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R C Woodruff

Publications and source records attributed to R C Woodruff.

At least 19 recordsLinked to original sources

Varied expression of a Y-linked P[w+] insert due to imprinting in Drosophila melanogaster.

During gametogenesis, a gene can become imprinted affecting its expression in progeny. We have used the expression of a Y-linked P[w+]YAL transposable DNA element as a reporter system to investigate the effect of parental origination on the expression of the w+ insert. Expression of w+ was greater in male progeny when the Y chromosome, harboring the insert, was inherited from the parental male rather than from the parental female. Imprinting was not due to a genetic background influence in the males, since the only difference among the males was the parental origin of the Y chromosome. It was also observed that the genetic background can affect imprinting, since w+ expression was also higher in males when the Y was derived from C(1)DX attached-X parental females rather than from C(1)RM attached-X parental females. Though the heterochromatic imprinting mechanism is unknown, a mutated Heterochromatin Protein 1 (HP1) gene, which is associated with suppression of position-effect variegation, increases expression of the w+ locus in the P[w+]YAL insert, indicating that HP1 may play a role in Y chromosome packaging.

Animals↗

The genetics and evolution of the mariner transposable element in Drosophila simulans: worldwide distribution and experimental population dynamics.

We have studied both the frequency and biogeographical distribution of the transposable DNA element mariner in natural populations of Drosophila simulans and the short-term evolutionary characteristics of mariner in experimental populations. The mariner element has been identified in natural populations of D. simulans from Africa, Europe, the Middle East, Japan, Australia, several Pacific islands, North America, and South America. Only four lines out of 296 were devoid of active mariner elements, as measured by the presence of functional mariner transposase. A slight correlation was found between the latitudinal coordinate of the collection sites and the level of mariner activity in the populations; this correlation became highly significant in Australia where a cline in mariner activity was observed along the eastern coast of the continent. We also observed that wild-type laboratory strains kept for several years as small populations might lose mariner activity over time. Using experimental populations, we modeled what might happen when naturally occurring populations exhibiting high and low levels of mariner activity encounter one another. We found that active mariner elements either will tend to lose their activity over time and gradually become inactive or possibly will be lost from the population; in either case, this will lead to the pattern seen in this experiment of a significant loss of mariner activity over time.

Animals↗

Mutation rate: a simple concept has become complex.

The factors that cause new mutations or affect the rate at which they occur have important implications for many areas of genetics. But recent work on phenomena such as premeiotic mutations, which yield a cluster of identical new mutants at the some time, led us to realize that researchers are using the term "mutation rate" in different, and sometimes contradictory, ways. One premeiotic genetic change may ultimately yield several new mutant offspring, but should this be considered one new mutation or many? The way the data are handled in analyses can have a significant effect on the results. How, then, does one handle clusters in the estimation of mutation rates? We explore this question and propose that geneticists begin to distinguish clearly between three different phenomena that to this point have been given the same name: the initial prerepair "genetic damage rate," the postrepair "mutational event rate," and the observed "mutation rate" as it is expressed in the proportion of new mutant offspring. We believe that all new mutant offspring should be counted when estimating mutation rate, irrespective of when in the developmental cycle it is believed that the initial mutational event occurred.

Animals↗

Origin and decay of the P element-associated latitudinal cline in Australian Drosophila melanogaster.

The latitudinal cline in P transposable element-associated characteristics in eastern Australian populations of Drosophila melanogaster has changed between 1986 and 1991-1994. New collections were made in 1991-1994 from localities along the eastern coast of Australia. P element-associated properties of 256 isofemale lines from 43 localities were evaluated using gonadal dysgenesis and/or singed-weak hypermutability assays. The overall results indicate that both P activity and P susceptibility have declined, with all populations showing a tendency towards a state with little P activity potential but with P repressor function (neutral or 'Q'). P repressor function is strong in all populations except some of the most southernly. P activity potential peaks at about 27 degrees SLat, and drops off to the south (as in 1983-1986 collections) and to the north (in contrast to 1983-1986 collections); thus the cline is no longer a simple P-to-Q-to-M pattern from north to south, but is now Q-P-Q-M. A mtDNA RFLP that putatively distinguishes North American and European populations varies in frequency among the populations but the frequency does not vary clinically with latitude, ruling out massive introductions from North America and Europe as causing the cline.

Animals↗

Clusters of new identical mutants and the fate of underdominant mutations.

