The Action of the Notch Locus in DROSOPHILA MELANOGASTER. II. Biochemical Effects of Recessive Lethals on Mitochondrial Enzymes

We show that six mapped recessive lethal point mutations of the Notch locus affect mitochondrial enzyme activities: NADH oxidase, NADH dehydrogenase, succinate dehydrogenase and α-glycerophosphate dehydrogenase. The mutant N^264-40, which has the same morphological and embryological effects as the Notch⁸ deletion, demonstrates the same biochemical effects and dosage relations as Notch⁸. The other five mapped recessive lethals also affect four enzymic activities. They show specific patterns of activity that depend in several cases on the wild-type chromosome in the heterozygous females. That effect occurs with mutants located in the extreme right part of the Notch locus where some mutations, according to other authors, show temperature-sensitive expression.     

Drosophila Alcohol Dehydrogenase Polymorphism and Carbon-13 Fluxes: Opportunities for Epistasis and Natural Selection

The influence of genetic variations in Drosophila alcohol dehydrogenase (ADH) on steady-state metabolic fluxes was studied by means of (13)C NMR spectroscopy. Four pathways were found to be operative during 8 hr of ethanol degradation in third instar larvae of Drosophila. Seven strains differed by 18-25% in the ratio between two major pathway fluxes, i.e., into glutamate-glutamine-proline vs. lactate-alanine-trehalose. In general, Adh genotypes with higher ADH activity exhibit a twofold difference in relative carbon flux from malate into lactate and alanine vs. α,α-trehalose compared to low ADH activity genotypes. Trehalose was degraded by the pentose-phosphate shunt. The pentose-phosphate shunt and malic enzyme could supply NADPH necessary for lipid synthesis from ethanol. Lactate and/or proline synthesis may maintain the NADH/NAD(+) balance during ethanol degradation. After 24 hr the flux into trehalose is increased, while the flux into lipids declines in Adh(F) larvae. In Adh(S) larvae the flux into lipids remains high. This co-ordinated nature of metabolism and the genotype-dependent differences in metabolic fluxes may form the basis for various epistatic interactions and ultimately for variations in organismal fitness.     

The function of anal papillae in salt adaptation of Drosophila melanogaster larvae

The anal papillae of Drosophila melanogaster larvae showed strong developmental plasticity in media with different osmotic pressure. The size of the papillae decreased with increasing salt content of the medium, which supports the hypothesis of its absorptive function. We showed that CI-transport at the site of the anal papillae decreased with increasing salt concentration, which also supports the hypothesis, that the function of anal papillae in osmoregulation is absorption at low salt content and not excretion at high salt content. The inactive area between the anal papillae increased with increased salt concentration, and thus contributed to the decreasing size of the anal papillae. Remnants of this area in pupae were mistakenly measured as anal papillae in a study on salt adaptation by Waddington (1959).     

Alcohol dehydrogenase controls the flux from ethanol into lipids in Drosophila larvae. A 13C NMR study

The dependence of the flux in the alcohol-degrading pathway on the activity of alcohol dehydrogenase was investigated in Drosophila larvae. Third-instar larvae were supplied with [2-13C]ethanol as a dietary carbon source. Specific carbon enrichments in de novo synthesized fatty acids were determined in vitro by means of 13C nuclear magnetic resonance spectroscopy. Carbon fluxes deduced from these enrichment patterns were correlated with the in vitro alcohol dehydrogenase activities in three different Adh genotypes in seven different strains. The flux control coefficient for alcohol dehydrogenase was shown to be approximately 1.0. This indicates that the alcohol dehydrogenase gene-enzyme system in Drosophila larvae can be a major target of natural selection.     

The effects of disruptive and stabilizing selection on body size inDrosophila melanogaster. III. Genetic analysis of two lines with different reactions to disruptive selection with mating of opposite extremes

A genetic analysis was made of two lines which when subjected to disruptive selection with compulsary mating of opposite extremes (D^?) showed a different response viz. one, D^?-1, showing predominantly an increase of environmental variance and possibly interaction variance, the other, D^?-2, showing an increase of genetic variance. Crosses between extreme flies within lines revealed that D^?-1 genomes from large flies are dominant to genomes from small individuals. In D^?-2 the genetic variation is predominantly additive variance. Tests for dominant chromosome effect in crosses with an inbred stock with recessive markers showed clear third chromosome differences in D^?-2 and not in D^?-1. Chromosome exchange between extreme flies corroborated the importance of genetic differences in D^?-2. A factor or complex of factors with large effect decreasing body size is located on third chromosomes from small flies in D^?-2. Interaction between chromosomes has a similar magnitude in the two lines. Crowding and temperature experiments did not reveal an increased general sensitivity to environmental factors in D^?-1, which was suggested by the enlarged environmental variance of this line.     

