Wild-type Canton-S Research Articles

Lithium affects sodium balance but not intestinal microbiota - studies in Drosophila melanogaster

BackgroundThe trace element lithium (Li) is known for its therapeutic mood-stabilizing application in humans, but also for its various bioactivities, which have been uncovered in model organisms. According to the literature, Li may interfere with the homeostasis of other minerals in mammals, namely sodium, calcium and magnesium. In addition, Li was found to influence the composition and diversity of the intestinal microbiota in vertebrates, an observation that may be related to the many bioactivities of Li. MethodsBased on these previous findings, we employed the model organism Drosophila melanogaster to decipher whether Li exhibits similar bioactivities in invertebrates. First, we examined the influence of increasing dietary Li supply (0 −100 mM LiCl) on the status of Li and ten other minerals via Inductively coupled plasma - mass spectrometry (ICP-MS) in heads and remaining body parts of the three wildtype strains w1118, Oregon-R-C and Canton-S. In addition, we investigated the potential impact of Li feeding (0, 0.1, 1 mM LiCl) on the total bacterial load, α- and β-diversity via real-time quantitative polymerase chain reaction (RT q-PCR) and 16S rDNA sequencing in the intestines of female w1118. ResultsOur observations revealed that Li accumulates linearly in both sexes and all body parts of the three Drosophila strains as the dietary Li supply increases. While the status of most elements remained unchanged, the sodium levels of the fly also correlated positively with the Li content of the diet. The intestinal microbiota, however, remained largely unaffected by Li feeding in terms of both, bacterial load and diversity. ConclusionThese findings support the hypothesis that elevating the Li supply affects sodium homeostasis in Drosophila, a finding coherent with observations in mammals. Furthermore, our data opposes a possible involvement of the bacterial intestinal colonization in the bioactivity of Li in Drosophila.

Whole-Genome and Poly(A)+Transcriptome Analysis of the Drosophila Mutant agnts3 with Cognitive Dysfunctions.

The temperature-sensitive Drosophila mutant agnts3 exhibits the restoration of learning defects both after heat shock (HS) and under hypomagnetic conditions (HMC). Previously, agnts3 was shown to have an increased level of LIM kinase 1 (LIMK1). However, its limk1 sequence did not significantly differ from that of the wild-type strain Canton-S (CS). Here, we performed whole-genome and poly(A)-enriched transcriptome sequencing of CS and agnts3 males normally, after HMC, and after HS. Several high-effect agnts3-specific mutations were identified, including MED23 (regulation of HS-dependent transcription) and Spn42De, the human orthologs of which are associated with intellectual disorders. Pronounced interstrain differences between the transcription profiles were revealed. Mainly, they included the genes of defense and stress response, long non-coding RNAs, and transposons. After HS, the differences between the transcriptomes became less pronounced. In agnts3, prosalpha1 was the only gene whose expression changed after both HS and HMC. The normal downregulation of prosalpha1 and Spn42De in agnts3 was confirmed by RT-PCR. Analysis of limk1 expression did not reveal any interstrain differences or changes after stress. Thus, behavioral differences between CS and agnts3 both under normal and stressed conditions are not due to differences in limk1 transcription. Instead, MED23, Spn42De, and prosalpha1 are more likely to contribute to the agnts3 phenotype.

Social setting interacts with hyper dopamine to boost the stimulant effect of ethanol.

Alcohol consumption occurring in a social or solitary setting often yields different behavioural responses in human subjects. For example, social drinking is associated with positive effects while solitary drinking is linked to negative effects. However, the neurobiological mechanism by which the social environment during alcohol intake impacts on behavioural responses remains poorly understood. We investigated whether distinct social environments affect behavioural responses to ethanol and the role of the dopamine system in this phenomenon in the fruit fly Drosophila melanogaster. The wild-type Canton-S (CS) flies showed higher locomotor response when exposed to ethanol in a group setting than a solitary setting, and there was no difference in females and males. Dopamine signalling is crucial for the locomotor stimulating effect of ethanol. When subjected to ethanol exposure alone, the dopamine transport mutant flies fumin (fmn) with hyper dopamine displayed the locomotor response similar to CS. When subjected to ethanol in a group setting, however, the fmn's response to the locomotor stimulating effect was substantially augmented compared with CS, indicating synergistic interaction of dopamine signalling and social setting. To identify the dopamine signalling pathway important for the social effect, we examined the flies defective in individual dopamine receptors and found that the D1 receptor dDA1/Dop1R1 is the major receptor mediating the social effect. Taken together, this study underscores the influence of social context on the neural and behavioural responses to ethanol.

