Increase In GABA Content Research Articles

Enhanced γ-aminobutyric acid production by Co-culture fermentation with Enterococcus faecium AB157 and Saccharomyces cerevisiae SC125

γ-aminobutyric acid (GABA), a non-protein amino acid, is a major inhibitory neurotransmitter within the human nervous system. Research on enhancing GABA production through the co-culture of yeast and lactic acid bacteria is limited, and the intricacies of their interactions and the mechanisms behind GABA enhancement are not fully elucidated. In this study, five yeast strains from Sichuan Pao Cai were co-cultured with Enterococcus faecium AB157 to increase GABA content, and it was found that combining E. faecium AB157 with Saccharomyces cerevisiae SC125 at a 4:1 ratio, along with 12 g/L MSG, at 35 °C, optimized GABA production to reach levels of 6.57 g/L. Within the co-culture system, there was a notable increase in both glutamate decarboxylase activity (GAD) and medium pH, while the activity of Autoinducer-2 (AI-2) remained stable. Furthermore, the cell-free supernatants (CFS) from S. cerevisiae SC125 were able to enhance GABA production by E. faecium AB157, up to 4.61 g/L. RNA sequencing revealed that the GABA biosynthetic pathways and quorum sensing (QS) in co-fermentation on E. faecium AB157 were upregulated, and the ATP synthesis and arginine deiminase pathway were downregulated compare to the pure culture of E. faecium AB157.

L-Glutamate treatment alleviates chilling injury of prune (Prunus domestica L.) fruit by regulating ROS homeostasis, GABA shunt, and energy metabolism

The impacts of L-glutamate (L-Glu) treatment on chilling injury (CI), Ca2+ signaling, mitochondrial ultrastructure, and metabolisms of reactive oxygen species (ROS), γ-aminobutyric acid (GABA), energy of prune fruit under chilling stress were studied. The results found that the optimal concentration of L-Glu to suppress CI occurrence and maintain quality in prune fruit was 0.1 g L−1, which also enhanced the PdGLRs expression, cytoplasmic Ca2+ concentration, the contents of CaM, and CML under cold stress. Moreover, L-Glu treatment could reduce ROS accumulation and increase GABA content, and energy level, contributing to maintaining the integrity of the mitochondrial structure in cold-stored prune fruit. More importantly, PdGLRs expression and CaM/CML content positively correlated with antioxidant enzyme activities, GABA shunt, and energy status in prune fruit. These results indicated that the enhanced cold resistance of L-Glu-treated prunes might be attributed to the activated Ca2+ signaling, thus improving the antioxidant capacity, GABA, and energy levels.

Screening and characterization of lactic acid bacteria and fermentation of gamma-aminobutyric acid-enriched bamboo shoots.

In order to produce fermented bamboo shoots with functional properties, two strains of lactic acid bacteria were selected for inoculation and fermentation. One strain, Lactiplantibacillus plantarum R1, exhibited prominent potential probiotic properties (including gastrointestinal condition tolerance, adhesion ability, antimicrobial ability, and antibiotic resistance), while the other, Levilactobacillus brevis R2, demonstrated the capability of high γ-aminobutyric acid (GABA) production (913.99 ± 14.2 mg/L). The synergistic inoculation of both strains during bamboo shoot fermentation led to a remarkable increase in GABA content (382.31 ± 12.17 mg/kg), surpassing that of naturally fermented bamboo shoots by more than 4.5 times and outperforming mono-inoculated fermentation. Simultaneously, the nitrite content was maintained at a safe level (5.96 ± 1.81 mg/kg). Besides, inoculated fermented bamboo shoots exhibited an increased crude fiber content (16.58 ± 0.04 g/100 g) and reduced fat content (0.39 ± 0.02 g/100 g). Sensory evaluation results indicated a high overall acceptability for the synergistically inoculated fermented bamboo shoots. This study may provide a strategy for the safe and rapid fermentation of bamboo shoots and lay the groundwork for the development of functional vegetable products enriched with GABA.

Enhancement of γ-Aminobutyric Acid and the Characteristics of Nutrition and Function in White Quinoa through Ultrasound Stress at the Pre-Germination Stage.

