Ecological differences among clones of Daphnia pulex Leydig

Summary Natural populations of Daphnia pulex that reproduce by obligate parthenogenesis include a number of clones. Studies on two common and two rare clones from southwestern Ontario revealed significant differences in their intrinsic rates of increase, competitive abilities, rates of ephippial egg production, and lifespans. Environmental factors such as temperature and food type had large inluences on the rate of increase of each clone. Differences in rates of increase among clones were most pronounced at temperatures higher than those encountered in nature. In general, the covariance of life history traits among clones was high. The outcome of competitive encounters between clones was deterministic and in most cases was unaffected by temperature. Clones with high rates of increase tended to be better competitors than those with low rates of increase.     

Filtering rate of the cladoceran,Daphnia pulex as a function of body size,light and acclimation

Daphnia pulex were raised under nine light intensities (0, 1.7, 3.5, 7, 14, 28, 55, and 110 ft-c), polarized light (6.6 ft-c), and four wavelength ranges.Light intensity significantly affected the relationship between filtering rate and body size of unacclimated animals. The b values were lowest at 7 and 14 ft-c and they increased above 7 ft-c as light intensity increased. There were significant differences among b values and adjusted means for unacclimated animals. Acclimation to their respective conditions resulted in some significant differences between b values and adjusted means. For acclimated animals there were no significant differences among b values but there were some differences among adjusted means. The filtering rates of unacclimated and acclimated animals were lower at 14 ft-c while 1.7 and 3.5 ft-c were generally stimulatory and the effect was more pronounced in larger animals. Light intensities above 28 ft-c tended to suppress the filtering rate of small unacclimated animals and stimulate filtering rate in larger unacclimated animals. After acclimation, intensities above 7 ft-c did not affect the filtering rate of either small or large animals.The effect of polarized light on filtering rate was inseparable from the effect of light intensity, however, acclimation to polarized light resulted in a significantly higher b value.There were no significant effects of wavelength among b values for unacclimated animals and the adjusted mean for blue wavelengths was significantly higher than for violet wavelengths. There were no significant differences among b values or adjusted means for acclimated animals. Acclimation to their respective wavelengths did result in some significant differences between b values and adjusted means. Except for the effects of blue wavelengths on unacclimated animals and red wavelengths on acclimated animals, the effects of wavelengths are inseparable from the effects of light intensity.This paper is part of a Ph.D. Thesis submitted to the Faculty of Graduate School of The University of Kansas.Supported in part by a NSF Summer Traineeship and a grant from the Research Corporation to St. Olaf College, Northfield, Minnesota 55057.     

High mutation rates in the mitochondrial genomes of Daphnia pulex

Despite the great utility of mitochondrial DNA (mtDNA) sequence data in population genetics and phylogenetics, key parameters describing the process of mitochondrial mutation (e.g., the rate and spectrum of mutational change) are based on few direct estimates. Furthermore, the variation in the mtDNA mutation process within species or between lineages with contrasting reproductive strategies remains poorly understood. In this study, we directly estimate the mtDNA mutation rate and spectrum using Daphnia pulex mutation-accumulation (MA) lines derived from sexual (cyclically parthenogenetic) and asexual (obligately parthenogenetic) lineages. The nearly complete mitochondrial genome sequences of 82 sexual and 47 asexual MA lines reveal high mtDNA mutation rate of 1.37 × 10(-7) and 1.73 × 10(-7) per nucleotide per generation, respectively. The Daphnia mtDNA mutation rate is among the highest in eukaryotes, and its spectrum is dominated by insertions and deletions (70%), largely due to the presence of mutational hotspots at homopolymeric nucleotide stretches. Maximum likelihood estimates of the Daphnia mitochondrial effective population size reveal that between five and ten copies of mitochondrial genomes are transmitted per female per generation. Comparison between sexual and asexual lineages reveals no statistically different mutation rates and highly similar mutation spectra.     

