Ether resistance in Drosophila melanogaster

Summary Strains set up from single inseminated females of D. melanogaster from the wild differ in their resistance to the anaesthetics, ether and chloroform. The main differences between four selected extreme strains could be explained by additive genes, which in the case of ether resistance were located to regions of chromosomes 2 and 3. The lack of correspondence between ether and chloroform resistance between strains indicates that although the type of genetic architecture controlling the traits is similar, the actual genes differ, which is reasonable in view of their differing chemical structures. Quite high heritabilities were found for resistance to ether based on five inbred strains. No significant associations between resistance to ether and body weight, developmental rate or longevity were found.It is clear that resistance to both anaesthetics would be amenable to more detailed genetic analyses. It is pointed out that the general conclusions reached from such studies will have implications with respect to the effect of chemicals such as insecticides, not naturally present in nature.     

Ecdysone signaling in adult Drosophila melanogaster

The steroid hormone 20-hydroxyecdysone and its EcR/USP receptor are vital during arthropod development for coordinating molting and metamorphosis. Traditionally, little attention has been given to potential post-developmental functions for this hormone signaling system. However, recent studies in Drosophila melanogaster indicate that the hormone and receptor are present and active in adults and that mutations decreasing hormone or receptor levels affect diverse processes such as reproduction, behavior, stress resistance, and lifespan. We review the current state of knowledge regarding adult hormone production and titers and discuss receptor expression and activity in order to identify potential mechanisms which explain the observed mutant phenotypes. Finally, we describe future research directions focused on identifying isoform-specific functions of EcR, distinguishing effects from EcR/USP gene activation and repression, and determining how ecdysone signaling impacts different tissue types.     

Inhibition of ADRP prevents diet-induced insulin resistance

Diets with high fat content induce steatosis, insulin resistance, and type 2 diabetes. The lipid droplet protein adipose differentiation-related protein (ADRP) mediates hepatic steatosis, but whether this affects insulin action in the liver or peripheral organs in diet-induced obesity is uncertain. We fed C57BL/6J mice a high-fat diet and simultaneously treated them with an antisense oligonucleotide (ASO) against ADRP for 4 wk. Glucose homeostasis was assessed with clamp and tracer techniques. ADRP ASO decreased the levels of triglycerides and diacylglycerol in the liver, but fatty acids, long-chain fatty acyl CoAs, ceramides, and cholesterol were unchanged. Insulin action in the liver was enhanced after ADRP ASO treatment, whereas muscle and adipose tissue were not affected. ADRP ASO increased the phosphorylation of insulin receptor substrate (IRS)1, IRS2, and Akt, and decreased gluconeogenic enzymes and PKCepsilon, consistent with its insulin-sensitizing action. These results demonstrate an important role for ADRP in the pathogenesis of diet-induced insulin resistance.     

The effects of age on radiation resistance and oxidative stress in adult Drosophila melanogaster

Drosophila melanogaster (fruit fly) is a well-established model organism for genetic studies of development and aging. We examined the effects of lethal ionizing radiation on male and female adult Drosophila of different ages, using doses of radiation from 200 to 1500 Gy. Fifty percent lethality 2 days postirradiation (LD(50/2)) in wild-type 1-day-old adult fruit flies was approximately 1238 Gy for males and 1339 Gy for females. We observed a significant age-dependent decline in the radiation resistance of both males and females. Radiation damage is postulated to occur by the generation of oxygen radicals. An age-related decline in the ability of flies to resist an agent that induces oxygen radicals, paraquat, was observed when comparing 10- and 20-day adults. Female flies are more resistant to paraquat than male flies. Oxidative stress mediated by paraquat was additive with sublethal exposures to radiation in young adults. Therefore, the ability to repair the damage caused by oxygen radicals seems to decline with the age of the flies. Because Drosophila adults are largely post-mitotic, our data suggest that adult Drosophila melanogaster can serve as an excellent model to study the factors responsible for radiation resistance in post-mitotic tissue and age-dependent changes in this resistance.     