The global production of quinoa has been increasing in recent years. In plant-based foods, ultrasound stress has received increasing attention, owing to its ability to enhance the production of primary and secondary metabolites. We studied the effects of ultrasonic stress at the pre-germination stage on the γ-aminobutyric acid (GABA) accumulation and characteristics of nutrition and function in quinoa. The results showed that ultrasonic conditions of 100 W for 4 min promoted an increase in GABA content by 9.15-fold, to 162.47 ± 6.69 mg/100 g·DW, compared to that of untreated quinoa, through promoting a 10.2% and 71.9% increase in the water absorption and glutamate decarboxylase activity of quinoa, respectively. Meanwhile, compared to untreated quinoa, ultrasonic stress at the pre-germination stage enhanced the total phenolic, total flavonoid, and total saponin contents of quinoa by 10.2%, 33.6%, and 90.7%, to 3.29 mg GA/g·DW, 104.0 mg RE/100 g·DW, and 7.13 mg/g, respectively, without decreasing its basic nutritional quality. Ultrasonic stress caused fissures on the surface of quinoa starch particles. Additionally, germination under ultrasonic stress increased the n3 polyunsaturated fatty acids by 14.4%. Furthermore, ultrasonic stress at the pre-germination stage promoted the scavenging of 2,2-diphenyl1-picrylhydrazyl radicals and inhibitions of α-amylase, α-glucosidase, and pancreatic lipase by 14.4%, 14.9%, 24.6%, and 20.0% in vitro, compared to untreated quinoa. The results indicated that the quinoa sprouted via ultrasonic stress could represent a promising method through which to develop nutritionally balanced whole grains rich in GABA, with hypoglycemic and hypolipidemic activities, which could provide theoretical support for the development of functional whole-grain foods based on quinoa.

CmCML11 interacts with CmCAMTA5 to enhance γ-aminobutyric acid (GABA) accumulation by regulating GABA shunt in fresh-cut cantaloupe

The effect of calcium chloride (CaCl2) treatment on γ-aminobutyric acid (GABA) accumulation in fresh-cut cantaloupe and the involved mechanisms were investigated. The result showed that 1% (w/v) CaCl2 treatment increased GABA content and activities of glutamate decarboxylase (GAD) and succinate semialdehyde dehydrogenase (SSADH), while decreased glutamate (Glu) content and GABA transaminase (GABA-T) activities in fresh-cut cantaloupe. CmCML11 and CmCAMTA5 expressions of CaCl2-treated fruit increased by 187.4% and 165.6% than control fruit in the initial 6 h. Besides, expressions of GABA shunt genes, including CmGAD1, CmGAD2, CmGABA-T and CmSSADH were also up-regulated by CaCl2 treatment during early storage. Moreover, acting as a transcriptional activator, CmCAMTA5 could bind to the CG-box in promoters of CmGAD1, CmGABA-T and CmSSADH and activate their transcription. Furthermore, the interaction between CmCML11 and CmCAMTA5 could enhance the transcriptional activation on GABA shunt genes which were regulated by CmCAMTA5. Collectively, our findings revealed that CaCl2 treatment promoted GABA accumulation in fresh-cut cantaloupe via the combined effect of CmCML11 and CmCAMTA5 in the regulation of expressions of CmGAD1, CmGABA-T, and CmSSADH in GABA shunt.

Melatonin mobilizes the metabolism of sugars, ascorbic acid and amino acids to cope with chilling injury in postharvest pear fruit

