Altered Metabolism during the Dark Period in Drosophila Short Sleep Mutants.

Sleep is regulated via circadian mechanisms, but effects of sleep disruption on physiological rhythms, in particular metabolic cycling, remain unclear. To examine this question, we probed diurnal metabolic alterations of two Drosophila short sleep mutants, fumin and sleepless. Samples were collected with high temporal sampling (every 2 h) over 24 h under a 12:12 light:dark cycle, and profiling was done using an ion-switching LCMS/MS method. Fewer metabolites with 24 h oscillations were noted with short sleep (50 and 46 in fumin and sleepless, BH. Q < 0.2 by RAIN analysis) compared to a wild-type control (iso31, 63 with BH. Q < 0.2), and peak phases of the sleep mutants were consolidated into two major phase peaks at mid-day and middle of night. Overall, altered nicotinate/nicotinamide, alanine/aspartate/glutamate, acetylcholine, glyoxylate/dicarboxylate, and TCA cycle metabolism were observed in the short sleep mutants, indicative of increased energetic demand and oxidative stress compared to wild type. Both changes in cycling and discriminant models suggest unique alterations in the dark period indicative of constrained metabolic networks. Thus, we conclude that sleep loss alters metabolic function uniquely throughout the day, and further examination of specific mechanisms is warranted.

Tracking Sugar-Elicited Local Searching Behavior in Drosophila.

Foraging behavior is essential for the survival of organisms as it enables them to locate and acquire essential food resources. In Drosophila, hunger triggers a distinct search behavior following the consumption of small quantities of a sugar solution. This report presents a simple experimental setup to study sugar-elicited search behavior with the aim of uncovering the underlying mechanisms. Minute quantities of concentrated sugar solution elicit sustained searching behavior in flies. The involvement of path integration in this behavior has been established, as flies utilize their trajectory to return to the sugar location. The most recent findings provide evidence of temporal modulation in the initiation and intensity of the search behavior after sugar intake. We have also used this setup for artificial activation of specific taste-receptor neurons in the pharynx, which elicits the search behavior. The Drosophila neurogenetic toolkit offers a diverse array of tools and techniques that can be combined with the sugar-elicited search behavior paradigm to study the neural and genetic mechanisms underlying foraging. Understanding the neural basis of hunger-driven searching behavior in flies contributes to the field of neurobiology as a whole, offering insights into the regulatory mechanisms that govern feeding behaviors not only in other organisms but also in humans.

Covid-19 Pandemic and Metabolic Aging.

In contrast to chronological aging, biological aging or metabolic aging is a relative age of cells and tissues and the damage they've accumulated over years. Comparison of one's basal metabolic rate (BMR) to the average BMR of one's chronological age group gives the Metabolic age. Higher metabolic age of younger population is a serious concern and a key factor that leads to metabolic disorders in all age groups. Slower metabolism is a symbol of older age. Higher metabolic age indicates poor metabolism and higher risk of getting diseases and health complications later in life. Therefore, aging faster metabolically can severely impact chronological age. People with obesity and diabetes suffered the most due to current ongoing Covid-19 pandemic. Data suggest that SARS-CoV-2 patients with concomitant metabolic diseases had higher risk of worse prognosis and mortality. Lowering metabolic age can thus reduce the risk of getting age related health conditions and mitigate morbidities caused by pandemic like Covid-19. Therefore, maintaining healthy metabolic age in all age groups is required in the current unprecedented times. The aim of the review is to raise the prime concerns and to improve the population health outcomes by reducing the metabolic age.

Nutrient Sensing via Gut in Drosophila melanogaster.

Nutrient-sensing mechanisms in animals' sense available nutrients to generate a physiological regulatory response involving absorption, digestion, and regulation of food intake and to maintain glucose and energy homeostasis. During nutrient sensing via the gastrointestinal tract, nutrients interact with receptors on the enteroendocrine cells in the gut, which in return respond by secreting various hormones. Sensing of nutrients by the gut plays a critical role in transmitting food-related signals to the brain and other tissues informing the composition of ingested food to digestive processes. These signals modulate feeding behaviors, food intake, metabolism, insulin secretion, and energy balance. The increasing significance of fly genetics with the availability of a vast toolbox for studying physiological function, expression of chemosensory receptors, and monitoring the gene expression in specific cells of the intestine makes the fly gut the most useful tissue for studying the nutrient-sensing mechanisms. In this review, we emphasize on the role of Drosophila gut in nutrient-sensing to maintain metabolic homeostasis and gut-brain cross talk using endocrine and neuronal signaling pathways stimulated by internal state or the consumption of various dietary nutrients. Overall, this review will be useful in understanding the post-ingestive nutrient-sensing mechanisms having a physiological and pathological impact on health and diseases.

