Sexual rest and post-meiotic sperm ageing in house mice.

Fertilization by aged sperm can result in adverse fitness consequences for both males and females. Sperm storage during male sexual rest could provide an environment for post-meiotic sperm senescence causing a deterioration in the quality of stored sperm, possibly impacting on both sperm performance (e.g. swimming ability) and DNA quality. Here, we compared the proportion of sperm with fragmented DNA, an indicator of structural damage of DNA within the sperm cell, among males that had been sexually rested for approximately 2 months, to that of males that had mated recently. We found no evidence of intra-epididymal sperm DNA damage or any impairment in sperm performance, and consequently no evidence of post-meiotic sperm senescence. Our results suggest that male house mice are likely to possess mechanisms that function to ensure that their sperm reserves remain stocked with 'young', viable sperm during periods of sexual inactivity. We also discuss the possibility that our experimental design leads to no difference in the age of sperm among males from the two mating treatments. Post-meiotic sperm senescence is especially relevant under sperm competition. Thus, we sourced mice from populations that differed in their levels of post-copulatory sexual selection, enabling us to gain insight into how selection for higher sperm production influences the rate of sperm ageing and levels of DNA fragmentation. We found that males from the population that produced the highest number of sperm also had the smallest proportion of DNA-fragmented sperm and discuss this outcome in relation to selection acting upon males to ensure that they produce ejaculates with high-quality sperm that are successful in achieving fertilizations under competitive conditions.

Studies of single-chain antibody expression in quiescent Escherichia coli.

Quiescent Escherichia coli cells are generated by overexpressing the Rcd transcript in an hns-205 mutant host. The resulting nongrowing, metabolically active cells were used here to express a single-chain antibody fragment (scFv) in shake flask and fermentor cultures. The expression system is based on two plasmids; one carries the product gene expressed from lambdaP(L) under the control of the cI857 temperature-sensitive repressor, while the second expresses Rcd from lambdaP(R). Shifting the culture from 30 to 42 degrees C induces Rcd expression and product expression simultaneously. Our scFv carried a PelB leader, and 90% of the protein was secreted into the culture supernatant. In a batch culture, the supernatant concentration of scFv in the quiescent-cell culture (optical density at 600 nm [OD(600)] of 3.5) was 37 mg x liter(-1), compared to a maximum of 13 mg x liter(-1) in the control culture (final OD(600) of 20). In a fed-batch fermentor culture, quiescent cells were held at an OD(600) of 20 for 24 h and the extracellular scFv concentration reached a maximum of 150 mg x liter(-1). A control culture with a similar feed reached an OD(600) of 80, but despite the higher density, the extracellular scFv concentration did not exceed 35 mg x liter(-1). Quiescent cells at an OD(600) of 50 exhibited a small decline in the specific product formation rate, but nevertheless, an extracellular scFv concentration of 160 mg x liter(-1) was achieved in 8 h. The rate of extracellular accumulation was 10-fold greater in the quiescent culture than in the control culture. This study demonstrates that it is possible to establish high-density quiescent E. coli cultures that are capable of efficient synthesis, folding, and export of proteins.

Association between a serotonin transporter promoter region polymorphism and mood response during tryptophan depletion.

This study investigated the relationship between depressive symptom response during tryptophan (TRP) depletion and a functional polymorphism of the promoter region of the serotonin (5-HT) transporter gene (SLC6A4).(1) Forty-three subjects in remission from a major depressive episode who underwent TRP depletion were genotyped. DNA was extracted from blood lymphocytes or from cheek cells.(2) The two common alleles are designated long (l) and short (s). Depressive symptoms were measured with the 25-item Hamilton Depression Rating Scale (HDRS).(3) There was a significant association between the l homozygous genotype and the depressive response to TRP depletion, with a significant main effect of time (F = 8.763, df = 3, 38, P = <0.001), and time x l homozygous allele interaction (F = 3.676, df = 3, 38, P = 0.02). Individuals whose genotype predicted increased 5-HT transporter activity may be more susceptible to depressive changes in response to transient 5-HT perturbations. The use of endophenotypic markers for affective disorders such as the mood response to TRP depletion may facilitate studies of complex genetic traits such as depression by decreasing its heterogeneity.

A step in another direction: looking for maternal genetic and environmental effects on racial differences in birth weight.

To advance research on birth weight differences between black and white infants, it may be useful to study maternal effects. These effects present a set of risk factors that are largely unrelated to those that are presently under investigation and fail to explain the gap in birth weight; empirical findings suggest their involvement. Although maternal effects can be environmental, as illustrated by recent findings, genetic effects could be important as well because gene frequencies are known to differ across the "racial" groups as studied by birth weight researchers, and maternal genes can exert a causal effect on birth weight.

