First-generation Offspring Research Articles

The effects of methyl-donor deficiency on mutation induction and transgenerational instability in mice

The results of recent human and animal studies have provided strong evidence for the epigenetic effects of a dietary deficiency of methyl donors such as folate, choline and methionine on cancer risk and some other common diseases. However, the mechanisms underlying the links between epigenetic alterations and disease remain elusive. To establish whether a methyl-donor deficient diet can result in long-term changes in mutation rate in treated animals and their offspring, BALB/c male mice were maintained for 8 weeks, from 4 weeks of age, on a synthetic diet lacking in choline and folic acid. Using single-molecule PCR, the frequency of mutation at the mouse expanded simple tandem repeat (ESTR) locus Ms6-hm was established in sperm samples of treated males, as well as in sperm and brain of their first-generation offspring. ESTR mutation frequency in the germline of males sacrificed immediately after treatment or sampled 6 and 10 weeks after the end of dietary restriction did not significantly differ from that in age-matched control groups. The frequency of ESTR mutation in DNA samples extracted from sperm and brain of the first-generation offspring of treated mice was also similar to that in controls. The results of our study suggest that the effects of a methyl-donor deficient diet on mutation induction and transgenerational instability in mice are likely to be negligible.

Transgenerational Glucose Intolerance With Igf2/H19 Epigenetic Alterations in Mouse Islet Induced by Intrauterine Hyperglycemia

Gestational diabetes mellitus (GDM) has been shown to be associated with high risk of diabetes in offspring. However, the mechanisms involved and the possibilities of transgenerational transmission are still unclear. We intercrossed male and female adult control and first-generation offspring of GDM (F1-GDM) mice to obtain the second-generation (F2) offspring in four groups: C♂-C♀, C♂-GDM♀, GDM♂-C♀, and GDM♂-GDM♀. We found that birth weight significantly increased in F2 offspring through the paternal line with impaired glucose tolerance (IGT). Regardless of birth from F1-GDM with or without IGT, high risk of IGT appeared as early as 3 weeks in F2 offspring and progressed through both parental lineages, especial the paternal line. IGT in male offspring was more obvious than that in females, with parental characteristics and sex-specific transmission. In both F1 and F2 offspring of GDM, the expression of imprinted genes Igf2 and H19 was downregulated in pancreatic islets, caused by abnormal methylation status of the differentially methylated region, which may be one of the mechanisms for impaired islet ultrastructure and function. Furthermore, altered Igf2 and H19 gene expression was found in sperm of adult F1-GDM, regardless of the presence of IGT, indicating that changes of epigenetics in germ cells contributed to transgenerational transmission.

Diet-induced paternal obesity in the absence of diabetes diminishes the reproductive health of two subsequent generations of mice

Obesity and related conditions, notably subfertility, are increasingly prevalent. Paternal influences are known to influence offspring health outcome, but the impact of paternal obesity and subfertility on the reproductive health of subsequent generations has been overlooked. A high-fat diet (HFD) was used to induce obesity but not diabetes in male C57Bl6 mice, which were subsequently mated to normal-weight females. First-generation offspring were raised on a control diet and their gametes were investigated for signs of subfertility. Second-generation offspring were generated from both first generation sexes and their gametes were similarly assessed. We demonstrate a HFD-induced paternal initiation of subfertility in both male and female offspring of two generations of mice. Furthermore, we have shown that diminished reproductive and gamete functions are transmitted through the first generation paternal line to both sexes of the second generation and via the first generation maternal line to second-generation males. Our previous findings that founder male obesity alters the epigenome of sperm, could provide a basis for the developmental programming of subfertility in subsequent generations. This is the first observation of paternal transmission of diminished reproductive health to future generations and could have significant implications for the transgenerational amplification of subfertility observed worldwide in humans.

