Benzylisoquinoline alkaloids (BIAs) are a structurally diverse group of plant specialized metabolites with a long history of investigation. Although the ecophysiological functions of most BIAs are unknown, the medicinal properties of many compounds have been exploited for centuries. These include the narcotic analgesics codeine and morphine, the antimicrobial agents sanguinarine and berberine, and the antitussive and anticancer drug noscapine. BIA biosynthesis involves a restricted number of enzyme types that catalyze landmark coupling reactions and subsequent functional group modifications. A pathogenesis-related (PR)10/Bet v1 'Pictet-Spenglerase', several O-methyl-, N-methyl- and O-acetyltransferases, cytochromes P450, FAD-dependent oxidases, non-heme dioxygenases and NADPH-dependent reductases have been implicated in the multistep pathways leading to structurally diverse alkaloids. A small number of plant species, including opium poppy (Papaver somniferum) and other members of the Ranunculales, have emerged as model systems to study BIA metabolism. The expansion of resources to include a wider range of plant species is creating an opportunity to investigate previously uncharacterized BIA pathways. Contemporary knowledge of BIA metabolism reflects over a century of research coupled with the development of key innovations such as radioactive tracing, enzyme isolation and molecular cloning, and functional genomics approaches such as virus-induced gene silencing. Recently, the emergence of transcriptomics, proteomics and metabolomics has expedited the discovery of new BIA biosynthetic genes. The growing repository of BIA biosynthetic genes is providing the parts required to apply emerging synthetic biology platforms to the development of production systems in microbes as an alternative to plants as a commecial source of valuable BIAs.

/pdf/benzylisoquinoline-alkaloid-metabolism-a-century-of-41gr2sigfl.pdf

Benzylisoquinoline Alkaloid Metabolism: A Century of Discovery and a Brave New World

https://www.cell.com/trends/genetics/pdf/S0168-9525(15)00165-1.pdf

Drosophila Chemoreceptors: A Molecular Interface Between the Chemical World and the Brain.

The ability to find and consume nutrient-rich diets for successful reproduction and survival is fundamental to animal life. Among the nutrients important for all animals are polyamines, a class of pungent smelling compounds required in numerous cellular and organismic processes. Polyamine deficiency or excess has detrimental effects on health, cognitive function, reproduction, and lifespan. Here, we show that a diet high in polyamine is beneficial and increases reproductive success of flies, and we unravel the sensory mechanisms that attract Drosophila to polyamine-rich food and egg-laying substrates. Using a combination of behavioral genetics and in vivo calcium imaging, we demonstrate that Drosophila uses multisensory detection to find and evaluate polyamines present in overripe and fermenting fruit, their favored feeding and egg-laying substrate. In the olfactory system, two coexpressed ionotropic receptors (IRs), IR76b and IR41a, mediate the long-range attraction to the odor. In the gustatory system, multimodal taste sensation by IR76b receptor and GR66a bitter receptor neurons is used to evaluate quality and valence of the polyamine providing a mechanism for the fly’s high attraction to polyamine-rich and sweet decaying fruit. Given their universal and highly conserved biological roles, we propose that the ability to evaluate food for polyamine content may impact health and reproductive success also of other animals including humans.

/pdf/ionotropic-chemosensory-receptors-mediate-the-taste-and-28r87xydtm.pdf

Ionotropic Chemosensory Receptors Mediate the Taste and Smell of Polyamines.

This Analysis compares four commonly used assays to measure food intake in flies and identifies radioisotope-labeling and the capillary feeder (CAFE) as the most reproducible and sensitive.

Quantifying Drosophila food intake: comparative analysis of current methodology

To survive and successfully reproduce animals need to maintain a balanced intake of nutrients and energy. The nervous system of insects has evolved multiple mechanisms to regulate feeding behaviour. When animals are faced with the choice to feed, several decisions must be made: whether or not to eat, how much to eat, what to eat and when to eat. Using Drosophila melanogaster substantial progress has been achieved in understanding the neuronal and molecular mechanisms controlling feeding decisions. These feeding decisions are implemented in the nervous system on multiple levels, from alterations in the sensitivity of peripheral sensory organs to the modulation of memory systems. This review discusses methodologies developed in order to study insect feeding, the effects of neuropeptides and neuromodulators on feeding behaviour, behavioural evidence supporting the existence of internal energy sensors, neuronal and molecular mechanisms controlling protein intake and finally the regulation of feeding by circadian rhythms and sleep. From the discussed data a conceptual framework starts to emerge which aims to explain the molecular and neuronal processes maintaining the stability of the internal milieu.

/pdf/the-dilemmas-of-the-gourmet-fly-the-molecular-and-neuronal-4gik406uh5.pdf

The dilemmas of the gourmet fly: the molecular and neuronal mechanisms of feeding and nutrient decision making in Drosophila.