Modulation of sugar feeding behavior by Gymnema sylvestre in Drosophila melanogaster.

INTRODUCTION: Sugar is the main source of energy for nearly all animals. However, consumption of a high amount of sugars can lead to many metabolic disorders hence, balancing calorie intake in the form of sugar is required. Various herbs are in use to control body weight, cure diabetes and control elevated blood sugar levels. One such herb is Gymnema sylvestre commonly called Gurmar (destroyer of sugar). Gurmar selectively inhibits sugar sensation by mechanisms that are still elusive. OBJECTIVES: The primary objective of this study is to understand the effect of gurmar on sweet taste feeding behaviour in insects using the invertebrate model system Drosophila melanogaster. METHODS: For this study, we used feeding assays, spectrophotometry and Proboscis Extension Reflex (PER) assay to determine how flies detect gurmar. Additionally, life span analysis, egg-laying behaviour and developmental profiles were used to probe the role of gurmar on the overall health of the flies. During the whole study, we used only the raw powdered form of gurmar (dried leaves) to examine its effect on sweet taste feeding behaviour. RESULTS: Our data demonstrate that whole gurmar in a raw powdered form is aversive to flies and inhibits sugar evoked PER and feeding responses. Also, we observed it takes at least 24 h of starvation time to reduce the consumption of sugar in flies pre-fed on gurmar. Flies lay a fewer number of eggs on gurmar media and show developmental defects. Our data suggest that flies detect gurmar using both taste and olfactory cues. CONCLUSION: Understanding how gurmar reshapes taste curves to promote reduced consumption of sugars in flies will open up avenues to help people with health issues related to high sugar consumption, but our data also highlights that its consumption should be carefully considered since gurmar is aversive to flies and has detrimental effects on development.

Starvation and activity dependent modulation of salt taste behavior in Drosophila.

Background: Sodium present in NaCl is a fundamental nutrient required for many physiological processes but high salt consumption in western world is contributing health risk to all age individuals. Although high salt detection pathways have been studied in great detail, the mechanisms that regulate high salt consumption in animals are largely unknown. To understand how pre-exposure to high NaCl diet modulates subsequent feeding behavior, we looked into the neural mechanisms of high NaCl consumption in adult Drosophila. Method: We used Neuro-Genetics, imaging and behavioral assays to determine how flies respond to high NaCl exposure. Result: We studied the neural mechanism by which flies modify their acceptance of high salt as a function of diet, where a long-term high-salt exposure increases taste sensitivities of pharyngeal LSO (Labral sense organ) neurons and enhances high salt intake. We discovered that exposing flies to high NaCl diet(200mM NaCl in fly food) for three days modify their feeding responses to high levels of salt. High NaCl fed flies show decline in high salt aversion under starvation. Genetic suppression of LSO pharyngeal neurons in high NaCl fed flies inhibits excessive salt intake. We found that this modulation requires functional LSO neurons and starvation state, and that multiple independent taste receptor neurons and pathways are involved in this process. Silencing any one of multiple LSO neuronal types inhibits excessive salt intake. Conclusion: Our data support the idea that high dietary salt modulates and reshapes salt and other taste curves to promote over consumption of food in flies. Our study suggest flies can adapt to the amount of salt ingested over several days, indicating the presence of a critical mechanism to reset the salt appetite and related neural circuits. Identification of new molecular sensors for salt and related neural controls such as hormones, neuropeptides, and neurotransmitters may yield insights into the coordination of processes in the nervous system.

Fluoresceinated Aminohexanol Tethered Inositol Hexakisphosphate: Studies on Arabidopsis thaliana and Drosophila melanogaster and Docking with 2P1M Receptor.

Inositol hexakisphosphate (InsP6; phytic acid) is considered as the second messenger and plays a very important role in plants, animals, and human beings. It is the principal storage form of phosphorus in many plant tissues, especially in dry fruits, bran, and seeds. The resulting anion is a colorless species that plays a critical role in nutrition and is believed to cure many diseases. A fluoresceinated aminohexanol tethered inositol hexakisphosphate (III) had been synthesized earlier involving many complicated steps. We describe here a simple two-step synthesis of (III) and its characterization using different techniques such as matrix-assisted laser desorption ionization mass spectrometry, tandem mass spectrometry, and Fourier transform infrared, ultraviolet-visible, ultraviolet-fluorescence, 1H nuclear magnetic resonance (NMR), and two-dimensional NMR spectroscopies. The effect of (III) has been investigated in the model systems, Arabidopsis thaliana and Drosophila melanogaster. Using Schrodinger software, computational studies on the binding of (III) with the protein 2P1M (Auxin-receptor TIR1-adaptor ASK1 complex) has revealed strong binding propensity with this compound. These studies on the fluoresceinated tethered phytic acid could have far reaching implications on its efficacy for human health and treatment of diseases (cancer/tumor and glioblastoma) and for understanding phosphorous recycling in the environment, especially for plant systems.

