Effects of caffeine on chromosomal loss and nondisjunction in Drosophila melanogaster

Three types of male larvae, normal X males and two types with structurally abnormal X chromosomes (ring X and short X sc ⁴ sc ³, y) were treated during the third instar with 0.5 per cent caffeine in nutrient medium. Upon eclosion, these males were mated to yellow and Oregon-R wild type females. The F₁ generation of each cross was scored for normal (XX and XY) and abnormal (XO and XXY) progeny. Statistical analyses of data demonstrate that caffeine increases chromosomal loss for all genotypes tested. The effect of caffeine on nondisjunction, however, is not clear. There are at least marginal increases in all cases when Oregon-R females are used. Slight increase and decreases noted for offspring of yellow females appear to be dependent upon the genotype of the inseminating male.     

Differential sexual survival of <Emphasis Type="Italic">Drosophila melanogaster</Emphasis> on copper sulfate

Based on studies of the influence of X-chromosomes on the viability of Drosophila melanogaster exposed to cadmium, and on the role of X-linked genes on copper homeostasis, we examined the effect of copper sulfate (CuSO₄) on offspring viability using three independent, inbred D. melanogaster crosses (ensuring identical autosomes for males and females within each cross). Each cross was performed with attached X-chromosome females and males with a single X-chromosome. As female D. melanogaster have less metallothionein RNA expression than males, we predicted fewer female offspring than male offspring in crosses exposed to CuSO₄, even though females have two copies of X-chromosome genes, possibly resulting in overdominant heterozygosity. In two of three crosses, CuSO₄ caused significantly higher numbers of male offspring compared to female offspring. We hypothesized that these gender-based viability differences to copper exposure are caused by X-chromosome ploidy and X-linked genetic variation affecting metallothionein expression. Observed differential offspring viability responses among crosses to copper exposure also showed that different genetic backgrounds (autosomal and/or X-chromosome) can result in significant differences in heavy metal and metallothionein regulation. These results suggest that the effect of copper on offspring viability depends on both genetic background and gender, as both factors can affect the regulation of metallothionein proteins as well as homeostasis of biologically necessary heavy metals.     

How multiple mating by females affects sexual selection

Multiple mating by females is widely thought to encourage post-mating sexual selection and enhance female fitness. We show that whether polyandrous mating has these effects depends on two conditions. Condition 1 is the pattern of sperm utilization by females; specifically, whether, among females, male mating number, m (i.e. the number of times a male mates with one or more females) covaries with male offspring number, o. Polyandrous mating enhances sexual selection only when males who are successful at multiple mating also sire most or all of each of their mates'' offspring, i.e. only when Cov_♂(m,o), is positive. Condition 2 is the pattern of female reproductive life-history; specifically, whether female mating number, m, covaries with female offspring number, o. Only semelparity does not erode sexual selection, whereas iteroparity (i.e. when Cov_♀(m,o), is positive) always increases the variance in offspring numbers among females, which always decreases the intensity of sexual selection on males. To document the covariance between mating number and offspring number for each sex, it is necessary to assign progeny to all parents, as well as identify mating and non-mating individuals. To document significant fitness gains by females through iteroparity, it is necessary to determine the relative magnitudes of male as well as female contributions to the total variance in relative fitness. We show how such data can be collected, how often they are collected, and we explain the circumstances in which selection favouring multiple mating by females can be strong or weak.     