Two Old Wild-Type Strains of Drosophila melanogaster Can Serve as an Animal Model of Faster and Slower Aging Processes.

Drosophila melanogaster provides a powerful platform to study the physiology and genetics of aging, i.e., the mechanisms underpinnings healthy aging, age-associated disorders, and acceleration of the aging process under adverse environmental conditions. Here, we tested the responses of daily rhythms to age-accelerated factors in two wild-type laboratory-adapted strains, Canton-S and Harwich. On the example of the 24 h patterns of locomotor activity and sleep, we documented the responses of these two strains to such factors as aging, high temperature, carbohydrate diet, and diet with different doses of caffeine-benzoate sodium. The strains demonstrated differential responses to these factors. Moreover, compared to Canton-S, Harwich showed a reduced locomotor activity, larger amount of sleep, faster rate of development, smaller body weight, lower concentrations of main sugars, lower fecundity, and shorter lifespan. It might be recommended to use at least two strains, one with a relatively fast and another with a relatively slow aging process, for the experimental elaboration of relationships between genes, environment, behavior, physiology, and health.

STARVATION DURING DEVELOPMENT AFFECTS METABOLISM IN DROSOPHILA

Aim. To investigate how starvation during early stage of fly development affects carbohydrate metabolism in imago flies and their progeny of F1 generation. Methods. Wild-type Canton-S strain Drosophila melanogaster flies were used in all experiments. Flies of parental and offspring generations were used for the determination of glycogen and glucose content using the diagnostic kit Glucose-Mono-400-P according to the manufacturer's instructions. Results represent as the mean ± SEM of 3-4 replicates per group. According Student's t-test significant difference between groups was P<0.05. Graphing and statistical analysis were performed by using GraphPad Prism. Results. Starvation during development significantly influenced the level of hemolymph and body glucose in imago flies of parental generation. Hemolymph glucose concentration was lower by 34% (P=0.008) and 32% (P=0.033) in experimental females and males, respectively, as compared to control groups. Starvation during development led to lower level of body glucose in adult parental flies of both sexes. Adult males F1, generated by parents that were starved during development, showed 3-fold lower glycogen content, as compared to control. Conclusions. Starvation at early stage of development led to lower hemolymph glucose and body glucose level in imago flies. Moreover, parental starvation decreased glycogen pool in F1 males.

Cd1 Mutation in Drosophila Affects Phenoxazinone Synthase Catalytic Site and Impairs Long-Term Memory.

Being involved in development of Huntington’s, Parkinson’s and Alzheimer’s diseases, kynurenine pathway (KP) of tryptophan metabolism plays a significant role in modulation of neuropathology. Accumulation of a prooxidant 3-hydroxykynurenine (3-HOK) leads to oxidative stress and neuronal cell apoptosis. Drosophila mutant cardinal (cd1) with 3-HOK excess shows age-dependent neurodegeneration and short-term memory impairments, thereby presenting a model for senile dementia. Although cd gene for phenoxazinone synthase (PHS) catalyzing 3-HOK dimerization has been presumed to harbor the cd1 mutation, its molecular nature remained obscure. Using next generation sequencing, we have shown that the cd gene in cd1 carries a long deletion leading to PHS active site destruction. Contrary to the wild type Canton-S (CS), cd1 males showed defective long-term memory (LTM) in conditioned courtship suppression paradigm (CCSP) at days 5–29 after eclosion. The number of dopaminergic neurons (DAN) regulating fly locomotor activity showed an age-dependent tendency to decrease in cd1 relative to CS. Thus, in accordance with the concept “from the gene to behavior” proclaimed by S. Benzer, we have shown that the aberrant PHS sequence in cd1 provokes drastic LTM impairments and DAN alterations.

Astragalus membranaceus Extract Prevents Calcium Oxalate Crystallization and Extends Lifespan in a Drosophila Urolithiasis Model.

