Deciphering the Possible Mechanism of GABA in Tobacco Pollen Tube Growth and Guidance

γ-Aminobutyric acid (GABA) is an inhibitory transmitter in animal central and peripheral nervous systems, and also plays an important role in pollen tube growth and guidance. However, the mechanisms underlying these effects in plants are poorly understood, mainly because the GABA receptor in plants has not been elucidated. To address this issue, we recently created quantum dot probes to identify possible GABA receptors on the membrane surfaces of pollen protoplasts. We found that GABA bound to cell membranes and regulated downstream Ca²⁺ oscillation in the cells. These results provide important clues to further specifying the nature of the binding sites and deciphering the role of GABA as a signal molecule in pollen tube growth and orientation.Key WordS: γ-aminobutyric acid, fertilization, GABA receptor, signal transduction and tobacco     

The emerging role of GABA as a transport regulator and physiological signal

While the proposal that γ-aminobutyric acid (GABA) acts a signal in plants is decades old, a signaling mode of action for plant GABA has been unveiled only relatively recently. Here, we review the recent research that demonstrates how GABA regulates anion transport through aluminum-activated malate transporters (ALMTs) and speculation that GABA also targets other proteins. The ALMT family of anion channels modulates multiple physiological processes in plants, with many members still to be characterized, opening up the possibility that GABA has broad regulatory roles in plants. We focus on the role of GABA in regulating pollen tube growth and stomatal pore aperture, and we speculate on its role in long-distance signaling and how it might be involved in cross talk with hormonal signals. We show that in barley (Hordeum vulgare), guard cell opening is regulated by GABA, as it is in Arabidopsis (Arabidopsis thaliana), to regulate water use efficiency, which impacts drought tolerance. We also discuss the links between glutamate and GABA in generating signals in plants, particularly related to pollen tube growth, wounding, and long-distance electrical signaling, and explore potential interactions of GABA signals with hormones, such as abscisic acid, jasmonic acid, and ethylene. We conclude by postulating that GABA encodes a signal that links plant primary metabolism to physiological status to fine tune plant responses to the environment.

γ-Aminobutyric acid (GABA) encodes a plant signal that links primary metabolism to physiological status to fine tune plant responses to the environment.     

Does GABA Act as a Signal in Plants?: Hints from Molecular Studies

GABA is a non-protein amino acid that accumulates rapidly in plant tissues in response to biotic and abiotic stress. There have been a number of suggestions as to the role that GABA might play in plants, ranging from a straightforward involvement in N metabolism to a signal mediating plant-animal and plant-microbe interactions. It has also been several proposed that it might function as an intracellular signalling molecule in plants. Here, we discuss recent evidence that plant cells respond at the molecular level to the presence of applied GABA. We argue that these data might serve as the basis for investigating the possible signalling role for GABA in plant development and stress responses in more detail.Key Words: 14-3-3 proteins, GABA, signalling, gene expression, stress, senescenceGABA (γ-aminobutyric acid), which comprises a significant fraction of the free amino acid pool in plant cells, was first identified in potato tubers over half a century ago, but its functions remained obscure for many years. In animal systems, GABA is present at high levels in the brain where it acts as an important neurotransmitter. GABA is synthesised in a pathway known as the GABA shunt, which operates not only in the animals, but in bacteria, fungi and plants too.^1,2 The function of GABA in plants has attracted renewed attention in the last decade following the discovery that intracellular and/or extracellular GABA concentrations increase rapidly in response to a range of stresses. Subsequently, a number of possible roles for GABA and the GABA shunt in plants have been suggested.^1,2 These include acting as a buffering mechanism in C and N metabolism, cytosolic pH regulation, protection against oxidative stress and defence against herbivorous pests. It has been proposed recently that one common function of GABA might be to mediate interactions between plants and other organisms, including bacterial and fungal pathogens, nematodes and insect pests.³ On the other hand, because of the rapid increases in GABA concentration in response to stress, it has sometimes been inferred that GABA might act as an intracellular signalling molecule in plants. However, few studies have adopted molecular approaches to GABA function, and molecular responses are largely unknown.     

Putative role of γ -aminobutyric acid (GABA) as a long-distance signal in up-regulation of nitrate uptake in Brassica napus L.

