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R Arking

Publications and source records attributed to R Arking.

35 records · Page 2Linked to original sources
PubMed1988

Metabolic rates in genetically based long lived strains of Drosophila.

The goal of these experiments was to determine if the increased longevity characteristics of our genetically selected long lived line of Drosophila could be attributed to metabolic differences. The data shows an inverse relationship between life span and temperature for both the long lived (L) and normal (R) strains; however, the higher longevity of the L strain relative to the R strain is not affected by these treatments. Therefore, the genetic factors unique to the L strain do not affect the same processes affected by the temperature treatments. A second set of experiments detected a linear relationship between the MDMR (mean daily metabolic rate) and the ambient adult temperature. However, at each temperature, the MDMR of either strain was statistically equivalent; a finding which demonstrates that an increased life span depends on something other than conservation of calories. A third set of experiments looked at the metabolic efficiency of the two strains and were not able to detect any statistically significant differences. The two strains appear to expend approximately equivalent numbers of calories per day in an approximately equivalent manner. These data are interpreted in the context both of a previously postulated genetic switch mechanism believed responsible for initiating the onset of senescence, and of contemporary reinterpretations of the "rate of living" theory which implicates the essential role of various anti-oxidant defense systems.

Aging↗
PubMed1987

Successful selection for increased longevity in Drosophila: analysis of the survival data and presentation of a hypothesis on the genetic regulation of longevity.

Long lived strains of Drosophila melanogaster have been generated via 25 generations of artificial selection. The mean and the maximum lifespans have been increased both absolutely as well as relative to the controls. The mean lifespan of the selected line now exceeds the maximum lifespan of the controls. The data shows that this increase is entirely accounted for by a genetically based delay in the onset of senescence. Identification and analysis of biomarker data involving reproductive functions supports this interpretation and leads to a suggestion of the processes involved in the lifespan extension. This increase in the duration of the pre-senescent period is under both genetic and environmental control. Senescence itself is not under genetic control and appears to occur stochastically. Selection for decreased longevity was unsuccessful, supporting the concept of a minimum species specific lifespan. A testable hypothesis regarding the biphasic mode of gene regulation of senescence is presented in which a gene-environment interaction takes place in larval life that results in a temporal reprogramming of other, presumably structural, genes which act in adult life at a time prior to the onset of senescence.

Aging↗
PubMed1984

Development profile of the heat shock response in early embryos of Drosophila.

Drosophila melanogaster embryos reared at 22 degrees C were subjected to a mild heat shock (40 min at 37 degrees C) at various ages in order to determine whether there are changes in the heat shock response during embryogenesis. The effects of the heat shock were measured by assaying (1), subsequent developmental abnormalities (2), developmental time (3), hatchability, and (4), the ability to synthesize the heat shock proteins as assayed by 35S-methionine pulse labeling followed by protein separations using both one-and two-dimensional polyacrylamide gel electrophoresis. Our data show that, first, proteins with molecular weights similar to those of six of the seven major heat shock proteins are normally found in the embryo at control temperatures (22 degrees C); second, that the pregastrula embryo (stages 2-6) is not capable of displaying any aspect of the heat shock response upon treatment, although it may possess all of the so-called heat shock proteins; third, that the complete heat shock response is acquired very rapidly by early gastrula embryos; and fourth, that the heat shock treatment brings about developmental delays and/or abnormalities, depending on the developmental stage of the embryo at the time of the treatment. These developmental abnormalities appear to stem from the failure of early embryos to completely inhibit their synthesis of non-heat-shock proteins. In the light of these findings, it becomes important not to base conclusions about the putative presence of a heat shock response in a particular tissue or developmental stage solely on the presence or absence of the heat shock proteins.

Animals↗
PubMed1980

Effects of RNA inhibitors on the development of Drosophila embryos permeabilized by a new technique.

