Demography of genotypes: failure of the limited life-span paradigm in Drosophila melanogaster.

Experimental systems that are amenable to genetic manipulation can be used to address fundamental questions about genetic and nongenetic determinants of longevity. Analysis of large cohorts of ten genotypes of Drosophila melanogaster raised under conditions that favored extended survival has revealed variation between genotypes in both the slope and location of age-specific mortality curves. More detailed examination of a single genotype showed that the mortality trajectory was best fit by a two-stage Gompertz model, with no age-specific increase in mortality rates beyond 30 days after emergence. These results are contrary to the limited life-span paradigm, which postulates well-defined, genotype-specific limits on life-span and brief periods of intense and rapidly accelerating mortality rates at the oldest age.

Biodemographic trajectories of longevity.

Old-age survival has increased substantially since 1950. Death rates decelerate with age for insects, worms, and yeast, as well as humans. This evidence of extended postreproductive survival is puzzling. Three biodemographic insights--concerning the correlation of death rates across age, individual differences in survival chances, and induced alterations in age patterns of fertility and mortality--offer clues and suggest research on the failure of complicated systems, on new demographic equations for evolutionary theory, and on fertility-longevity interactions. Nongenetic changes account for increases in human life-spans to date. Explication of these causes and the genetic license for extended survival, as well as discovery of genes and other survival attributes affecting longevity, will lead to even longer lives.

Components of selection in X chromosome lines of Drosophila melanogaster: sex ratio modification by meiotic drive and viability selection.

Selection coefficients and segregation parameters have been estimated in 18 randomly chosen lines carrying wild X chromosomes on the cn bw genetic background. Each line was studied in replicated crosses of four types, with approximately 100 replications per line per cross. Crosses in which male X chromosomes differed exhibited significant sex ratio heterogeneity. Maximum likelihood estimation of segregation parameters revealed two lines in which the proportion of X-bearing gametes produced by males was significantly different from Mendelian expectations. These observations suggest that segregation distortion is a common feature of naturally occurring genetic variation. Non-Mendelian segregation has important evolutionary implications.

On the opportunity for polymorphism with sex-linkage or haplodiploidy.

This paper addresses the assertion that X-linked and haplodiploid genetic systems are inherently limited with respect to the potential for selectively maintained genetic polymorphisms. Using a variation of Haldane and Jayakar's (1964) parameterization of selection on an X-linked locus, analytical expressions are derived for the proportion of the total parameter space (P) in which stable diallelic polymorphism is attained. P is a function of the ratio of selection coefficients (r) associated with homozygous and hemizygous genotypes, and the intensity of selection (s). Analytical expressions for the opportunity for polymorphism at an autosomal locus (P(a)) are also derived for comparison to the X-linked case. P and P(a) are maximal and equal if the ratios of selection coefficients are -1 and selection is intense. Otherwise, P is slightly less than P(a), but the difference between autosomal and sex-linked loci is less than the range of values of P obtained over the range of r. Several arguments are presented suggesting that polymorphism arising from differential selection in the sexes (r < 0) is probabilistically and biologically feasible.

Experimental and Theoretical Analysis of the "Sex-Ratio" Polymorphism in DROSOPHILA PSEUDOOBSCURA.

The Sex-ratio chromosome (SR) is a widespread, multiply inverted rearrangement of the X chromosome present in several species of Drosophila. Male carriers transmit mostly X-bearing sperm. In the absence of strong counteracting selection, SR is expected to increase rapidly to fixation, causing extinction. The present study incorporates a selection-components analysis of SR in laboratory populations, using the closely linked Esterase-5 locus as a marker. Estimated fitnesses show directional viability selection against SR in both males and females, heterosis for fertility and no significant effects on virility, the male adult component of fitness. Estimated fitnesses satisfy conditions for protected polymorphism and accurately predict gene-frequency trajectories in experimental populations. A model of SR gene-frequency evolution is developed, which incorporates sex-linkage, meiotic drive, viability, fertility and virility selecton. We show that conditions for protected polymorphisms are not unduly restrictive and that differential fitness among males is not sufficient for protected polymorphism, irrespective of the degree of meiotic drive.