Aldehyde dehydrogenase is essential for both adult and larval ethanol resistance in Drosophila melanogaster

The enzyme aldehyde dehydrogenase (ALDH) is essential for ethanol metabolism in mammals, converting the highly toxic intermediate acetaldehyde to acetate. The role of ALDH in Drosophila has been debated, with some authors arguing that, at least in larvae, acetaldehyde detoxification is carried out mainly by alcohol dehydrogenase (ADH), the enzyme responsible for converting ethanol to acetaldehyde. Here, we report the creation and characterization of four null mutants of Aldh, the putative structural locus for ALDH. Aldh null larvae and adults are poisoned by ethanol concentrations easily tolerated by wild-types; their ethanol sensitivity is in fact comparable to that of Adh nulls. The results refute the view that ALDH plays only a minor role in ethanol detoxification in larvae, and suggest that Aldh and Adh may be equally important players in the evolution of ethanol resistance in fruit-breeding Drosophila.     

Selection for adult desiccation resistance in Drosophila melanogaster: fitness components, larval resistance and stress correlations

In previous experiments we found that Drosophila melanogaster lines selected for increased adult desiccation resistance had increased resistance to other environmental stresses at the adult stage including starvation, intense ⁶⁰Co-γ radiation and a toxic ethanol level. In further studies on these lines, we now show that selection did not alter resistance to desiccation and ethanol at the larval stage. As well as having a lower early fecundity, selected lines showed increased adult male longevity and increased viability at high larval densities compared with control lines. There were no changes in development time or mating success. The increased male longevity is consistent with the reduced metabolic rate of the selected lines.
A genetic correlation between resistance to different stresses was confirmed by an analysis of isofemale lines derived from a population founded by flies from a stress-resistant line and an unselected line. The results are consistent with the existence of genes segregating in natural populations conferring increased general stress resistance.     

Developmental stage modifies diet-induced peripheral insulin resistance in rats

In the present study, the effects of age and diet on glucose disappearance and tissue-specific glucose uptake (R'g) were examined under basal or hyperinsulinemic, euglycemic conditions in male Sprague-Dawley rats. Rats were equicalorically fed either a high-starch diet (68% of kcal), high-fat diet (HFD; 45% of kcal), or high-sucrose diet (68% of kcal), beginning at either 5 (W; weanling), 10 (Y; young), 18 (M; mature), or 58 wk (O; older) of age for 5 wks (n = 6-9. group(-1) x diet(-1)). Body weight gain was not significantly different among dietary groups within a given age. Significant (P< 0.05) age effects were observed on basal and clamp free fatty acid concentrations. Significant diet effects were observed on basal and clamp triglyceride concentrations. There were significant diet and age effects on basal skeletal muscle R'g. This interaction was primarily due to an age-associated increase in basal R'g microg x g(-1). min(-1)) in HFD (gastrocnemius R'g: 0.9+/-0.2 in W, 1.1+/-0.2 in Y, 1.8+/-0.2 in M, 2.5+/-0.2 in O). Both age and diet significantly decreased insulin-stimulated muscle R'g. However, whereas age-associated reductions in both glucose-6-phosphate concentration and glycogen synthase activity were observed, significant diet effects were observed on glucose-6-phosphate concentrations only. Age significantly reduced basal and clamp adipose tissue R'g when expressed per gram of tissue but significantly increased R'g when expressed per total fat pad mass. These data suggest that diet-induced changes in peripheral glucose metabolism are modulated by age.     