Approximately 1 in 20 people develops kidney stones at some point in their life. Although the surgical removal of stones is common, the recurrence rate remains high and it is therefore important to prevent the occurrence of kidney stones. We chose Astragalus membranaceus (AM), which is a traditional Chinese medicine, to study the prevention of urolithiasis using a Drosophila model based on our previous screening of traditional Chinese herbs. Wild-type Drosophila melanogaster Canton-S adult fruit flies were used in this study. Ethylene glycol (EG, 0.5%) was added to food as a lithogenic agent. The positive control agent (2% potassium citrate (K-citrate)) was then compared with AM (2, 8, and 16 mg/mL). After 21 days, the fruit flies were sacrificed under carbon dioxide narcotization, and the Malpighian tubules were dissected, removed, and processed for polarized light microscopy examination to observe calcium oxalate (CaOx) crystallization. Then, the ex vivo dissolution of crystals in the Malpighian tubules was compared between K-citrate and AM. Survival analysis of the EG, K-citrate, and AM groups was also performed. Both 2% K-citrate and AM (16 mg/mL) significantly inhibited EG-induced CaOx crystal formation. Mean lifespan was significantly reduced by the administration of EG, and the results were significantly reversed in the AM (8 and 16 mg/mL) groups. However, AM extract did not directly dissolve CaOx crystals in Drosophila Malpighian tubules ex vivo. In conclusion, AM extract decreased the ratio of CaOx crystallization in the Malpighian tubules and significantly ameliorated EG-induced reduction of lifespan. AM prevented CaOx crystal formation in the Drosophila model.

Impact of social environment on alcohol‐induced behavior

Alcohol consumption occurring in social or solitary settings often yield different behavioral responses. For example, drinking of alcoholic beverages in a social setting is associated with enhanced positive effects including euphoria and increased sociality while drinking in a solitary setting is linked to augmented negative effects such as depressive symptoms and anxiety. The neurobiological mechanism by which the social environment impacts on behavioral responses to alcohol, however, remains understudied. Using the Drosophilamodel, we investigated whether distinct social environments affect behavioral responses to ethanol. In this study, we focused on the hyper‐locomotor activity that is typically used as a proxy for euphoria induced by ethanol. Wild‐type Canton‐S (CS) flies were exposed to ethanol in either socially‐isolated (1 fly per chamber) or socially‐enriched (13 flies per chamber) settings and their locomotor activities were analyzed. We found that the socially‐enriched flies showed significantly more hyperactivity under the influence of ethanol compared to the singly exposed flies. This suggests that the presence of other flies amplifies ethanol‐induced euphoria. Dopamine is known to be important for social interactions thus we explored the role of dopamine in the social effect on the ethanol response by examining the dopamine transporter mutant fumin with hyperdopaminergic tone. When exposed to ethanol in a social setting, fuminmutants showed the augmented hyperactivity compared to the singly exposed fumin as well as single‐ and group‐ exposed CSflies. Together, these findings suggest that hyperdopaminergic signaling augments the effect of social enrichment on the ethanol response. The findings of this study provide important information regarding the mechanism by which genetic (dopamine transporter) and non‐genetic (social context) factors interact for the locomotor response to ethanol.

Drosophila functional screening of de novo variants in autism uncovers damaging variants and facilitates discovery of rare neurodevelopmental diseases.

SUMMARYIndividuals with autism spectrum disorder (ASD) exhibit an increased burden of de novo mutations (DNMs) in a broadening range of genes. While these studies have implicated hundreds of genes in ASD pathogenesis, which DNMs cause functional consequences in vivo remains unclear. We functionally test the effects of ASD missense DNMs using Drosophila through “humanization” rescue and overexpression-based strategies. We examine 79 ASD variants in 74 genes identified in the Simons Simplex Collection and find 38% of them to cause functional alterations. Moreover, we identify GLRA2 as the cause of a spectrum of neurodevelopmental phenotypes beyond ASD in 13 previously undiagnosed subjects. Functional characterization of variants in ASD candidate genes points to conserved neurobiological mechanisms and facilitates gene discovery for rare neurodevelopmental diseases.

Chromatin Structure and "DNA Sequence View": The Role of Satellite DNA in Ectopic Pairing of the Drosophila X Polytene Chromosome.