The evolution of age-specific mortality rates in Drosophila melanogaster: genetic divergence among unselected lines.

Age-specific effects of spontaneous mutations on mortality rates in Drosophila are inferred from three large demographic experiments. Data were collected from inbred lines that were allowed to accumulate spontaneous mutations for 10, 19, and 47 generations. Estimates of age-specific mutational variance for mortality were based on data from all three experiments, totalling approximately 225,000 flies, using a model developed for genetic analysis of age-dependent traits (the character process model). Both within- and among-generation analyses suggest that the input of genetic variance is greater for early life mortality rates than for mortality at older ages. In females, age-specific mutational variances ranged over an order of magnitude from 5.96 x 10(-3) at 2 wk posteclosion to 0.02 x 10(-3) at 7 wk. The male data show a similar pattern. Age-specific genetic variances were substantially less at generation 47 than at generation 19-an unexplained observation that is likely due to block effects. Mutational correlations among mortality rates at different ages tend to increase with the accumulation of new mutations. Comparison of the mutation-accumulation lines at generations 19 and 47 with their respective control lines suggests little age-specific mutational bias.

Genetic Variability of Flight Metabolism in DROSOPHILA MELANOGASTER. I. Characterization of Power Output during Tethered Flight.

The mechanical power imparted to the wings during tethered flight of Drosophila melanogaster is estimated from wing-beat frequency, wing-stroke amplitude and various aspects of wing morphology by applying the steady-state aerodynamics model of insect flight developed by Weis-Fogh (1972, 1973). Wing-beat frequency, the major determinant of power output, is highly correlated with the rate of oxygen consumption. Estimates of power generated during flight should closely reflect rates of ATP production in the flight muscles, since flies do not acquire an oxygen debt or accumulate ATP during flight. In an experiment using 21 chromosome 2 substitution lines, lines were a significant source of variation for all flight parameters measured. Broadsense heritabilities ranged from 0.16 for wing-stroke amplitude to 0.44 for inertial power. The variation among lines is not explained by variation in total body size (i.e., live weight). Line differences in flight parameters are robust with respect to age, ambient temperature and duration of flight. These results indicate that characterization of the power output during tethered flight will provide a sensitive experimental system for detecting the physiological effects of variation in the structure or quantity of the enzymes involved in flight metabolism.

Age-specific properties of spontaneous mutations affecting mortality in Drosophila melanogaster.

An analysis of the effects of spontaneous mutations affecting age-specific mortality was conducted using 29 lines of Drosophila melanogaster that had accumulated spontaneous mutations for 19 generations. Divergence among the lines was used to estimate the mutational variance for weekly mortality rates and the covariance between weekly mortality rates at different ages. Significant mutational variance was observed in both males and females early in life (up to approximately 30 days of age). Mutational variance was not significantly different from zero for mortality rates at older ages. Mutational correlations between ages separated by 1 or 2 wk were generally positive, but they declined monotonically with increasing separation such that mutational effects on early-age mortality were uncorrelated with effects at later ages. Analyses of individual lines revealed several instances of mutation-induced changes in mortality over a limited range of ages. Significant age-specific effects of mutations were identified in early and middle ages, but surprisingly, mortality rates at older ages were essentially unaffected by the accumulation procedure. Our results provide strong evidence for the existence of a class of polygenic mutations that affect mortality rates on an age-specific basis. The patterns of mutational effects measured here relate directly to recently published estimates of standing genetic variance for mortality in Drosophila, and they support mutation accumulation as a viable mechanism for the evolution of senescence.

Age-specific patterns of genetic variance in Drosophila melanogaster. I. Mortality.