Chilling injury manifesting as internal browning is an irreversible physiological disorder in cold-stored ‘Yali’ pear fruit, which seriously restricts postharvest handling and commercialization of pear fruit. This study investigated the impact of melatonin (MN) treatment on chilling injury and the metabolism of sugars, ascorbic acid (AsA), proline, and γ-aminobutyric acid (GABA) during chilling injury development in ‘Yali’ pears. The findings revealed that MN-treated pear fruit exhibited considerably improved tolerance to chilling stress compared with untreated control fruit. MN application augmented proline accumulation, likely owing to increased activities and transcriptional concentrations of Δ1-pyrroline-5-carboxylate synthase and ornithine-δ-aminotransferase as well as diminished activity and transcriptional level of proline dehydrogenase. MN immersion increased enzyme activities and gene expression related to the GABA shunt pathway, leading to increased GABA concentrations in cold-stored pear fruit. Furthermore, MN treatment elevated AsA concentrations in the fruit, exerting protective effects against cell membrane damage and mitigating oxidative stress damage caused by chilling. Compared with the control group, MN-treated fruit exhibited reduced neutral invertase enzyme activity and gene expression. Additionally, MN treatment increased both sucrose-phosphate synthase and sucrose synthase activities and gene expression in pear fruit. These observations indicated that the enhanced chilling tolerance of ‘Yali’ pear fruit following MN treatment may be associated with the regulation of proline, GABA, AsA, and soluble sugars metabolism.

Treatment modalities for infantile spasms: current considerations and evolving strategies in clinical practice.

Infantile spasms, newly classified as infantile epileptic spasm syndrome (IESS), occur in children under 2 years of age and present as an occur as brief, symmetrical, contractions of the musculature of the neck, trunk, and extremities. When infantile spasms occur with a concomitant hypsarrhythmia on electroencephalogram (EEG) and developmental regression, it is known as West Syndrome. There is no universally accepted mainstay of treatment for this condition, but some options include synthetic adrenocorticotropic hormone (ACTH), repository corticotropin injection (RCI/Acthar Gel), corticosteroids, valproic acid, vigabatrin, and surgery. Without effective treatment, infantile spasms can cause an impairment of psychomotor development and/or cognitive and behavioral functions. The first-line treatment in the USA is ACTH related to high efficacy for cessation of infantile spasms long-term and low-cost profile. Acthar Gel is a repository corticotropin intramuscular injection that became FDA-approved for the treatment of IESS in 2010. Though it is believed that ACTH, Acthar Gel, and corticosteroids all work via a negative feedback pathway to decrease corticotropin-releasing hormone (CRH) release, their safety and efficacy profiles all vary. Vigabatrin and valproic acid are both anti-seizure medications that work by increasing GABA concentrations in the CNS and decreasing excitatory activity. Acthar Gel has been shown to have superior efficacy and a diminished side effect profile when compared with other treatment modalities.

Controlled germination of grain crops is an effective way to processing low-quality raw materials

Relevance. The development of technologies of cereal crops germination and obtaining of raw ingredients with increased antioxidant properties, containing in their composition biologically active substances of natural origin, will allow to form a modern domestic market of useful food products to maintain the health of the country in the long term. One such approach could be controlled germination of grain crops by ultrasonic treatment during the soaking stage. This technology will maximise the yield of finished products per unit of processed raw material, process low-quality raw materials and create products with increased nutritional value.Methods. The objects of the study were three samples of grain crops: wheat, barley, oats. At the initial stage, incoming quality control of raw materials and their disinfection by physical method of exposure were carried out. For experimental samples of cereal crops at the stage of soaking the impact of ultrasound (22 ± 1.25 kHz) 245 W/l for 5 min, then the grain was soaked for 8 hours (wheat grain) and 12 hours (barley and oats grain), the process of germination was carried out until reaching the sprout size 1.5–2 mm in more than 90% of grains. All operations with control samples were performed in the same sequence, excluding the process of ultrasound exposure. The total content of flavonoids and polyphenolic compounds was determined in all tested samples using spectrophotometric method, total antioxidant activity using DPPH-method, and γ-aminobutyric acid content using automated liquid chromatography system.Results. The studies showed that all germinated samples of cereal crops are characterized by sufficiently high values of flavonoid and polyphenolic compounds. At the same time, the samples obtained by ultrasound exposure showed an increase in the content of flavonoids by an average of 7.3–8.9 times, polyphenols by 2–5.6 times. In the process of intensified controlled germination, the total antioxidant activity increased by 31.6–40.0%, relative to the control samples of grain crops. The increase in GABA content in samples after ultrasound exposure averaged 360–490%. The results confirmed the possibility and feasibility of using the proposed technology of controlled germination in obtaining raw ingredients from germinated cereal crops. The study was carried out with the financial support of the Russian Science Foundation Grant 23-26-00290.

