Francisco J. Sánchez

Epigenetic Predictor of Age

From the moment of conception, we begin to age. A decay of cellular structures, gene regulation, and DNA sequence ages cells and organisms. DNA methylation patterns change with increasing age and contribute to age related disease. Here we identify 88 sites in or near 80 genes for which the degree of cytosine methylation is significantly correlated with age in saliva of 34 male identical twin pairs between 21 and 55 years of age. Furthermore, we validated sites in the promoters of three genes and replicated our results in a general population sample of 31 males and 29 females between 18 and 70 years of age. The methylation of three sites—in the promoters of the EDARADD, TOM1L1, and NPTX2 genes—is linear with age over a range of five decades. Using just two cytosines from these loci, we built a regression model that explained 73% of the variance in age, and is able to predict the age of an individual with an average accuracy of 5.2 years. In forensic science, such a model could estimate the age of a person, based on a biological sample alone. Furthermore, a measurement of relevant sites in the genome could be a tool in routine medical screening to predict the risk of age-related diseases and to tailor interventions based on the epigenetic bio-age instead of the chronological age.

Androgen Receptor Repeat Length Polymorphism Associated with Male-to-Female Transsexualism

BACKGROUND There is a likely genetic component to transsexualism, and genes involved in sex steroidogenesis are good candidates. We explored the specific hypothesis that male-to-female transsexualism is associated with gene variants responsible for undermasculinization and/or feminization. Specifically, we assessed the role of disease-associated repeat length polymorphisms in the androgen receptor (AR), estrogen receptor beta (ERbeta), and aromatase (CYP19) genes. METHODS Subject-control analysis included 112 male-to-female transsexuals and 258 non-transsexual males. Associations and interactions were investigated between CAG repeat length in the AR gene, CA repeat length in the ERbeta gene, and TTTA repeat length in the CYP19 gene and male-to-female transsexualism. RESULTS A significant association was identified between transsexualism and the AR allele, with transsexuals having longer AR repeat lengths than non-transsexual male control subjects (p=.04). No associations for transsexualism were evident in repeat lengths for CYP19 or ERbeta genes. Individuals were then classified as short or long for each gene polymorphism on the basis of control median polymorphism lengths in order to further elucidate possible combined effects. No interaction associations between the three genes and transsexualism were identified. CONCLUSIONS This study provides evidence that male gender identity might be partly mediated through the androgen receptor.

Regional gray matter variation in male-to-female transsexualism.

Gender identity-ones sense of being a man or a woman-is a fundamental perception experienced by all individuals that extends beyond biological sex. Yet, what contributes to our sense of gender remains uncertain. Since individuals who identify as transsexual report strong feelings of being the opposite sex and a belief that their sexual characteristics do not reflect their true gender, they constitute an invaluable model to understand the biological underpinnings of gender identity. We analyzed MRI data of 24 male-to-female (MTF) transsexuals not yet treated with cross-sex hormones in order to determine whether gray matter volumes in MTF transsexuals more closely resemble people who share their biological sex (30 control men), or people who share their gender identity (30 control women). Results revealed that regional gray matter variation in MTF transsexuals is more similar to the pattern found in men than in women. However, MTF transsexuals show a significantly larger volume of regional gray matter in the right putamen compared to men. These findings provide new evidence that transsexualism is associated with distinct cerebral pattern, which supports the assumption that brain anatomy plays a role in gender identity.

Puberty in the corpus callosum

Adolescence is an important period for brain development. White matter growth is influenced by sex hormones such as testosterone, and the corpus callosum-the largest white matter structure in the human brain-may change structurally during the hormone-laden period of adolescence. Little is known about pubertys relationship to structural brain development, even though pubertal stage may better predict cognitive and behavioral maturity than chronological age. We therefore aimed to establish the presence and direction of pubertal effects on callosal anatomy. For this purpose, we applied advanced surface-based mesh-modeling to map correlations between callosal thickness and pubertal stage in a large and well-matched sample of 124 children and adolescents (62 female and 62 male) aged 5-18years from a normative database. When linking callosal anatomy to pubertal status, only positive correlations reached statistical significance, indicating that callosal growth advances with puberty. In tests of differences in callosal anatomy at different stages of puberty, callosal growth was concentrated in different locations depending on the pubertal stage. Changing levels of circulating sex hormones during different phases of puberty likely contributed to the observed effects, and further research is clearly needed. Direct quantification of sex hormone levels and regional fiber connectivity-ideally using fiber tractography-will reveal whether hormones are the main drivers of callosal change during puberty. These callosal findings may lead to hypotheses regarding cortical changes during puberty, which may promote or result from changes in inter-hemispheric connectivity.

Genes and brain sex differences

Throughout development, numerous biological events occur that differentially affect males and females. Specifically, sex-determining genes that are triggered by the sex-chromosome complement initiate a series of events that determine an organisms sex and lead to the differentiation of the body in sex-specific ways. Such events contribute to many unique sex differences, including the susceptibility to different diseases. Although it was believed that sex hormones singularly differentiated the brain and body, there is emerging research showing that genes also play a direct role. In this chapter, we review this line of work and focus on the use of a unique mouse model that separates the effect of gonadal hormones and sex chromosomes. As genetic technology continues to advance, our understanding of the role that hormones and genes play in sex differences can be used to advance the physical and mental health of both men and women.

