change neutrophil activation. Both altered surface expression of adhesion molecules on neutrophils and reduced release of interleukin-6 (IL-6). The increased α1 subunit expression on lung epithelium and endothelium after inflammation was abolished by Muscimol and Bicuculline. In GABAα1-knock-in mice the protective effects of both agents were no longer observed. Only Muscimol lowered protein extravasation, improved blood gas analysis and lung function. A multiplex assay ascribed these anti-inflammatory effects to the influence of the IL-6 and phosphoinositide 3-kinase signaling pathways. In conclusion, Muscimol and Bicuculline exert various protective effects in two murine models of acute pulmonary inflammation. The multiple effects of Muscimol were linked to the inhibition of the proinflammatory signaling pathways IL-6 and PI3K.

Altered GABAergic Homeostasis in the Striatum of Dopamine Transporter Knockout Rats.

It is now widely established that dopamine, despite its nature as a slowacting biogenic monoamine, modulates fast neurotransmitters such as GABA. However, the mechanism through which this occurs still needs to be fully elucidated. The dopamine transporter (DAT) is the primary regulator of dopamine homeostasis, controlling extracellular levels of dopamine as well as its storage in vesicles. Here, we took advantage of the availability of dopamine transporter knockout (DAT-/-) rats, which provide a unique opportunity to investigate the response of the GABAergic system under hyperactivity of the dopaminergic system, a condition found in different disorders of the Central Nervous System. The expression levels of GABAergic markers have been evaluated by means of western blot in the whole homogenate, cytosolic fraction, and post-synaptic density of the striatum of male DAT-/- rats. We found a widespread down-regulation of GABAergic markers in the striatum of DAT-/- rats. Our data show that DA overactivity critically reorganizes the striatal GABAergic synapse in a way that GABA neurotransmission appears to be toned down. Such changes are equally distributed among proteins regulating GABA synthesis (GAD67), release (vGAT) and reuptake (GAT1, GAT3). It also involve the main subunits of GABA receptors (GABA-A a1, a2, b1; GABA-B R1), their anchoring proteins (Gephyrin) and adhesion molecules (Neuroligin-2). Taken together, such changes paint a picture showing a compromised integrity of the striatal GABAergic system under conditions of functional hyperdopaminergia, which may be of interest for several disorders of the central nervous system.

Early signs of neuron autonomous and non-autonomous hyperexcitability in locus coeruleus noradrenergic neurons of a mouse model of tauopathy and Alzheimer's disease.

The locus coeruleus (LC) is one of the earliest brain regions affected by phosphorylated tau (p-tau) in Alzheimer's disease (AD). Using the P301S mouse model, we investigated the temporal progression of tau pathology and its functional consequences. Immunohistochemistry was used to assess p-tau deposition in LC noradrenergic neurons at 2-3 and 5-6 months. Electrophysiological recordings evaluated neuronal hyperexcitability, measuring membrane potential, rheobase, and spontaneous action potential (AP) frequency in P301S and wild-type (WT) mice. Fast-scan cyclic voltammetry (FSCV) was used to measure norepinephrine (NE) release. GABA(A) receptor subunit expression was analyzed via immunoblotting. P-tau was detected in LC neurons as early as 2-3 months, with a rostral-to-caudal gradient, and by 5-6 months, nearly all LC neurons exhibited p-tau immunoreactivity. P301S neurons showed hyperexcitability, characterized by depolarized membrane potentials, a more negative rheobase, and increased spontaneous AP frequency. Synaptic blockade elicited a reduced increase in AP frequency, suggesting diminished inhibitory tone. GABA(A) α2 subunit expression significantly declined with age in P301S mice, whereas α3 remained unchanged. FSCV showed significantly elevated NE release in P301S mice at 3 and 6 months compared to WT. The findings highlight early LC dysfunction in tauopathies, characterized by increased excitability, reduced inhibitory tone, and exaggerated NE release. This hyperactivity may contribute to excitotoxicity and downstream dysfunction in LC-regulated brain regions. Targeting LC hyperactivity and restoring inhibitory signaling could be promising therapeutic strategies for mitigating AD progression.

Dysregulation of miR‑24‑3p and miR‑186‑5p and GABAA receptor expression in focal cortical dysplasia.