Alcohol dehydrogenase polymorphism inDrosophila: Enzyme kinetics of product inhibition

Summary Because natural populations ofDrosophila melanogaster are polymorphic for different allozymes of alcohol dehydrogenase (ADH) and becauseD. melanogaster is more tolerant to the toxic effects of ethanol than its sibling speciesD. simulans, information regarding the sensitivities of the different forms of ADH to the products of ethanol degradation are of ecological importance. ADH-F, ADH-S, ADH-71k ofD. melanogaster and the ADH ofD. simulans were inhibited by NADH, but the inhibition was relieved by NAD⁺. The order of sensitivity of NADH was ADH-F<ADH-71k, ADH-S<ADH-simulans with ADH-F being about four times less sensitive than theD. melanogaster enzymes and 12 times less sensitive than theD. simulans enzyme. Acetaldehyde inhibited the ethanolto-acetaldehyde activity of the ADHs, but at low acetaldehyde concentrations ethanol and NAD⁺ reduced the inhibition. ADH-71k and ADH-F were more subject to the inhibitory action of acetaldehyde than ADH-S and ADH-simulans, with ADH-71k being seven times more sensitive than ADH-S. The pattern of product inhibition of ADH-71k suggests a rapid equilibrium random mechanism for ethanol oxidation. Thus, although the ADH variants only differ by a few amino acids, these differences exert a far larger impact on their intrinsic properties than previously thought. How differences in product inhibition may be of significance in the evolution of the ADHs is discussed.     

A MULTILEVEL APPROACH TO THE SIGNIFICANCE OF GENETIC VARIATION IN ALCOHOL DEHYDROGENASE OF DROSOPHILA

Prior studies showed that differences in alcohol dehydrogenase (ADH) activity across genotypes of Drosophila are decisive for the outcome of selection by ethanol. In the present paper, the effect on ADH activity and egg-to-adult survival of combinations of ethanol, propan-2-ol, and acetone in naturally occurring concentrations is examined. Propan-2-ol is converted into acetone by ADH in vitro. Acetone is considered a competitive inhibitor of ethanol for the ADH enzymes. The melanogaster-ADH-S allozyme is two times more sensitive towards inhibition by acetone than either simulans-ADH or melanogaster-ADH-F. The physiological implications of these in vitro differences for larvae were studied in short-term in vivo and long-term exposure experiments. No major differences in acetone accumulation or fitness parameters were found between the strains in response to ecologically relevant concentrations of acetone or propan-2-ol. Ethanol, however, strongly decreased egg-to-pupal survival in both Drosophila simulans strains and increased developmental time in four out of the five strains tested. Therefore, under physiological conditions only ethanol was shown to act as a selective agent on the ADH polymorphism during egg-to-pupa development in Drosophila.     

THE GENETIC BASIS OF SEXUAL ISOLATION BETWEEN DROSOPHILA MELANOGASTER AND D. SIMULANS

The genetic analysis of sexual isolation between the closely-related species Drosophila melanogaster and Drosophila simulans involved two experiments with no-choice tests. The efficiency of sexual isolation was measured by the frequency of courtship initiation and interspecific mating. We first surveyed the variation in sexual isolation between D. melanogaster strains and D. simulans strains of different geographic origin. Then, to investigate variation in sexual isolation within strains, we made F₁ diallel sets of reciprocal crosses within strains of D. melanogaster and D. simulans. The F₁ diallel progeny of one sex were paired with the opposite sex of the other species. The first experiment showed significant differences in the frequency of interspecific mating between geographic strains. There were more matings between D. simulans females and D. melanogaster males than between D. melanogaster females and D. simulans males. The second experiment uncovered that the male genotypes in the D. melanogaster diallel significantly differed in interspecific mating frequency, but not in courtship initiation frequency. The female genotypes in the D. simulans diallel were not significantly different in courtship initiation and interspecific mating frequency. Genetic analysis reveals that in D. melanogaster males sexual isolation was not affected by either maternal cytoplasmic effects, sex-linked effects, or epistatic interaction. The main genetic components were directional dominance and overdominance. The F₁ males achieved more matings with D. simulans females than the inbred males. The genetic architecture of sexual isolation in D. melanogaster males argues for a history of weak or no selection for lower interspecific mating propensity. The behavioral causes of variation in sexual isolation between the two species are discussed.     

Stabilizing and Disruptive Selection on a Mutant Character in Drosophila. IV. Selection on Sensitivity to Temperature

Individual selection on temperature sensitivity was applied to the relative length of the 4th vein of the mutant ci^D-G in four selection lines according to different selection schemes indicated in Figure 1. These selection systems include, besides the selection on temperature, disruptive selection. In all lines the disruptive component causes an increase of the phenotypic variance, but there are large differences in its composition. The selection changed temperature sensitivity in the expected direction in all lines. In one line (C-D^-) however, the change in temperature sensitivity and the increase in variance were only small because the disruptive component and the canalizing component of the selection scheme restricted each others' effect. The results are discussed in relation to earlier results obtained by disruptive selection at one temperature.