Chromatin 3D structure plays a crucial role in regulation of gene activity. Previous studies have envisioned spatial contact formations between chromatin domains with different epigenetic properties, protein compositions and transcription activity. This leaves specific DNA sequences that affect chromosome interactions. The Drosophila melanogaster polytene chromosomes are involved in non-allelic ectopic pairing. The mutant strain agnts3, a Drosophila model for Williams–Beuren syndrome, has an increased frequency of ectopic contacts (FEC) compared to the wild-type strain Canton-S (CS). Ectopic pairing can be mediated by some specific DNA sequences. In this study, using our Homology Segment Analysis software, we estimated the correlation between FEC and frequency of short matching DNA fragments (FMF) for all sections of the X chromosome of Drosophila CS and agnts3 strains. With fragment lengths of 50 nucleotides (nt), CS showed a specific FEC–FMF correlation for 20% of the sections involved in ectopic contacts. The correlation was unspecific in agnts3, which may indicate the alternative epigenetic mechanisms affecting FEC in the mutant strain. Most of the fragments that specifically contributed to FMF were related to 1.688 or 372-bp middle repeats. Thus, middle repetitive DNA may serve as an organizer of ectopic pairing.

Melatonin Increases Life Span, Restores the Locomotor Activity, and Reduces Lipid Peroxidation (LPO) in Transgenic Knockdown Parkin Drosophila melanogaster Exposed to Paraquat or Paraquat/Iron.

Parkinson's disease (PD) is a complex progressive neurodegenerative disorder involving impairment of bodily movement caused by the specific destruction of dopaminergic (DAergic) neurons. Mounting evidence suggests that PD might be triggered by an interplay between environmental neurotoxicants (e.g., paraquat, PQ), heavy metals (e.g., iron), and gene alterations (e.g., PARKIN gene). Unfortunately, there are no therapies currently available that protect, slow, delay, or prevent the progression of PD. Melatonin (Mel, N-acetyl-5-methoxy tryptamine) is a natural hormone with pleiotropic functions including receptor-independent pathways which might be useful in the treatment of PD. Therefore, as a chemical molecule, it has been shown that Mel prolonged the lifespan and locomotor activity, and reduced lipid peroxidation (LPO) in wild-type Canton-S flies exposed to PQ, suggesting antioxidant and neuroprotective properties. However, it is not yet known whether Mel can protect or prevent the genetic model parkin deficient in flies against oxidative stress (OS) stimuli. Here, we show that Mel (0.5, 1, 3mM) significantly extends the life span and locomotor activity of TH > parkin-RNAi/ + Drosophila melanogaster flies (> 15days) compared to untreated flies. Knock-down (K-D) parkin flies treated with PQ (1mM) or PQ (1mM)/iron (1mM) significantly diminished the survival index and climbing abilities (e.g., 50% of flies were dead and locomotor impairment by days 4 and 3, respectively). Remarkably, Mel reverted the noxious effect of PQ or PQ/iron combination in K-D parkin. Indeed, Mel protects TH > parkin-RNAi/ + Drosophila melanogaster flies against PQ- or PQ/iron-induced diminish survival, locomotor impairment, and LPO (e.g., 50% of flies were death and locomotor impairment by days 6 and 9, respectively). Similarly, Mel prevented K-D parkin flies against both PQ and PQ/iron. Taken together, these findings suggest that Mel can be safely used as an antioxidant and neuroprotectant agent against OS-stimuli in selective individuals at risk to suffer early-onset Parkinsonism and PD.

Variation in resistance to oxidative stress in Oregon-(R)R-flare and Canton-S strains of Drosophila melanogaster.

All aerobic organisms are susceptible to damage by reactive oxygen species (ROS). ROS-induced damage has been associated with aging and diseases such as metabolic syndrome and cancer. However, not all organisms develop these diseases, nor do they age at the same rate; this is partially due to resistance to oxidative stress, a quantitative trait attributable to the interaction of factors including genetics and environmental. Drosophila melanogaster represents an ideal system to study how genetic variation can affect resistance to oxidative stress. In this work, oxidative stress (total and mitochondrial ROS), antioxidant response, and Cap 'n' collar isoform C and Spineless gene expression, one pesticide resistant (Oregon R(R)-flare) and wild-type (Canton-S) strains of D. melanogaster, were analyzed to test resistance to basal oxidative stress. ROS, catalase, and superoxide dismutase were determined by flow cytometry, and Cap 'n' collar isoform C and Spineless expression by qRT-PCR. The intensity of oxidative stress due to the pro-oxidant zearalenone in both was evaluated by flow cytometry. Data confirm expected differences in oxidative stress between strains that differ in Cyp450s levels. The Oregon (R)R-flare showed greater ROS, total and mitochondrial, compared to Canton-S. Regarding oxidative stress genes expression Cap 'n' collar isoform C and Spineless (Ss), Oregon R(R)-flare strain showed higher expression. In terms of response to zearalenone mycotoxin, Canton-S showed higher ROS concentration. Our data show variation in the resistance to oxidative stress among these strains of D. melanogaster.