Two dopamine genes related to reports of childhood retrospective inattention and conduct disorder symptoms.

The 7-repeat allele of the dopamine receptor D4 gene (DRD4) and the 10 repeat allele of the dopamine transporter gene (DAT1) have shown association and linkage with symptoms of attention deficit hyperactivity disorder (ADHD) in childhood. The parents of ADHD children (clinic group, n = 80 fathers and 107 mothers) and control children (control group, n = 42 fathers and 51 mothers) were the focus of this study. These parents reported retrospectively on their level of ADHD Inattention and Conduct Disorder symptoms in adolescence. In analyses of the relation of symptom levels to the DRD4 and DAT1 genotypes, fathers possessing the 7 repeat DRD4 allele had greater levels of both inattention and conduct disorder symptoms. Mothers with the 10/10 genotype had higher levels of inattention symptoms. Thus, genetic associations found in children may be replicable in their parents.

Do people make environments or do environments make people?

This article discusses the influence of people's genetic make-up on their mental states of happiness and depression. Contrary to the conventional wisdom, great fortune does not guarantee happiness; neither does great misfortune assure depression. Emotional states are surprisingly immune to "objective" social circumstances. A biological basis for this relative immunity is that people possess biological set points for these emotional states, rendering the effects of most life events transitory. Genotypes also have indirect effects. People react differently to psychological stressors depending on their genotypes. A susceptible person may succumb to depression, whereas a resilient person may remain unaffected. People also expose themselves to different social environments. Exposure to controllable life events is partly a result of genetic predispositions. Consilience requires that this biological individuality be considered in any understanding of human behavior, including the pursuit of happiness.

Racial differences in birth health risk: a quantitative genetic approach.

In the United States the gap between black and white babies' birth weights has remained largely unexplained. Rather than trying to measure all relevant variables, we used a genetically informative design to study the relative importance of genetic and environmental factors. Employing multiple indicators of "birth health risk," we found that the racial differences increased with the magnitude of the shared environmental effects. This suggested that possible genetic effects would not pertain to fetal genes, although genes affecting the mother's physical or physiological characteristics could be important because they contribute to shared environment in our analysis.

Behavior problems among children from different family structures: the influence of genetic self-selection.

To examine both genetic and environmental influences on children's behavior problems in households defined by marital status and sibling relatedness, this study applied behavioral genetic methodology to four groups totalling 1524 sibling pairs drawn from 796 households: (1) two-parent full siblings, (2) two-parent half siblings, (3) mother-only full siblings, and (4) mother-only half siblings. Model-fitting procedures found that within-group variation on four subscales from the Behavior Problems Index was best explained by a model including both genetic and shared environmental factors. This model was then fit to the behavior problems means of the four groups. Its successful fit to these mean structures suggested that mean-level differences between groups were explained with the same influences that accounted for within-group variation. Genetic influences accounted for 81% to 94% of the mean-level difference in behavior problems between the two-parent, full sibling and the mother-only, half sibling groups. In contrast, shared environmental influences accounted for 67% to 88% of the mean-level difference in behavior problems between the two-parent, full sibling and mother-only, full sibling groups. The genetic influences are interpreted in terms of genetic self-selection into family structures.

Resolving the debate over birth order, family size, and intelligence.

Hundreds of research articles have addressed the relationship between birth order and intelligence. Virtually all have used cross-sectional data, which are fundamentally flawed in the assessment of within-family (including birth order) processes. Although within-family models have been based on patterns in cross-sectional data, a number of equally plausible between-family explanations also exist. Within-family (preferably intact-family) data are prerequisite for separating within- and between-family causal processes. This observation reframes an old issue in a way that can be easily addressed by studying graphical patterns. Sibling data from the National Longitudinal Survey of Youth are evaluated, and the results are compared with those from other studies using within-family data. It appears that although low-IQ parents have been making large families, large families do not make low-IQ children in modern U.S. society. The apparent relation between birth order and intelligence has been a methodological illusion.

Parental education and child's verbal IQ in adoptive and biological families in the National Longitudinal Study of Adolescent Health.

This study compared adoptive children and matched, biological children to estimate the genetic and environmental effect of years of mothers' and fathers' education on children's verbal intelligence (VIQ), as assessed by knowledge of vocabulary words. Adoptive and biological adolescent children in the National Longitudinal Study of Adolescent Health (Add Health) were matched for sex, age, parental education, and ethnicity. The adolescents all resided with two parents. Structural equation modeling was employed using Mx to estimate the genetic and transmissible environmental components of the correlation between parental education and children's VIQ. The mother-child and father-child correlations in biological families were .41 and .36, respectively, vs .16 and .18 in adoptive families. As suggested by these correlations, both genetic and shared environmental influences were statistically significant in the Mx models. We conclude that parental education exerts a modest shared environmental effect, explaining no more than 3 to 4% of the variation in verbal intelligence.

