Toxic and signaling effects of the anaesthetic lidocaine on rice cultured cells.

Most studies on anesthesia focus on the nervous system of mammals due to their interest in medicine. The fact that any life form can be anaesthetised is often overlooked although anesthesia targets ion channel activities that exist in all living beings. This study examines the impact of lidocaine on rice (Oryza sativa). It reveals that the cellular responses observed in rice are analogous to those documented in animals, encompassing direct effects, the inhibition of cellular responses, and the long-distance transmission of electrical signals. We show that in rice cells, lidocaine has a cytotoxic effect at a concentration of 1%, since it induces programmed reactive oxygen species (ROS) and caspase-like-dependent cell death, as already demonstrated in animal cells. Additionally, lidocaine causes changes in membrane ion conductance and induces a sharp reduction in electrical long-distance signaling following seedlings leaves burning. Finally, lidocaine was shown to inhibit osmotic stress-induced cell death and the regulation of Ca2+ homeostasis. Thus, lidocaine treatment in rice and tobacco (Nicotiana benthamiana) seedlings induces not only cellular but also systemic effects similar to those induced in mammals.

[Anaesthesia, a process common to all living organisms]. / L'anesthésie, un processus commun à tout le vivant.

Because of their interest in medicine, most studies of anaesthesia focus on the nervous system of metazoans, and the fact that any life form can be anaesthetised is often underlooked. If electrical signalling is an essential phenomenon for the success of animals, it appears to be widespread beyond metazoans. Indeed, anaesthesia targets Na+/Ca2+ voltage-gated channels that exist in a wide variety of species and originate from ancestral channels that predate eukaryotes in the course of evolution. The fact that the anaesthetic capacity that leads to loss of sensitivity is common to all phyla may lead to two hypotheses: to be investigated is the evolutionary maintenance of the ability to be anaesthetised due to an adaptive advantage or to a simple intrinsic defect in ion channels? The study of anaesthesia in organisms phylogenetically distant from animals opens up promising prospects for the discovery of new anaesthetic treatments. Moreover, it should also lead to a better understanding of a still poorly understood phenomenon that yet unifies all living organisms. We hope that this new understanding of the unity of life will help humans to assume their responsibilities towards all species, at a time when we are threatening biodiversity with mass extinction.

Title: L'anesthésie, un processus commun à tout le vivant. Abstract: Du fait de leur intérêt en médecine, la majeure partie des études actuelles sur les anesthésiques se concentrent sur le système nerveux des animaux et négligent le fait que toute forme de vie peut être anesthésiée. En effet, l'anesthésie cible des canaux dépendants du voltage, canaux qui existent dans un grand nombre d'espèces diverses et qui proviennent de canaux ancestraux antérieurs à l'apparition même des eucaryotes. La question demeure : le maintien au cours de l'évolution de la capacité à être anesthésié est-il dû à un avantage adaptatif ou à un simple défaut intrinsèque des canaux ioniques ? Le regain d'intérêt actuel pour les modèles non animaux ouvre l'espoir non seulement de découvrir de nouvelles molécules anesthésiantes, mais aussi de progresser dans notre connaissance fondamentale de ce phénomène encore mal compris.

Our sisters the plants? notes from phylogenetics and botany on plant kinship blindness.

Before the upheaval brought about by phylogenetic classification, classical taxonomy separated living beings into two distinct kingdoms, animals and plants. Rooted in 'naturalist' cosmology, Western science has built its theoretical apparatus on this dichotomy mostly based on ancient Aristotelian ideas. Nowadays, despite the adoption of the Darwinian paradigm that unifies living organisms as a kinship, the concept of the "scale of beings" continues to structure our analysis and understanding of living species. Our aim is to combine developments in phylogeny, recent advances in biology, and renewed interest in plant agency to craft an interdisciplinary stance on the living realm. The lines at the origin of plant or animal have a common evolutionary history dating back to about 3.9 Ga, separating only 1.6 Ga ago. From a phylogenetic perspective of living species history, plants and animals belong to sister groups. With recent data related to the field of Plant Neurobiology, our aim is to discuss some socio-cultural obstacles, mainly in Western naturalist epistemology, that have prevented the integration of living organisms as relatives, while suggesting a few avenues inspired by practices principally from other ontologies that could help overcome these obstacles and build bridges between different ways of connecting to life.

Erratum: Tran et al. Early Cellular Responses Induced by Sedimentary Calcite-Processed Particles in Bright Yellow 2 Tobacco Cultured Cells. Int. J. Mol. Sci. 2020, 21, 4279.

The authors would like to remove the scientific consortium 'Camille Nous' from the author list and the Author Contributions section in the published paper [...].