Maternal preconception PFOS exposure of Drosophila melanogaster alters reproductive capacity, development, morphology and nutrient regulation

Perfluorooctanesulfonic acid (PFOS) is a persistent synthetic surfactant widely detected in the environment. Developmental PFOS exposures are associated with low birth weight and chronic exposures increase risk for obesity and type 2 diabetes. As an obesogen, PFOS poses a major public health exposure risk and much remains to be understood about the critical windows of exposure and mechanisms impacted, especially during preconception. Here, we leverage evolutionarily conserved pathways and processes in the fruit fly Drosophila melanogaster (wild-type Canton-S and megalin-UAS RNAi transgenic fly lines) to investigate the window of maternal preconception exposure to PFOS on reproductive and developmental toxicity, and examine receptor (megalin)-mediated endocytosis of nutrients and PFOS into the oocyte as a potential mechanism.Preconception exposure to 2 ng PFOS/female resulted in an internal concentration of 0.081 ng/fly over two days post exposure, no mortality and reduced megalin transcription. The number of eggs laid 1–3 days post exposure was reduced and contained 0.018 ng PFOS/egg. Following heat shock, PFOS was significantly reduced in eggs from megalin-knockdown transgenic females. Cholesterol and triglycerides were increased in eggs laid immediately following PFOS exposure by non-heat shocked transgenic females whereas decreased cholesterol and increased protein levels were found in eggs laid by heat shocked transgenic females. Preconception exposure likewise increased cholesterol in early emerging wildtype F1 adults and also resulted in progeny with a substantial developmental delay, a reduction in adult weights, and altered transcription of Drosophila insulin-like peptide genes. These findings support an interaction between PFOS and megalin that interferes with normal nutrient transport during oocyte maturation and embryogenesis, which may be associated with later in life developmental delay and reduced weight.

Long-Term Dietary Restriction Leads to Development of Alternative Fighting Strategies.

In competition for food, mates and territory, most animal species display aggressive behavior through visual threats and/or physical attacks. Such naturally-complex social behaviors have been shaped by evolution. Environmental pressure, such as the one imposed by dietary regimes, forces animals to adapt to specific conditions and ultimately to develop alternative behavioral strategies. The quality of the food resource during contests influence animals' aggression levels. However, little is known regarding the effects of a long-term dietary restriction-based environmental pressure on the development of alternative fighting strategies. To address this, we employed two lines of the wild-type Drosophila melanogaster Canton-S (CS) which originated from the same population but raised under two distinct diets for years. One diet contained both proteins and sugar, while the second one was sugar-free. We set up male-male aggression assays using both CS lines and found differences in aggression levels and the fighting strategies employed to establish dominance relationships. CS males raised on a sugar-containing diet started fights with a physical attack and employed a high number of lunges for establishing dominance but displayed few wing threats throughout the fight. In contrast, the sugar-free-raised males favored wing threats as an initial aggressive demonstration and used fewer lunges to establish dominance, but displayed a higher number of wing threats. This study demonstrates that fruit flies that have been raised under different dietary conditions have adapted their patterns of aggressive behavior and developed distinct fighting strategies: one favoring physical attacks, while the other one favoring visual threats.

Comparative transcriptome analyses of the Drosophila pupal eye.