Salt an Essential Nutrient: Advances in Understanding Salt Taste Detection Using Drosophila as a Model System.

Taste modalities are conserved in insects and mammals. Sweet gustatory signals evoke attractive behaviors while bitter gustatory information drive aversive behaviors. Salt (NaCl) is an essential nutrient required for various physiological processes, including electrolyte homeostasis, neuronal activity, nutrient absorption, and muscle contraction. Not only mammals, even in Drosophila melanogaster, the detection of NaCl induces two different behaviors: Low concentrations of NaCl act as an attractant, whereas high concentrations act as repellant. The fruit fly is an excellent model system for studying the underlying mechanisms of salt taste due to its relatively simple neuroanatomical organization of the brain and peripheral taste system, the availability of powerful genetic tools and transgenic strains. In this review, we have revisited the literature and the information provided by various laboratories using invertebrate model system Drosophila that has helped us to understand NaCl salt taste so far. We hope that this compiled information from Drosophila will be of general significance and interest for forthcoming studies of the structure, function, and behavioral role of NaCl-sensitive (low and high concentrations) gustatory circuitry for understanding NaCl salt taste in all animals.

Secondary taste neurons that convey sweet taste and starvation in the Drosophila brain.

The gustatory system provides vital sensory information to determine feeding and appetitive learning behaviors. Very little is known, however, about higher-order gustatory circuits in the highly tractable model for neurobiology, Drosophila melanogaster. Here we report second-order sweet gustatory projection neurons (sGPNs) in the Drosophila brain using a powerful behavioral screen. Silencing neuronal activity reduces appetitive behaviors, whereas inducible activation results in food acceptance via proboscis extensions. sGPNs show functional connectivity with Gr5a(+) sweet taste neurons and are activated upon sucrose application to the labellum. By tracing sGPN axons, we identify the antennal mechanosensory and motor center (AMMC) as an immediate higher-order processing center for sweet taste. Interestingly, starvation increases sucrose sensitivity of the sGPNs in the AMMC, suggesting that hunger modulates the responsiveness of the secondary sweet taste relay. Together, our results provide a foundation for studying gustatory processing and its modulation by the internal nutrient state.

Odour receptors and neurons for DEET and new insect repellents.

There are major impediments to finding improved DEET alternatives because the receptors causing olfactory repellency are unknown, and new chemicals require exorbitant costs to determine safety for human use. Here we identify DEET-sensitive neurons in a pit-like structure in the Drosophila melanogaster antenna called the sacculus. They express a highly conserved receptor, Ir40a, and flies in which these neurons are silenced or Ir40a is knocked down lose avoidance to DEET. We used a computational structure-activity screen of >400,000 compounds that identified >100 natural compounds as candidate repellents. We tested several and found that most activate Ir40a(+) neurons and are repellents for Drosophila. These compounds are also strong repellents for mosquitoes. The candidates contain chemicals that do not dissolve plastic, are affordable and smell mildly like grapes, with three considered safe in human foods. Our findings pave the way to discover new generations of repellents that will help fight deadly insect-borne diseases worldwide.

Mutants in Drosophila TRPC channels reduce olfactory sensitivity to carbon dioxide.

BACKGROUND: Members of the canonical Transient Receptor Potential (TRPC) class of cationic channels function downstream of Gαq and PLCß in Drosophila photoreceptors for transducing visual stimuli. Gαq has recently been implicated in olfactory sensing of carbon dioxide (CO(2)) and other odorants. Here we investigated the role of PLCß and TRPC channels for sensing CO(2) in Drosophila. METHODOLOGY/PRINCIPAL FINDINGS: Through behavioral assays it was demonstrated that Drosophila mutants for plc21c, trp and trpl have a reduced sensitivity for CO(2). Immuno-histochemical staining for TRP, TRPL and TRPγ indicates that all three channels are expressed in Drosophila antennae including the sensory neurons that express CO(2) receptors. Electrophysiological recordings obtained from the antennae of protein null alleles of TRP (trp(343)) and TRPL (trpl(302)), showed that the sensory response to multiple concentrations of CO(2) was reduced. However, trpl(302); trp(343) double mutants still have a residual response to CO(2). Down-regulation of TRPC channels specifically in CO(2) sensing olfactory neurons reduced the response to CO(2) and this reduction was obtained even upon down-regulation of the TRPCs in adult olfactory sensory neurons. Thus the reduced response to CO(2) obtained from the antennae of TRPC RNAi strains is not due to a developmental defect. CONCLUSION: These observations show that reduction in TRPC channel function significantly reduces the sensitivity of the olfactory response to CO(2) concentrations of 5% or less in adult Drosophila. It is possible that the CO(2) receptors Gr63a and Gr21a activate the TRPC channels through Gαq and PLC21C.