Dopamine enhances mtNOS activity: Implications in mitochondrial function

Dopamine and nitric oxide systems can interact in different processes in the central nervous system. Dopamine and oxidation products have been related to mitochondrial dysfunction. In the present study, intact mitochondria and submitochondrial membranes were incubated with different DA concentrations for 5 min. Dopamine (1 mM) increased nitric oxide production in submitochondrial membranes and this effect was partially prevented in the presence of both DA and NOS inhibitor N_ω-nitro-l-arginine (l-NNA). A 46% decrease in state 3 oxygen uptake (active respiration state) was found after 15 mM dopamine incubation. When mitochondria were incubated with 15 mM dopamine in the presence of l-NNA, state 3 respiratory rate was decreased by only 17% showing the involvement of NO. As shown for O₂ consumption, the inhibition of cytochrome oxidase by 1 mM DA was mediated by NO. Hydrogen peroxide production significantly increased after 15 mM DA incubation, being mainly due to its metabolism by MAO. Also, DA-induced depolarization was prevented by the addition of l-NNA showing the involvement of nitric oxide in this process too. This work provides evidence that in the studied conditions, dopamine modifies mitochondrial function by a nitric oxide-dependent pathway.     

Life history variation among low-arctic clones of obligately parthenogenetic Daphnia pulex: a diploid-polyploid complex

Summary Laboratory life table experiments were conducted using nine clones of obligately parthenogenetic Daphnia pulex that were collected from a site in the Canadian low-arctic. Two of the nine clones were diploids, while the other seven clones were polyploids. Significant clonal differences in age at first reproduction, size at first reproduction, number of offspring in each of the first three broods, offsrring sizes for the first two broods, and intrinsic rates of natural increase were detected. Differences in life histories were evident between polyploids and diploids. Generally, polyploid clones reached maturity at later ages, matured at larger sizes, produced smaller broods, and larger offspring than the diploid clones. The data are discussed in reference to potential biotic (i.e. invertebrate predation) and abiotic factors (i.e. physicochemical gradients) that may influence life history variation in this clonal assemblage.     

A population of sexual Daphnia pulex resists invasion by asexual clones

Asexual reproduction avoids the costs associated with sex, predicting that invading asexual clones can quickly replace sexual populations. Daphnia pulex populations in the Great Lakes area are predominately asexual, but the elimination of sexual populations by invading clones is poorly understood. Asexual clones were detected at low frequency in one rare sexual population in 1995, with some increase in frequency during 2003 and 2004. However, these clones remained at low frequency during further yearly sampling (2005–2013) with no evidence that the resident sexual population was in danger of elimination. There was evidence for hybridization between rare males produced by asexual clones and sexual females with the potential to produce new asexual genotypes and spread the genetic factors for asexuality. In a short-term laboratory competition experiment, the two most common asexual clones did not increase in frequency relative to a genetically diverse sexual population due in part to a greater investment in diapausing eggs that trades-off current population growth for increased contribution to the egg bank. Our results suggest that a successful invasion can be prolonged, requiring a combination of clonal genotypes with high fitness, persistence of clones in the egg bank and negative factors affecting the sexual population such as inbreeding depression resulting from population bottlenecks.     

Thermal sensitivity of mitochondrial function in the Antarctic Notothenioid Lepidonotothen nudifrons

The thermal sensitivity of mitochondrial function was investigated in the stenothermal Antarctic fish Lepidonotothen nudifrons. State 3 respiration increases with increasing temperature between 0 °C and 18 °C with a Q ₁₀ of 2.43–2.63. State 4 respiration in the presence of oligomycin, an inhibitor of mitochondrial ATP synthase, quantifies the leakage of protons through the inner mitochondrial membrane, which causes oxygen consumption without concomitant ATP production. This parameter shows an unusually high Q ₁₀ of 4.21 ± 0.42 (0–18 °C), which indicates that proton leakage does not depend merely on ion diffusion but is an enzyme-catalysed process. The differential thermal sensitivity of oxidative phosphorylation (=state 3) and proton leakage (=state 4 in the presence of oligomycin) leads to progressive uncoupling of the mitochondria and decreased efficiency of oxidative phosphorylation under in vivo conditions if the body temperature of L. nudifrons increases. Accepted: 2 September 1999     