PETER MEDAWAR proposed that senescence arises from an age-related decline in the force of selection, which allows late-acting deleterious mutations to accumulate. Subsequent workers have suggested that mutation accumulation could produce an age-related increase in additive genetic variance (VA) for fitness traits, as recently found in Drosophila melanogaster. Here we report results from a genetic analysis of mortality in 65,134 D. melanogaster. Additive genetic variance for female mortality rates increases from 0.007 in the first week of life to 0.325 by the third week, and then declines to 0.002 by the seventh week. Males show a similar pattern, though total variance is lower than in females. In contrast to a predicted divergence in mortality curves, mortality curves of different genotypes are roughly parallel. Using a three-parameter model, we find significant VA for the slope and constant term of the curve describing age-specific mortality rates, and also for the rate at which mortality decelerates late in life. These results fail to support a prediction derived from MEDAWAR's "mutation accumulation" theory for the evolution of senescence. However, our results could be consistent with alternative interpretations of evolutionary models of aging.

Age-specific patterns of genetic variance in Drosophila melanogaster. II. Fecundity and its genetic covariance with age-specific mortality.

Under the mutation accumulation model of senescence, it was predicted that the additive genetic variance (VA) for fitness traits will increase with age. We measured age-specific mortality and fecundity from 65,134 Drosophila melanogaster and estimated genetic variance components, based on reciprocal crosses of extracted second chromosome lines. Elsewhere we report the results for mortality. Here, for fecundity, we report a bimodal pattern for VA with peaks at 3 days and at 17-31 days. Under the antagonistic pleiotropy model of senescence, it was predicted that negative correlations will exist between early and late life history traits. For fecundity itself we find positive genetic correlations among age classes > 3 days but negative nonsignificant correlations between fecundity at 3 days and at older age classes. For fecundity vs. age-specific mortality, we find positive fitness correlations (negative genetic correlations) among the traits at all ages > 3 days but a negative fitness correlation between fecundity at 3 days and mortality at the oldest ages (positive genetic correlations). For age-specific mortality itself we find overwhelmingly positive genetic correlations among all age classes. The data suggest that mutation accumulation may be a major source of standing genetic variance for senescence.

Naturally occurring enzyme activity variation in Drosophila melanogaster. I. Sources of variation for 23 enzymes.

The genetic component of variation of enzyme activity levels in Drosophila melanogaster was investigated by using 48 second- and 48 third-chromosome isogenic substitution lines derived from natural populations. The results confirm those of our earlier experiments with the same lines and extend them to a number of additional enzymes. All 23 enzymes show a significant genetic component to the variation in one or both sets of lines and only a small part of this variation is accounted for by variation among the lines in the amount of tissue per fly. The magnitude of line effects is, in most cases, considerably larger than the magnitude of environmental and measurement error effects, and the line effects are approximately continuous in distribution. Variation in the geographic origin and karyotype of the chromosomes generally does not contribute to the line component of variation, but allozymes provide an important source of variation for a few of the enzymes. Many of the enzymes show evidence for variation of activity modifiers that are not linked to the structural locus of the enzyme.

Naturally occurring enzyme activity variation in Drosophila melanogaster. II. Relationships among enzymes.

This report describes an investigation of the specificities of the genetic effects, caused by whole chromosome substitution, on the activities of 23 enzymes in Drosophila melanogaster. Two types of correlation estimates are examined, the product-moment correlation over the chromosome substitution line means and the corresponding correlation of line effects, which is a standardized covariance component estimate. The two types of correlations give very similar results. Although there is ample evidence for specific line effects on individual enzyme activities, there are extensive intercorrelations among many of the enzymes for both second- and third-chromosome substitution lines. The pattern of correlations with respect to the metabolic functions or other properties of the enzymes is difficult to visualize by inspection of the correlation matrix, so a multivariate graphical technique, the biplot (GABRIEL 1971), was employed to obtain a two-dimensional view of relationships among the enzyme activities. The second and third chromosome lines show similar patterns. Four of the five mitochondrial enzymes form one highly intercorrelated group whereas another highly intercorrelated group contains several cytosolic enzymes. Within the cytosolic group, particularly high correlations are observed between enzymes that have glucose 6-phosphate as a substrate or product and between enzymes that are NADP-dependent. Although the pattern of intercorrelations is not clearly explicable in terms of metabolic relationships among the enzymes, there is some tendency for enzymes that catalyze sequential reactions or share a substrate or product to have correlated activity levels.