Spontaneous synaptic activity mediated by GABAA receptor is associated with epileptogenicity in focal cortical dysplasia (FCD). miRNAs are potentially involved in the regulation of GABAA receptor subunit expression and activity. This study aimed to determine the expression of miRNAs in FCD and correlate their expression level with mRNA levels of GABAA receptor subunits. Expression of GABAA receptor subunits (α1 and α4) and miRNAs (miR‑155‑5p, miR‑186‑5p, and miR‑24‑3p) were evaluated using real‑time PCR in resected brain samples from FCD patients. miRNA levels were also determined in the serum of FCD patients. Spontaneous GABAA receptor‑mediated synaptic activity was measured using patch clamp technique. Significant increase in α1 and α4 subunit expression and miR‑155‑5p levels, while decrease in miR‑24‑3p and miR‑186‑5p levels, was observed in the brain samples of FCD. In the serum of FCD patients, miR‑155‑5p levels were increased, whereas miR‑24‑3p and miR‑186‑5p levels remained unaltered. Increased α4 subunit expression in FCD might be due to reduced levels of miR‑24‑3p and miR‑186‑5p. In addition, reduced miR‑186‑5p levels might be responsible for increased expression of α1 subunit. We also observed an increase in the spontaneous GABAA receptor‑mediated synaptic transmission in FCD. In conclusion, dysregulation of miRNAs and GABAA receptor expression suggest that these miRNAs may contribute to altered GABAA receptor‑mediated synaptic activity in FCD.

Anxiogenic-like effects of coumarin, possibly through the GABAkine interaction pathway: Animal studies with in silico approaches.

Anxiogenic-like effects of coumarin, possibly through the GABAkine interaction pathway: Animal studies with in silico approaches.

PRECLINICAL EVIDENCE SUPPORTING THE ROLE OF ALOGABAT, A GABAA-α5 POSITIVE ALLOSTERIC MODULATOR, AS A NOVEL THERAPEUTIC APPROACH FOR NEURODEVELOPMENTAL DISORDERS

BackgroundThere is compelling evidence that dysfunction of the GABAergic system, the main inhibitory neurotransmitter system in the brain, contributes to the pathophysiology of neurodevelopmental disorders including autism spectrum disorder and deletion Angelman syndrome (1, 2, 3, 4). Genetic studies highlight the critical role of GABAA receptor gene dosage and function within the 15q11-13 chromosomal region in the etiology of these conditions (5, 6, 7, 8). This region encompasses several genes including UBE3A and GABRB3, GABRA5, GABRG3 encoding the β3, α5, and g3 GABAA receptor subunits, respectively. These subunits together with the g2 subunit co-assemble to form the GABAA- α5 receptor subtype (9). Therefore, positive modulation of GABAA-α5 receptors may provide a novel therapeutic approach to restore deficient GABAergic signaling without the typical side effects of non- selective GABAA positive allosteric modulators (PAM) i.e., diazepam.Aims & ObjectivesAssess the in vitro and in vivo pharmacological profile of alogabat, a novel small molecule, selective GABAA-α5 receptor positive allosteric modulator. Provide preclinical proof of concept data to support clinical studies in ASD and Angelman syndrome.MethodsAlogabat was evaluated for selective binding and functional activity in vitro at GABAA-α5β3g2 receptors, including electrophysiological studies in hippocampal slices. In vivo studies included receptor occupancy (RO) using a selective GABAA-α5 tracer (autoradiography), pharmacological magnetic resonance imaging (phMRI) and EEG in rodents. Alogabat was assessed on the repetitive behavior phenotype in BTBR and contactin-associated protein-like 2 knockout (Cntnap2-/-) mice, on seizure models and cognitive performance in rats, and on rotarod performance following a combination treatment with diazepam.ResultsAlogabat is a potent PAM of the GABAA-α5 receptor with binding and functional selectivity. RO studies provided direct proof of dose-dependent target engagement. Functional circuit modulation was demonstrated by dose-dependent regional perfusion changes in phMRI and change in EEG theta and beta band power. Alogabat at >50% RO at GABAA-α5 normalized elevated self-grooming in both Cntnap2-/- and BTBR mice and exhibited antiepileptic activity in rats. Alogabat did not impair cognition in rats at RO up to 88%, although impairment occurred at higher doses probably due to decreased α-subunit selectivity. Alogabat did not worsen diazepam-induced rotarod impairment.ConclusionsAlogabat showed beneficial effects in mouse models relevant for NDD, as well as anti- seizure activity, at doses without cognitive, sedative, and motoric side effects. In addition, translational biomarkers were identified to guide clinical studies: PET, phMRI, EEG.