Inter-genotype variation in reproductive response to crowding among Daphnia pulex

Crowding is known to have a major influence on reproduction in the freshwater microcrustacean Daphnia pulex. We analyzed reproductive output of six different D. pulex genotypes under two different density regimes in the laboratory. Four of these genotypes reproduce via obligate parthenogenesis, allowing thorough analysis of the life history strategies of some asexual lines. Among 30,109 neonate offspring and 1041 resting egg ephippia collected, several trends were evident. Crowding induced increased resting egg production and reduced neonate offspring production among all genotypes. Offspring sex ratios grew more male-biased with maternal age. The extent, but not direction, of each of these trends varied among genotypes. Offspring sex ratios, and the very direction in which they changed in response to crowding, differed significantly among genotypes with some genotypes producing more and others fewer males in response to crowding. Obligately parthenogenetic genotypes seemed to respond to the crowding stimulus in similar ways as the facultatively parthenogenetic genotypes, as expected from the sexual origins of their genomes. The inter-genotype variation in life-history traits observed in this and other investigations calls into question the common practice of extrapolating results from a single Daphnia genotype to an entire species. Our findings are considered in the context of other research in the field of environmental influences on Daphnia reproduction with a review of representative literature.     

Variation in allocation to sexual and asexual reproduction among clones of cyclically parthenogenetic Daphnia pulex (Crustacea: Gladocera)

Organisms reproducing by cyclical parthenogenesis combine the benefits of both sexual and asexual reproduction within the same life cycle. Few studies have examined the evolution of variation in the pattern of investment in parthenogenetic compared to sexual reproduction. Seven clones of Daphnia pulex (Crustacea: Cladocera) varying in allocation to sexual reproduction, as measured by the production of males, were raised in isolation and together in a microcosm to study the pattern of sexual reproduction and the effect of this variation on clone fitness. Sex allocation for clones raised together a microcosm was similar to their allocation when raised in isolation, suggesting a genetic basis to the variation. Three clones showed a cost of producing males that lead to their extinction after about 30 days due to the lack of females required for the clones to persist by parthenogenetic reproduction. The remaining four clones persisted until the end of the 72-day experiment. Clones with little or no allocation to males showed no increased allocation to sexual females. The seven clones showed a greater variation in estimated fitness through male and female function than in total estimated fitness. The clone with the greatest total fitness gained most of its fitness through male function but also had a relatively high fitness through female function. Although one clone produced only females it had the next highest fitness. The three clones that went extinct because of a high investment in males had estimated fitness as high as some clones that persisted in the microcosm because of a higher investment in parthenogenetic reproduction. The similarity in total fitness among clones suggests that Daphnia pulex populations in temporary habitats maintain a sex polymorphism where different genotypes vary-in functional gender ranging from female to primarily male.     

Genetic differentiation among Oregon lake populations of the Daphnia pulex species complex

Gene flow among invertebrate populations inhabiting bodies of nonflowing freshwater such as ponds or lakes must at some stage involve transport across habitat unsuitable for adult stages. Consequently the potential for interpopulational differentiation is high in these species, yet empirical studies of lake populations of Cladocerans such as Daphnia have failed to reveal high levels of genetic distinctiveness among populations and have led to much speculation about how these populations exchange genes and remain cohesive evolutionary units. In this study we surveyed 42 Oregon lake populations of Daphnia from the D. pulex species complex for genetic variation within the mitochondrial DNA control region. We have used this data to test the relative abilities of various ecological factors to explain the observed patterns in genetic differentiation among lakes. Despite limited genetic variation detected among our samples--11 very similar RFLP-defined mtDNA genotypes from 388 individuals--analyses of nucleotide variance using analogs to Wright's F statistics indicate that when multilake populations are defined in terms of the river drainage basin to which they belong, strong and significant amounts of among-population genetic variation can be detected at this locus (F(ST) estimates between 0.5 and 0.6). In contrast, we fail to detect consistent significant among-population variation when populations are defined on the basis of regional physical geography, bird migratory flyways, or lake trophic status. The manner in which the data are compiled, that is, whether RFLPs or nucleotide sequences are used, has little effect on the overall conclusions, yet it is clear that nucleotide sequence data would lower the standard errors of F(ST) estimates. We propose that periodic widescale flooding during the late Pleistocene may be an important mechanism to homogenize genetic differences among lake Daphnia continent-wide south of the southern-most extent of Pleistocene glaciation.     