Genetics of dioecy and causal sex chromosomes in plants

Dioecy (separate male and female individuals) ensures outcrossing and is more prevalent in animals than in plants. Although it is common in bryophytes and gymnosperms, only 5% of angiosperms are dioecious. In dioecious higher plants, flowers borne on male and female individuals are, respectively deficient in functional gynoecium and roecium. Dioecy is inherited via three sex chromosome systems: XX/XY, XX/X0 and WZ/ZZ, such that XX or WZ is female and XY, X0 or ZZ are males. The XX/XY system generates the rarer XX/X0 and WZ/ZZ systems. An autosome pair begets XY chromosomes. A recessive loss-of-androecium mutation (ana) creates X chromosome and a dominant gynoecium-suppressing (GYS) mutation creates Y chromosome. The ana/ANA and gys/GYS loci are in the sex-determining region (SDR) of the XY pair. Accumulation of inversions, deleterious mutations and repeat elements, especially transposons, in the SDR of Y suppresses recombination between X and Y in SDR, making Y labile and increasingly degenerate and heteromorphic from X. Continued recombination between X and Y in their pseudoautosomal region located at the ends of chromosomal arms allows survival of the degenerated Y and of the species. Dioecy is presumably a component of the evolutionary cycle for the origin of new species. Inbred hermaphrodite species assume dioecy. Later they suffer degenerate-Y-led population regression. Cross-hybridization between such extinguishing species and heterologous species, followed by genome duplication of segregants from hybrids, give rise to new species.     

Molecular Spectrum of Spontaneous de Novo Mutations in Male and Female Germline Cells of Drosophila melanogaster

We carried out mutation screen experiments to understand the rate and molecular nature of spontaneous de novo mutations in Drosophila melanogaster, which are crucial for many evolutionary issues, but still poorly understood. We screened for eye-color and body-color mutations that occurred in the germline cells of the first generation offspring of wild-caught females. The offspring were from matings that had occurred in the field and therefore had a genetic composition close to that of flies in natural populations. We employed 1554 F₁ individuals from 374 wild-caught females for the experiments to avoid biased contributions of any particular genotype. From ~8.6 million alleles screened, we obtained 10 independent mutants: two point mutations (one for each sex), a single deletion of ~6 kb in a male, a single transposable element insertion in a female, five large deletions ranging in size from 40 to 500 kb in females, and a single mutation of unknown nature in a male. The five large deletions were presumably generated by nonallelic homologous recombination (NAHR) between transposable elements at different locations, illustrating the mutagenic nature of recombination. The high occurrence of NAHR that we observed has important consequences for genome evolution through the production of segmental duplications.     

Chromosomal differentiation in two species of Aedes and their hybrids revealed by giemsa C-banding

Giemsa C-banding patterns in two species of mosquitoes, Aedes aegypti and Aedes mascarensis, their hybrids and backcross progeny revealed differences in the sex chromosome pair. In A. aegypti, the female determining or the m chromosome in both males and females shows a conspicuous band in the centromere region and another band in one arm. The male determining or the M chromosome is devoid of any bands. Progeny of crosses involving A. aegypti females and A. mascarensis males showed interesting albeit unexpected results. The intercalary band was suppressed in both sons and daughters. When such F₁ sons were backcrossed to A. aegypti females, a proportion of males developed into intersexes. These intersex progeny differed from the normal males in terms of their banding pattern. In the reciprocal cross (A. mascarensis female × A. aegypti male), the F₁ and the backcross progeny yielded the expected C-banding patterns. The implications of the reversible expression of the intercalary band on the A. aegypti m chromosome and its relevance to genetic regulation are discussed.     

Sex determination and phenotype in wood lemmings with XXY and related karyotypic anomalies

Summary The wood lemming, Myopus schisticolor, possesses a unique sex determining system comprising both XX and XY females. Normal female development in the presence of XY is guaranteed by a mutation on the X, apparently associated with a structural rearrangement in Xp. This mutation inactivates the testis-inducing and male-determining factor on the Y and distinguishes X^* from X, and X^*Y females from XY males. Normal fertility of X^*Y females is ensured by a mitotic (double) nondisjunction mechanism which, at an early fetal stage, eliminates the Y from the germ line and replaces it by a copy of the X^*.Numerical sex chromosome aberrations are not infrequent and the trisomics XXY and X^*XY are relatively common. XXY individuals are sterile males with severe suppression of spermatogenesis. Among X^*XY animals, both males and females, as well as a true lateral hermaphrodite have been observed. Primary deficiency of germ cells, impairment of spermatogenesis and sterility are characteristic traits of the X^*XY males, whereas X^*XY females have normal oogenesis and are fertile. Both these extremes (except female fertility) coexist in the true hermaphrodite described in the present study. These apparently contradictory observations are explainable under the assumption that X^* and X in X^*XY individuals are inactivated non-randomly or that the cells are distributed unequally. Inactivation of the X or X^* determines whether or not the H-Y antigen will be expressed. When comparing conditions in Myopus and in man, an additional assumption has to be made in relation to the gene(s) involved in sex determination, located in Xp:In Myopus they do not escape inactivation, whereas in man they have been claimed to remain active.     