The DRD2 TaqI polymorphism and symptoms of attention deficit hyperactivity disorder.

The relationship of the DRD2 TaqI-A1 allele to hyperactive/impulsive and inattentive symptoms of attention deficit hyperactivity disorder (ADHD) in children and adolescents was examined in a sample of clinic-referred children and their siblings, and control children and their siblings (n = 236). The contribution of genetic dominance and additivity to mean differences among the A2A2, A1A2, and A1A1 genotypes was estimated using structural equation modeling. The effect of genetic additivity was statistically significant for both traits in an analysis of all children. The heritability from the DRD2 locus was estimated at 4.27% for hyperactive-impulsive symptoms and 2.12% for inattentive symptoms. Children with the A2A2 genotype had the highest mean level of symptoms. To control for any possible effects of population stratification, this analysis was repeated with parental genotypes as controls. In this smaller sample, although the direction of the effect was the same as that in the whole sample, the genotypic differences failed to reach conventional significance levels and the effect sizes were smaller (h2 = 1.62% and 0.79%, respectively). Furthermore, a genotype relative risk test of children who had questionnaire-based diagnoses of ADHD also failed to yield evidence of either association or linkage. Given that the A1 allele was expected to be the high risk allele, and that results were non-significant in tests that controlled for population heterogeneity, we doubt that this DRD2 polymorphism influences symptoms of ADHD in childhood.

Genetic and environmental influences on vocabulary IQ: parental education level as moderator.

This article examines how parental education level moderates the genetic and environmental contributions to variation in verbal IQ. Data are from 1909 non-Hispanic Whites and African American sibling pairs from the National Longitudinal Study of Adolescent Health, which obtained nationally-based samples of identical (MZ) twins, fraternal (DZ) twins, full and half siblings, cousins (in the same household), and biologically unrelated siblings. In the whole sample, the variance estimate for heritability (h2 = .57, SE = .08) was greater than that for shared environment (c2 = .13, SE = .04). Both heritability and the shared environmental estimate were moderated, however, by level of parental education. Specifically, among more highly educated families, the average h2 = .74 (SE = .10) and the average c2 = .00 (SE = .05). Conversely, among less well-educated families, heritability decreased and shared environmental influences increased, yielding similar proportions of variance explained by genetic and environmental factors, average h2 = .26 (SE = .15), and average c2 = .23 (SE = .07).

Genetic and environmental influences on the relationships between family connectedness, school connectedness, and adolescent depressed mood: sex differences.

This study investigated (a) genetic and environmental contributions to the relationship between family and school environment and depressed mood and (b) potential sex differences in genetic and environmental contributions to both variation in and covariation between family connectedness, school connectedness, and adolescent depressed mood. Data are from 2,302 adolescent sibling pairs (mean age = 16 years) who were part of the National Longitudinal Study of Adolescent Health. Although genetic factors appeared to be important overall, model-fitting analyses revealed that the best-fitting model was a model that allowed for different parameters for male and female adolescents. Genetic contributions to variation in all 3 variables were greater among female adolescents than male adolescents, especially for depressed mood. Genetic factors also contributed to the correlations between family and school environment and adolescent depressed mood, although, again, these factors were stronger for female than for male adolescents.

The quiescent-cell expression system for protein synthesis in Escherichia coli.

The quiescent-cell expression system is a radical alternative to conventional fermentation for protein overproduction in Escherichia coli. It is dependent on the controlled overexpression of a small RNA called Rcd in hns mutant strains to generate nongrowing, quiescent cells which are not nutrient limited. Quiescent cells no longer produce biomass and have their metabolic resources channelled toward the expression of plasmid-based genes. The biosynthetic capacity of the system is demonstrated by its ability to express chloramphenicol acetyltransferase to more than 40% of total cell protein. Quiescent cells may provide an ideal environment for the expression of toxic as well as benign proteins.

Nature, nurture and first sexual intercourse in the USA: fitting behavioural genetic models to NLSY kinship data.

Fisher (1930) presented both theoretical and empirical results concerning genetic influences on fertility. Since then, only sparse research has been done on the genetics of fertility, although more sophisticated methodogy and data now exist than were available to Fisher. This paper presents a behavioural genetic analysis of age at first intercourse, accounting for genetic, shared environmental, and selected non-shared environmental influences. The data came from the nationally representative National Longitudinal Survey of Youth (NLSY). A newly developed kinship linking procedure was used that identifies links for cousins, half-siblings, full-siblings and twins in the NLSY. The results suggest a genetic influence in the overall dataset, and also among whites and in male-male and opposite-sex pairs. Genetic influences were extremely small or non-existent for blacks and for female-female pairs. Shared environmental influences were small for most subsets of the data, but moderate for female-female pairs. Two specific non-shared environmental influences--self-esteem and locus of control--were ruled out as accounting for any meaningful variance, although other general sources of non-shared environmental influence appear potentially important. Analysis of selected samples from upper and lower tails suggested that genetic influences are important in accounting for both early and late non-virginity. These findings are consistent with work reported by Miller et al. (1999), who used molecular genetic methods. Generally, these findings support the existence of genetic influences and implicate non-shared environmental influences as being important determinants of the timing of loss of virginity among US adolescents and young adults.