The relationship between nitrate influx, BnNrt2 nitrate transporter gene expression and amino acid composition of phloem exudate was investigated during N‐deprivation (short‐term experiment) and over a growth cycle (long‐term experiment) in Brassica napus L. The data showed a positive correlation between γ‐aminobutyric acid (GABA) in phloem exudate and nitrate uptake in the short‐ and the long‐term experiments. The hypothesis that this non‐protein amino acid could up‐regulate nitrate uptake via a long‐distance signalling pathway was tested by providing an exogenous GABA supply to the roots. The effect of GABA was compared with the effects of Gln, Glu and Asn, each known to be inhibitors of nitrate uptake. The results showed that GABA treatment induced a significant increase of BnNrt2 mRNA expression, but had less effect on nitrate influx. By contrast, Gln, Glu and Asn significantly reduced nitrate influx and BnNrt2 mRNA expression compared with the control plants. This study provides the first evidence that GABA may act as a putative long‐distance inter‐organ signal molecule in plants in conjunction with negative control exerted by Gln. The up‐regulation effect of GABA on nitrate uptake is discussed in the context of its role in N metabolism, nutritional stress and the recent discovery of a putative role of GABA as a signal molecule in plant development.     

Receptor modifiers indicate that 4-aminobutyric acid (GABA) is a potential modulator of ion transport in plants

GABA (4-aminobutyric acid) is a ubiquitousnon-protein amino acid that accumulates rapidly inplants in response to stress. GABA was firstidentified in plants (potato tubers) and animals(brain tissue) 50 years ago. Although GABA is nowrecognized as the most important inhibitoryneurotransmitter in the mammalian central nervoussystem (CNS), the role of GABA in plants remainsunclear. Studies were performed using Lemna toinvestigate the possibility that GABA elicits aresponse in plants that may be related to that of asignaling molecule as described for GABA effects onthe CNS. Lemna growth was increased 2 to 3-foldby 5 mM GABA, but growth was strongly inhibited by 0.5mM of the isomers 3-aminobutyric acid and2-aminobutyric acid. Growth promotion by GABA wasrapidly terminated by addition of 2-aminobutyric acidto the culture medium, but inhibitory effects of2-aminobutyric acid were not reversed by GABAregardless of amounts added. Promotion of Lemnagrowth by GABA was associated with an increase inmineral content of treated plants in a dose dependentmanner. Results support the hypothesis that GABAactivity in plants involves an effect on ion transportand an interaction with a receptor. Evidence for GABAreceptors in Lemna was obtained from experimentswith pharmacological agents that have been used toidentify GABA receptors in animals. GABA mediatedpromotion of Lemna growth was inhibited bybicuculline and picrotoxin, which are respectivelycompetitive and non-competitive antagonists of GABAreceptors in the CNS. Growth inhibition bybicuculline was not relieved by increasing the amountsof GABA in the medium, indicating that the alkaloid isnot acting, as in the CNS, by competitive antagonismof GABA at GABA receptor sites. Baclofen, a GABAagonist that promotes GABA activity in animalssignificantly increased GABA mediated promotion ofLemna growth. These findings and the knownaction of GABA in regulating ion channels in animalssuggests a way that GABA could amplify the stressresponse in plants.     

Therapeutically leveraging GABAA receptors in cancer

γ-aminobutyric acid or GABA is an amino acid that functionally acts as a neurotransmitter and is critical to neurotransmission. GABA is also a metabolite in the Krebs cycle. It is therefore unsurprising that GABA and its receptors are also present outside of the central nervous system, including in immune cells. This observation suggests that GABAergic signaling impacts events beyond brain function and possibly human health beyond neurological disorders. Indeed, GABA receptor subunits are expressed in pathological disease states, including in disparate cancers. The role that GABA and its receptors may play in cancer development and progression remains unclear. If, however, those cancers have functional GABA receptors that participate in GABAergic signaling, it raises an important question whether these signaling pathways might be targetable for therapeutic benefit. Herein we summarize the effects of modulating Type-A GABA receptor signaling in various cancers and highlight how Type-A GABA receptors could emerge as a novel therapeutic target in cancer.     

Peroxynitrite formation and function in plants

Peroxynitrite (ONOO⁻) is a reactive nitrogen species formed when nitric oxide (NO) reacts with the superoxide anion (O₂⁻). It was first identified as a mediator of cell death in animals but was later shown to act as a positive regulator of cell signaling, mainly through the posttranslational modification of proteins by tyrosine nitration. In plants, peroxynitrite is not involved in NO-mediated cell death and its physiological function is poorly understood. However, it is emerging as a potential signaling molecule during the induction of defense responses against pathogens and this could be mediated by the selective nitration of tyrosine residues in a small number of proteins. In this review we discuss the general role of tyrosine nitration in plants and evaluate recent evidence suggesting that peroxynitrite is an effector of NO-mediated signaling following pathogen infection.     

