Non-Mendelian Female Sterility in DROSOPHILA MELANOGASTER: Influence of Aging and Thermic Treatments. III. Cumulative Effects Induced by These Factors

Crosses between various strains of Drosophila melanogaster may give rise to a female sterility of non-Mendelian determination. Reduced fertility is observed in females, known as SF females, bred from crosses between females of "reactive" strains and males of "inducer" strains. The reduced fertility of the SF females is the result of an interaction between an extrachromosomal property varies considerably in its ability to reduce fertility. The fertility reduction of the SF females corresponds to what is known as the reactivity level of their reactive mothers. Two nongenetic factors can modify the level of reactivity: aging and temperature. The action of aging is cumulative. When the flies of a reactive strain are submitted at each generation to the action of this factor, the level of reactivity of this strain is gradually modified. The modifications induced are reversible. Indeed, when such a modified strain is returned to standard breeding conditions, the reactivity returns progressively to its initial level. The effect of thermic treatments also seems to be cumulative and reversible.     

Germ-line expression of the I factor,a functional LINE from the fruit fly Drosophila melanogaster,is positively regulated by reactivity,a peculiar cellular state

I factor is a functional LINE (long interspersed nucleotidic element) which is mobilized in the germ-line of dysgenic SF females during I-R hybrid dysgenesis. Such females are obtained when an oocyte from a reactive stock, devoid of I factors but characterized by a level of reactivity, i.e. its potential for hybrid dysgenesis, is fertilized by a spermatozoon from an I factor-containing inducer stock. In a previous paper we described the expression of an I factor-lacZ fusion. Expression was detected in the ovaries of reactive and dysgenic flies only. In this paper we show that this transgenic activity can be quantified and depends upon the maternally inherited reactivity. Reactivity is not just a permissive state and modifiers of the reactivity level such as heat treatment and ageing change the level of expression of our transgenic fusion accordingly. Moreover, ageing through generations has the same cumulative and reversible effect on both reactivity and I factor expression. Using our fusion as a test for reactivity we show that the silencing of I factor after its introduction into a reactive genome may not be established in a single generation.     

Germ-line expression of the I factor, a functional LINE from the fruit fly Drosophila melanogaster, is positively regulated by reactivity, a peculiar cellular state

I factor is a functional LINE (long interspersed nucleotidic element) which is mobilized in the germ-line of dysgenic SF females during I-R hybrid dysgenesis. Such females are obtained when an oocyte from a reactive stock, devoid of I factors but characterized by a level of reactivity, i.e. its potential for hybrid dysgenesis, is fertilized by a spermatozoon from an I factor-containing inducer stock. In a previous paper we described the expression of an I factor-lacZ fusion. Expression was detected in the ovaries of reactive and dysgenic flies only. In this paper we show that this transgenic activity can be quantified and depends upon the maternally inherited reactivity. Reactivity is not just a permissive state and modifiers of the reactivity level such as heat treatment and ageing change the level of expression of our transgenic fusion accordingly. Moreover, ageing through generations has the same cumulative and reversible effect on both reactivity and I factor expression. Using our fusion as a test for reactivity we show that the silencing of I factor after its introduction into a reactive genome may not be established in a single generation.     

Evidence for an Inducible Repair-Recombination System in the Female Germ Line of Drosophila Melanogaster. II. Differential Sensitivity to Gamma Rays

In a previous paper, we reported that the reactivity level, which regulates the frequency of transposition of I factor, a LINE element-like retrotransposon, is enhanced by the same agents that induce the SOS response in Escherichia coli. In this report, we describe experimental evidence that, for identical genotypes, the reactivity levels correlate with the sensitivity of oogenesis to gamma rays, measured by the number of eggs laid and by frequency of dominant lethals. This strongly supports the hypothesis that the reactivity level is one manifestation of an inducible DNA repair system taking place in the female germ line of Drosophila melanogaster. The implications of this finding for the understanding of the regulation of I factor are discussed and some other possible biological roles of this system are outlined.     

