Aging In Drosophila Research Articles

The dopaminergic system in the aging brain of Drosophila.

Drosophila models of Parkinson's disease are characterized by two principal phenotypes: the specific loss of dopaminergic (DA) neurons in the aging brain and defects in motor behavior. However, an age-related analysis of these baseline parameters in wildtype Drosophila is lacking. Here we analyzed the DA system and motor behavior in aging Drosophila. DA neurons in the adult brain can be grouped into bilateral symmetric clusters, each comprising a stereotypical number of cells. Analysis of TH > mCD8::GFP and cell type-specific MARCM clones revealed that DA neurons show cluster-specific, stereotypical projection patterns with terminal arborization in target regions that represent distinct functional areas of the adult brain. Target areas include the mushroom bodies, involved in memory formation and motivation, and the central complex, involved in the control of motor behavior, indicating that similar to the mammalian brain, DA neurons in the fly brain are involved in the regulation of specific behaviors. Behavioral analysis revealed that Drosophila show an age-related decline in startle-induced locomotion and negative geotaxis. Motion tracking however, revealed that walking activity, and exploration behavior, but not centrophobism increase at late stages of life. Analysis of TH > Dcr2, mCD8::GFP revealed a specific effect of Dcr2 expression on walking activity but not on exploratory or centrophobic behavior, indicating that the siRNA pathway may modulate distinct DA behaviors in Drosophila. Moreover, DA neurons were maintained between early- and late life, as quantified by TH > mCD8::GFP and anti-TH labeling, indicating that adult onset, age-related degeneration of DA neurons does not occur in the aging brain of Drosophila. Taken together, our data establish baseline parameters in Drosophila for the study of Parkinson's disease as well as other disorders affecting DA neurons and movement control.

A Single Amino Acid Mutation in the Drosophila Myosin SH1 Domain Severely Affects Muscle Function, Myofibril Structure, Myosin Enzymatic Activity, and Actin Sliding Velocity

Hereditary inclusion-body myopathy type III (IBM-3) is caused by a single amino acid Glu706Lys substitution in the SH1 helix of the myosin head. The SH1 domain has been proposed to play a key role in the conformational changes that occur in the myosin head during force generation which is coupled to ATP hydrolysis. We are using an integrative approach to study the structure-function relationship of the myosin SH1 domain in the Drosophila model system. We constructed a gene encoding myosin with the single amino acid mutation and expressed it in place of wild-type myosin heavy chain by germline transformation and crossing into a line that lacks myosin in its flight and jump muscles. The homozygous flies are flightless and their jump abilities are also greatly reduced. The indirect flight muscle fibers of young flies show considerable ultrastructural disarray, with some regions of missing thick and thin filaments, and myofibrils that are not uniform in width. Our initial study showed that actin sliding velocity and basal and actin stimulated ATPase were reduced more than 70% compared to wild-type indirect flight muscle myosin. Homology models indicate that the surface charge change of the substitution in the highly conversed SH1 region could destabilize the helix, which is critical for the converter domain to rotate to its full range of movement during the power-stroke. This structural change would affect the lever arm swing, resulting in dysfunctional myosin. Given that human IBM-3 is mild in childhood but severe during aging, with the accumulation of inclusion bodies, we are investigating whether inclusion bodies or aggregates appear in aged mutant Drosophila muscle tissues.

Aging Predisposes Oocytes to Meiotic Nondisjunction When the Cohesin Subunit SMC1 Is Reduced

In humans, meiotic chromosome segregation errors increase dramatically as women age, but the molecular defects responsible are largely unknown. Cohesion along the arms of meiotic sister chromatids provides an evolutionarily conserved mechanism to keep recombinant chromosomes associated until anaphase I. One attractive hypothesis to explain age-dependent nondisjunction (NDJ) is that loss of cohesion over time causes recombinant homologues to dissociate prematurely and segregate randomly during the first meiotic division. Using Drosophila as a model system, we have tested this hypothesis and observe a significant increase in meiosis I NDJ in experimentally aged Drosophila oocytes when the cohesin protein SMC1 is reduced. Our finding that missegregation of recombinant homologues increases with age supports the model that chiasmata are destabilized by gradual loss of cohesion over time. Moreover, the stage at which Drosophila oocytes are most vulnerable to age-related defects is analogous to that at which human oocytes remain arrested for decades. Our data provide the first demonstration in any organism that, when meiotic cohesion begins intact, the aging process can weaken it sufficiently and cause missegregation of recombinant chromosomes. One major advantage of these studies is that we have reduced but not eliminated the SMC1 subunit. Therefore, we have been able to investigate how aging affects normal meiotic cohesion. Our findings that recombinant chromosomes are at highest risk for loss of chiasmata during diplotene argue that human oocytes are most vulnerable to age-induced loss of meiotic cohesion at the stage at which they remain arrested for several years.