Biphasic activation of survival and death pathways in Arabidopsis thaliana cultured cells by sorbitol-induced hyperosmotic stress.

Hyperosmotic stresses represent some of the most serious abiotic factors that adversely affect plants growth, development and fitness. Despite their central role, the early cellular events that lead to plant adaptive responses remain largely unknown. In this study, using Arabidopsis thaliana cultured cells we analyzed early cellular responses to sorbitol-induced hyperosmotic stress. We observed biphasic and dual responses of A. thaliana cultured cells to sorbitol-induced hyperosmotic stress. A first set of events, namely singlet oxygen (1O2) production and cell hyperpolarization due to a decrease in anion channel activity could participate to signaling and osmotic adjustment allowing cell adaptation and survival. A second set of events, namely superoxide anion (O2-) production by RBOHD-NADPH-oxidases and SLAC1 anion channel activation could participate in programmed cell death (PCD) of a part of the cell population. This set of events raises the question of how a survival pathway and a death pathway could be induced by the same hyperosmotic condition and what could be the meaning of the induction of two different behaviors in response to hyperosmotic stress.

Early Cellular Responses Induced by Sedimentary Calcite-Processed Particles in Bright Yellow 2 Tobacco Cultured Cells.

Calcite processed particles (CaPPs, Megagreen®) elaborated from sedimentary limestone rock, and finned by tribomecanic process were found to increase photosynthetic CO2 fixation grapevines and stimulate growth of various cultured plants. Due to their processing, the CaPPs present a jagged shape with some invaginations below the micrometer size. We hypothesised that CaPPs could have a nanoparticle (NP)-like effects on plants. Our data show that CaPPs spontaneously induced reactive oxygen species (ROS) in liquid medium. These ROS could in turn induce well-known cellular events such as increase in cytosolic Ca2+, biotic ROS generation and activation of anion channels indicating that these CaPPs could activate various signalling pathways in a NP-like manner.

Activation of plasma membrane H+-ATPases participates in dormancy alleviation in sunflower seeds.

Using various inhibitors and scavengers we took advantage of the size of sunflower (Helianthus annuus) seeds to investigate in vivo the effects of hormones, namely abscisic acid (ABA) and ethylene (ET), and reactive oxygen species (ROS) on the polarization of dormant (D) and non-dormant (ND) embryonic seed cells using microelectrodes. Our data show that D and ND seed cells present different polarization likely due to the regulation of plasma membrane (PM) H+-ATPase activity. The data obtained after addition of hormones or ROS scavengers further suggest that ABA dependent inhibition of PM H+-ATPases could participate in dormancy maintenance and that ET-and ROS-dependent PM H+-ATPase stimulation could participate in dormancy release in sunflower seeds.

Cellular mechanisms to survive salt in the halophyte Cakile maritima.

We recently identified two behaviours in cultured cells of the salt accumulating halophyte Cakile maritima: one related to a sustained depolarization due to Na+ influx through the non-selective cation channels leading to programmed cell death of these cells, a second one related to a transient depolarization allowing cells to survive (Ben Hamed-Laouti, 2016). In this study, we considered at the cellular level mechanisms that could participate to the exclusion of Na+ out of the cell and thus participate in the regulation of the internal contents of Na+ and cell survival. Upon addition of NaCl in the culture medium of suspension cells of C. maritima, we observed a rapid influx of Na+ followed by an efflux dependent of the activity of plasma membrane H+-ATPases, in accordance with the functioning of a Na+/H+ antiporter and the ability of some cells to repolarize. The Na+ efflux was shown to be dependent on Na+-dependent on Ca2+ influx like the SOS1 Na+/H+ antiporter. We further could observe in response to salt addition, an early production of singlet oxygen (1O2) probably due to peroxidase activities. This early 1O2 production seemed to be a prerequisite to the Na+ efflux. Our findings suggest that in addition to the pathway leading to PCD (Ben Hamed-Laouti, 2016), a second pathway comprising an SOS-like system could participate to the survival of a part of the C. maritima cultured cells challenged by salt stress.

Methanol induces cytosolic calcium variations, membrane depolarization and ethylene production in arabidopsis and tobacco.

Background and Aims: Methanol is a volatile organic compound released from plants through the action of pectin methylesterases (PMEs), which demethylesterify cell wall pectins. Plant PMEs play a role in developmental processes but also in responses to herbivory and infection by fungal or bacterial pathogens. However, molecular mechanisms that explain how methanol could affect plant defences remain poorly understood. Methods: Using cultured cells and seedlings from Arabidopsis thaliana and tobacco BY2 expressing the apoaequorin gene, allowing quantification of cytosolic Ca2+, a reactive oxygen species (ROS) probe (CLA, Cypridina luciferin analogue) and electrophysiological techniques, we followed early plant cell responses to exogenously supplied methanol applied as a liquid or as volatile. Key Results: Methanol induces cytosolic Ca2+ variations that involve Ca2+ influx through the plasma membrane and Ca2+ release from internal stores. Our data further suggest that these Ca2+ variations could interact with different ROS and support a signalling pathway leading to well known plant responses to pathogens such as plasma membrane depolarization through anion channel regulation and ethylene synthesis. Conclusions: Methanol is not only a by-product of PME activities, and our data suggest that [Ca2+]cyt variations could participate in signalling processes induced by methanol upstream of plant defence responses.