Evidence for an Inducible Repair-Recombination System in the Female Germ Line of Drosophila Melanogaster. III. Correlation between Reactivity Levels, Crossover Frequency and Repair Efficiency

We previously reported evidence that the so-called reactivity level, a peculiar cellular state of oocytes that regulates the frequency of transposition of I factor, a LINE element-like retrotransposon, might be one manifestation of a DNA repair system. In this article, we report data showing that the reactivity level is correlated with the frequency of crossing over, at least on the X chromosome and on the pericentromeric region of the third chromosome. Moreover, a check for X-chromosome losses and recessive lethals produced after gamma irradiation in flies with different reactivity levels, but common genetic backgrounds, brings more precise evidence for the relationship between reactivity levels and DNA repair. Those results support the existence of a repair-recombination system whose efficiency is modulated by endogenous and environmental factors. The implications of this biological system in connecting genomic variability and environment may shed new lights on adaptative mechanisms. We propose to call it VAMOS for variability modulation system.     

Rosette formation by insect macrophages inhibition by cytochalasin B.

Macrophages from the insect Spodoptera eridania possess membrane receptors for unmodified avian and mammalian erythrocytes, with which they form spontaneous rosettes. Rosette formation occurs in the absence of serum proteins and divalent cations. Individual macrophages bear receptors for several types of red cells. The level of naturally-occurring hemagglutinins against a particular test erythrocyte is not correlated with macrophage reactivity against that red cell. In contrast with mammalian macrophages, neuraminidase treatment of either hemocytes or erythrocytes does not cause a marked enhancement of binding. Pretreatment of macrophages or erythrocytes with cytochalasin B causes reversible inhibition of resetting probably by interfering with normal microfilament function, suggesting that optimal binding occurs when membranes are functioning normally on both macrophages and red cells. Colchicine and vinblastine do not influence resetting; therefore, microtubules are probably not involved in erythrocyte binding.     

Defective I elements introduced intoDrosophila as transgenes can regulate reactivity and prevent I-R hybrid dysgenesis

The I-R hybrid dysgenesis syndrome is characterized by a high level of sterility and I element transposition, occurring in the female offspring of crosses between males of inducer (I) strains, which contain full-length transposable I elements, and females of reactive (R) strains, devoid of functional I elements. The intensity of the syndrome in the dysgenic cross is essentially dependent on the reactivity level of the R females, which is ultimately controlled by still unresolved polygenic chromosomal determinants. In the work reported here, we have introduced a transposition-defective I element with a 2.6 kb deletion within its second open reading frame into a highly reactive R strain, by P-mediated transgenesis. We demonstrate that this defective I element gradually alters the level of reactivity in the three independent transgenic lines that were obtained, over several generations. After > 15 generations, the transgenicDrosophila show strongly reduced reactivity, and finally become refractory to hybrid dysgenesis, without, however, acquiring the inducer phenotype. Induction of a low reactivity level is reversible reactivity again increases upon transgene removal and is maternally inherited, as observed for the control of reactivity in natural R strains. These results demonstrate that defective I elements introduced as single-copy transgenes can act as regulators of reactivity, and suggest that some of the ancestral defective pericentromeric I elements that can be found in all reactive strains could be the molecular determinants of reactivity.     

Defective I elements introduced intoDrosophila as transgenes can regulate reactivity and prevent I-R hybrid dysgenesis