The novel mutant scl of the medaka fish, Oryzias latipes, shows no secondary sex characters

A new mutant that has neither male nor female secondary sex characters was found in the medaka, Oryzias latipes. Both XX and XY mature mutants had gonads with many spermatozoa, but spawning did not occur when the mutants were paired with normal males or normal females. F1 progeny were successfully obtained by artificial insemination using unfertilized eggs from wild-type females and spermatozoa of the XY mutant. The mutant phenotype did not occur in the F1 progeny from this cross. Incrossing among the F1 progeny produced 17 mutant offspring out of 68 progeny (25%), demonstrating that the mutant phenotype is caused by a single recessive mutation. This mutant was named scl (sex character-less). Because papillary processes, a male secondary sex character, were induced in the XY mutants by androgen administration, it seems that the androgen receptor is functioning normally. We found a loss-of-function type mutation in the P450c17 gene of the mutant; this gene encodes a steroidogenic enzyme required for the production of estrogen and androgen. The scl phenotype was completely linked to the mutant genotype of P450c17, strongly suggesting that mutation at the P450c17 locus is responsible for the scl mutant phenotype.     

Demonstration of viability and fertility and development of a molecular tool to identify YY supermales in a fish with both genotypic and environmental sex determination

The pejerrey possesses a genotypic sex determination system driven by the amhy gene and yet shows marked temperature‐dependent sex determination. Sex‐reversed XY females have been found in a naturally breeding population established in Lake Kasumigaura, Japan. These females could mate with normal XY males and generate YY “supermale” individuals that, if viable and fertile, would sire only genotypic male offspring. This study was conducted to verify the viability, gender, and fertility of YY pejerrey and to develop a molecular method for their identification. Production of YY fish was attempted by crossing a thermally sex‐reversed XY female and an XY male, and rearing the progeny until sexual maturation. To identify the presumable YY individuals, we first conducted a PCR analysis using amhy‐specific primers to screen only amhy‐positive (XY and YY) fish. This screening showed that 60.6% of the progeny was amhy‐positive, which suggested the presence of YY fish. We then conducted a second screening by qPCR in order to identify the individuals with two amhy copies in their genome. This screening revealed 13 individuals, all males, with values twice higher than the other 30 amhy‐positive fishes, suggesting they have a YY complement. This assumption as well as the viability, fertility, and “supermale” nature of these individuals was confirmed in progeny tests with XX females that yielded 100% amhy‐positive offspring. These results demonstrate that qPCR can obviate progeny test as a means to identify the genotypic sex and therefore may be useful for the survey of all three possible genotypes in wild populations.     

Effect of Age,Duration of Exposure,and Dose of Atrazine on Sexual Maturation and the Luteinizing Hormone Surge in the Female Sprague–Dawley Rat