Gut-joint axis: Oral Probiotic ameliorates Osteoarthritis

Osteoarthritis (OA) etiology is multifactorial, and its prevalence is growing globally. The Gut microbiota shapes our immune system and impacts all aspects of health and disease. The idea of utilizing probiotics to treat different conditions prevails. Concerning musculoskeletal illness and health, current data lack the link to understand the interactions between the host and microbiome. We report that S. thermophilus, L. pentosus (as probiotics), and γ-aminobutyric acid (GABA) harbour against osteoarthritis in vivo and alleviate IL-1β induced changes in chondrocytes in vitro. We examined the increased GABA concentration in mice's serum and small intestine content followed by bacterial treatment. The treatment inhibited the catabolism of cartilage and rescued mice joints from degradation. Furthermore, the anabolic markers upregulated and decreased inflammatory markers in mice knee joints and chondrocytes. This study is the first to represent GABA's chondrogenic and chondroprotective effects on joints and human chondrocytes. This data provides a foundation for future studies to elucidate the role of GABA in regulating chondrocyte cell proliferation. These findings opened future horizons to understanding the gut-joint axis and OA treatment. Thus, probiotic/GABA therapy shields OA joints in mice and could at least serve as adjuvant therapy to treat osteoarthritis.

Impact of microwave and germination on physicochemical properties, functional properties, and solubility of black rice powder

AbstractBackground and ObjectivesBlack rice is limited due to factors such as low solubility, difficulty to disperse, and high viscosity. The aim of the study is to overcome these shortcomings and improve their practicability in various industrial applications. The effects of germination and microwave on the physicochemical properties, functional properties, and solubility of black rice powder were compared.FindingsThe effect of germination improved the GABA contents, increased the solubility and starch digestibility, which was consistent with the decrease in relative crystallinity and porous surface structure, and the increase in the gelatinization enthalpy. Microwave increased the solubility and starch digestibility of black rice powder. Total polyphenol content was increased by microwave compared to untreated and germinated black rice. The type of starch showed no significant effect in single and dual treatment. The combined treatment enhanced the GABA content, reduced the loss of polyphenols, anthocyanins and improved its solubility, physicochemical properties characteristics.ConclusionThe results might help understand the mechanism of germination and microwave treatments influencing the structural properties of starches and contribute to better‐developing products based on black rice flour.Significance and NoveltyThe association treatment increased GABA content, the solubility, and nutrition of black rice flour, but reduced the digestibility of black rice flour.

Influence of different lactic acid bacteria strains and milling process on the solid-state fermented green and red lentils (Lens culinaris L.) properties including gamma-aminobutyric acid formation.

The aim of this study was to evaluate the influence of lactic acid bacteria (LAB) strains (Lactiplantibacillus plantarum No.122 and Lacticaseibacillus casei No.210) and milling process on the solid-state fermented (for 24 h, at 30°C) green and red lentils (Lens culinaris L.) properties, chiefly pH, LAB viable counts, color coordinates, free amino acid (FAA) profile, γ-aminobutyric acid (GABA) and biogenic amine (BA) concentrations, fatty acid (FA) and volatile compound (VC) profiles. Results showed that both of the tested LAB strains are suitable for the fermentation of lentils: pH of fermented lentils was <4.5 and LAB viable counts >8.0 log10 colony-forming units (CFU)/g. A very strong negative correlation was found (r = -0.973, p ≤ 0.0001) between LAB counts and pH of the samples. Also, fermentation and milling process were significant factors toward color coordinates of the lentils. In most of the cases, solid-state fermentation (SSF) increased essential FAA content in lentils; however, some of the non-essential FAA content was reduced. SSF significantly increased GABA concentration in lentils and milling process was a significant factor on GABA content of the samples (p ≤ 0.05). The main BA in lentils was spermidine, and SSF decreased their total BA content (34.8% on average in red lentils and 39.9% on average in green lentils). The main FA in lentils were linoleic and oleic. The main VC in lentils were hexanal, 1-hexanol, hexanoic acid, D-limonene and (E)-2-nonen-1-ol. Furthermore, most of the VC showed significant correlations with pH of lentil samples, LAB counts and FA content. Finally, the LAB strain used for fermentation and the milling process of lentils are significant factors for most of the analyzed parameters in lentil. Moreover, despite the higher GABA concentration found in green non-milled SSF lentils, application of combined milling and SSF is recommended because they showed the lowest BA content in addition to higher essential FAA and GABA concentrations.