Serotonin neurons integrate GABA and dopamine inputs to regulate meal initiation.

Serotonin neurons integrate GABA and dopamine inputs to regulate meal initiation.

Multifactorial approach is needed to unravel the maturation phases of human neurons derived from induced pluripotent stem cells

Neurons derived from induced pluripotent stem cells (h-iPSC-Ns) provide an invaluable model for studying the physiological aspects of human neuronal development under healthy and pathological conditions. However, multiple studies have demonstrated that h-iPSC-Ns exhibit a high degree of functional and epigenetic diversity. Due to the imprecise characterization and significant variation among the currently available maturation protocols, it is essential to establish a set of criteria to standardize models and accurately characterize and define the developmental properties of human neurons derived from iPSCs. In this study, we conducted comprehensive cellular and network level analysis of the functional development of human neurons, generated from iPSCs obtained from healthy young female peripheral blood mononuclear cells by BDNF and GDNF treatment. We provide a thorough description of the maturation process of h-iPSC-Ns over a 10-week in vitro period using conventional whole-cell patch clamp and dynamic clamp techniques, alongside with morphometry and immunocytochemistry. Additionally, we utilized calcium imaging to monitor the progression of synaptic activity and network communication. At the single cell level, human neurons exhibited gradually decreasing membrane resistance in parallel with improved excitability. By the fifth week of maturation, firing profiles were consistent with those of mature regular firing type of neurons. At the network level, fast glutamatergic and depolarizing GABAergic synaptic connections were abundant together with synchronized network activity from the sixth week of maturation. Alterations in the expression of GABAA receptor subunits were also observed during the process of maturation. The sequence of differentiation events was consistent, providing a robust temporal framework to execute experiments at defined stages of neuronal maturation as well as to use a specific set of experiments to assess a culture’s maturation. The uncovered progression of differentiation events provides a powerful tool to aid the planning and designing of targeted experiments during defined stages of neuronal maturation.

Therapeutic potential of rutin in premenstrual depression: evidence from in vivo and in vitro studies.

Premenstrual dysphoric disorder (PMDD) is a cyclical mood disorder that severely affects the daily life of women of reproductive age. Most of the medications being used clinically have limitations such as low efficacy, side effects, and high cost, so there is an urgent need to discover safer and more effective medications. Rutin is a natural flavonol glycoside with various pharmacological properties including antidepressant. The study of the efficacy and mechanism of action of rutin in PMDD-depressed subtype model rats plays an important role in the discovery of new drugs for the treatment of PMDD. Binding of rutin to gamma-aminobutyric acid type A receptors (GABAA receptors) was probed using molecular docking, microscale thermophoresis, radioactive receptor ligand binding assay and cell membrane clamp experiment. Behavioral tests in mice were performed to screen the optimal dose of rutin. Behavioral tests were performed to evaluate the effects of rutin on depressed mood, memory impairment, and social impairment in PMDD-depressed subtype model rats. HE staining and Golgi staining were performed to observe the neuronal damage in rat hippocampus. UHPLC-MS/MS targeted metabolomics was performed to detect the changes of neurotransmitter content in rat hippocampus. PCR array to detect the effect of rutin on mRNA expression of GABAA receptor partial subunits in rat hippocampus. The docking score of rutin with the GABAA receptor benzodiazepine site was -11.442 and the gliding score was -11.470. The Kd of rutin with the GABAA receptor (α1β2γ2) was 1.17 ± 0.89μM. Rutin competed with [H3]-flunitrazepam for the GABAA receptor benzodiazepine site and inhibited the inward flow of chloride ions (P < 0.05). In PMDD-depressed subtype rats, rutin alleviated depressed mood, memory impairment and social impairment, ameliorated hippocampal neuronal damage and reduces gamma-aminobutyric acid (GABA) and acetylcholine (ACh) levels (P < 0.05). Moreover, we found that rutin did not affect the relative mRNA expression of GABAA receptor subunits in rat hippocampus. Overall, rutin alleviated depressed mood, memory impairment and social impairment in PMDD-depressed subtype rats, which may be related to binding to GABAA receptor benzodiazepine sites, inhibiting chloride ions inward flow, ameliorating hippocampal neuronal damage and reducing GABA and ACh levels. The results of this study provide an experimental basis and scientific evidence for the development of new drugs for the treatment of PMDD.