The molecular analysis of Drosophila embryogenesis has been hindered by the impermeable nature of the vitelline membrane, which has made it difficult to introduce exogenous substances into the developing embryo. We have developed a modification of the permeabilization technique of Limbourg and Zalokar ('73) in which octane is used to permeabilize the vitelline membrane and dimethylsulfoxide is used to facilitate the transport of exogenous substance across the cell membranes. The procedure is highly effective (ca. 95%) and is consistent with a high frequency of normal development. We have used this technique to analyze the effect in vivo of four inhibitors of RNA synthesis (alpha-amanitin, actinomycin-D, rifampin, and rifamycin SV) on the embryogenesis of Drosophila. We have found that there are characteristic stage-specific alterations in the sensitivity of the embryo to these inhibitors which is reflected both by changes in the ID50 dosages and by changes in the developmental abnormalities caused by the drugs. Embryos aged 2-12 hours old undergo a developmental arrest within 30 minutes after application of the inhibitors. Embryos older than 12 hours are able to develop for 140+ minutes after treatment before arresting. The effects of these drugs are consistent with the idea that there exists a definite sequential program of gene activity that is necessary to the normal embryonic development of Drosophila.

Age Factors↗
PubMed1980

Effect of DNA inhibitors upon DNA synthesis and development of Drosophila embryos.

Staged wildtype embryos of Drosophila melanogaster were permeabilized and then subjected to a short pulse of either methyl-3H-thymidine, one of four different inhibitors of DNA synthesis (mitomycin C, 5-fluorouracil, nalidixic acid, or 1-beta-D-arabino-furanosylcytosine-5'-monophosphate), or a combination of both. The incorporation of methyl-3H-thymidine into acid insoluble material was at a maximum during the first half-hour of embryogenesis, after which the incorporation dropped to half the initial value and remained constant throughout the remainder of development. There was no correlation between the rate of incorporation of methyl-3H-thymidine into DNA and the known periods of high mitotic activity. The time course of the estimated specific activity of the DNA newly synthesized in vivo closely paralleled the known changes in the DNA polymerase activity determined in vitro. The known periods of high mitotic activity in the embryo (0-3 hours, 5-12 hours) agree with the periods of maximal sensitivity of the embryo to the inhibitors of DNA synthesis. All four DNA inhibitors affected the incorporation of methyl-3H-thymidine into DNA, although they did not affect it in any simplistic manner. Inhibitor treatment during early cleavage stages resulted in arrested development, treatment during late cleavage and blastoderm stages resulted in abnormal development, and treatment during late blastoderm and early gastrula resulted in normal development. The major phenotypic abnormality caused by the inhibitors is an abnormal distribution of blastoderm cells. As judged by the ID50 values, the embryos remained very sensitive to the effects of the inhibitors until the stages of head and body segmentation, when they then very rapidly became insensitive.

Age Factors↗
PubMed1975

Phenogenetics of the eyeless-dominant mutant of Drosophila melanogaster.

The eyeless-Dominant (eyD) mutation is a fourth chromosome insertional translocation which affects the eyes, antennae, ocelli, and sexcombs when heterozygous but is a larval-pupal lethal when homozygous. By use of a crowding technique, it was possible to separate eyD homozygotes and heterozygotes with 100% accuracy at an early stage of larval development. Under these conditions, the eyD homozygotes had a biphasic lethal period since 45% died as first or second instar larvae and 55% died as pupae. The eyDleyD pupal lethal, isolated by this technique, was able to form all the adult structures with the exception of the eye-antennal disc derivatives. The brain was present but abnormal. In testing the competence of the eyDleyD larval eye discs by means of transplantation experiments, it has been demonstrated that the mutant discs develop non-autonomously. Histological studies have revealed the existence of neurological defects in both eyD heterozygotes and homozygotes. The eyDleyD lethal larvae lacked detectable optic formation centers and showed an extreme reduction in the number of cells present in the cortex of the brain. The ey"dleyD lethal pupae possessed partial formation centers and also had severe reduction in the number of cortical cells. The eyD heterozygotes possessed normal appearing formation centers but they did exhibit a moderate reduction in the number of their cortical cells as compared to wild-type. These studies have shown that there is a direct correlation between the extent of neurological damage and the time of developmental arrest. It appears that the eyD mutation must adversely affect the neuroblasts at a very early stage of development. As a working hypothesis, it is suggested that the eyD mutation operates via the same basic mechanism of cell death in heterozygous and homozygous animals.