Biochemical and Enzymatic Analyses to Understand the Accumulation of γ-Aminobutyric Acid in Wheat Grown under Flooding Stress

Flooding induces low-oxygen stress, which reduces plant growth. The activity of the γ-aminobutyric acid (GABA) shunt is crucial for plant stress adaptation, in which it acts by changing cytosolic pH, limiting reactive oxygen species production, regulating nitrogen metabolism, and bypassing steps in the tricarboxylic acid cycle. GABA accumulates under osmotic stress as well as flooding stress. To clarify the dynamic roles of GABA accumulation in wheat under flooding stress, biochemical and enzymatic analyses were performed using a plant-derived smoke solution (PDSS), which rescued wheat growth from flooding stress. Alcohol dehydrogenase abundance increased under flooding stress; however, under the same conditions, pyruvic acid content increased only following PDSS application. Glutamic acid content increased under flooding stress, but decreased following the application of PDSS after 2 days of flooding. Glutamate decarboxylase abundance and GABA content increased under flooding stress, and further increased after 1 day of application of PDSS. Succinyl semialdehyde dehydrogenase accumulated after 2 days of flooding. These results suggest that flooding stress increases GABA content along with the increase and decrease of glutamate decarboxylase and succinyl semialdehyde dehydrogenase, respectively. Additionally, PDSS increased GABA content along with the increase of glutamate decarboxylase abundance at the initial stage of application.

Social deficits via dysregulated Rac1-dependent excitability control of prefrontal cortical neurons and increased GABA/glutamate ratios.

Identifying symptom-specific convergent mechanisms for neurodevelopmental disorders is a promising strategy in advancing therapies. Here, we show that bidirectional dysregulation of Rac1 activity in the medial prefrontal cortex (mPFC) dictates shared social deficits in mice. Selective upregulation or downregulation of Rac1 activity in glutamatergic or fast-spiking GABAergic neurons results in excessive or inadequate control of excitability combined with a decrease in glutamate or an increase in GABA concentrations and an increase in the GABA/glutamate ratio, which is responsible for social deficits. Notably, the autism model of Shank3B knockout mice exhibits aberrantly enhanced Rac1 activity, reduced glutamate concentrations, and pyramidal neuron excitability in mPFC accompanied with social deficits, which were corrected by either excitatory-neuron-specific downregulation of Rac1 activity or upregulation of neuronal excitability. Thus, this work shows a convergence between genetic autism risk factors, dysregulation of Rac1 signaling, and excitation-inhibition imbalance, enabling mechanism-based stratification of patients with social deficits.

Increased seizure sensitivity in pregnant mice with genetic knockdown of acid sensing ion channel 2a is associated with impaired hippocampal inflammatory response.

Acid sensing ion channels (ASICs) are mechano- and chemo-receptor channels that are activated by drops in extracellular pH as occurs after neurotransmission. In our previous study, we demonstrated that mice subjected to reduced utero-placental perfusion pressure during pregnancy, to mimic the pregnancy complication of preeclampsia, have reduced hippocampal expression of ASIC2a protein. We also showed that pregnant mice with heterozygous expression of ASIC2a (+/-) had increased sensitivity and severity to pentylenetetrazol-induced seizures; however, the mechanisms by which this occurs remain unclear. The purpose of this study was to investigate key molecular targets involving neurotransmission and inflammation that are differentially changed following seizure exposure in pregnant ASIC2a +/- mice. On gestational day 18.5, ASIC2a wild-type (+/+, n = 7) and +/- (n = 14) mice were injected with 40 mg/kg pentylenetetrazol and monitored for 30 min. Western blot and ELISA analysis revealed no difference in hippocampal synaptosome glutamate-related proteins but an increase in GABA concentration in pregnant +/- mice. Using ELISA and multiplex assays, we found a significant decrease in serum TNFα, and a decreased concentration of pro-inflammatory cytokines and chemokines in hippocampal cytosolic fraction. Significant reductions in IL-1β, IL-3, IL-12 (p70), eotaxin, interferon gamma, and macrophage inflammatory protein (MIP-1β), in the hippocampal cytosolic fractions of +/- mice were observed compared to +/+ mice. Additionally, there was no difference in hippocampal microglia density or activation in pregnant ASIC2a+/+ vs. +/- mice. These results support the hypothesis that pregnant mice with reduced ASIC2a may not be able to mount an inflammatory response following acute seizure exposure.