Nitric oxide: comparative synthesis and signaling in animal and plant cells

Since its identification as an endothelium-derived relaxing factor in the 1980s, nitric oxide has become the source of intensive and exciting research in animals. Nitric oxide is now considered to be a widespread signaling molecule involved in the regulation of an impressive spectrum of mammalian cellular functions. Its diverse effects have been attributed to an ability to chemically react with dioxygen and its redox forms and with specific iron- and thiol-containing proteins. Moreover, the effects of nitric oxide are dependent on the dynamic regulation of its biosynthetic enzyme nitric oxide synthase. Recently, the role of nitric oxide in plants has received much attention. Plants not only respond to atmospheric nitric oxide, but also possess the capacity to produce nitric oxide enzymatically. Initial investigations into nitric oxide functions suggested that plants use nitric oxide as a signaling molecule via pathways remarkably similar to those found in mammals. These findings complement an emerging body of evidence indicating that many signal transduction pathways are shared between plants and animals.     

Molecular mechanism of salicylic acid-induced abiotic stress tolerance in higher plants

Salicylic acid (SA), a key signaling molecule in higher plants, has been found to play a role in the response to a diverse range of phytopathogens and is essential for the establishment of both local and systemic-acquired resistance. Recent studies have indicated that SA also plays an important role in abiotic stress-induced signaling, and studies on SA-modulated abiotic tolerance have mainly focused on the antioxidant capacity of plants by altering the activity of anti-oxidative enzymes. However, little information is available about the molecular mechanisms of SA-induced abiotic stress tolerance. Here, we review recent progress toward characterizing the SA-regulated genes and proteins, the SA signaling pathway, the connections and differences between SA-induced tolerances to biotic and abiotic stresses, and the interaction of SA with other plant hormones under conditions of abiotic stress. The future prospects related to molecular tolerance of SA in response to abiotic stresses are also further summarized.     

Sucrose signaling in plants: A world yet to be explored

The role of sucrose as a signaling molecule in plants was originally proposed several decades ago. However, recognition of sucrose as a true signal has been largely debated and only recently this role has been fully accepted. The best-studied cases of sucrose signaling involve metabolic processes, such as the induction of fructan or anthocyanin synthesis, but a large volume of scattered information suggests that sucrose signals may control a vast array of developmental processes along the whole life cycle of the plant. Also, wide gaps exist in our current understanding of the intracellular steps that mediate sucrose action. Sucrose concentration in plant tissues tends to be directly related to light intensity, and inversely related to temperature, and accordingly, exogenous sucrose supply often mimics the effect of high light and cold. However, many exceptions to this rule seem to occur due to interactions with other signaling pathways. In conclusion, the sucrose role as a signal molecule in plants is starting to be unveiled and much research is still needed to have a complete map of its significance in plant function.     

Indole-3-acetic acid in microbial and microorganism-plant signaling

Diverse bacterial species possess the ability to produce the auxin phytohormone indole-3-acetic acid (IAA). Different biosynthesis pathways have been identified and redundancy for IAA biosynthesis is widespread among plant-associated bacteria. Interactions between IAA-producing bacteria and plants lead to diverse outcomes on the plant side, varying from pathogenesis to phyto-stimulation. Reviewing the role of bacterial IAA in different microorganism-plant interactions highlights the fact that bacteria use this phytohormone to interact with plants as part of their colonization strategy, including phyto-stimulation and circumvention of basal plant defense mechanisms. Moreover, several recent reports indicate that IAA can also be a signaling molecule in bacteria and therefore can have a direct effect on bacterial physiology. This review discusses past and recent data, and emerging views on IAA, a well-known phytohormone, as a microbial metabolic and signaling molecule.     