Calcium homeostasis in larval and adult Drosophila melanogaster

Calcium homeostasis in Drosophila melanogaster was examined in response to the challenges imposed by growth, reproduction and variations in dietary calcium content. Turnover time for calcium, calculated as the time for (45)Ca(2+)to accumulate to half the steady state value of 3.46 nmol/fly, was 3.3 days. Although larvae weighed 2x as much as adults, they contained 3-4x as much calcium. Anterior Malpighian tubules (Mts) contain much more calcium than posterior Mts, accounting for 25-30% of the calcium content of the whole fly. In response to a 6.2-fold increase in dietary calcium level, calcium content of whole flies increased only 10%. Hemolymph calcium concentration ( approximately 0.5 mM) was similar in males and females and in animals raised on diets differing in calcium content. Fluid secretion rate, secreted fluid calcium concentration, and transepithelial calcium flux in tubules isolated from flies raised on high and low calcium diets did not differ significantly. Malpighian tubules secrete calcium at rates sufficient to eliminate whole body calcium content in 0.5 and 3 days for tubules secreting fluid at basal and maximal rates, respectively. It is suggested that flies absorb high quantities of calcium from the diet and maintain homeostasis through the combined effects of elimination of calcium in fluid secreted by the Malpighian tubules and the sequestration of calcium in granules, especially within the distal segment of the anterior pair of Malpighian tubules.     

Persistent larval sensory neurons in adult Drosophila melanogaster

Using a combination of lineage tracing and laser ablation, we have identified a segmentally repeated array of embryonically produced sensory neurons that persist through metamorphosis into adult stages of Drosophila development. The persistent sensory neurons are found in all unfused abdominal segments, but there is segment-specific variation in the number of neurons observed. There are 12 persistent neurons in the first abdominal segment (A1), 18 in the second (A2), and 16 in segments A3-A7. Most are internal sensory neurons (dendritic arborization neurons and bipolar dendrite neurons), but two are associated with external sensilla on the sternite. All of these neurons and their axons define specific adult sensory pathways in the periphery and their locations and persistence through metamorphosis suggest a role in guiding the growth of adult sensory and motor axons.     

Genomic imprinting absent in Drosophila melanogaster adult females

Genomic imprinting occurs when expression of an allele differs based on the sex of the parent that transmitted the allele. In D. melanogaster, imprinting can occur, but its impact on allelic expression genome-wide is unclear. Here, we search for imprinted genes in D. melanogaster using RNA-seq to compare allele-specific expression between pools of 7- to 10-day-old adult female progeny from reciprocal crosses. We identified 119 genes with allelic expression consistent with imprinting, and these genes showed significant clustering within the genome. Surprisingly, additional analysis of several of these genes showed that either genomic heterogeneity or high levels of intrinsic noise caused imprinting-like allelic expression. Consequently, our data provide no convincing evidence of imprinting for D. melanogaster genes in their native genomic context. Elucidating sources of false-positive signals for imprinting in allele-specific RNA-seq data, as done here, is critical given the growing popularity of this method for identifying imprinted genes.     

Molecular thermal telemetry of free-ranging adult Drosophila melanogaster

The expression of two temperature-sensitive reporter genes, hsp70 and an hsp70-LacZ fusion, in free-ranging adult Drosophila melanogaster indicates that natural thermal stress experienced by such small and mobile insects may be either infrequent or not severe. Levels of the heat-shock protein Hsp70, the major inducible Hsp of Drosophila, were similar in most wild Droso- phila captured after warm days to levels previously reported for unstressed flies in the laboratory. In a transgenic strain transformed with an hsp70-LacZ fusion (i.e., the structural gene encoding bacterial β-galactosidase under control of a heat shock promoter), exposure to temperatures ≥32°C in the laboratory typically resulted in β-galactosidase activities exceeding 140 mOD₄₅₀ h^–1μg^–1 soluble protein. Flies caged in sun frequently had β-galactosidase activities in excess of this level, whereas flies caged in shade and flies released and recaptured on cool days did not. Most flies (>80%) released on warm, sunny days had low β-galactosidase activities upon recapture. Although the balance of recaptured flies had elevated β-galactosidase activities on these days, their β-galactosidase activities were <50% of levels for flies caged in direct sunlight or exposed to laboratory heat shock. These data suggest that even on warm days most flies may avoid thermal stress, presumably through microhabitat selection, but that a minority of adult D. melanogaster undergo mild thermal stress in nature. Both temperature-sensitive reporter genes, however, are limited in their ability to infer thermal stress and demonstrate its absence. Received: 14 July 1999 / Accepted: 21 December 1999     