Effect of fermented oyster (Crassostrea gigas) extracts and regular walking on muscle strength and mass in older adults with relatively low muscle mass: A randomized controlled trial.

IntroductionOysters possess an excellent nutritional profile containing γ-aminobutyric acid (GABA). Previous data suggest that GABA is a potent bioactive compound for improving muscle health. Lactic acid fermentation is thought to increase GABA content. However, the effect of oral supplementation of fermented oyster extracts (FO) on human muscle strength and mass is unclear. Therefore, we tested the effects and safety of consumption of FO combined with regular walking for 12 weeks on muscle strength and mass in older adults with relatively low muscle mass.Materials and methodsA randomized controlled trial was conducted on 54 adults between 50 and 78 years of age. Participants were randomized to receive either placebo or 1,200 mg FO daily for 12 weeks. By fermentation with Lactobacillus brevis BJ20, FO was prepared from Crassostrea gigas. At baseline and at 12 weeks after treatment, the following parameters of the participants were examined: knee strengths, handgrip strengths, body composition, blood tests, and 24-h dietary recall. All participants were required to walk for 30–60 min/day for >3 days/week during the trial period. Physical activity was assessed using an exercise log during the study.ResultsOf the 54 participants, 49 completed the trial without reporting adverse effects. FO supplementation over 12 weeks did not cause any increase in knee or grip strength compared to the control group. Also, no differences were observed in the muscle mass, growth hormone, muscle biomarkers, anti-inflammatory markers, and antioxidative markers between the two groups. None of the participants experienced adverse events. Application of FO was well tolerated, and no notable adverse effect was reported in both groups.DiscussionFO supplementation with regular walking did not improve remarkably muscle function compared to regular walking alone in adults with relatively low muscle mass.Clinical Trial Registration[www.ClinicalTrials.gov], identifier [NCT04109911].

Green Light Partial Replacement of Red and Blue Light Improved Drought Tolerance by Regulating Water Use Efficiency in Cucumber Seedlings

Light is one of the most important environmental signals in plant growth, development, and stress response. Green light has been proved to enhance plant defense against biotic and/or abiotic stress. To illustrate the effects of green light partially replaced red light and blue light on the plant under drought condition, cucumber (Cucumis sativus L. cv. Xinchun No. 4) seedlings were treated with short-term drought stress and were concomitantly exposed to four treatments, which were set up by adjusting the relative amount of green light as 0 (RB), 25 (RBG25), 50 (RBG50), and 75 (RBG75) μmol m−2 s−1, respectively, with a total photosynthetic photon flux density of 250 μmol m−2 s−1 and a fixed red-to-blue ratio of 4:1. The results showed that compared with RB, RBG50 significantly increased shoot fresh weight (FW) and dry weight (DW), root DW, plant height, stem diameter, leaf area, and leaf dry mass per unit area (LMA) by 10.61, 7.69, 66.13, 6.22, 10.02, 4.10, and 12.41%, respectively. Also, the addition of green light significantly increased the root volume and root tip number. Moreover, green light partial replacement of red light and blue light increased total water content, especially free water content, improved leaf water status, and alleviated water loss in plants caused by drought stress. Also, the addition of green light increased net photosynthetic rate (Pn), reduced both stomata conductance (gs) and transpiration rate (E), enhanced the intrinsic water-use efficiency (WUE) and instantaneous water-use efficiency (iWUE) of leaves, and increased the content of chlorophylls a and b. Green light substituting a proportion of blue and red light regulated stomatal aperture by significantly increasing abscisic acid (ABA) and γ-aminobutyric acid (GABA) content. In addition, the increase of GABA was resulted from the upregulation of Glutamate Decarboxylase 2 (CsGAD2). However, the relative electrolytic leakage and contents of malondialdehyde (MDA), superoxide anion (), and hydrogen peroxide (H2O2) vigorously decreased as the intensity of green light was added to the spectrum under drought. Conclusively, green light partially replaced red light and blue light and improved drought tolerance of cucumber seedlings by upregulating the expression of CsGAD2 gene and promoting the synthesis of GABA. The increase in GABA content further downregulated the expression of aluminum-activated malate transporter 9 (CsALMT9) gene, induced stomata to close, improved water utilization, and alleviated damage caused by drought. This study highlights a role of green light in plant physiological processes. Moreover, analyzing the function of green light on improving drought tolerance of plants could open alternative avenues for improving plant stress resilience.