Plasticity of the responses to chronic hypoxia and dietary restriction in an aged organism: Evidence from the Drosophila model

Major cardiovascular diseases arise as consequences of a reduced blood flow to oxygen consuming organs such as the brain and the heart. Their incidence and consequences increase with ageing and inappropriate nutrition conditions. In this study we compared the responses of young (1 day old) or aged (1 month old) Drosophila to dietary restriction and chronic hypoxia. Changing the two major diet components (sucrose and yeast) had complex and non linear consequences on longevity. Ageing and hypoxia completely remodelled the longevity/nutrient reaction norm. Aged flies required more sucrose and less yeast than young flies and became insensitive to dietary restriction by food dilution. The responses to chronic hypoxia were largely age independent and strongly diet dependent. For instance rich diets sensitized Drosophila to chronic hypoxia and hypoxia increased the life span of starving flies (aged or young). The results highlight the importance of nutrition conditions when assessing ageing and hypoxic tolerance in the Drosophila system.

Regulation of Drosophila Life Span by Olfaction and Food-Derived Odors

Smell is an ancient sensory system present in organisms from bacteria to humans. In the nematode Caenorhabditis elegans, gustatory and olfactory neurons regulate aging and longevity. Using the fruit fly, Drosophila melanogaster, we showed that exposure to nutrient-derived odorants can modulate life span and partially reverse the longevity-extending effects of dietary restriction. Furthermore, mutation of odorant receptor Or83b resulted in severe olfactory defects, altered adult metabolism, enhanced stress resistance, and extended life span. Our findings indicate that olfaction affects adult physiology and aging in Drosophila, possibly through the perceived availability of nutritional resources, and that olfactory regulation of life span is evolutionarily conserved.

A Drosophila model for age-associated changes in sleep:wake cycles.

One of the most consistent behavioral changes that occurs with age in humans is the loss of sleep consolidation. This can be quite disruptive and yet little is known about its underlying basis. To better understand the effects of aging on sleep:wake cycles, we sought to study this problem in Drosophila melanogaster, a powerful system for research on aging and behavior. By assaying flies of different ages as well as monitoring individual flies constantly over the course of their lifetime, we found that the strength of sleep:wake cycles decreased and that sleep became more fragmented with age in Drosophila. These changes in sleep:wake cycles became faster or slower with manipulations of ambient temperature that decreased or increased lifespan, respectively, demonstrating that they are a function of physiological rather than chronological age. The effect of temperature on lifespan was not mediated by changes in overall activity level or sleep amount. Flies treated with the oxidative stress-producing reagent paraquat showed a breakdown of sleep:wake cycles similar to that seen with aging, leading us to propose that the accumulation of oxidative damage with age contributes to the changes in rhythm and sleep. Together, these findings establish Drosophila as a valuable model for studying age-associated sleep fragmentation and breakdown of rhythm strength, and indicate that these changes in sleep:wake cycles are an integral part of the physiological aging process.

Distinct genetic influences on locomotor senescence in Drosophila revealed by a series of metrical analyses

Decline of locomotor function is a prominent feature of aging in most species. We previously described a series of metrical analyses for quantifying functional declines associated with aging. Here, we have applied these metrics to elucidate differences in the nature of locomotor senescence between Drosophila cohorts from four different genetic backgrounds (Canton-S, Oregon-R, Samarkand and Lausanne-S) and two long-lived mutant strains, Indy 206 and chico 1 . These analyses demonstrate that senescence of locomotor function in Oregon-R and Canton-S proceeds more rapidly than in Samarkand and Lausanne-S flies. Our analyses also show that while chico 1 and Indy mutants both exhibit attenuated locomotor declines, the impact of chico loss of function on locomotor senescence is substantially greater. Our studies illustrate how important differences in functional senescence characteristics can be unveiled by in-depth analysis on functional senescence data sets.