Atrazine (ATZ) was administered daily by gavage to pregnant female Sprague Dawley rats at doses of 0, 6.25, 25 or 50 mg/kg/day, either during gestation, lactation and post‐weaning (G/L/PW cohort) to F₁ generation female offspring or only from postnatal day (PND 21) until five days after sexual maturation (vaginal opening) when the estrogen‐primed, luteinizing hormone (LH) surge was evaluated (PW cohort). Additional subgroups of F₁ females received the vehicle or ATZ from PND 21–133 or from PND 120–133. Slight reductions in fertility and the percentage of F₁ generation pups surviving to PND 21 in the gestationally exposed 50 mg/kg dose group were accompanied by decreased food intake and body weight of dams and F₁ generation offspring. The onset of puberty was delayed in of the F₁ generation G/L/PW females at doses of 25 and 50 mg/kg/day. F₁ generation females in the PW high‐dose ATZ group also experienced a delay in the onset of puberty. ATZ had no effect on peak LH or LH AUC in ovariectomized rats 5 days after sexual maturation, irrespective of whether the F₁ generation females were treated from gestation onward or only peripubertally. There was no effect of ATZ treatment on the estrous cycle, peak LH or LH AUC of F₁ generation females exposed from gestation through to PND 133 or only for two weeks from PND 120–133. These results indicate that developing females exposed to ATZ are not more sensitive compared to animals exposed to ATZ as young adults     

Genetic analysis of the contributions of individual chromosomes to the expression of mating behavior traits in Drosophila melanogaster

The model of isogenous strains with chromosome substitutions has been used to estimate the relative contributions of the X chromosome and autosomes (chromosomes 2 and 3) to the control of some mating behavior traits in Drosophila melanogaster. It has been found that the male sexual activity (SA), female sexual receptivity (SR), and copulation latency (CL) are determined by interaction between X-chromosome and autosomal genes, whereas the copulation duration (CD) is mainly controlled by the X-chromosome genes. The synthesized isogenous strains have been shown to be more similar to hybrids than to the original strains. In the offspring with hybrid genotypes, the relationships between all traits are less stable, which may be related with an increase in the heterozygosity level and changes in genetic homeostasis.     

Implications of maternal nutrient restriction in transgenerational programming of hypertension and endothelial dysfunction across F1-F3 offspring

AimsAn extensive variety of prenatal insults are associated with an increased incidence of metabolic and cardiovascular disorders in adult life. We previously demonstrated that maternal global nutrient restriction during pregnancy leads to increased blood pressure and endothelial dysfunction in the adult offspring. This study aimed to assess whether prenatal exposure to nutritional insult has transgenerational effects in F₂ and F₃ offspring.Main methodsFor this, female Wistar rats were randomly divided into two groups on day 1 of pregnancy: a control group fed standard chow ad libitum and a restricted group fed 50% of the ad libitum intake throughout gestation. At delivery, all animals were fed a standard laboratory chow diet. At 11 weeks of age, one female and one male from each restricted litter were randomly selected and mated with rats from another restricted litters in order to generate the F₂ offspring. The same procedure produced F₃ generation. Similarly, the rats in the control group were bred for each generation.Key FindingsOur findings show that the deleterious effects of maternal nutrient restriction to which the F₀ mothers were exposed may not be limited to the male first generation. In fact, we found that elevated blood pressure, an impaired vasodilatory response to acetylcholine and alterations in NO production were all transferred to the subsequent males from F₂ and F₃ generations.SignificanceOur data show that global nutrient restriction during pregnancy results in a specific phenotype that can be passed transgenerationally to a second and third generation.     

Phenotype-dependent mate choice in Propylea dissecta and its fitness consequences

Sexual selection in relation to male or female polymorphism has been investigated across the taxa. However, how polymorphism influences sexual selection has been investigated less in most organisms including ladybirds, with most studies in the latter on mating incidences in the field. Laboratory studies on morph-dependent sexual selection in ladybirds have assessed resultant reproductive performance but not consequences on offspring attributes, as well as maintenance of polymorphism. In the current study, we assessed mate choice in a dimorphic ladybird, Propylea dissecta, and its effects on fitness in terms of reproductive performance and offspring development. Females mate significantly more readily with typical darker morphs than with pale ones. Whether this is female choice or male–male competition requires more investigation. Better reproductive performance and increased offspring development and survival in monomorphic typical pairs indicated fitness benefits for individuals obtaining typical mates. Typical females had higher fecundity and their offspring developed faster and survived better than those of pale females. Females paired with typical males had higher egg hatchability than those paired with pale ones. Significantly more offspring of dimorphic pairs were of typical phenotypes whereas offspring of monomorphic pairs expressed the same phenotype as their parents. Female preference for typical males and the dominance of the typical trait likely explain the preponderance of typical morphs in the field.     