CsCuAOs and CsAMADH1 Are Required for Putrescine-Derived γ-Aminobutyric Acid Accumulation in Tea.

Polyamines are a potential source of γ-aminobutyric acid (GABA) in plants under abiotic stress. However, studies on GABA enrichment in tea mostly focus on the GABA shunt, while the correlation between polyamine degradation and GABA formation in tea is largely unknown. In this study, tea plants responded to exogenous putrescine, resulting in a significant increase in GABA content, while the glutamate level did not change. At the same time, five copper-containing amine oxidase (CuAO) and eight aminoaldehyde dehydrogenase (AMADH) genes involved in the putrescine-derived GABA pathway were identified from the Tea Plant Information Archive. Expression analysis indicated that CsCuAO1, CsCuAO3 as well as CsAMADH1 were induced to play an important function in response to exogenous putrescine. Thus, the three genes were cloned and the catalytic efficiency of soluble recombinant proteins was determined. CsCuAOs and CsAMADH1 exhibited indispensable functions in the GABA production from putrescine in vitro. Subcellular localization assays indicated that CsAMADH1 was localized in plastid, while both CsCuAO1 and CsCuAO3 were localized in peroxisome. In addition, the synergistic effects of CsCuAOs and CsAMADH1 were investigated by a transient co-expression system in Nicotiana benthamiana. Our data suggest that these three genes regulate the accumulation of GABA in tea by participating in the polyamine degradation pathway and improve the content of GABA in tea to a certain extent. The results will greatly contribute to the production of GABA tea.

An aqueous extract of Khaya senegalensis (Desv.) A. Juss. (Meliaceae) prevents seizures and reduces anxiety in kainate-treated rats: modulation of GABA neurotransmission, oxidative stress, and neuronal loss in the hippocampus

Ethnopharmacological relevanceTemporal lobe epilepsy is the most common form of drug-resistant epilepsy. Therefore, medicinal plants provide an alternative source for the discovery of new antiepileptic drugs. Aim of the studyThis study was aimed at investigating the antiepileptic- and anxiolytic-like effects of an aqueous extract of Khaya senegalensis (K. senegalensis) in kainate-treated rats. MethodsSeventy-two rats received a single dose of kainate (12 mg/kg) intraperitoneally. Those that exhibited two hours of status epilepticus were selected and monitored for the first spontaneous seizure. Then, animals that developed seizures were divided into 6 groups of 8 rats each and treated twice daily for 14 days as follows: negative control group received per os (p.o.) distilled water (10 ml/kg); two positive control groups received either sodium valproate (300 mg/kg, p.o.) or phenobarbital (20 mg/kg, p.o.); and three test groups received different doses of the extract (50, 100, and 200 mg/kg, p.o.). In addition, a group of 8 normal rats (normal control group) received distilled water (10 ml/kg, p.o.). During the treatment period, the animals were video-monitored 12 h/day for behavioral seizures. At the end of the treatment period, animals were subjected to elevated plus-maze and open field tests. Thereafter, rats were euthanized for the analysis of γ-aminobutyric acid (GABA) concentration, oxidative stress status, and neuronal loss in the hippocampus. ResultsThe aqueous extract of K. senegalensis significantly reduced spontaneous recurrent seizures (generalized tonic-clonic seizures) and anxiety-like behavior compared to the negative control group. These effects were more marked than those of sodium valproate or phenobarbital. Furthermore, the extract significantly increased GABA concentration, alleviated oxidative stress, and mitigated neuronal loss in the dentate gyrus of the hippocampus. ConclusionThese findings suggest that the aqueous extract of K. senegalensis possesses antiepileptic- and anxiolytic-like effects. These effects were greater than those of sodium valproate or phenobarbital, standard antiepileptic drugs. Furthermore, these effects are accompanied by neuromodulatory and antioxidant activities that may be related to their behavioral effects. These data justify further studies to identify the bioactive molecules present in the extract for possible future therapeutic development and to unravel their mechanisms of action.