Given favorable environmental and demographic conditions, premeiotic clusters of identical mutations can produce a broad distribution of the initial frequency of underdominant alleles. Because of these clusters, new underdominant mutations may not necessarily be as rare in a population as previously assumed. The fixation of underdominant mutations, especially those with low heterozygous fitness, is increased when mutations appear in a cluster due to a genetic change that occurred before germline differentiation. Most restrictions on the fixation of underdominant mutations in a single population, such as strong genetic drift, weak selection against mutant heterozygotes, isolated population structure, inbreeding, meiotic drive, and selection in favor of mutant homozygotes can be relaxed or even dropped. Instead, the fate of strong underdominant mutations is determined mainly by ecological and genetic factors that affect the cluster size distribution of new premeiotic mutations. Accumulation of reproductive isolation by the fixation of underdominant mutations becomes more feasible with clusters, and mutation is not always the weakest force during this evolutionary process. The large mean and variance of reproductive success in many multicellular species make it possible that even underdominant mutations with very low heterozygous fitness could contribute substantially to reproductive isolation.

Alleles↗

Clusters of identical new mutations can account for the "overdispersed" molecular clock.

Germ-cell mutations may occur during meiosis, giving rise to independent mutant gametes in a Poisson process, or before meiosis, giving rise to multiple copies of identical mutant gametes at a much higher probability than the Poisson expectation. We report that the occurrence of these early premeiotic clusters of new identical mutant alleles increases the variance-to-mean ratio of mutation rate (R(u) > 1). This leads to an expected variance-to-mean ratio (R(t)) of the molecular clock that is always greater than one and may cover the observed range of R(t) values. Hence, the molecular clock may not be over-dispersed based on this new mutational model that includes clusters. To get a better estimation of R(u) and R(t), one needs measurements of the intrageneration variation of reproductive success (Nt/Ne(i)), population dynamics (ki), and the proportion of new mutations that occur in clusters (rc), especially those formed before germ-cell differentiation.

Animals↗

Clusters of identical new mutation in the evolutionary landscape.

In contrast to the common assumption that each new mutant results from a unique, independent mutation event, clusters of identical premeiotic mutant alleles are common. Clusters can produce large numbers of related individuals carrying identical copies of the same new genetic change. By entering the gene pool in multiple copies at one time, clusters can influence fundamental processes of population genetics. Here we report evidence that clusters can increase the arrival and fixation probabilities and can lengthen the average time to extinction of new mutations. We also suggest it may be necessary to reconsider other fundamental elements of population genetic theory.

Animals↗

P DNA element movement in somatic cells reduces lifespan in Drosophila melanogaster: evidence in support of the somatic mutation theory of aging.

Evidence is presented in support of the hypothesis that P DNA element movement in somatic cells of Drosophila melanogaster induces genetic damage that significantly reduces lifespan. The lifespan of D. melanogaster males was significantly reduced by the somatic movement of a single P element in the presence of P[ry+ delta 2-3](99B) transposase. In addition, the P[ry+ SalI](89D) repressor of P[ry+ delta 2-3](99B) somatic transposase was observed to reduce the effect of P element movement on lifespan. Finally, the frequency of somatic-cell chromosome breakage was significantly increased in neuroblasts of males with somatically active P elements. These results show that lifespan in D. melanogaster is decreased with increased somatic genetic damage from DNA-element movement. Although this conclusion does not confirm that transposable element movement is a cause of natural senescence, this conclusion is clear evidence in support of a close relationship between somatic genetic damage and aging.

Aging↗

Somatic movement of the mariner transposable element and lifespan of Drosophila species.

The effect of somatic movement of the mariner transposable element on lifespan was measured in Drosophila simulans and Drosophila melanogaster males at 25 degrees C. In D. simulans this movement significantly decreased lifespan, whereas in D. melanogaster no correlation between transposon movement and lifespan was found. The results in D. simulans support the hypothesis that somatic genetic damage induced by DNA element movement can reduce lifespan.

Aging↗

P element mediated germ line transformation of Drosophila melanogaster with the Tc1 transposable DNA element from Caenorhabditis elegans.