Hierarchical analysis of population genetic variation in mitochondrial and nuclear genes of Daphnia pulex

The geographic structure of Daphnia pulex populations from the central United States is analyzed with respect to isozyme and mitochondrial DNA variation. The species complex consists of cyclic and obligate parthenogens. A hierarchical analysis of population structure in the cyclic parthenogens by using a fixation-index approach indicates that this is one of the most extremely subdivided species yet studied. This genetic structure, much of which accrues within 100 km, is certainly due in part to the limited dispersal ability of Daphnia. However, previous work has shown that fluctuating selection can account for the spatial heterogeneity in isozyme frequencies in these populations. This may explain why the population subdivision for the mitochondrial genome increases approximately three times as rapidly with distance as does that for nuclear genes, which is slower than the neutral expectation. The obligate parthenogens are shown to be polyphyletic in origin, evolutionarily young, and, in some cases, geographically widespread.     

The role of mitochondrial electron transport in tumorigenesis and metastasis

Background

Tumor formation and spread via the circulatory and lymphatic drainage systems is associated with metabolic reprogramming that often includes increased glycolytic metabolism relative to mitochondrial energy production. However, cells within a tumor are not identical due to genetic change, clonal evolution and layers of epigenetic reprogramming. In addition, cell hierarchy impinges on metabolic status while tumor cell phenotype and metabolic status will be influenced by the local microenvironment including stromal cells, developing blood and lymphatic vessels and innate and adaptive immune cells. Mitochondrial mutations and changes in mitochondrial electron transport contribute to metabolic remodeling in cancer in ways that are poorly understood.

Scope of Review

This review concerns the role of mitochondria, mitochondrial mutations and mitochondrial electron transport function in tumorigenesis and metastasis.

Major Conclusions

It is concluded that mitochondrial electron transport is required for tumor initiation, growth and metastasis. Nevertheless, defects in mitochondrial electron transport that compromise mitochondrial energy metabolism can contribute to tumor formation and spread. These apparently contradictory phenomena can be reconciled by cells in individual tumors in a particular environment adapting dynamically to optimally balance mitochondrial genome changes and bioenergetic status.

General Significance

Tumors are complex evolving biological systems characterized by genetic and adaptive epigenetic changes. Understanding the complexity of these changes in terms of bioenergetics and metabolic changes will permit the development of better combination anticancer therapies. This article is part of a Special Issue entitled Frontiers of Mitochondrial Research.     

Induction of male production in clones of Daphnia pulex by the juvenoid hormone methyl farnesoate under short photoperiod

The juvenile hormone, methyl farnesoate (MF), and its analog insecticides have been used successfully to induce the production of males in cladocerans under long-day conditions in the laboratory. However, without hormone addition Daphnia do not usually produce male offspring under long photoperiods, while short photoperiods are a stimulus for the induction of males. We used 21 clones of Daphnia pulex differing in their propensity to produce males under short-day conditions to test whether the treatment with MF would result in an additive effect of shifting the sex ratio towards males. Contrary to our expectations, clones with a high tendency of male production showed a reduced sex ratio in response to MF treatment under short-day conditions, but clones that produced normally few males or did not produce males were stimulated by 700 nM MF to produce up to 40% males. We suggest that the endocrine disruptive effect of MF or juvenile hormone analogs in the field may depend on the clonal composition of the cladoceran population and on the natural photoperiod. This may affect the seasonal occurrence of sexual reproduction and eventually cause a mismatch between the presence of males and ephippial females.     