Specific age related signatures in Drosophila body parts transcriptome

BackgroundDuring the last two decades progress in the genetics of aging in invertebrate models such as C. elegans and D. melanogaster has clearly demonstrated the existence of regulatory pathways that control the rate of aging in these organisms, such as the insulin-like pathway, the Jun kinase pathway and the Sir2 deacetylase pathway. Moreover, it was rapidly shown that some of these pathways are conserved from yeast to humans.In parallel to genetic studies, genomic expression approches have given us significant information on the gene expression modifications that occur during aging either in wild type or long-lived mutant animals. But most of the genomic studies of invertebrate models have been performed so far on whole animals, while several recent studies in mammals have shown that the effects of aging are tissue specific.ResultsWe used oligonucleotide microarrays to address the specificities of transcriptional responses in aging Drosophila in head, thorax or whole body. These fly parts are enriched in transcripts that represent different and complementary sets of genes. We present evidence for both specific and common transcriptional responses during the aging process in these tissues. About half of the genes described as downregulated with age are linked to reproduction and enriched in gonads. Greater downregulation of mitochondrial genes, activation of the JNK pathway and upregulation of proteasome subunits in the thorax of aged flies all suggest that muscle may be particularly sensitive to aging. Simultaneous age-related impairment of synaptic transmission gene expression is observed in fly heads. In addition, a detailed comparison with other microarray data indicates that in aged flies there are significant deviations from the canonical responses to oxidative stress and immune stress.ConclusionOur data demonstrates the advantages and value of regionalized and comparative analysis of gene expression in aging animals. Adding to the age-regulated genes already identified in whole animal studies, it provides lists of new regionalized genes to be studied for their functional role in the aging process. This work also emphasizes the need for such experiments to reveal in greater detail the consequences of the transcriptional modifications induced by aging regulatory pathways.

Detection of deletions flanked by short direct repeats in mitochondrial DNA of aging Drosophila

Cumulative damage due to reactive oxygen species (ROS) in mitochondria, especially in mitochondrial DNA (mtDNA), would result in a decrease in mitochondrial respiratory function and contributes to the age-related decline in the physiological functioning of organisms. Previously, we reported the tissue-specific accumulation of deleted mtDNA with age in Drosophila melanogaster. In the present study, to understand the mechanism by which mtDNA deletion is generated with age, nucleotide sequences of deleted mtDNA were determined. Consequently, 33 different sequences each containing a deletion were obtained from flies that were more than 55-day-old. Most of the deletions were found to be flanked by short direct repeats. The present results, together with those from other animals, suggest that there is a common mechanism generating mtDNA deletions through direct repeats.

Individual Fecundity and Senescence in Drosophila and Medfly

Evolutionary theory postulates that there should be a robust relationship between fecundity and longevity. Prior work has generally supported this concept, but has not shed much light on the mechanisms at play. In preceding work, we have developed and verified a mathematical model of Drosophila melanogaster female fecundity based on the analysis of empirical studies independently done by several different laboratories. Then we applied this technique to Mediterranean fruit fly (medfly) populations. In this article we analyze associations between individual longevity and the parameters of individual fecundity pattern in Drosophila and medfly. We cluster both Drosophila and medfly individuals by life span and discuss the differences. It allows us to demonstrate that only one fecundity-related parameter is associated with longevity in Drosophila, whereas two such parameters can be found in medflies. This difference demonstrates different ways of aging in various Diptera species. Finally, we discuss the possible implications of this finding.

Functional senescence in Drosophila melanogaster

The fruit fly Drosophila melanogaster is one of the principal model organisms used for studying the biology of aging. Flies are well suited for such studies for a number of reasons. Flies develop to adulthood quickly, have a relatively short life span, and are inexpensive to house. Most of the fly genome has been sequenced, powerful genetic tools are available to manipulate it, and most fly genes have obvious homologues in mammals. While the majority of aging studies in flies have focused on regulation of life span, the fly is emerging as a powerful model system for investigating the biology that underlies age-related functional decline. Key to the use of flies in this way is the striking number of parallels between functional senescence in Drosophila and humans. Here, we review age-related functional declines in Drosophila, human correlates of these age-related declines, and common mechanisms that influence longevity and specific aspects of functional senescence in flies.

Aging of the innate immune response in Drosophila melanogaster

Increased activation of the innate immune system is a common feature of aging animals, including mammals and Drosophila melanogaster. With age, D. melanogaster progressively express higher levels of many antimicrobial peptides. It is unknown, however, whether this pattern reflects age-dependent changes in the function of the immune system itself or arises simply because aged adults have greater cumulative exposure to pathogens. Here we demonstrate that aged D. melanogaster transcribe more antimicrobial diptericin when experimentally exposed to septic bacterial infections. This strong net response in older females is the result of persistent diptericin transcription upon septic exposure, whereas young females rapidly terminate this induction. In contrast to their response to septic exposure, when exposed to killed bacteria aged females have less capacity to induce diptericin. Because this functional capacity of innate immunity declines with age, we conclude that female Drosophila undergo immune senescence. Furthermore, we show that fecundity is reduced by induction of innate immunity via the immune deficiency pathway. Consequently, maximum reproduction will occur when the immune response is tightly controlled in young females, even if this increases infection risk at later ages.