Nitric oxide as a key component in hormone-regulated processes

Nitric oxide (NO) is a small gaseous molecule, with a free radical nature that allows it to participate in a wide spectrum of biologically important reactions. NO is an endogenous product in plants, where different biosynthetic pathways have been proposed. First known in animals as a signaling molecule in cardiovascular and nervous systems, it has turned up to be an essential component for a wide variety of hormone-regulated processes in plants. Adaptation of plants to a changing environment involves a panoply of processes, which include the control of CO₂ fixation and water loss through stomatal closure, rearrangements of root architecture as well as growth restriction. The regulation of these processes requires the concerted action of several phytohormones, as well as the participation of the ubiquitous molecule NO. This review analyzes the role of NO in relation to the signaling pathways involved in stomatal movement, plant growth and senescence, in the frame of its interaction with abscisic acid, auxins, gibberellins, and ethylene.     

Oxylipin metabolism in response to stress

Oxylipins comprise a group of biologically active compounds whose structural diversity is generated by the coordinate action of lipases, lipoxygenases, and a group of cytochromes P450 that are specialized for the metabolism of hydroperoxy fatty acids. Research on oxylipins has focused mainly on the biosynthesis of the plant signaling molecule jasmonic acid, and its role in the regulation of developmental and defense-related processes. Recent genetic studies indicate that metabolic precursors of jasmonate are active as signals in their own right, and that the synthesis and perception of jasmonates is critical for wound-induced systemic defense responses. Increasing evidence indicates that the collective biological importance of oxylipins in plants is comparable to that of the eicosanoid family of lipid mediators in animals.     

The Uptake of GABA in Trypanosoma cruzi

Gamma aminobutyric acid (GABA) is widely known as a neurotransmitter and signal transduction molecule found in vertebrates, plants, and some protozoan organisms. However, the presence of GABA and its role in trypanosomatids is unknown. Here, we report the presence of intracellular GABA and the biochemical characterization of its uptake in Trypanosoma cruzi, the etiological agent of Chagas' disease. Kinetic parameters indicated that GABA is taken up by a single transport system in pathogenic and nonpathogenic forms. Temperature dependence assays showed a profile similar to glutamate transport, but the effect of extracellular cations Na⁺, K⁺, and H⁺ on GABA uptake differed, suggesting a different uptake mechanism. In contrast to reports for other amino acid transporters in T. cruzi, GABA uptake was Na⁺ dependent and increased with pH, with a maximum activity at pH 8.5. The sensitivity to oligomycin showed that GABA uptake is dependent on ATP synthesis. These data point to a secondary active Na⁺/GABA symporter energized by Na⁺‐exporting ATPase. Finally, we show that GABA occurs in the parasite's cytoplasm under normal culture conditions, indicating that it is regularly taken up from the culture medium or synthesized through an still undescribed metabolic pathway.     

Cyanide action in plants — from toxic to regulatory

Recent biochemical and genetic studies on hydrogen cyanide (HCN) metabolism and function in plants were reviewed. The potential sources of endogenous cyanide and the pathways of its detoxification are outlined and the possible signaling routes by which cyanide exerts its physiological effects are discussed. Cyanide is produced in plant tissues as the result of hydrolysis of cyanogenic compounds and is also released as a co-product of ethylene biosynthesis. Most cyanide produced in plants is detoxified primarily by the key enzyme β-cyanoalanine synthase. The remaining HCN at non-toxic concentration may play a role of signaling molecule involved in the control of some metabolic processes in plants. So, HCN may play a dual role in plants, depending on its concentration. It may be used in defense against herbivores at high toxic concentration and may have a regulatory function at lower concentration. Special attention is given to the action of HCN during biotic and abiotic stresses, nitrate assimilation and seed germination. Intracellular signaling responses to HCN involve enhancement of reactive oxygen species (ROS) generation and the expression of cyanide-insensitive alternative oxidase (AOX) and ACC synthase (ACS) genes. The biochemical and cellular mechanisms of these responses are, however, not completely understood.     