The effects of maternal irradiation during adulthood on mutation induction and transgenerational instability in mice

The long-term genetic effects of maternal irradiation remain poorly understood. To establish the effects of radiation exposure on mutation induction in the germline of directly exposed females and the possibility of transgenerational effects in their non-exposed offspring, adult female BALB/c and CBA/Ca mice were given 1Gy of acute X-rays and mated with control males. The frequency of mutation at expanded simple tandem repeat (ESTR) loci in the germline of directly exposed females did not differ from that of controls. Using a single-molecule PCR approach, ESTR mutation frequency was also established for both germline and somatic tissues in the first-generation offspring of irradiated parents. While the frequency of ESTR mutation in the offspring of irradiated males was significantly elevated, maternal irradiation did not affect stability in their F1 offspring. Considering these data and the results of our previous study, we propose that, in sharp contrast to paternal exposure to ionising radiation, the transgenerational effects of maternal high-dose acute irradiation are likely to be negligible.

On S.N. Bernstein's derivation of Mendel's Law and 'rediscovery' of the Hardy-Weinberg distribution

Around 1923 the soon-to-be famous Soviet mathematician and probabilist Sergei N. Bernstein started to construct an axiomatic foundation of a theory of heredity. He began from the premise of stationarity (constancy of type proportions) from the first generation of offspring. This led him to derive the Mendelian coefficients of heredity. It appears that he had no direct influence on the subsequent development of population genetics. A basic assumption of Bernstein was that parents coupled randomly to produce offspring. This paper shows that a simple model of non-random mating, which nevertheless embodies a feature of the Hardy-Weinberg Law, can produce Mendelian coefficients of heredity while maintaining the population distribution. How W. Johannsen’s monograph influenced Bernstein is discussed.

European Regulations for Consumption of Cloned Animals and First-Generation Offspring Still Unwritten

European Regulations for Consumption of Cloned Animals and First-Generation Offspring Still Unwritten

Potential Maternal Effects of Elevated Atmospheric CO2 on Development and Disease Severity in a Mediterranean Legume

Global change can greatly affect plant populations both directly by influencing growing conditions and indirectly by maternal effects on development of offspring. More information is needed on transgenerational effects of global change on plants and their interactions with pathogens. The current study assessed potential maternal effects of atmospheric CO2 enrichment on performance and disease susceptibility of first-generation offspring of the Mediterranean legume Onobrychis crista-galli. Mother plants were grown at three CO2 concentrations, and the study focused on their offspring that were raised under common ambient climate and CO2. In addition, progeny were exposed to natural infection by the fungal pathogen powdery mildew. In one out of 3 years, offspring of high-CO2 treatments (440 and 600 ppm) had lower shoot biomass and reproductive output than offspring of low-CO2 treatment (280 ppm). Disease severity in a heavy-infection year was higher in high-CO2 than in low-CO2 offspring. However, some of the findings on maternal effects changed when the population was divided into two functionally diverging plant types distinguishable by flower color (pink, Type P; white, Type W). Disease severity in a heavy-infection year was higher in high-CO2 than in low-CO2 progeny in the more disease-resistant (Type P), but not in the more susceptible plant type (Type W). In a low-infection year, maternal CO2 treatments did not differ in disease severity. Mother plants of Type P exposed to low CO2 produced larger seeds than all other combinations of CO2 and plant type, which might contribute to higher offspring performance. This study showed that elevated CO2 potentially exerts environmental maternal effects on performance of progeny and, notably, also on their susceptibility to natural infection by a pathogen. Maternal effects of global change might differently affect functionally divergent plant types, which could impact population fitness and alter plant communities.

Radiation in low doses reduces life span of Daphnia magna

The effects of low-dose γ-irradiation on the crustacean Daphnia magna have been analyzed. Given that the LD50 for this species is approximately 100 Gy, one-day-old daphnids were exposed to 10, 100 and 1000 mGy of acute γ-rays. The survival of irradiated animals was evaluated over the maximum lifespan of 90 days. Increased mortality among irradiated at doses of 100 and 1000 mGy daphnids was found. Our data also showed that the observed decrease in survival of irradiated animals was attributed to their early aging, thus providing an important evidence for the effects of low-dose irradiation on early senescence. Overall, the results of our studies have shown that low-dose irradiation can result in a significant reduction in life span of exposed daphnids. As the majority of irradiated Daphnia showed reduced viability regardless the dose, our data therefore imply the presence of a threshold of dose of low-LET exposure capable of impairing the viability of this species. We have also found that parental irradiation significantly affected the survival of first-generation offspring, the magnitude of which cannot be attributed to the segregation of radiation-induced deleterious mutations. Potential implications of the results of our studies for the risk assessment of low-dose exposure have been discussed.