Kinesin heavy chain function in Drosophila glial cells controls neuronal activity.

Kinesin heavy chain (Khc) is crucially required for axonal transport and khc mutants show axonal swellings and paralysis. Here, we demonstrate that in Drosophila khc is equally important in glial cells. Glial-specific downregulation of khc by RNA interference suppresses neuronal excitability and results in spastic flies. The specificity of the phenotype was verified by interspecies rescue experiments and further mutant analyses. Khc is mostly required in the subperineurial glia forming the blood-brain barrier. Following glial-specific knockdown, peripheral nerves are swollen with maldistributed mitochondria. To better understand khc function, we determined Khc-dependent Rab proteins in glia and present evidence that Neurexin IV, a well known blood-brain barrier constituent, is one of the relevant cargo proteins. Our work shows that the role of Khc for neuronal excitability must be considered in the light of its necessity for directed transport in glia.

Mutants in phospholipid signaling attenuate the behavioral response of adult Drosophila to trehalose.

In Drosophila melanogaster, gustatory receptor genes (Grs) encode putative G-protein-coupled receptors (GPCRs) that are expressed in gustatory receptor neurons (GRNs). One of the Gr genes, Gr5a, encodes a receptor for trehalose that is expressed in a subset of GRNs. Although a role for the G protein, Gsα, has been shown in Gr5a-expressing taste neurons, there is the residual responses to trehalose in Gsα mutants which could suggest additional transduction mechanisms. Expression and genetic analysis of the heterotrimeric G-protein subunit, Gq, shown here suggest involvement of this Gα subunit in trehalose perception in Drosophila. A green fluorescent protein reporter of Gq expression is detected in gustatory neurons in the labellum, tarsal segments, and wing margins. Animals heterozygous for dgq mutations and RNA interference-mediated knockdown of dgq showed reduced responses to trehalose in the proboscis extension reflex assay and feeding behavior assay. These defects were rescued by targeted expression of the wild-type dgqα transgene in the GRNs. These data together with observations from other mutants in phospholipid signaling provide insights into the mechanisms of taste transduction in Drosophila.

Drosophila mutants in phospholipid signaling have reduced olfactory responses as adults and larvae.

In this paper, we show that mutants in the gene stambhA (stmA), which encodes a putative phosphatidylinositol 4,5 bisphosphate-diacylglycerol lipase, exhibit a significant reduction in the amplitudes of odor-evoked responses recorded from the antennal surface of adult Drosophila. This lends support to previously published findings that olfactory transduction in Drosophila requires a phospholipid intermediate. Mutations in stmA also affect the olfactory behavior response of larvae. Moreover, there is a requirement for G(q)alpha and phospholipase Cbeta function in larval olfaction. The results suggest that larval olfactory transduction, like that of the adult, utilizes a phospholipid second messenger, generated by the activation of G(q)alpha and Plcbeta21c, and modulated by the stmA gene product.

Reduced odor responses from antennal neurons of G(q)alpha, phospholipase Cbeta, and rdgA mutants in Drosophila support a role for a phospholipid intermediate in insect olfactory transduction.

Mechanisms by which G-protein-coupled odorant receptors transduce information in insects still need elucidation. We show that mutations in the Drosophila gene for G(q)alpha (dgq) significantly reduce both the amplitude of the field potentials recorded from the whole antenna in responses to odorants as well as the frequency of evoked responses of individual sensory neurons. This requirement for G(q)alpha is for adult function and not during antennal development. Conversely, brief expression of a dominant-active form of G(q)alpha in adults leads to enhanced odor responses. To understand signaling downstream of G(q)alpha in olfactory sensory neurons, genetic interactions of dgq were tested with mutants in genes known to affect phospholipid signaling. dgq mutant phenotypes were further enhanced by mutants in a PLCbeta (phospholipase Cbeta) gene, plc21C. Interestingly although, the olfactory phenotype of mutant alleles of diacylglycerol kinase (rdgA) was rescued by dgq mutant alleles. Our results suggest that G(q)alpha-mediated olfactory transduction in Drosophila requires a phospholipid second messenger the levels of which are regulated by a cycle of phosphatidylinositol 1,4-bisphosphate breakdown and regeneration.