Embryology of Chaoborus-induced spines in Daphnia pulex

Daphnia pulex (Crustacea: Cladocera) embryos were found to be sensitive to a chemical cue (kairomone) in an extract of the predator Chaoborus americanus (Insecta:Diptera). Sensitivity of embryos to the kairomone remains throughout embryonic development. Apparently declining sensitivity as development proceeds may be due to the amount of time the embryos are exposed to the kairomone. Male embryos were also found to be sensitive to the kairomone. The smallest eggs within a brood produced small offspring, which showed the antipredator morphology to a significantly lower degree than largest eggs. The production of the neckteeth is described, at the developmental stage in the maturation of the Daphnia coinciding approximately with the escape of the embryos from the brood chamber.     

The complete sequence of the mitochondrial genome of Daphnia pulex (Cladocera: Crustacea).

The sequence of the mitochondrial DNA (mtDNA) of the branchiopod crustacean Daphnia pulex has been completed. It is 15333bp with an A+T content of 62.3%, and contains the typical complement of 13 protein-coding, 22 transfer RNA (tRNA) and two ribosomal RNA (rRNA) genes. Comparison of this sequence with the sequences of the other eight completely sequenced arthropod mtDNAs showed that gene order and orientation are identical to that of Drosophila but different from Artemia due to the rearrangement of two tRNA genes. Nucleotide composition, codon usage, and amino acid composition are very similar in the crustaceans, but divergent from insects and chelicerates which show a much higher bias towards A+T. However, with few exceptions, the mitochondrial proteins of Daphnia are more similar to those of the dipteran insects (Drosophila and Anopheles) than to those of Artemia, at both the nucleotide and amino acid levels, suggesting that Artemia mtDNA is evolving at an accelerated rate. These results also show that sequence evolution and the evolution of nucleotide composition can be decoupled. Analysis of nucleotide substitution patterns in COII showed that there has been an unbiased acceleration of the overall substitution rate in Artemia. In contrast, the accelerated substitution rate in Apis is due partly to extreme A+T mutation pressure. Secondary structures are proposed for the Daphnia tRNAs and rRNAs. The tRNAs are similar to those of other arthropods but tend to have TPsiC arms that are only 4bp long. The rRNA secondary structures are similar to those proposed for insects except for the absence of a small number of helices in Daphnia. Phylogenetic analysis of second codon positions grouped Daphnia with Artemia, as expected, despite the latter's accelerated divergence rate. In contrast, the unusual pattern of mtDNA divergence in Apis led to a topology in which the holometabolous insects (Anopheles, Drosophila, Apis) appeared to be paraphyletic with respect to the hemimetabolous insect, Locusta, due to the early branching of Apis.     

UCP1 ectopically expressed in murine muscle displays native function and mitigates mitochondrial superoxide production

Mitochondrial uncoupling in skeletal muscle has raised a major interest as a therapeutic target for treatment of obesity, insulin sensitivity, and age-related disease. These physiological effects could be demonstrated in several mouse models ectopically expressing uncoupling protein 1 (UCP1). Here, we investigated whether UCP1 expressed under the control of the human skeletal actin (HSA) promoter in mouse skeletal muscle can be regulated, and whether it affects mitochondrial superoxide production. We show that the skeletal muscle UCP1 can be fully inhibited by a purine nucleotide (GDP) and reactivated by fatty acids (palmitate). During mitochondrial resting state (State 4), mitochondrial superoxide production is about 76% lower in transgenic mice. We suggest that this reduction is due to uncoupling activity as the administration of GDP restores superoxide production to wildtype levels. Our study confirms native behaviour of UCP1 in skeletal muscle and demonstrates beneficial effects on prevention of mitochondrial reactive oxygen species production which may reduce age-related deleterious processes.     