Tissue function is dependent on correct cellular organization and behavior. As a result, the identification and study of genes that contribute to tissue morphogenesis is of paramount importance to the fields of cell and developmental biology. Many of the genes required for tissue patterning and organization are highly conserved between phyla. This has led to the emergence of several model organisms and developmental systems that are used to study tissue morphogenesis. One such model is the Drosophila melanogaster pupal eye that has a highly stereotyped arrangement of cells. In addition, the pupal eye is postmitotic that allows for the study of tissue morphogenesis independent from any effects of proliferation. While the changes in cell morphology and organization that occur throughout pupal eye development are well documented, less is known about the corresponding transcriptional changes that choreograph these processes. To identify these transcriptional changes, we dissected wild-type Canton S pupal eyes and performed RNA-sequencing. Our analyses identified differential expression of many loci that are documented regulators of pupal eye morphogenesis and contribute to multiple biological processes including signaling, axon projection, adhesion, and cell survival. We also identified differential expression of genes not previously implicated in pupal eye morphogenesis such as components of the Toll pathway, several non-classical cadherins, and components of the muscle sarcomere, which could suggest these loci function as novel patterning factors.

Mating experience modifies locomotor performance and promotes episodic motor activity in Drosophila melanogaster

Sexual behavior is a routine among animal species. Sexual experience has several behavioral consequences in insects, but its physiological basis is less well-understood. The episodic motor activity with a periodicity around 19 s was unintentionally observed in the wildtype Canton-S flies and was greatly reduced in the white-eyed mutant w1118 flies. Episodic motor activity co-exists with several consistent locomotor performances in Canton-S flies whereas reduced episodic motor activity is accompanied by neural or behavioral abnormalities in w1118 flies. The improvements of both episodic motor activity and locomotor performance are co-inducible by a pulsed light illumination in w1118. Here we show that mating experience of w1118 males promoted fast and consistent locomotor activities and increased the power of episodic motor activities. Compared with virgin males, mated ones showed significant increases of boundary preference, travel distance over 60 s, and increased path increments per 0.2 s. In contrast, mated males of Canton-S showed decreased boundary preference, increased travel distance over 60 s, and increased path increments per 0.2 s. Additionally, mated males of w1118 displayed increased power amplitude of periodic motor activities at 0.03-0.1 Hz. These data indicated that mating experience promoted fast and consistent locomotion and improved episodic motor activities in w1118 male flies.

Frequency-specific modification of locomotor components by the white gene in Drosophila melanogaster adult flies.

The classic eye-color gene white (w) in Drosophila melanogaster (fruitfly) has unexpected behavioral consequences. How w affects locomotion of adult flies is largely unknown. Here, we show that a mutant allele (w1118 ) selectively increases locomotor components at relatively high frequencies (> 0.1 Hz). The w1118 flies had reduced transcripts of w+ from the 5' end of the gene. Male flies of w1118 walked continuously in circular arenas while the wildtype Canton-S walked intermittently. Through careful control of genetic and cytoplasmic backgrounds, we found that the w1118 locus was associated with continuous walking. w1118 -carrying male flies showed increased median values of path length per second (PPS) and 5-min path length compared with w+ -carrying males. Additionally, flies carrying 2-4 genomic copies of mini-white+ (mw+ ) in the w1118 background showed suppressed median PPSs and decreased 5-min path length compared with controls, and the suppression was dependent on the copy number of mw+ . Analysis of the time-series (i.e., PPSs over time) by Fourier transform indicated that w1118 was associated with increased locomotor components at relatively high frequencies (> 0.1 Hz). The addition of multiple genomic copies of mw+ (2-4 copies) suppressed the high-frequency components. Lastly, the downregulation of w+ in neurons but not glial cells resulted in increased high-frequency components. We concluded that mutation of w modified the locomotion in adult flies by selectively increasing high-frequency locomotor components.

Adaptation of the sulfophosphovanillin method of analysis of total lipids for various biological objects as exemplified by <i>Drosophila melanogaster</i>

Lipid metabolism is crucial in physiology. In recent decades the model object Drosophila melanogaster has been actively used in the study of the fundamental issues of lipid metabolism and its disorders, including obesity, as well as in the search for therapeutic goals for the treatment of metabolic disorders in humans. Quick and accurate quantification of lipid content is an important step in solving these problems. For the first time the method of quantitative measurement of total lipids with the use of the sulfophosphovanillin (SPV) method was described by Zollner and colleagues in 1962, and adapted for insects by Van Handel on females of the yellow fever mosquito Aedes aegypti . The advantages of this method compared to traditional gravimetric and chromatographic methods of analysis are the use of a small amount of biological material, lack of need for complex manipulations with the sample, its high sensitivity, reproducibility and simplicity of implementation with a minimum set of equipment. Here, a modification of the Van Handel protocol is described, which allows the method to be adapted for quantitative determination of total lipids for various organisms as exemplified a widely used model, D. melanogaster . To test the effectiveness of the modified method, we measured the content of total lipids in D. melanogaster females carrying hypomorphic mutations of the dilp6 and dfoxo insulin signaling pathway genes compared to the wild-type Canton-S line, and showed that dilp6 took part in the regulation of fat metabolism, while dfoxo did not. The results obtained emphasize the effectiveness of the colorimetric method with the use of SPV reaction and spectrophotometry for the quantitative analysis of total lipids.