Localization of genes for V+LDL plasma cholesterol levels on two diets in the opossum Monodelphis domestica

Plasma cholesterol levels among individuals vary considerably in response to diet. However, the genes that influence this response are largely unknown. Non-HDL (V+LDL) cholesterol levels vary dramatically among gray, short-tailed opossums fed an atherogenic diet, and we previously reported that two quantitative trait loci (QTLs) influenced V+LDL cholesterol on two diets. We used hypothesis-free, genome-wide linkage analyses on data from 325 pedigreed opossums and located one QTL for V+LDL cholesterol on the basal diet on opossum chromosome 1q [logarithm of the odds (LOD) = 3.11, genomic P = 0.019] and another QTL for V+LDL on the atherogenic diet (i.e., high levels of cholesterol and fat) on chromosome 8 (LOD = 9.88, genomic P = 5 x 10(-9)). We then employed a novel strategy involving combined analyses of genomic resources, expression analysis, sequencing, and genotyping to identify candidate genes for the chromosome 8 QTL. A polymorphism in ABCB4 was strongly associated (P = 9 x 10(-14)) with the plasma V+LDL cholesterol concentrations on the high-cholesterol, high-fat diet. The results of this study indicate that genetic variation in ABCB4, or closely linked genes, is responsible for the dramatic differences among opossums in their V+LDL cholesterol response to an atherogenic diet.

A novel herbicide‐inducible male sterility system

Heterosis is a phenomenon that first-generation offspring perform better than their parents. Conventional breeding methods have their shortcomings. It would be optimal to construct inducible male sterile plants. We developed a novel system for creating male sterile transgenic plants by downregulating the specific expression of the glyphosate tolerance CP4 EPSPS gene in male reproductive tissues. Transcriptional repression was achieved by manipulating DNA binding proteins with their specific corresponding sites. We transferred the CP4 EPSPS gene driven by a modified CaMV 35S promoter with three tetracycline operator copies in the vicinity of the TATA box and tetracycline repressor gene under the control of an anther-specific promoter Osg6B to Arabidopsis thaliana. As a result, we successfully obtained controllable transgenic plants: the whole plant could tolerate exposure of glyphosate but the male tissue was sensitive. The novel inducible male sterility system is applied and easy to handle, so it might be applicable to a wide range of crop plants.

Urinary Volatile Molecules Vary in Males of the 2 European Subspecies of the House Mouse and Their Hybrids

Mice recognize other mice by identifying chemicals that confer a molecular signature to urinary marks. Such molecules may be involved in species recognition, and previous behavioral studies have related divergence of sexual preference between 2 subspecies of the house mouse (Mus musculus musculus and Mus musculus domesticus) to urinary odors. To characterize the differences between odors of males of the 2 subspecies and their first-generation offspring, the urinary volatile molecules were examined via gas chromatography coupled to mass spectrometry. Seven molecules were present in the samples from mice of at least one group. Their quantity varied among groups: M. m. domesticus showed a quantitatively richer panel of odorants in their urine when compared with M. m. musculus. The hybrids showed a more complex picture that was not directly related to one or the other parental subspecies. These quantitative differences may contribute to the specificity of the odorant bouquet of the 2 subspecies.