Safety of a novel oxygen-coordinated niacin-bound chromium(III) complex (NBC): I. Two-generation reproduction toxicity study

The objective of this study was to evaluate the effects of a novel oxygen-coordinated niacin-bound chromium(III) complex (NBC) on the reproductive systems of male and female rats, the postnatal maturation and reproductive capacity of their offspring, and possible cumulative effects through multiple generations. Sprague-Dawley rats were maintained on feed containing NBC at dose levels of 0, 4, 15, or 60 ppm for 10 weeks prior to mating, during mating, and, for females through gestation and lactation, across two generations. For the parents (F₀ and F₁) and the offspring (F₁ and F_2a), reproductive parameters such as fertility and mating, gestation, parturition, litters, lactation, sexual maturity and development of offspring were assessed. Results from the current study indicated that dietary exposure of NBC to parental male and female rats of both (F₀ and F₁) the generations during the premating and mating periods, for both sexes, and during gestation and lactation in case of female rats, did not cause any significant incidence of mortality or abnormal clinical signs. Compared to respective controls, NBC exposure did not affect reproductive performance as evaluated by sexual maturity, fertility and mating, gestation, parturition, litter properties, lactation and development of the offspring. Based on the findings of this study, the parental as well as the offspring no-observed-adverse-effect level for NBC was determined to be greater than 60 ppm in diet or equivalent to 7.80 and 8.31 mg/kg body weight/day in male and female rats, respectively.     

Hybrid immune response to parental liver tissue grafts

Parental-to-F₁-hybrid liver tissue grafts in like-sex donor-recipient combinations survive indefinitely, although several F₁ recipients demonstrate an immunological response to the parental graft. Female F₁ recipients, particularly those carrying theH-2 ^b haplotype, respond vigorously to male parental liver grafts. However F₁ female responses to male parental liver tissue grafts differ substantively from the responses of parental females to syngeneic male grafts. C3H male liver grafts are rejected vigorously by F₁ females as long as the F₁ carries theH-2 ^b haplotype. These findings support previous reports of strong immunological responses to C3H H-Y antigen in female F₁ and C3H.SW animals, a response which is absent in C3H females. Female F₁ hybrids carrying theH-2 ^b haplotype do not reject grafts of B10 or B6 male liver as rapidly as do B10 or B6 parental females. This reduced F₁ response may be related to the formation of hybrid antigens and consequent alteration of the anti-H-Y response. Alternatively, cells that specifically suppress the anti-H-Y response may be present in F₁ hybrids. Factors responsible for suppression appear to be controlled by non-MHC antigens, at least in (OH x B6 or B10) F₁ hybrids.     

Hybridization of Australian Haemonchus placei (place, 1893), Haemonchus contortus cayugensis (Das & Whitlock, 1960) and Haemonchus contortus (Rudolphi, 1803) from Louisiana

There were distinct ecotypie differences in the ability to develop to third stage larvae at a constant 11 or 13°C for two weeks. H. conforms cayugensis could develop at both 11 and 13°C; H. contortus from Louisiana could develop at 13°C but not 11°C and H. placei could not complete development at either temperature. Eggs produced from the first cross between ecotypes had the same cold tolerance as their maternal parent. F₁ eggs were intermediate between parental ecotypes, while F₂ eggs, when fertile, developed at the same temperature as the most cold-tolerant parent in the first cross. The H. placei knobbed vulvar morph type was dominant to H. contortus cayugensis smooth morph type and recessive to the H. contortus (Louisiana) smooth morph type. Knobbed morph type of H. contortus (Louisiana) was dominant to smooth of H. contortus cayugensis. Dominance of morph type was enhanced by a matroclinous effect in the between species matings but not in the within species matings. No fertile eggs were produced by the F1 of the mating between female H. placei × male H. contortus (Louisiana) or by the F₂ of the reciprocal mating. Reciprocal matings between H. placei × H. contortus cayugensis produced an F₁ and F₂ that had reduced fertility but were not completely sterile. No decrease in fertility was observed in the F₂ and F₁ from the H. contortus cayugensis × H. contortus(Louisiana)matings. Cytological studies revealed several kinds of meiotic disturbances in the between species F₁ and F₂. The most frequent were aneuploidy, failure of chromosome pairing, and pairing between non-homologous chromosomes. Males were more seriously affected than females by meiotic disturbances. The H. placei X chromosome appeared to be selected against by the hybrid genetic background. Within species hybrids showed no meiotic disturbances and the percentage of male offspring from these crosses was the same as for female offspring.     