A logistic regression based extension of the TDT for continuous and categorical traits.

The transmission disequilibrium test (TDT), designed as a test of linkage in the presence of association (i.e. linkage disequilibrium), has received considerable attention in the recent statistical genetics literature due to its advantages over other within-family analytic methods. One limitation of the conventional TDT is its application solely to linkage disequilibrium between a genetic marker and a single categorical trait (e.g. presence or absence of a disease). In this paper, we present an extension of the TDT using logistic regression to examine the relation between a candidate gene or genetic marker and one or more continuous or categorical explanatory variables. This logistic regression extension of the TDT possesses all of the desirable features of the conventional TDT, as well as many advantages associated with traditional regression analysis. We describe the model and its properties, as well as a number of its possible applications, and apply it to examine linkage disequilibrium between the dopamine receptor D2 gene (DRD2) and symptoms of childhood attention deficit hyperactivity disorder (ADHD). We also briefly compare the logistic regression TDT to other quantitative TDTs that have been proposed in the literature, and highlight the advantages of a regression-based approach for examining the relation between a candidate gene and one or more continuous or categorical traits. Given its features, we regard the logistic regression extension of the TDT as a flexible new data analytic method with extensive potential applications to problems in medical, psychiatric, and behavioral genetics.

Association and linkage of the dopamine transporter gene and attention-deficit hyperactivity disorder in children: heterogeneity owing to diagnostic subtype and severity.

Attention-deficit hyperactivity disorder (ADHD) affects approximately 3%-5% of children in the United States. In the current psychiatric nomenclature, ADHD comprises three subtypes: inattentive, hyperactive-impulsive, and combined. In this study, we used four analytic strategies to examine the association and linkage of the dopamine transporter gene (DAT1) and ADHD. Our sample included 122 children referred to psychiatric clinics for behavioral and learning problems that included but were not limited to ADHD, as well as their parents and siblings. Within-family analyses of linkage disequilibrium, using the transmission disequilibrium test (TDT), confirmed the 480-bp allele as the high-risk allele. In between-family association analyses, levels of hyperactive-impulsive symptoms but not inattentive symptoms were related to the number of DAT1 high-risk alleles. Siblings discordant for the number of DAT1 high-risk alleles differed markedly in their levels of both hyperactive-impulsive and inattentive symptoms, such that the sibling with the higher number of high-risk alleles had much higher symptom levels. Within-family analyses of linkage disequilibrium, using the TDT, suggested association and linkage of ADHD with DAT1 and that this relation was especially strong with the combined but not the inattentive subtype. The relation of DAT1 to ADHD increased monotonically, from low to medium to high levels of symptom severity. Our results replicate and extend previous findings of the association between the DAT1 gene and childhood ADHD. This represents one of the first replicated relations of a candidate gene and a psychiatric disorder in children.

Genetic and shared environmental influences on adolescent BMI: interactions with race and sex.

The present study uses a behavioral genetic design to investigate the genetic and environmental influences on variation in adolescent body mass index (BMI) and to determine whether the relative influences of genetic and environmental factors on variation in BMI are similar across racial groups and sexes. Data for the present study come from the National Longitudinal Study on Adolescent Health (Add Health), a large, nationally representative study of adolescent health and health-related behaviors. The Add Health sample contains a subset of sibling pairs that differs in levels of genetic relatedness, making it well suited for behavioral genetics analyses. The present study examines whether genetic and environmental influences on adolescent BMI are the same for males and females and for Black and White adolescents. Results indicate that genetic factors contribute substantially to individual differences in adolescent BMI, explaining between 45 and 85% of the variance in BMI. Furthermore, based on an analysis of opposite-sex sibling pairs, the genes that influence variation in adolescent BMI are similar for males and females. However, the relative importance of genetic and environmental influences on variation in BMI differs for males and females and for Blacks and Whites. Although parameter estimates could be constrained to be equal for Black and White males, they could not be constrained to be equal for Black and White females. Moreover, the best-fitting model for Black females was an ADE model, for White females it was an ACE model, and for males it was an AE model. Thus, shared environmental influences are significant for White female adolescents, but not for Black females or males. Likewise, nonadditive genetic influences are indicated for Black females, but not for White females or males. Implications of these results are discussed.