Ecdysone receptor expression and activity in adult Drosophila melanogaster

Disrupting components of the ecdysone/EcR/USP signaling pathway in insects leads to morphological defects and developmental arrest. In adult Drosophila melanogaster decreased EcR function affects fertility, lifespan, behavior, learning, and memory; however we lack a clear understanding of how EcR/USP expression and activity impacts these phenotypes. To shed light on this issue, we characterized the wild-type expression patterns and activity of EcR/USP in individual tissues during early adult life. EcR and usp were expressed in numerous adult tissues, but receptor activity varied depending on tissue type and adult age. Receptor activity did not detectably change in response to mating status, environmental stress, ecdysone treatment or gender but is reduced when a constitutively inactive ecdysone receptor is present. Since only a subset of adult tissues expressing EcR and usp contain active receptors, it appears that an important adult function of EcR/USP in some tissues may be repression of genes containing EcRE's.     

Ceramide kinase deficiency improves diet-induced obesity and insulin resistance

Ceramide kinase (CERK) is an enzyme that phosphorylates ceramide to produce ceramide 1-phosphate. Recently, evidence has emerged that CERK has a role in inflammatory signaling of immune cells. Since obesity is accompanied by chronic, low-grade inflammation, we examined whether CERK might be involved using CERK-null mice. We determined that CERK deficiency suppresses diet-induced increases in body weight, and improves glucose intolerance. Furthermore, we demonstrated that CERK deficiency attenuates MCP-1/CCR2 signaling in macrophages infiltrating the adipose tissue, resulting in the suppression of inflammation in adipocytes, which might otherwise lead to obesity and diabetes.     

SOCS-1 deficiency does not prevent diet-induced insulin resistance

Obesity is associated with inflammation and increased expression of suppressor of cytokine signaling (SOCS) proteins, which inhibit cytokine and insulin signaling. Thus, reducing SOCS expression could prevent the development of obesity-induced insulin resistance. Using SOCS-1 knockout mice, we investigated the contribution of SOCS-1 in the development of insulin resistance induced by a high-fat diet (HFD). SOCS-1 knockout mice on HFD gained 70% more weight, displayed a 2.3-fold increase in epididymal fat pads mass and increased hepatic lipid content. This was accompanied by increased mRNA expression of leptin and the macrophage marker CD68 in white adipose tissue and of SREBP1c and FAS in liver. HFD also induced hyperglycemia in SOCS-1 deficient mice with impairment of glucose and insulin tolerance tests. Thus, despite the role of SOCS proteins in obesity-related insulin resistance, SOCS-1 deficiency alone is not able to prevent insulin resistance induced by a diet rich in fat.     

Development of the genitalia in Drosophila melanogaster

The imaginal discs of Drosophila melanogaster are an excellent material with which to analyze how signaling pathways and Hox genes control growth and pattern formation. The study of one of these discs, the genital disc, offers, in addition, the possibility of integrating the sex determination pathway into this analysis. This disc, whose growth and shape are sexually dimorphic, gives rise to the genitalia and analia, the more posterior structures of the fruit fly. Male genitalia, which develop from the ninth abdominal segment, and female genitalia, which develop mostly from the eighth one, display a characteristic array of structures. We will review here some recent findings about the development of these organs. As in other discs, different signaling pathways establish the positional information in the genital primordia. The Hox and sex determination genes modify these signaling routes at different levels to specify the particular growth and differentiation of male and female genitalia.     