Design, synthesis, and pharmacological evaluation of aryl oxadiazole linked 1,2,4-triazine derivatives as anticonvulsant agents

A series of new clubbed aryl oxadiazole-1,2,4-triazine derivatives (6a-l) were designed and synthesized using appropriate chemical routes. The structures were designed to have the required structural elements for any compounds to be potential anticonvulsant. Preliminary screening of anticonvulsant activity was performed using maximal electroshock seizure (MES), subcutaneous pentylenetetrazole-induced seizure (scPTZ) and behavioral activity were assessed by motor impairment test and actophotometer test. The derivatives 6-((5-(4-methoxyphenyl)-1,3,4-oxadiazol-2-yl)amino)-1,2,4-triazine-3(2H)-thione (6f) and 6-((5-(4-hydroxyphenyl)-1,3,4-oxadiazol-2-yl)amino)-1,2,4-triazine-3(2H)-thione (6g) revealed significant activity against both MES and scPTZ indicating that the compounds are effective against both generalized tonic-clonic and absence seizure. The lead compound (6g) was further evaluated for quantitative evaluation and emerged as the most effective anticonvulsant with median effective dose of 28.5 mg/kg (MES ED50), 76.6 mg/kg (scPTZ) and toxic dose (TD50) was found to be >500 mg/kg. In the GABA estimation study results showed significantly increased GABA concentration. Graphical abstract

Metabotropic glutamate receptor 5 upregulation of γ-aminobutyric acid transporter 3 expression ameliorates cognitive impairment after traumatic brain injury in mice

Traumatic brain injury (TBI) causes neurotransmitter disturbances contributing to neuronal cell death and neurological deficits. In humans, brain injuries impair γ-aminobutyric acid (GABA) uptake and ultimately result in cognitive impairment. GABA transporter 3 (GAT3) is a vital approach of GABA reuptake and catabolism. The contribution of GAT3 in TBI-induced cognitive impairment and its underlying mechanisms remain unknown, which were explored in the present study. Here, we found that expression of GAT3 was downregulated to increase GABA concentration in the mouse brain after TBI. And GAT3 was detected in neurons and astrocytes after TBI unexpectedly, instead of merely expressed on astrocytes in physiological states. Subsequently, activated metabotropic glutamate receptor 5 (mGluR5) reduced GABA content by elevating the expression levels of GAT3. Then, increased mGluR5 activity obviously improved cognitive impairment. Mechanistically, mGluR5 was activated to evidently induce the expression of p-ERK, CREB, and p-CREB after TBI. The inhibition of CREB decreased the expression of CREB, p-CREB, and GAT3 elevated by active mGluR5. However, the CREB inhibitor increased GABA content. Furthermore, Rab11a regulating GAT3 trafficking by endocytosis was elevated after TBI. And Rab11a downregulated by active mGluR5 was reversed by CREB inhibitor. In summary our findings elucidated that activated mGluR5 ameliorated cognitive function by upregulating GAT3 in TBI. And mGluR5 possibly regulated GAT3 by ERK/CREB/Rab11a pathway. GAT3 could serve as a potential target for TBI cognitive treatment.