Animals↗
PubMed1975

Temperature-sensitive cell-lethal mutants of drosophila: isolation and characterization.

One hundred and twenty-one sensitive (ts) sex-linked lethals were screened by means of X-ray-induced somatic crossing over to determine if any were ts cell-lethal mutants. Cell-lethal mutations were identified by their ability to block the development of homozygous clones when raised under restrictive conditions (29degrees). Twenty-two ts cell-lethal mutants were isolated and categorized into three classes, depending upon the patterns of damage observed in larval and imaginal tissues. The phenotypes produced by these mutations ranged from those which affected only a limited set of structures (i.e., genital discs only) to those which affected diverse tissues at all stages of the life cycle. Each mutation has its own characteristic time-dependent pattern, frequency, and type of damage. All the mutations affect imaginal tissue, but only one-third of the mutations affect both larval and imaginal tissue. The fastest-acting lethals need 15 hours at the restrictive temperature to kill the cells and the slowest-acting lethals require at least 48 hours. By choosing the appropriate mutant and by manipulating the times of exposure to the restrictive temperature, it has proven possible to produce duplications and deficiencies in specific structures of the adult. A mechanism by which lethality might yield such structures is suggested. In addition, 15 of the mutants are ts female sterile mutants. Only one of these 15 mutants can recover its fertility when shifted back down to the permissive temperature (22degrees).

Animals↗
PubMedDate not supplied

The expression of the EF1 alpha genes of Drosophila is not associated with the extended longevity phenotype in a selected long-lived strain.

A quantitative dot blot analysis was performed to determine whether the expression of the EF-1 alpha genes of Drosophila melanogaster are associated with the extended longevity phenotype characteristic of our genetically selected long-lived strain. These data were compared to that obtained from two normal-lived strains and from two iso-chromosomal strains with an intermediate life span. The relative mRNA levels of both EF-1 alpha genes (EF-1 alpha F1 and EF-1 alpha F2) for all five strains were measured through the larval, pupal, and early adult stages, and statistically analyzed. Our findings from these studies indicate that the F2 mRNA tracks with the extended longevity; however, the F1 mRNA is the major component and, thus, the relative total expression of these genes at the mRNA level is approximately equivalent for all five strains. These conclusions suggest that the expression of the EF-1 alpha genes is not associated with the extended longevity phenotype.

Animals↗
PubMedDate not supplied

Factors contributing to the plasticity of the extended longevity phenotypes of Drosophila.

A number of laboratories have constructed independently derived long-lived strains of Drosophila, each of which have similar but not identical patterns of variability in their adult longevity. Given the observed plasticity of longevity within each of these strains, it would be useful to review the operational and environmental factors that give rise to this phenotypic plasticity and ascertain whether they are common or strain specific. Our review of the more extensively analyzed strains suggests that the allelic composition of the initial genomes and the selection/transgene strategy employed yield extended longevity strains with superficially similar phenotypes but which are probably each the result of different proximal genetic mechanisms. This then offers a plausible explanation for the differential effects of various environmental factors on each strain's particular pattern of phenotypic plasticity. It also illustrates that the species has the potential to employ any one of a number of different proximal mechanisms, each of which give rise to a similar longevity phenotype.

Animals↗
PubMedDate not supplied

Genetic analyses of aging processes in Drosophila.

Genetic investigations into the aging processes of Drosophila have a long history. Much of the earlier work attempted the analysis of longevity in already existing and (usually) short-lived strains and mutants, but was unsuccessful because there was no way of assuring that the genes involved actually affected the normal aging processes. Success was achieved only when procedures were devised to specifically select for mutants and/or strains affecting the normal aging processes. Recent work has shown that the life span may be genetically altered either via an acceleration of the normal aging rate or via the stage-specific lengthening of certain portions of the adult life span. A variety of evidence suggests that aging is best viewed as a genetically determined, environmentally modulated, event dependent process. The evidence underlying these observations is discussed, a possible genetic model is presented and future directions are suggested.

Aging↗