A biochemical and functional characterization of diet-induced brain insulin resistance

While considerable research has examined diminished insulin responses within peripheral tissues, comparatively little has been done to examine the effects of this metabolic disruption upon the CNS. The present study employed biochemical and electrophysiological assays of acutely prepared brain slices to determine whether neural insulin resistance is a component of the metabolic syndrome observed within the fructose-fed (FF) hamster. The tyrosine phosphorylation levels of the insulin receptor (IR) and insulin receptor substrate 1 (IRS-1) in response to insulin were significantly reduced within FF hamsters. Also, insulin-mediated phosphorylation of both residues necessary for activation of the serine-threonine kinase Akt/PKB, a key effector of insulin signaling, was markedly decreased. Elevated levels of the protein tyrosine phosphatase 1B, which dephosphorylates the IR and IRS-1, were also observed within the cerebral cortex and hippocampus of FF hamsters. Examination of whether a nutritionally induced compromise of neural insulin signaling altered synaptic function revealed a significant attenuation of insulin-induced long-term depression, but no effect upon either paired-pulse facilitation or electrically induced long-term potentiation. Collectively, our results demonstrate, for the first time, that nutritionally induced insulin resistance significantly affects the neural insulin signaling pathway, and suggest that brain insulin resistance may contribute to cognitive impairment.     

Quantitative trait loci affecting starvation resistance in Drosophila melanogaster

The ability to withstand periods of scarce food resources is an important fitness trait. Starvation resistance is a quantitative trait controlled by multiple interacting genes and exhibits considerable genetic variation in natural populations. This genetic variation could be maintained in the face of strong selection due to a trade-off in resource allocation between reproductive activity and individual survival. Knowledge of the genes affecting starvation tolerance and the subset of genes that affect variation in starvation resistance in natural populations would enable us to evaluate this hypothesis from a quantitative genetic perspective. We screened 933 co-isogenic P-element insertion lines to identify candidate genes affecting starvation tolerance. A total of 383 P-element insertions induced highly significant and often sex-specific mutational variance in starvation resistance. We also used deficiency complementation mapping followed by complementation to mutations to identify 12 genes contributing to variation in starvation resistance between two wild-type strains. The genes we identified are involved in oogenesis, metabolism, and feeding behaviors, indicating a possible link to reproduction and survival. However, we also found genes with cell fate specification and cell proliferation phenotypes, which implies that resource allocation during development and at the cellular level may also influence the phenotypic response to starvation.     

Nobiletin improves obesity and insulin resistance in high-fat diet-induced obese mice

Nobiletin (NOB) is a polymethoxylated flavone present in citrus fruits and has been reported to have antitumor and anti-inflammatory effects. However, little is known about the effects of NOB on obesity and insulin resistance. In this study, we examined the effects of NOB on obesity and insulin resistance, and the underlying mechanisms, in high-fat diet (HFD)-induced obese mice. Obese mice were fed a HFD for 8 weeks and then treated without (HFD control group) or with NOB at 10 or 100 mg/kg. NOB decreased body weight gain, white adipose tissue (WAT) weight and plasma triglyceride. Plasma glucose levels tended to decrease compared with the HFD group and improved plasma adiponectin levels and glucose tolerance. Furthermore, NOB altered the expression levels of several lipid metabolism-related and adipokine genes. NOB increased the mRNA expression of peroxisome proliferator-activated receptor (PPAR)-γ, sterol regulatory element-binding protein-1c, fatty acid synthase, stearoyl-CoA desaturase-1, PPAR-α, carnitine palmitoyltransferase-1, uncoupling protein-2 and adiponectin, and decreased the mRNA expression of tumor necrosis factor-α and monocyte chemoattractant protein-1 in WAT. NOB also up-regulated glucose transporter-4 protein expression and Akt phosphorylation and suppressed IκBα degradation in WAT. Taken together, these results suggest that NOB improves adiposity, dyslipidemia, hyperglycemia and insulin resistance. These effects may be elicited by regulating the expression of lipid metabolism-related and adipokine genes, and by regulating the expression of inflammatory makers and activity of the insulin signaling pathway.