The Arabidopsis her1 mutant implicates GABA in E-2-hexenal responsiveness

When wounded or attacked by herbivores or pathogens, plants produce a blend of six-carbon alcohols, aldehydes and esters, known as C6-volatiles. Undamaged plants, when exposed to C6-volatiles, respond by inducing defense-related genes and secondary metabolites, suggesting that C6-volatiles can act as signaling molecules regulating plant defense responses. However, to date, the molecular mechanisms by which plants perceive and respond to these volatiles are unknown. To elucidate such mechanisms, we decided to isolate Arabidopsis thaliana mutants in which responses to C6-volatiles were altered. We observed that treatment of Arabidopsis seedlings with the C6-volatile E-2-hexenal inhibits root elongation. Among C6-volatiles this response is specific to E-2-hexenal, and is not dependent on ethylene, jasmonic and salicylic acid. Using this bioassay, we isolated 18 E-2-hexenal-response (her) mutants that showed sustained root growth after E-2-hexenal treatment. Here, we focused on the molecular characterization of one of these mutants, her1. Microarray and map-based cloning revealed that her1 encodes a gamma-amino butyric acid transaminase (GABA-TP), an enzyme that degrades GABA. As a consequence of the mutation, her1 plants accumulate high GABA levels in all their organs. Based on the observation that E-2-hexenal treatment induces GABA accumulation, and that high GABA levels confer resistance to E-2-hexenal, we propose a role for GABA in mediating E-2-hexenal responses.     

Benzodiazepines: electron affinity,receptors and cell signaling – a multifaceted approach

Abstract

This report entails a multifaceted approach to benzodiazepine (BZ) action, involving electron affinity, receptors, cell signaling and other aspects. Computations of the electron affinities (EAs) of different BZs have been carried out to establish the effect of various substituents on their EA. These computations were undertaken to serve as a first step in determining what role electron transfer (ET) plays in BZ activity. The calculations were conducted on the premise that the nature of the substituent will either decrease or increase the electron density of the benzene ring, thus altering the ability of the molecule to accept an electron. Investigations were performed on the effect of drug protonation on EA. Similarities involving substituent effects in prior electrochemical studies are also discussed. As part of the multifaceted approach, EA is linked to ET, which appears to play a role in therapeutic activity and toxicity. There is extensive literature dealing with the role of receptors in BZ activity. Significant information on receptor involvement was reported more than 40 years ago. Gamma-aminobutyric acid (GABA) is known to be importantly involved. GABA is a probable mediator of BZ effects. BZ and GABA receptors, although not identical, are physiologically linked. Cell signaling is known to play a part in the biochemistry of BZ action. Various factors participated, such as gene expression, allosteric influence, toxic effects and therapeutic action. Evidence points to involvement of EA and ET in the mode of action in cell signaling. Oxidative stress and antioxidant effects are also addressed.     

Regulation of Arabidopsis thaliana 14-3-3 gene expression by gamma-aminobutyric acid

The function in plants of the non-protein amino acid, gamma-aminobutyric acid (GABA) is poorly understood. In this study, we show that GABA down-regulates the expression of a large subset of 14-3-3 gene family members in Arabidopsis thaliana seedlings in a calcium, ethylene and abscisic acid (ABA)-dependent manner. Gene expression is not affected when seedlings are supplied with glutamate (GLU), a precursor of GABA. The repression of 14-3-3 gene expression by GABA is dependent on functional ethylene and ABA signalling pathways, because the response is lost in the etr1-1, abi1-1 and abi2-1 mutants. Calcium measurements show that in contrast to GLU, GABA does not elicit a cytoplasmic calcium elevation, suggesting that the GABA response is unlikely to be mediated by GLU receptors (GLRs), as has been suggested previously. We suggest that in addition to its role as a stress-related metabolite, GABA may regulate gene expression in A. thaliana, including members of the 14-3-3 gene family.     

GABA Enhances Thermotolerance of Seeds Germination by Attenuating the ROS Damage in Arabidopsis

Seeds germination is strictly controlled by environment factor such as high temperature (HT) through altering the balance between gibberellin acid (GA) and abscisic acid (ABA). Gama-aminobutyric acid (GABA) is a small molecule with four-carbon amino acid, which plays a crucial role during plant physiological process associated with pollination, wounding or abiotic stress, but its role in seeds germination under HT remains elusive. In this study we found that HT induced the overaccumulation of ROS, mainly H₂O₂ and O₂^- , to suppress seeds germination, meanwhile, HT also activated the enzyme activity of GAD for the rapid accumulation of GABA, hinting the regulatory function of GABA in controlling seeds germination against HT stress. Applying GABA directly attenuated HT-induced ROS accumulation, upregulated GA biosynthesis and downregulated ABA biosynthesis, ultimately enhanced seeds germination. Consistently, genetic analysis using the gad1/2 mutant defective in GABA biosynthesis, or pop2-5 mutant with high endogenous GABA content supported the potential function of GABA in improving seeds germination tolerance to HT through scavenging ROS overaccumulation. Based on these data, we propose that GABA acts as a novel signal to enhance thermotolerance of seeds germination through alleviating the ROS damage to seeds viability.