Longevity-promoting efficacies of rutin in high fat diet fed Drosophila melanogaster.

Composition of diet significantly impacts lifespan in Drosophila melanogaster. Diet-composition becomes even more crucial while assessing a phytocompound for probable pro-longevity effects in flies. Rutin is a flavonol glycoside present in apple, buckwheat, black tea and green tea. Our previous study had reported hormetic efficacy of rutin to improve longevity and other physiological parameters in Drosophila melanogaster fed with standard diet. This study aimed to understand whether rutin could exhibit similar longevity promoting effects in flies fed with a high fat diet (HFD). In this study, wild type Canton-S males and females were reared on high fat diet (HFD) treated with or without rutin at different doses (100-800 µM) and assessed for survival, food intake, fecundity, locomotion, development, resistance to various forms of stresses and relative mRNA expression of specific genes associated with ageing, namely dFoxO, MnSod, Cat, dTsc1, dTsc2, Thor, dAtg1, dAtg5, dAtg7 and dTor. Rutin at only 400 µM significantly improved survival in males fed with HFD; while at 200 µM and 400 µM it significantly improved survival in females. Doses beyond 400 µM proved detrimental for both sexes. Rutin at 200 µM and 400 µM significantly reduced average food intake in both males and females fed with HFD. A significant reduction in number of eggs laid per female per day was observed in females treated with rutin at 400 µM. Rutin at 200 µM and 400 µM significantly improved climbing efficiency in males and females. A significant reduction in eclosion time was observed in larvae fed with HFD and treated with rutin at 400 µM. Rutin at 400 µM significantly improved resistance of males and females to different stresses namely heat shock, cold shock and starvation stresses. Interestingly, rutin at 400 µM significantly reduced survival of males and females exposed to oxidative stress. In males fed with HFD, rutin at 200 µM showed significantly increased relative expression of dFoxo, MnSod, Cat, dAtg1, dAtg5 and dAtg7; at 400 µM it significantly increased the relative expression of dFoxO, MnSod, Cat, dTsc1, dTsc2, Thor, dAtg1, dAtg5, dAtg7 while decreasing relative expression of dTor. Thus, data from this study collectively showed that rutin at 400 µM and to an extent 200 µM positively impacted lifespan and modulated other physiological parameters in males and females fed with HFD.

The Role of Parental Origin of Chromosomes in the Instability of the Somatic Genome in Drosophila Brain Cells and Memory Trace Formation in Norm and Stress

It is impossible to imagine the nervous system without controllable genome instability, resulting in brain somatic mosaicism, one of the basic mechanisms of structural and functional heterogeneity of neurons. The source of such an instability is the presence of “hot spots” (repeating DNA sequences, provoking nonallelic recombination) and double-strand (DS) DNA breaks in the genome that occur in the matrix processes and physiological activity of neurons and are involved in memory formation and learning. The realization of the “norm–pathology” scenario is under epigenetic control; in particular, it depends on the parental effect of genome origin and stress. In this work, using the Williams model of drosophila carrying the agnts3 mutation in the gene for LIMK1 (the key enzyme of actin remodeling) using reciprocal hybrids with wild-type Canton-S line, we studied the contribution of maternal and paternal genomes in processes of learning and memory, as well as the formation of chromosome rearrangement in neuroblasts, determined by DS breaks and disorders of mitotic apparatus in norm and in exposure to stress impact via weak static magnetic field. The prevalent role of paternal genome in memory trace formation is shown. A patroclinous inheritance is established for frequencies of chromosome rearrangements and DS breaks, as well as chromatid bridges in anaphase neuroblasts at stress in the case of paternal agnts3 line. In the breed of agnts3 females, mitosis disorders are inherited via the maternal type. Based on previous research, we revealed microRNA contexts that can make patroclinous effects possible.