Mitochondrial functional complementation in mitochondrial DNA-based diseases

Mitochondria exist in networks that are continuously remodeled through fusion and fission. Why do individual mitochondria in living cells fuse and divide continuously? Protein machinery and molecular mechanism for the dynamic nature of mitochondria have been almost clarified. However, the biological significance of the mitochondrial fusion and fission events has been poorly understood, although there is a possibility that mitochondrial fusion and fission are concerned with quality controls of mitochondria. trans-mitochondrial cell and mouse models possessing heteroplasmic populations of mitochondrial DNA (mtDNA) haplotypes are quite efficient for answering this question, and one of the answers is “mitochondrial functional complementation” that is able to regulate respiratory function of individual mitochondria according to “one for all, all for one” principle. In this review, we summarize the observations about mitochondrial functional complementation in mammals and discuss its biological significance in pathogeneses of mtDNA-based diseases.     

Cardiac mitochondrial biogenesis in endotoxemia is not accompanied by mitochondrial function recovery

Mitochondrial biogenesis emerges as a compensatory mechanism involved in the recovery process in endotoxemia and sepsis. The aim of this work was to analyze the time course of the cardiac mitochondrial biogenesis process occurring during endotoxemia, with emphasis on the quantitative analysis of mitochondrial function. Female Sprague-Dawley rats (45 days old) were ip injected with LPS (10 mg/kg). Measurements were performed at 0–24 h after LPS administration. PGC-1α and mtTFA expression for biogenesis and p62 and LC3 expression for autophagy were analyzed by Western blot; mitochondrial DNA levels by qPCR, and mitochondrial morphology by transmission electron microscopy. Mitochondrial function was evaluated as oxygen consumption and respiratory chain complex activity. PGC-1α and mtTFA expression significantly increased in every time point analyzed, and mitochondrial mass was increased by 20% (P<0.05) at 24 h. p62 expression was significantly decreased in a time-dependent manner. LC3-II expression was significantly increased at all time points analyzed. Ultrastructurally, mitochondria displayed several abnormalities (internal vesicles, cristae disruption, and swelling) at 6 and 18 h. Structures compatible with fusion/fission processes were observed at 24 h. A significant decrease in state 3 respiration was observed in every time point analyzed (LPS 6 h: 20%, P<0.05). Mitochondrial complex I activity was found decreased by 30% in LPS-treated animals at 6 and 24 h. Complex II and complex IV showed decreased activity only at 24 h. The present results show that partial restoration of cardiac mitochondrial architecture is not accompanied by improvement of mitochondrial function in acute endotoxemia. The key implication of our study is that cardiac failure due to bioenergetic dysfunction will be overcome by therapeutic interventions aimed to restore cardiac mitochondrial function.     

Predator-induced phenotypic plasticity in the exotic cladoceran Daphnia lumholtzi

1. The exotic cladoceran Daphnia lumholtzi has recently invaded freshwater systems throughout the United States. Daphnia lumholtzi possesses extravagant head spines that are longer than those found on any other North American Daphnia. These spines are effective at reducing predation from many of the predators that are native to newly invaded habitats; however, they are plastic both in nature and in laboratory cultures. The purpose of this experiment was to better understand what environmental cues induce and maintain these effective predator‐deterrent spines. We conducted life‐table experiments on individual D. lumholtzi grown in water conditioned with an invertebrate insect predator, Chaoborus punctipennis, and water conditioned with a vertebrate fish predator, Lepomis macrochirus. 2. Daphnia lumholtzi exhibited morphological plasticity in response to kairomones released by both predators. However, direct exposure to predator kairomones during postembryonic development did not induce long spines in D. lumholtzi. In contrast, neonates produced from individuals exposed to Lepomis kairomones had significantly longer head and tail spines than neonates produced from control and Chaoborus individuals. These results suggest that there may be a maternal, or pre‐embryonic, effect of kairomone exposure on spine development in D. lumholtzi. 3. Independent of these morphological shifts, D. lumholtzi also exhibited plasticity in life history characteristics in response to predator kairomones. For example, D. lumholtzi exhibited delayed reproduction in response to Chaoborus kairomones, and significantly more individuals produced resting eggs, or ephippia, in the presence of Lepomis kairomones.