Strong sexual selection in males against a mutation load that reduces offspring production in seed beetles

Theory predicts that sexual reproduction can increase population viability relative to asexual reproduction by allowing sexual selection in males to remove deleterious mutations from the population without large demographic costs. This requires that selection acts more strongly in males than females and that mutations affecting male reproductive success have pleiotropic effects on population productivity, but empirical support for these assumptions is mixed. We used the seed beetle Callosobruchus maculatus to implement a three‐generation breeding design where we induced mutations via ionizing radiation (IR) in the F₀ generation and measured mutational effects (relative to nonirradiated controls) on an estimate of population productivity in the F₁ and effects on sex‐specific competitive lifetime reproductive success (LRS) in the F₂. Regardless of whether mutations were induced via F₀ males or females, they had strong negative effects on male LRS, but a nonsignificant influence on female LRS, suggesting that selection is more efficient in removing deleterious alleles in males. Moreover, mutations had seemingly shared effects on population productivity and competitive LRS in both sexes. Thus, our results lend support to the hypothesis that strong sexual selection on males can act to remove the mutation load on population viability, thereby offering a benefit to sexual reproduction.     

Identifizierung der genetischen Geschlechtschromosomen bei der monogenen Schmeißfliege Chrysomya rufifacies (Calliphoridae,Diptera)

Previous investigations have shown the sex determination in the monogenic blowfly Chrysomya rufifacies to be controlled by a cytologically not discernible homogamety-heterogamety mechanism in the female. Female-producing (thelygenic) females are assumed to be heterozygous for a dominant female sex realizer (F′) with sex-predetermining properties, while male-producing (arrhenogenic) females as well as males are supposed to be homozygous for the recessive allele (f). In order to identify the genetic sex chromosomes of C. rufifacies among its five pairs of long euchromatic chromosomes (nos.1–5) plus one pair of small heterochromatic ones (no. 6), all chromosomes were marked by reciprocal translocations induced by X-ray treatment of adult males. The inheritance of thirteen different heterozygous translocations has been analyzed. All of the translocations (eleven) between two of the four longer chromosomes did not show sex-linked inheritance, thus demonstrating the autosomal character of the chromosomes nos 1, 2, 3 and 4. The same is true for the translocation T6 (2/6). Therefore the small heterochromatic chromosome no. 6, corresponding to the morphologically differentiated sex chromosomes within the amphogenic calliphorid species, remains without sex determining function in the monogenic fly. This could be confirmed by the analysis of monosomic (monosomy-6) and trisomic (trisomy-6) individuals, which resulted from meiotic non-disjunction in T6/+ translocation heterozygotes. Contrary to these translocations, the heterozygous 5/2 translocation (T14) exhibited sex-linked inheritance: There was but a very low frequency (0,76 per cent) of recombinants resulting from crossing-over between F′/f and the translocation breakage point in thelygenic F₁ T14/+ females. The sex-linked inheritance of T14 was confirmed by the progeny of a thelygenic F₁ T14/+ female crossed to a homozygous T14/T14 translocation male. Among the offspring of that F₁ T14/+ female, which had received the translocation from its father, all of the F₂ T14/+ females were thelygenic compared to their arrhenogenic T14/T14 sisters. These results prove that the chromosomes of pair no. 5 genetically act as X′X-XX sex chromosomes in C. rufifacies.