Genetic variability of flight metabolism in Drosophila melanogaster. II. Relationship between power output and enzyme activity levels.

The major goal of the studies reported here was to determine the extent to which genetic variation in the activities of the enzymes participating in flight metabolism contributes to variation in the mechanical power output of the flight muscles in Drosophila melanogaster. Isogenic chromosome substitution lines were used to partition the variance of both types of quantitative trait into genetic and environmental components. The mechanical power output was estimated from the wingbeat frequency, wing amplitude and wing morphology of tethered flies by applying the aerodynamic models of Weis-Fogh and Ellington. There were three major results. (1) Chromosomes sampled from natural populations provide a large and repeatable genetic component to the variation in the activities of most of the 15 flight metabolism enzymes investigated and to the variation in the mechanical power output of the flight muscles. (2) The mechanical power output is a sensitive indicator of the rate of flight metabolism (i.e., rate of oxygen consumption during tethered flight). (3) In spite of (1) and (2), no convincing cases of individual enzyme effects on power output were detected, although the number and sign of the significant enzyme-power correlations suggests that such effects are not totally lacking.

Nucleotide variations in the lxd region of Drosophila melanogaster: characterization of a candidate modifier of lifespan.

We have investigated the structure and function of several proteins that might influence adult lifespans in Drosophila melanogaster. The present report focuses on the gene lxd ('low xanthine dehydrogenase'), which lies in a region of chromosome III identified by QTL-mapping as potentially important for lifespan. DNA sequence of a 3780 bp genomic fragment containing the lxd locus reveals differences between long-lived and control inbred lines. In order to determine the importance of nucleotide replacements, the intron/exon boundaries have been determined, based on peptide alignment and conserved amino acids. We identified four exons in the lxd coding region. The deduced amino acid sequence of exon 4 shows 46.5% identity with Escherichia coli MoaC sequences. There are eight nucleotide substitutions in exons differentiating the inbred lines, three in exon 3 and five in exon 4. One of the exon 4 substitutions has resulted in a Thr-Ile replacement at the protein surface, but not entirely solvent exposed. This substitution is potentially a modifier of lifespan via oxygen defense, but since the activities of three molybdoenzymes are unaffected in inbred lines, this possibility seems remote.

The fractionation experiment: reducing heterogeneity to investigate age-specific mortality in Drosophila.

Age-specific mortality rates decelerate at older ages in both genetically homogenous and heterogeneous populations of Drosophila. One explanation proposed for deceleration is population heterogeneity. This hypothesis suggests that a population consists of sub-populations that differ in mortality characteristics and that the deceleration is the result of selective survival of stronger individuals. Here we describe an experiment that fractionates populations into several sub-populations without changing the physiological characteristics of the post-fractionated populations. Through a careful process of selection of Drosophila eggs, larvae, pupae and adults, we attempt to reduce as much as possible the degree of pre-adult, environmentally induced heterogeneity among individuals of a genetically identical cohort. We then ask whether such cohorts, when compared to non-fractionated populations, exhibit a lesser degree of mortality deceleration at advanced ages. From a total of 106 fractionated and control populations, consisting of 51331 individuals, 101 populations (93% of the fractionated populations and 100% of the control populations) exhibit a significant amount of mortality deceleration late in life. These observations suggest that environmental heterogeneity accrued during larval development is not a major factor contributing to mortality deceleration at older ages.

Slowing of age-specific mortality rates in Drosophila melanogaster.