The I-R hybrid dysgenesis syndrome is characterized by a high level of sterility and I element transposition, occurring in the female offspring of crosses between males of inducer (I) strains, which contain full-length transposable I elements, and females of reactive (R) strains, devoid of functional I elements. The intensity of the syndrome in the dysgenic cross is essentially dependent on the reactivity level of the R females, which is ultimately controlled by still unresolved polygenic chromosomal determinants. In the work reported here, we have introduced a transposition-defective I element with a 2.6 kb deletion within its second open reading frame into a highly reactive R strain, by P-mediated transgenesis. We demonstrate that this defective I element gradually alters the level of reactivity in the three independent transgenic lines that were obtained, over several generations. After > 15 generations, the transgenicDrosophila show strongly reduced reactivity, and finally become refractory to hybrid dysgenesis, without, however, acquiring the inducer phenotype. Induction of a low reactivity level is reversible reactivity again increases upon transgene removal and is maternally inherited, as observed for the control of reactivity in natural R strains. These results demonstrate that defective I elements introduced as single-copy transgenes can act as regulators of reactivity, and suggest that some of the ancestral defective pericentromeric I elements that can be found in all reactive strains could be the molecular determinants of reactivity.     

Epigenetic inheritance, genetic assimilation and speciation.

Epigenetic inheritance systems enable the environmentally induced phenotypes to be transmitted between generations. Jablonka and Lamb (1991, 1995) proposed that these systems have a substantial role during speciation. They argued that divergence of isolated populations may be first triggered by the accumulation of (heritable) phenotypic differences that are later followed and strengthened by genetic changes. The plausibility of this idea is examined in this paper. At first, we discuss the "exploratory" behaviour of an epigenetic inheritance system on a one peak adaptive landscape. If a quantitative trait is far from the optimum, then it is advantageous to induce heritable phenotypic variation. Conversely, if the genotypes get closer to the peak, it is more favorable to canalize the phenotypic expression of the character. This process would lead to genetic assimilation. Next we show that the divergence of heritable epigenetic marks acts to reduce or to eliminate the genetic barrier between two adaptive peaks. Therefore, an epigenetic inheritance system can increase the probability of transition from one adaptive state to another. Peak shift might be initiated by (i) slight changes in the inducing environment or by (ii) genetic drift of the genes controlling epigenetic variability. Remarkably, drift-induced transition is facilitated even if phenotypic variation is not heritable. A corollary of our thesis is that evolution can proceed through suboptimal phenotypic states, without passing through a deep adaptive valley of the genotype. We also consider the consequences of this finding on the dynamics and mode of reproductive isolation.     

Bistability and phase variation in Salmonella enterica

Cell-to-cell differences in bacterial gene expression can merely reflect the occurrence of noise. In certain cases, however, heterogeneous gene expression is a programmed event that results in bistable expression. If bistability is heritable, bacterial lineages are formed. When programmed bistability is reversible, the phenomenon is known as phase variation. In certain cases, bistability is controlled by genetic mechanisms (e. g., DNA rearrangement). In other cases, bistability has epigenetic origin. A robust epigenetic mechanism for the formation of bacterial lineages is the formation of heritable DNA methylation patterns. However, bistability can also arise upon propagation of gene expression patterns by feedback loops that are stable upon cell division. This review describes examples of bistability and phase variation in Salmonella enterica and discusses their adaptive value, sometimes in a speculative manner.     

Short-term heritable variation overwhelms 200 generations of mutational variance for metabolic traits in Caenorhabditis elegans

Metabolic disorders have a large heritable component, and have increased markedly in human populations over the past few generations. Genome-wide association studies of metabolic traits typically find a substantial unexplained fraction of total heritability, suggesting an important role of spontaneous mutation. An alternative explanation is that epigenetic effects contribute significantly to the heritable variation. Here, we report a study designed to quantify the cumulative effects of spontaneous mutation on adenosine metabolism in the nematode Caenorhabditis elegans, including both the activity and concentration of two metabolic enzymes and the standing pools of their associated metabolites. The only prior studies on the effects of mutation on metabolic enzyme activity, in Drosophila melanogaster, found that total enzyme activity presents a mutational target similar to that of morphological and life-history traits. However, those studies were not designed to account for short-term heritable effects. We find that the short-term heritable variance for most traits is of similar magnitude as the variance among MA lines. This result suggests that the potential heritable effects of epigenetic variation in metabolic disease warrant additional scrutiny.     