Minireview: Transgenerational Inheritance of the Stress Response: A New Frontier in Stress Research

It is well established in animal models that the prenatal environment can have a major impact on stress axis function throughout life. These changes can predispose to various metabolic, cardiovascular, and neurobiological pathophysiologies. Emerging evidence indicates that the same programming effects occur in humans. It is now becoming clear that the pathophysiological effects are not confined to the first-generation offspring and that there is transgenerational memory of fetal experience that can extend across multiple generations. The complex mechanisms by which transgenerational transmission of stress responsiveness occur are rapidly becoming a focus of investigation. Understanding these fundamental biological processes will allow for development of intervention strategies that prevent or reverse adverse programming of the stress response.

Intergenerational transmission of glucose intolerance associated with gestational diabetes mellitus

OBJECTIVE: To investigate the mechanism of intergenerational transmission of glucose intolerance by in utero high glucose associated with gestational diabetes mellitus (GDM). DESIGN: We established a mouse model of GDM and observed the phenotype of GDM offspring (F1-GDM). Furthermore, we intercrossed male and female F1 adult control (C) and GDM mice and investigated phenotype in F2 offspring. MATERIALS AND METHODS: Mouse model of GDM was induced by a single intraperitoneal injection of streptozotocin on the day of pregnancy. F2 offspring were obtained from 4 groups including (1) C♂-C♀ (2) C♂-GDM♀ (3) GDM♂-C♀ and (4) GDM♂-GDM♀. Intraperitoneal glucose tolerance tests were performed on 3-week-old and 8-week-old mice. Gene expression was detected by real-time quantitative PCR. T test was used to compare with control group. A P<0.05 was considered statistically different. RESULTS: (1) Compared with control group, birth weight decreased in F1-GDM (1.49g, p<0.05) and increased in F2 offspring through the paternal line (C♂-GDM♀=1.65g; GDM♂-C♀=1.72g, p<0.05; GDM♂-GDM♀=1.79g, p<0.05); (2) in F1-GDM, impaired glucose tolerance (IGT) appeared at adult (glucose tolerance test area under curve (AUC)= 167.17, p<0.05), however, IGT in F2 offspring was observed as early as 3-week-old through both parental lineages (AUC of C♂-GDM♀= 226.35, p<0.05; GDM♂-C♀= 255.96, p<0.05; GDM♂-GDM♀= 518.55, p<0.05). IGT in both F1 and F2 generations is linked to impaired function of pancreas, partly because of abnormal Igf2 expression. CONCLUSIONS: During pregnancy, in utero significant high glucose environment of GDM lead to reduced birth weight in first-generation offspring. However, birth weight increased in second first-generation offspring. GDM induced IGT in both first- and second-generation offspring. Sex-specific transmission of phenotypes may be related to epigenetic regulation.

Transgenerational changes in somatic and germ line genetic integrity of first-generation offspring derived from the DNA damaged sperm

To report a more quantitative approach to study the influence of varying levels of sperm DNA damage on transgenerational changes in genomic instability in a mouse model. Experimental prospective study. Embryology research laboratory. Swiss albino mice. The sperm DNA damage was induced by different doses of gamma-irradiation to male mice followed by mating with healthy female mice. Genomic integrity in embryos, fetus, and spermatozoa of F1 mice derived from the DNA-damaged sperm. The transgenerational changes in genetic integrity were attributed by a dose-dependent increase in the frequency of micronuclei in preimplantation embryos and a concomitant increase in genomic instability in fetal liver cells and sperm chromatin modifications in F1 males. A strong positive correlation was observed between the extent of sperm DNA damage and somatic and germ-line genomic instability. Sperm-mediated transgenerational genomic instability is dependent on the amount of DNA damage present in the sire's sperm at the time of fertilization.

Amplified Fragment Length Polymorphism and Parentage Analysis of a Feral Barberry (Berberis thunbergii DC.) Population to Determine the Contribution of an Ornamental Landscape Genotype

We investigated the role of an ornamental, purple-leaved specimen of Japanese barberry in a local invasion using parentage analysis. We focused on a landscape plant of B. thunbergii var. atropurpurea in Willington, CT, that was first established at least 30 years ago. We genotyped every barberry plant found within a 92-m radius of this individual. In contrast to feral populations that are distant from residential or commercial plantings, 14% of the 43 feral plants in our sample had purple foliage and 30% were found growing within 16.5 m of the focal individual. Parentage analysis identified seven plants (five purple-leaved and two green-leaved) as descendants of the focal individual. Five of these descendants are likely first-generation offspring and two are likely second-generation seedlings. In addition, one plant was identified as a backcross between the focal plant and one of its offspring. Our results show that purple-leaved Japanese barberry used in residential landscapes can contribute to plant invasions, at least under some circumstances.