Questions relating to the origin and regulation of mobile genetic elements are currently of considerable interest. Since it is now possible to address more precisely issues concerning the entry, dispersion, and regulation of elements within a virgin genome, one approach that may afford a better understanding of transposable elements in general could be provided by interspecific DNA transformation. Therefore, the Tc1 transposable DNA element from Caenorhabditis elegans was chosen as a proposed invading element of the Drosophila melanogaster genome. The basis for this selection resided in the inherent structural and functional similarities, as well as sequence identities, between the Caenorhabditis element and elements innate to Drosophila (e.g., P, HB1, and Uhu). Initial investigations were carried out to define a clone carrying an intact Tc1 element. This Tc1 element was inserted into a P transposon vector and two P-Tc1-ry+ constructs, differing only in insert orientation, were identified. P element mediated germ line transfer was then used to generate a transformant that was genetically and molecularly identified as containing a single, structurally intact Tc1 element at cytological location 64C4-5 on the third chromosome. The single P[(Tc1,ry+)]SAS-B insertion was thereafter mobilized by using a P[ry+ delta 2-3] element as a transposase source, and the genetic and molecular data suggested that the insertion had been successfully reintegrated to a variety of genomic locations. On the basis of genetic and molecular analyses, the Tc1 element in the P[Tc1,ry+)] transformed stock is not highly unstable in germ line and somatic tissues.

Animals↗

A difference between front-loaded streptokinase and standard-dose recombinant tissue-type plasminogen activator in preserving left ventricular function after acute myocardial infarction (the Central Illinois Thrombolytic Therapy Study).

A blinded, randomized trial compared the effects of front-loaded streptokinase with those of the conventional dose of intravenous recombinant tissue-type plasminogen activator (rt-PA) on left ventricular (LV) function after acute myocardial infarction (AMI). Thrombolytic therapy was administered in the emergency departments of 30 community hospitals in central Illinois, and subsequent studies were performed at 1 tertiary referral center. Patients aged < or = 75 years with a first AMI who could be treated within 4 hours of the onset of chest pain were randomly assigned to receive either streptokinase (375,000 IU bolus, followed by 1,125,000 IU over 1 hour) or rt-PA (10 mg bolus, followed by 50 mg in the first hour, and 20 mg/hour for the next 2 hours). All patients were treated with aspirin (325 mg) and intravenous heparin. Patients were transferred for angiography within 24 hours. During the 30-month study, 253 patients were treated with intravenous thrombolytic therapy 2.4 +/- 1.0 hour after the onset of AMI. In patients with anterior wall AMI (n = 90), global LV ejection fraction measured by angiography within 24 hours was 45 +/- 12% with rt-PA, and 39 +/- 13% with streptokinase (p < 0.03). Convalescent radionuclide angiography documented a persistent beneficial effect of rt-PA on LV regional wall contractility, but not global ejection fraction. There were no differences between rt-PA and streptokinase in preserving global or regional LV function in patients with inferior wall AMI.

Aged↗

Nephrotoxicity of nonionic low-osmolality versus ionic high-osmolality contrast media: a prospective double-blind randomized comparison in human beings.

To test whether a nonionic, low-osmolality contrast medium (iopamidol) administered for coronary angiography was less harmful to renal function than ionic, high-osmolality medium (sodium diatrizoate), a prospective, double-blind randomized study of 70 patients with normal or mildly depressed renal function (serum creatinine < or = 2.0 mg/dL (175 mumol/L) was performed. Creatinine clearance was determined before coronary angiography and 24 and 48 hours after. There were no significant differences between the low- and high-osmolality groups with regard to age, baseline creatinine clearance, or dose of contrast medium given. In patients receiving low-osmolality medium (n = 35), creatinine clearance decreased by 19% +/- 13 (1 standard deviation) at 24 hours and recovered by 48 hours. In patients receiving high-osmolality medium (n = 35), creatinine clearance decreased by 40% +/- 16 at 24 hours and remained depressed by 47% +/- 14 at 48 hours. In patients with normal or mildly depressed renal function, use of a non-ionic, low-osmolality contrast medium minimized nephrotoxicity as measured by reductions in creatinine clearance after coronary angiography.

Adolescent↗

Transposable DNA elements and life history traits. I. Transposition of P DNA elements in somatic cells reduces the lifespan of Drosophila melanogaster.

As an initial study of the influence of transposable DNA elements on life history traits, and as a model system for estimating the impact of somatic genetic damage on longevity, the effect of P DNA element movement in somatic cells on adult lifespan was measured in Drosophila melanogaster males. Lifespan was significantly reduced in males that contained the somatically active P[ry+ delta 2-3](99B) element and 17, 4, 3, but not just a single P element. Furthermore, there appears to be a direct correlation between the number of transposing P elements and the amount of lifespan reduction. This reduction in lifespan observed in males with somatically active P elements is probably due to genetic damage in embryos, larvae and pupae from P-element excisions and insertions, leading to changes in gene structure and regulation, chromosome breakage, and subsequent cell death in adults. This hypothesis is supported in this study by a significant increase in recessive sex-linked lethal mutations in the same males that had reduced lifespans and by the previous observation of chromosome breakage in somatic cells of similar males. The evolutionary implications of these results are discussed, including the possible influence of somatic DNA transpositions on fitness and other life history traits.