Comparison of NaCl-induced programmed cell death in the obligate halophyte Cakile maritima and the glycophyte Arabidopsis thaliana.

Salinity represents one of the most important constraints that adversely affect plants growth and productivity. In this study, we aimed at determining possible differences between salt tolerant and salt sensitive species in early salt stress response. To this purpose, we subjected suspension-cultured cells from the halophyte Cakile maritima and the glycophyte Arabidopsis thaliana, two Brassicaceae, to salt stress and compared their behavior. In both species we could observe a time and dose dependent programmed cell death requiring an active metabolism, a dysfunction of mitochondria and caspase-like activation although C. maritima cells appeared less sensitive than A. thaliana cells. This capacity to mitigate salt stress could be due to a higher ascorbate pool that could allow C. maritima reducing the oxidative stress generated in response to NaCl. It further appeared that a higher number of C. maritima cultured cells when compared to A. thaliana could efficiently manage the Na(+) accumulation into the cytoplasm through non selective cation channels allowing also reducing the ROS generation and the subsequent cell death.

Early events induced by the toxin deoxynivalenol lead to programmed cell death in Nicotiana tabacum cells.

Deoxynivalenol (DON) is a mycotoxin affecting animals and plants. This toxin synthesized by Fusarium culmorum and Fusarium graminearum is currently believed to play a decisive role in the fungal phytopathogenesis as a virulence factor. Using cultured cells of Nicotiana tabacum BY2, we showed that DON-induced programmed cell death (PCD) could require transcription and translation processes, in contrast to what was observed in animal cells. DON could induce different cross-linked pathways involving (i) reactive oxygen species (ROS) generation linked, at least partly, to a mitochondrial dysfunction and a transcriptional down-regulation of the alternative oxidase (Aox1) gene and (ii) regulation of ion channel activities participating in cell shrinkage, to achieve PCD.

Deciphering the dual effect of lipopolysaccharides from plant pathogenic Pectobacterium.

Lipopolysaccharides (LPS) are a component of the outer cell surface of almost all Gram-negative bacteria and play an essential role for bacterial growth and survival. Lipopolysaccharides represent typical microbe-associated molecular pattern (MAMP) molecules and have been reported to induce defense-related responses, including the expression of defense genes and the suppression of the hypersensitive response in plants. However, depending on their origin and the challenged plant, LPS were shown to have complex and different roles. In this study we showed that LPS from plant pathogens Pectobacterium atrosepticum and Pectobacterium carotovorum subsp. carotovorum induce common and different responses in A. thaliana cells when compared to those induced by LPS from non-phytopathogens Escherichia coli and Pseudomonas aeruginosa. Among common responses to both types of LPS are the transcription of defense genes and their ability to limit of cell death induced by Pectobacterium carotovorum subsp carotovorum. However, the differential kinetics and amplitude in reactive oxygen species (ROS) generation seemed to regulate defense gene transcription and be determinant to induce programmed cell death in response to LPS from the plant pathogenic Pectobacterium. These data suggest that different signaling pathways could be activated by LPS in A. thaliana cells.

Deciphering early events involved in hyperosmotic stress-induced programmed cell death in tobacco BY-2 cells.

Hyperosmotic stresses represent one of the major constraints that adversely affect plants growth, development, and productivity. In this study, the focus was on early responses to hyperosmotic stress- (NaCl and sorbitol) induced reactive oxygen species (ROS) generation, cytosolic Ca(2+) concentration ([Ca(2+)]cyt) increase, ion fluxes, and mitochondrial potential variations, and on their links in pathways leading to programmed cell death (PCD). By using BY-2 tobacco cells, it was shown that both NaCl- and sorbitol-induced PCD seemed to be dependent on superoxide anion (O2·(-)) generation by NADPH-oxidase. In the case of NaCl, an early influx of sodium through non-selective cation channels participates in the development of PCD through mitochondrial dysfunction and NADPH-oxidase-dependent O2·(-) generation. This supports the hypothesis of different pathways in NaCl- and sorbitol-induced cell death. Surprisingly, other shared early responses, such as [Ca(2+)]cyt increase and singlet oxygen production, do not seem to be involved in PCD.