In flies and humans, bitter chemicals are known to inhibit sugar detection, but the adaptive role of this inhibition is often overlooked. At best, this inhibition is described as contributing to the rejection of potentially toxic food, but no studies have addressed the relative importance of the direct pathway that involves activating bitter-sensitive cells versus the indirect pathway represented by the inhibition of sugar detection. Using toxins to selectively ablate or inactivate populations of bitter-sensitive cells, we assessed the behavioral responses of flies to sucrose mixed with strychnine (which activates bitter-sensitive cells and inhibits sugar detection) or with L-canavanine (which only activates bitter-sensitive cells). As expected, flies with ablated bitter-sensitive cells failed to detect L-canavanine mixed with sucrose in three different feeding assays (proboscis extension responses, capillary feeding, and two-choice assays). However, such flies were still able to avoid strychnine mixed with sucrose. By means of electrophysiological recordings, we established that bitter molecules differ in their potency to inhibit sucrose detection and that sugar-sensing inhibition affects taste cells on the proboscis and the legs. The optogenetic response of sugar-sensitive cells was not reduced by strychnine, thus suggesting that this inhibition is linked directly to sugar transduction. We postulate that sugar-sensing inhibition represents a mechanism in insects to prevent ingesting harmful substances occurring within mixtures.

https://www.jneurosci.org/content/jneuro/35/9/3990.full.pdf

Dual mechanism for bitter avoidance in Drosophila.

Drosophila melanogaster adapt their food consumption to their internal needs and avoid ingesting noxious molecules. Defects in the genes involved in these decisions induce behavioral alterations that are usually screened by monitoring flies feeding in 2-choice or in no-choice situations. Here, we introduce a new behavioral test in which groups of flies are given access to 6 capillary feeders (MultiCAFE) containing fructose mixed with a serial dilution of a test substance. Using quinine, we first showed that fly density, distance between capillaries, and order of presentation have a minor impact on the discrimination performances of the flies. Fly discrimination was also only marginally affected by the type of test (no-choice, binary, or multiple-choice). Interestingly, the feeding reduction was well correlated with a reduction of the firing elicited by the mixture in sugar-sensitive gustatory receptor neurons, suggesting that several mechanisms concur to allow flies to make their choices. In addition to quinine, flies exhibited marked dose-dependent aversions to the consumption of berberine, caffeine, lobeline, nicotine, papaverine, strychnine, and theophylline, which all taste bitter to humans. Thus, despite of the multiplicity of choices available, flies consistently avoid alkaloids mixed with a sugar solution, and their choices are strongly dependent on their taste system. The MultiCAFE assay represents an interesting alternative to other feeding tests, in that it allows monitoring of the absolute consumption while also requiring less flies and time to run than other assays.

Consumption of bitter alkaloids in Drosophila melanogaster in multiple-choice test conditions.

Most animals possess taste receptors neurons detecting potentially noxious compounds. In humans, the ligands which activate these neurons define a sensory space called “bitter”. By extension, this term has been used in animals and insects to define molecules which induce aversive responses. In this review, based on our observations carried out in Drosophila, we examine how bitter compounds are detected and if the activation of bitter-sensitive neurons respond only to molecules bitter to humans. Like most animals, flies detect bitter chemicals through a specific population of taste neurons, distinct from those responding to sugars or to other modalities. Activating bitter-sensitive taste neurons induce aversive reactions and inhibits feeding. Bitter molecules also contribute to the suppression of sugar-neuron responses and can lead to a complete inhibition of the responses to sugar at the periphery. Since some bitter molecules activate bitter-sensitive neurons and some inhibit sugar detection, bitter molecules are represented by two sensory spaces which are only partially congruent. In addition to molecules which impact feeding, we recently discovered that the activation of bitter-sensitive neurons also induces grooming. Bitter-sensitive neurons of the wings and of the legs can sense chemicals from the gram negative bacteria, Escherichia coli, thus adding another biological function to these receptors. Bitter-sensitive neurons of the proboscis also respond to inhibitory pheromones such as 7-tricosene. Activating these neurons by bitter molecules in the context of sexual encounter inhibits courting and sexual reproduction, while activating these neurons with 7-tricosene in a feeding context will inhibit feeding. The picture that emerges from these observations is that the taste system is composed of detectors which monitor different “categories” of ligands, which facilitate or inhibit behaviors depending on the context (feeding, sexual reproduction, hygienic behavior), thus considerably extending the initial definition of “bitter” tasting.

/pdf/drosophila-bitter-taste-s-4tky8uj390.pdf

Marie-Jeanne Sellier

Papers

Dual mechanism for bitter avoidance in Drosophila.

Consumption of bitter alkaloids in Drosophila melanogaster in multiple-choice test conditions.

Drosophila Bitter Taste(s).

Mixture interactions in taste sensilla of [i]Drosophila melanogaster[/i]