Animals↗

Visible mutations induced by P-M hybrid dysgenesis in Drosophila melanogaster result predominantly from P element insertions.

This study supplied no evidence that P-M hybrid dysgenesis is a general release mechanisms for transposon movement. Newly induced mutations (23 singed, three yellow, and one white) were generated by hybrid dysgenesis and were molecularly analyzed for the presence or absence of P element insertions. Only one dysgenically-induced insertion mutation out of 27 analyzed was the result of a non-P insert; this frequency is not statistically above the non-dysgenic control level. Thus, it appears that individual transposable elements families are independently regulated.

Animals↗

Turnover of R1 (type I) and R2 (type II) retrotransposable elements in the ribosomal DNA of Drosophila melanogaster.

R1 and R2 are distantly related non-long terminal repeat retrotransposable elements each of which inserts into a specific site in the 28S rRNA genes of most insects. We have analyzed aspects of R1 and R2 abundance and sequence variation in 27 geographical isolates of Drosophila melanogaster. The fraction of 28S rRNA genes containing these elements varied greatly between strains, 17-67% for R1 elements and 2-28% for R2 elements. The total percentage of the rDNA repeats inserted ranged from 32 to 77%. The fraction of the rDNA repeats that contained both of these elements suggested that R1 and R2 exhibit neither an inhibition of nor preference for insertion into a 28S gene already containing the other type of element. Based on the conservation of restriction sites in the elements of all strains, and sequence analysis of individual elements from three strains, nucleotide divergence is very low for R1 and R2 elements within or between strains (less than 0.6%). This sequence uniformity is the expected result of the forces of concerted evolution (unequal crossovers and gene conversion) which act on the rRNA genes themselves. Evidence for the role of retrotransposition in the turnover of R1 and R2 was obtained by using naturally occurring 5' length polymorphisms of the elements as markers for independent transposition events. The pattern of these different length 5' truncations of R1 and R2 was found to be diverse and unique to most strains analyzed. Because recombination can only, with time, amplify or eliminate those length variants already present, the diversity found in each strain suggests that retrotransposition has played a critical role in maintaining these elements in the rDNA repeats of D. melanogaster.

Animals↗

Test of a semiselective screen for induced aneuploidy in germ cells of Drosophila melanogaster females with structurally normal chromosomes.

A new semiselective screen (only female progeny survive) for induced aneuploidy in germ cells of Drosophila melanogaster (referred to as 20/Q56 for the X-chromosome mutation markers in the parental females) has been validated by recovering cold, colchicine and N,N-dimethylnitrosamine (DMN) induced chromosome gain and loss events in females that contain structurally normal chromosomes. In addition, the spontaneous and induced results from the 20/Q56 assay, which identifies gain events at division I and loss events at divisions I and II of meiosis, were compared with a nonselective (all progeny survive) modified mating scheme that identifies gains and losses at both divisions of oogenesis. Females with the same genotypes are treated in the two mating schemes and are then mated with males that contain different marked Y chromosomes. The spontaneous rates of chromosome gains and losses were not significantly different in the two mating schemes (these rates ranged from 0.008 to 0.022%), supporting previous reports that spontaneous aneuploidy occurs at a higher frequency at division I of meiosis in females of D. melanogaster than at division II. Both the 20/Q56 and modified screens were able to identify significant increases in aneuploidy after adult treatments with cold shock (10 degrees C and 5 degrees C), colchicine (5 ppm and 10 ppm), and DMN (100 ppm). Brood analysis (five 2-day or five 3-day broods) showed that the largest increases in aneuploidy after cold treatment occurred in the first brood, which contains a high proportion of stage 14 oocytes, whereas colchicine induced the highest frequencies in the latter broods and DMN was effective in all but the last brood. Although the 20/Q56 mating scheme identifies gain events only in division I of meiosis whereas the modified mating scheme identifies gains in both divisions, the 20/Q56 scheme is just as effective in identifying induced aneuploidy as is the modified scheme. There were no significant differences in the frequencies of induced gains or losses in the two schemes. These results also suggest that the 3 treatments induced chromosome gain events mainly at division I of oogenesis. Taken together, the results from this study suggest that the 20/Q56 mating scheme in D. melanogaster, which is semiselective and therefore less expensive and time-consuming to perform, is an appropriate test system to screen for chemical induced aneuploidy in germ cells of a higher organism.

Aneuploidy↗