An Age-Structured Approach to Modelling Behavioural Variation Maintained by Life-History Trade-Offs

There have been numerous empirical studies on the fitness consequences of behavioural syndromes in various animal taxa; however, the ecological and evolutionary implications on a population level are still poorly understood. To better understand these implications, we develop a non-linear age-structured mathematical model to qualitatively examine the evolutionary consequences of a heritable boldness personality trait within an animal population. We assume that this heritable boldness trait is positively correlated with boldness towards predators and intraspecific aggressiveness. This assumption leads to a growth/reproductive success versus mortality trade-off, which is thoroughly investigated and documented in the literature. Another life-history trade-off we include in the model is future versus current reproduction, which was shown by Wolf et al. [1] to be a possible mechanism for the evolution of behavioural syndromes within an animal population. The stability of the system is analysed, whereby the characteristic equation is in the form of a homogeneous Fredholm equation of the second kind which depends on both the perturbation and equilibrium solution. The system is found to be stable due to the competition between individuals of similar boldness acting as a negative feedback mechanism. Using numerical simulations we examine the qualitative features of the solution to the system. In particular, we investigate the interplay between the mutation and competition strength between two individuals with different boldness, whereby we find that an increasing competition range acts to push individuals to both extremes of the shy-bold axis, while an increasing mutation range counteracts this effect. This qualitative trait of aggregation of individuals around the shy and bold extremes is also found when examining different birth, mortality and competition functions.     

A general definition of the heritable variation that determines the potential of a population to respond to selection

Genetic selection is a major force shaping life on earth. In classical genetic theory, response to selection is the product of the strength of selection and the additive genetic variance in a trait. The additive genetic variance reflects a population's intrinsic potential to respond to selection. The ordinary additive genetic variance, however, ignores the social organization of life. With social interactions among individuals, individual trait values may depend on genes in others, a phenomenon known as indirect genetic effects. Models accounting for indirect genetic effects, however, lack a general definition of heritable variation. Here I propose a general definition of the heritable variation that determines the potential of a population to respond to selection. This generalizes the concept of heritable variance to any inheritance model and level of organization. The result shows that heritable variance determining potential response to selection is the variance among individuals in the heritable quantity that determines the population mean trait value, rather than the usual additive genetic component of phenotypic variance. It follows, therefore, that heritable variance may exceed phenotypic variance among individuals, which is impossible in classical theory. This work also provides a measure of the utilization of heritable variation for response to selection and integrates two well-known models of maternal genetic effects. The result shows that relatedness between the focal individual and the individuals affecting its fitness is a key determinant of the utilization of heritable variance for response to selection.     

Enhancement of solubility by temporary dimethoxybenzyl-substitution of peptide bonds. Towards the synthesis of defined oligomers of alanine and of lysyl-glutamyl-glycine

The synthesis of the model compound Aloc-Ala-Ala-Dma-Ala-Ala-OMe has been described as an illustration of the fact that a large group reversibly alkylating the amido group of an oligomer can disturb the regularity of a peptide backbone, oppose its aggregation and thus enhance its solubility greatly, affording synthons for further oligomerization. Application of such a group not only affects the solubility, but alters also the properties of the intermediates. The concomitant change in reactivity may run to such an extent that N-alkylation of oligomers has to be abandoned (this was encountered in the attempted synthesis of Lys-Glu-Dmg). Consequently, the solubility of the growing protected peptide chain will become progressively less and in the mentioned example the oligomerization had to be terminated at the dodecapeptide level, indicating the severe need for reversible "structure-breaking" functions.     

Interaction of ethidium bromide with DNA as studied by kinetic spectrophotometry.

The interaction of ethidium bromide (EB) with DNA has been investigated using the pulse radiolysis technique. In particular, the absolute rate constant for the reaction of hydrated electrons, generated by single pulses of high-energy electrons, with EB is shown to drop dramatically in the presence of DNA. This drop in diffusion-limited reactivity results from the interaction of EB with DNA, effectively immobilising it, thus lowering the reaction cross-section or probability. Analysis of the resulting kinetic spectrophotometric data shows that they are consistent with a reversible interaction of EB with DNA as described by the law of mass action. The Scatchard-type plots obtained are linear, and give quantitative information on the extent and degree of association, comparable with that obtained by more conventional methods. The potential of the pulse radiolysis technique for studying different types of interactions between small molecules and various biopolymers has been demonstrated.     