Effect of Weather onIps typographus(Coleoptera Curculionidae) Phenology, Voltinism, and Associated Spruce Mortality in the Southeastern Alps

Summer drought associated with high temperatures recorded in the last few years has given rise to outbreaks of bark beetles developing in weakened host trees. The aim of this study was to investigate the possible weather effect on the biology of and damage caused by Ips typographus L. in the southeastern Alps. The study was carried out recording temperature (1962-2007), precipitation (1922-2007), and the damage caused by I. typographus (1993-2007). In addition, data from pheromone-baited traps (1996-2005) provided information on the main periods of flight activity of I. typographus. From 1922 to 2007, precipitation during March-July has decreased approximately 200 mm (-22%), whereas since 1962-2007, mean temperatures during March-July increased approximately 2 degrees C (+13%). Damage caused by I. typographus was inversely correlated with March-July precipitation from the previous year but not correlated with temperature. Increases in spring temperature did not affect the development timing of the first generation, but only changed its onset. Earlier swarming of both overwintering beetles and first-generation offspring ( approximately 20 d sooner over 10 yr), and the early start of the second generation permitted more complete development of the second brood. Voltinism in this species is discussed in relation to thermal and photoperiodic thresholds, indicating that the occurrence of a third generation is limited by the summer photoperiod rather than by temperature. In conclusion, results suggest that spring drought increases damage caused by I. typographus in the following year, whereas warmer spring affects insect phenology.

The effects of in utero irradiation on mutation induction and transgenerational instability in mice

Epidemiological evidence suggests that the deleterious effects of prenatal irradiation can manifest during childhood, resulting in an increased risk of leukaemia and solid cancers after birth. However, the mechanisms underlying the long-term effects of foetal irradiation remain poorly understood. This study was designed to analyse the impact of in utero irradiation on mutation rates at expanded simple tandem repeat (ESTR) DNA loci in directly exposed mice and their first-generation (F 1) offspring. ESTR mutation frequencies in the germline and somatic tissues of male and female mice irradiated at 12 days of gestation remained highly elevated during adulthood, which was mainly attributed to a significant increase in the frequency of singleton mutations. The prevalence of singleton mutations in directly exposed mice suggests that foetal irradiation results in genomic instability manifested both in utero and during adulthood. The frequency of ESTR mutation in the F 1 offspring of prenatally irradiated male mice was equally elevated across all tissues, which suggests that foetal exposure results in transgenerational genomic instability. In contrast, maternal in utero exposure did not affect the F 1 stability. Our data imply that the passive erasure of epigenetic marks in the maternal genome can diminish the transgenerational effects of foetal irradiation and therefore provide important clues to the still unknown mechanisms of radiation-induced genomic instability. The results of this study offer a plausible explanation for the effects of in utero irradiation on the risk of leukaemia and solid cancers after birth.

Morphological evidence suggests homoploid hybridization as a possible mode of speciation in the Triatominae (Hemiptera, Heteroptera, Reduviidae)

All known significant insect vectors of Trypanosoma cruzi are members of the Reduviidae, subfamily Triatominae. Infections with this parasite are the cause of Chagas disease, the single most costly parasitic disease in the Western Hemisphere. The Triatominae are almost completely restricted to the Americas, with >130 species distributed in several foci of species richness and endemism; nevertheless, the processes involved in the diversification of this group remain poorly understood. The Triatoma brasiliensis species complex was recently proposed based on geography, morphology, ecology, and molecular data, and is believed to comprise two species and two subspecies. Here, we report results from a broad series of studies, in which first-generation offspring of experimental crosses were studied in terms of wing morphometry and phylogenetic position. Morphometrics, morphological, ecological and geographic analyses were consistent with the hypothesis of T. brasiliensis macromelasoma as a product of hybridization between two others (T. brasiliensis brasiliensis and T. juazeirensis). Although evidence is supportive of the hypothesis of speciation via hybridization as a mode of triatomine diversification, the case is not as-yet conclusive, and confirmation via molecular markers is necessary.