Drosophila mutants of the kynurenine pathway as a model for ageing studies.

A search for Drosophila mutants with phenotypes similar to human diseases might help to unravel evolutionary conserved genes implicated in polygenic human disorders. Among these are neurodegenerative diseases, characterized by a late onset disturbance of memory, structural brain impairments and altered content of the intermediates of the kynurenine pathway. The ratio between kynurenate (KYNA) and 3-hydroxykynurenine (3-HOK) in the brain is a critical determinant of neuronal viability. Therefore, the Drosophila mutants cinnabar (KYNA excess) and cardinal (3-HOK excess) allow an evaluation of the specific roles of these metabolites which present in physiologic concentrations and mimic systemic administration. Previously we have demonstrated that the mutant cardinal can serve as a model for dementia and can help to unravel the earliest manifestations of brain dysfunction. Here we show that a state of the brain control of locomotor coordination characterized by the parameters of sound production in males results from the neuroprotective and neurotoxic effects of KYNA and 3-HOK accumulated in young and aged Drosophila mutants. The high instability of 1) cycle form and number in pulses; 2) of pulse amplitude and 3) rhythm in the courtship song of aged cardinal males are similar to the alterations in mutants with defective central complex of the brain. The cardinal mutants demonstrate apoptosis in the brain after stress treatment. This might reflect the misbalance in the content of excitatory amino acids' and the glycine site agonists revealed by HPLC-determination. The mutant cinnabar proved to be normal in respect of the parameters studied.

RNA interference-mediated silencing of Sod2 in Drosophila leads to early adult-onset mortality and elevated endogenous oxidative stress.

Oxidative stress has been widely implicated as an important factor in the aging process. Because mitochondrial respiration is the principal source of reactive oxygen within cells, the mitochondrially localized superoxide dismutase (SOD) 2 is thought to play an important front-line defensive role against aging-related oxidative stress. Although genetic studies with mutants deficient in SOD1, the predominantly cytosolic isoform of SOD, have been instrumental in elucidating the role of reactive oxygen metabolism in aging in Drosophila, the lack of available mutations in the Sod2 gene has hampered an equivalent analysis of the participation of this important antioxidant enzyme in the Drosophila aging model. Here we report that ablation of mitochondrial SOD2 through expression of a GAL4-regulated, inverted-repeat Sod2 RNA-interference transgene in an otherwise normal animal causes increased endogenous oxidative stress, resulting in loss of essential enzymatic components of the mitochondrial respiratory chain and the tricarboxylic acid cycle, enhances sensitivity to applied oxidative stress, and causes early-onset mortality in young adults. In sharp contrast, ablation of SOD2 has no overt effect on the development of larvae and pupae, which may reflect a fundamental transition in oxygen utilization andor reactive oxygen metabolism that occurs during metamorphosis from larval to adult life.

Dissociation between functional senescence and oxidative stress resistance in Drosophila

Many studies strongly suggest a causal link between oxidative stress and determination of life span. The relationship between oxidative stress and age-related functional declines, however, is less clear. Additionally, the full spectrum of functional declines associated with aging has not been systematically evaluated in the fruit fly, Drosophila melanogaster, one of the leading models for aging research. Toward a more comprehensive assessment of functional senescence in Drosophila, we evaluated a series of behaviors in control flies of increasing ages. Our studies reveal a novel age-dependent functional decline in the olfactory system and confirm previous reports of age-related locomotor defects in flies. Behavioral responses to electric shock and light are maintained in aged flies. Thus, some sensory systems senesce during the first several weeks of life while others do not. Interestingly, the age-dependent functional declines in olfactory and locomotor systems are indistinguishable in control flies and methuselah, a mutant with enhanced resistance to oxidative stress and increased life span. Our results indicate that enhanced resistance to oxidative stress and extension of life span do not necessarily confer protection from age-related functional declines.