Variation in the Competence of Tobacco Pith Cells for Cytokinin-Habituation in Culture

Pith parenchyma tissues of tobacco sometimes lose their exogenous requirement for a cell division factor such as the cytokinin, kinetin. This process, known as cytokinin habituation, appears to involve epigenetic changes since it is a heritable change in cell phenotype which is directed, regularly reversible, and leaves the cell totipotent. In this report, we show that pith cells in culture consist of at least two types of cytokinin-requiring cells. The first type habituates rapidly under inductive conditions. The second type continues to express the cytokinin-requiring phenotype for many cell generations in culture but retains the capacity for habituation. These findings suggest that pith cells differ in their competence to habituate and that different states of competence are inherited by individual cells.     

Heritable Epigenetic Changes Alter Transgenerational Waveforms Maintained by Cycling Stores of Information

Our view of heredity can potentially be distorted by the ease of introducing heritable changes in the replicating gene sequences but not in the cycling assembly of regulators around gene sequences. Here, key experiments that have informed the understanding of heredity are reinterpreted to highlight this distortion and the possible variety of heritable changes are considered. Unlike heritable genetic changes, which are always associated with mutations in gene sequence, heritable epigenetic changes can be associated with physical or chemical changes in molecules or only changes in the system. The transmission of cycling stores along the continuous lineage of cells that connects successive generations creates waves of activity and localization of the molecules that together form the cell code for development in each generation. As a result, heritable epigenetic changes can include any that can alter a wave such as changes in form, midline, frequency, amplitude, or phase. Testing this integrated view of all heritable information will require the concerted application of multiple experimental approaches across generations.     

Cell surface expression of C1qRP/CD93 is stabilized by O-glycosylation

C1qRP/CD93 is a cell surface receptor predominantly expressed on monocytes, neutrophils, endothelial cells, and early stem cell precursors. In phagocytic cells, it has been characterized as contributing to the enhancement of FcR- and CR1-induced phagocytosis triggered by innate immune system defense collagens such as C1q and mannose binding lectin (MBL). Previously, we demonstrated a high level of glycosylation on C1qRP/CD93 that was predominantly O-linked. In this study, we investigate the role of glycosylation in C1qRP/CD93 stability first by inhibiting O-glycosylation by addition of benzyl 2-acetamido-2-deoxy-alpha-D-galactopyranoside (BAG) to the human histiocytic cell line U937, and secondly, by expression of C1qRP/CD93 in the CHO-derived cell line ldlD which has a reversible defect in protein glycosylation. In both U937 cells and in ldlD cells transfected to express C1qRP/CD93, glycosylation deficiency caused cell surface expression levels of C1qRP/CD93 to decrease, concomitant with the detection of C1qRP/CD93 reactivity in the culture media. Metabolic labeling studies show that when glycosylation is absent, C1qRP/CD93 is synthesized and rapidly released into the culture supernatant or degraded. These studies demonstrate that O-glycosylation is important in the stable cell surface expression of C1qRP/CD93 .     

植被-大气相互作用中的气孔导度及其尺度转换

气孔导度是衡量植物和大气间水分、能量及CO2平衡和循环的重要指标,探讨气孔导度在叶片、冠层及区域尺度间的尺度转换及累积效应,对更好地认识植被与大气间的水热运移过程,合理评价植被在陆面过程中的地位和作用具有重要意义.本文着重从叶片尺度气孔导度模拟、气孔导度在冠层尺度的累积表现、冠层到区域尺度转换研究及气孔导度累积效应在陆面过程模型中的作用等4个层次总结了近期国内外研究状况,指出其中存在的异质性等问题,并就今后应加强多尺度间的同步观测提出了展望.