Genetic improvement on Chinese shrimp (Fenneropenaeus chinensis): growth and viability performance in F1 hybrids of different populations

Fenneropenaeus chinensis distributed in the Yellow Sea and Bohai Sea of China and the west coast of the Korean Peninsula. Different geographical populations represent potentially different genetic resources. To learn further the characteristics of different geographical population, crosses among two wild and three farmed populations were produced. The two wild populations were from the Yellow Sea and Bohai Sea (WYP), and the west coast of the Korean Peninsula and coast (WKN). The three farmed populations included the offspring of first generation of wild shrimp from coast in Korea (FKN), the Huang Hai (the Yellow Sea in Chinese) No.1 (HH1), and JK98. The phenotypes growth and survival rates of these populations were compared to confirm the feasibility for crossbreeding. The body length (BL), carapace length (CL), carapace width (CW), height of the second and third abdominal segment (HST), width of the second and third abdominal segment (WST), length of the first abdominal segment (LF), length of the last abdominal segment (LL), live body weight (BW), and survival rate were measured. Different combinations were statistically performed with ANOVA and Duncan’s Multiple Range Test. The results show that the survival rate of JK98(♀)×WKN(♂) was the highest, followed by WYP(♀)×WKN(♂), FKN(♀)×WYP(♂), FKN(♀)×HH1(♂) and WYP(♀)×FKN(♂); the body weight of FKN(♀)sxHH1(♂) was the highest, followed by FKN(♀)×WYP(♂), WYP(♀)×WKN(♂), WYP(♀)×FKN(♂) and JK98(♀)×WKN(♂); the total length had the same ranking as the body weight. All growth traits in hybrids JK98(♀)×WKN(♂) were the lowest among all combinations. F1 hybrids had significant difference (P 0.05) in other growth traits and survival rate. The results of Duncan’s Multiple Range Test are that BL and CL of JK98(♀)×WKN(♂) were significantly different from the other combinations; HST different from the combination of FKN(♀)×WYP(♂), FKN(♀)×HH1(♂) and WYP(♀)×WKN(♂); and BW different from FKN(♀)×WYP(♂) and FKN(♀)×HH1(♂). As a whole, the results indicate that the FKN(♀)×HH1(♂) was the best combination in all growth traits. Therefore, hybridization can introduce the variation to base populations. The systematic selection program based on additive genetic performance may be more effective than crossbreeding.

Paternal exposure to ethylnitrosourea results in transgenerational genomic instability in mice

Recent data shows that the effects of ionizing radiation are not restricted to the directly exposed parental germ cells, but can also manifest in their nonexposed offspring, resulting in elevated mutation rates and cancer predisposition. The mechanisms underlying these transgenerational changes remain poorly understood. One of the most important steps in elucidating these mechanisms is to investigate the initial cellular events that trigger genomic instability. Here we have analyzed the effects of paternal treatment by ethylnitrosourea, an alkylating agent which is known to form specific types of DNA adducts, on the transgenerational effects in the first-generation (F1) offspring of exposed CBA/Ca and BALB/c male mice. Mutation rates at two expanded simple tandem repeat loci were significantly elevated in the F1 germline of both strains. Pre and postmeiotic exposures resulted in similar increases in mutation rate in the F1 germline. Within each strain mutation rates were equally elevated in the germline of male and female F1 offspring of the directly exposed males. The results of our study suggest that transgenerational instability is not attributed to a specific sub-set of DNA lesions, such as double strand breaks, and is most probably triggered by a stress-like response to a generalized DNA damage.