Laboratory Evolution of Postponed Senescence in Drosophila melanogaster

A general corollary of population genetic theories of senescence is that the culture of outbred laboratory populations using females of later ages should lead to the evolution of postponed senescence. This has been tested before, but the predicted results have not been consistently reproducible. An experiment of this kind was both repeated and replicated, using a previously studied Drosophila melanogaster population. The results uniformly corroborated the evolutionary theory of senescence. Senescence was consistently postponed. Other, anomalous, results are explained in terms of experimental artifact. Reproduced by permission. Michael R. Rose, Laboratory Evolution of Postponed Senescence in Drosophila Melanogaster. Evolution 38 , 1004-1010 (1984).

Selection for Delayed Senescence in Drosophila melanogaster

Although theories of the evolution of senescence differ in the mode of action by which genes are thought to control the aging process, they all view the rate of senescence as an adaptable feature of life history and therefore modifiable by natural selection. Tests of theory, however, have generally yielded mixed results. The life-span of Drosophila melanogaster increased strongly in populations selected for late reproduction in these experiments. Delayed senescence was accompanied by a reduced early fecundity, suggesting pleiotropic genetic control of the aging process. Reproduced by permission. Leo S. Luckinbill, Robert Arking, Michael J. Clare, William C. Cirocco, Steven A. Buck, Selection for Delayed Senescence in Drosophila melanogaster. Evolution 38 , 996-1003 (1984).

Age-associated cardiac dysfunction in Drosophila melanogaster.

The fruit fly, Drosophila melanogaster, has served as a valuable model/organism for the study of aging and was the first organism possessing a circulatory system to have its genome completely sequenced. However, little is known about the function of the heartlike organ of flies during the aging process. We have developed methods for studying cardiac function in vivo in adult flies. Using 2 different cardiovascular stress methods (elevated ambient temperature and external electrical pacing), we found that maximal heart rate is significantly and reproducibly reduced with aging in Drosophila, analogous to observations in elderly humans. We also describe for the first time several other aspects of the cardiac physiology of young adult and aging Drosophila, including an age-associated increase in rhythm disturbances. These observations suggest that the study of declining cardiac function in aging flies may serve as a genetically tractable model for genome-wide mutational screening for genes that participate in or protect against cardiac aging and disease.

Genome-wide study of aging and oxidative stress response in Drosophila melanogaster.

Aging is a universal but poorly understood biological process. Free radicals accumulate with age and have been proposed to be a major cause of aging. We measured genome-wide changes in transcript levels as a function of age in Drosophila melanogaster and compared these changes with those caused by paraquat, a free-radical generator. A number of genes exhibited changes in transcript levels with both age and paraquat treatment. We also found genes whose transcript levels changed with age but not with paraquat treatment. This study suggests that free radicals play an important role in regulating transcript levels in aging but that they are not the only factors. This genome-wide survey also identifies candidates for molecular markers of aging.

Centrosome and microtubule instability in aging Drosophila cells

Several cytoskeletal changes are associated with aging which includes alterations in muscle structure leading to muscular atrophy, and weakening of the microtubule network which affects cellular secretion and maintenance of cell shape. Weakening of the microtubule network during meiosis in aging oocytes can result in aneuploidy or trisomic zygotes with increasing maternal age. Imbalances of cytoskeletal organization can lead to disease such as Alzheimer's, muscular disorders, and cancer. Because many cytoskeletal diseases are related to age we investigated the effects of aging on microtubule organization in cell cultures of the Drosophila cell model system (Schneider S-1 and Kc23 cell lines). This cell model is increasingly being used as an alternative system to mammalian cell cultures. Drosophila cells are amenable to genetic manipulations and can be used to identify and manipulate genes which are involved in the aging processes. Immunofluorescence, scanning, and transmission electron microscopy were employed for the analysis of microtubule organizing centers (centrosomes) and microtubules at various times after subculturing cells in fresh medium. Our results reveal that centrosomes and the microtubule network becomes significantly affected in aging cells after 5 days of subculture. At 5–14 days of subculture, 1% abnormal out of 3% mitoses were noted which were clearly distinguishable from freshly subcultured control cells in which 3% of cells undergo normal mitosis with bipolar configurations. Microtubules are also affected in the midbody during cell division. The midbody in aging cells becomes up to 10 times longer when compared with midbodies in freshly subcultured cells. During interphase, microtubules are often disrupted and disorganized, which may indicate improper function related to transport of cell organelles along microtubules. These results are likely to help explain some cytoskeletal disorders and diseases related to aging. J. Cell. Biochem. 74:229–241, 1999. © 1999 Wiley-Liss, Inc.