We have studied age-dependent mortality in large cohorts of male and female D. melanogaster from four inbred lines. Average longevity varies substantially between genotypes (broad-sense heritability = 22%). Contrary to the predictions of the Gompertz model, mortality rates tend to decelerate at the most advanced ages. Fitting Gompertz, Weibull, Logistic, and Two-stage Gompertz mortality models to the data, we find that the best fit is obtained with the two-stage model, with exponentially increasing mortality at early ages, and zero or nearly zero increase at older ages. There is little microenvironmental effect from cage to cage. There is a sex-dependent mortality crossover: males and females differ in initial mortality rate and degree of acceleration of mortality rate, but the ordering of the sexes according to mortality parameters depends on genotype. Model fitting can be affected by gaps between deaths in the tail of the survivorship distribution. The observations are inconsistent with the limited life-span paradigm, which predicts sudden and well-defined drops in survivorship and corresponding sharp increases in mortality at advanced ages for large cohorts of genetically identical individuals.

Locomotor activity as a function of age and life span in Drosophila melanogaster overexpressing hsp70.

Heat shock protein induction might be responsible for the longevity increase conferred by exposure to non-lethal stresses. To test this hypothesis, we studied in transgenic Drosophila melanogaster overexpressing hsp70 and controls, two behavioral variables (spontaneous locomotor activity and climbing activity) to evaluate the rate of aging, and life span. The results showed that in flies kept in groups, life span was decreased in transgenic flies compared to the parental line, but the contrary was observed in individually kept flies. Hsp70 overexpression had no dramatic effect on life span. Furthermore, we did not detect any advantage of Drosophila overexpressing hsp70 on the two measurements of locomotor activity. These results indicate that the rate of aging in transgenic flies is not different than in non-transgenic lines and that they are not more able to cope with the effects of aging on locomotor activity.

Deceleration of age-specific mortality rates in chromosomal homozygotes and heterozygotes of Drosophila melanogaster.

Age-specific mortality trajectories were estimated in mixed-sex cohorts of D. melanogaster. We studied 22,000 flies that were either second-chromosome homozygotes or heterozygotes with a randomized genetic background. Broad-sense heritabilities for longevity were estimated to be 6% for males and 9% for females. Heterozygotes lived longer than homozygotes on average, but there were exceptions to the usual heterotic pattern; in several crosses parental homozygotes had average life spans as long as that of their F1 heterozygotes. Estimated age-specific mortality rates were found to decelerate at advanced ages in both homozygotes and heterozygotes. The mortality models that best fit the data are the logistic model and the two-stage Gompertz model, both of which produce mortality trajectories that level off at advanced ages. Old-age mortality deceleration is not peculiar to inbred Drosophila.

Stress experiments as a means of investigating age-specific mortality in Drosophila melanogaster.

Age-specific mortality rates level off at older ages in genetically homogeneous experimental populations of Drosophila. Here we describe an experiment that is informative about the causes of mortality rate changes. By applying a brief, nondebilitating stress that increases mortality early in life and then observing subsequent mortality trajectories, it is possible to determine whether populations are heterogeneous for factors influencing mortality. We show that 24-h exposure to a desiccating air flow causes a spike and then a decrease in mortality rates in experimental populations compared to controls. If there is no stress-induced enhancement of vitality, then the results demonstrate the existence of heterogeneity for mortality rates in genetically homogeneous populations.

Effect of adult cohort density on age-specific mortality in Drosophila melanogaster.

Mortality rates decelerate at older ages in experimental populations of Drosophila. It is unclear whether this reflects a real slow-down in the aging process, or an artifact of declining density. Mortality was studied in age-synchronized cohorts of four inbred lines at three initial densities that varied 10-fold. A total of 70,000 flies of both sexes were studied. There were large line x density, line, and sex effects, but no systematic relationship between density and life span was detected. Mortality curves level off at older ages in 23 out of 24 sex-genotype combinations, irrespective of initial cohort density. Density has only second-order effects on the pattern of oldest-old mortality over the range of densities studied here. The dramatic departure from Gompertz-type mortality dynamics at older ages is not an artifact of declining density in Drosophila.