Transient cholesterol effects on nicotinic acetylcholine receptor cell-surface mobility.

To what extent do cholesterol-rich lipid platforms modulate the supramolecular organization of the nicotinic acetylcholine receptor (AChR)? To address this question, the dynamics of AChR particles at high density and its cholesterol dependence at the surface of mammalian cells were studied by combining total internal reflection fluorescence microscopy and single-particle tracking. AChR particles tagged with a monovalent ligand, fluorescent α-bungarotoxin (αBTX), exhibited two mobile pools: i) a highly mobile one undergoing simple Brownian motion (16%) and ii) one with restricted motion (∼50%), the rest being relatively immobile (∼44%). Depletion of membrane cholesterol by methyl-α-cyclodextrin increased the fraction of the first pool to 22% and 33% after 15 and 40 min, respectively; the pool undergoing restricted motion diminished from 50% to 44% and 37%, respectively. Monoclonal antibody binding results in AChR crosslinking-internalization after 2 h; here, antibody binding immobilized within minutes ∼20% of the totally mobile AChR. This proportion dramatically increased upon cholesterol depletion, especially during the initial 10 min (83.3%). Thus, antibody crosslinking and cholesterol depletion exhibited a mutually synergistic effect, increasing the average lifetime of cell-surface AChRs∼10 s to ∼20 s. The instantaneous (microscopic) diffusion coefficient D 2–4 of the AChR obtained from the MSD analysis diminished from ∼0.001 µm2 s−1 to ∼0.0001–0.00033 µm2 s−1 upon cholesterol depletion, ∼30% of all particles falling into the stationary mode. Thus, muscle-type AChR exhibits heterogeneous motional regimes at the cell surface, modulated by the combination of intrinsic (its supramolecular organization) and extrinsic (membrane cholesterol content) factors.

Feminized Behavior and Brain Gene Expression in a Novel Mouse Model of Klinefelter Syndrome

Klinefelter Syndrome (KS) is the most common sex chromosome aneuploidy in men and is characterized by the presence of an additional X chromosome (XXY). In some Klinefelter males, certain traits may be feminized or shifted from the male-typical pattern towards a more female-typical one. Among them might be partner choice, one of the most sexually dimorphic traits in the animal kingdom. We investigated the extent of feminization in XXY male mice (XXYM) in partner preference and gene expression in the bed nucleus of the stria terminalis/preoptic area and the striatum in mice from the Sex Chromosome Trisomy model. We tested for partner preference using a three-chambered apparatus in which the test mouse was free to choose between stimulus animals of either sex. We found that partner preference in XXYM was feminized. These differences were likely due to interactions of the additional X chromosome with the Y. We also discovered genes that differed in expression in XXYM versus XYM. Some of these genes are feminized in their expression pattern. Lastly, we also identified genes that differed only between XXYM versus XYM and not XXM versus XYM. Genes that are both feminized and unique to XXYM versus XYM represent strong candidates for dissecting the molecular pathways responsible for phenotypes present in KS/XXYM but not XXM. In sum, our results demonstrated that investigating behavioral and molecular feminization in XXY males can provide crucial information about the pathophysiology of KS and may aid our understanding of sex differences in brain and behavior.

The New Policy on Hyperandrogenism in Elite Female Athletes is Not About “Sex Testing”

In April 2011, the International Olympic Committee (IOC) and the International Association of Athletics Federations (IAAF) updated their regulations regarding elite female athletes with hyperandrogenism: Women whose testosterone levels crossed into the male range could not compete with other women unless it was shown that they are resistant to the effects of testosterone. Although the new rule is a marked improvement over past attempts to ensure that men were not trying to compete as women in elite competition, several criticisms have been leveled against the new regulations. Here we offer our reactions to claims that the new regulation promotes a sex-verification test, claims that intersex athletes will automatically be disqualified from competition, and proposals to either divide athletes based on variables beyond sex or completely eliminate sex groupings. Although elite sports can never achieve a perfectly level playing field, there should be parameters to which athletes must conform for a given sport. Yet elite athletes themselves should play a decisive role in what is best for their sport.

The Relationship Between Help-Seeking Attitudes and Masculine Norms Among Monozygotic Male Twins Discordant for Sexual Orientation

OBJECTIVE In general, heterosexual men are less favorable to asking for help compared to women and gay men. This can be problematic if a man avoids professional help when he is experiencing significant psychological distress. Yet, it is unclear to what degree such attitudes among men are due to innate differences or social environments. Studying twins provides one avenue for teasing apart these relationships. METHOD We recruited 38 pairs of monozygotic male twins (Mage = 35.87 years, SD = 9.52) raised together and who were discordant for sexual orientation. They completed measures of psychological distress (Symptom Checklist-90-Revised), positive attitudes toward psychological help-seeking behavior, and emphasis with fulfilling traditional masculine norms. RESULTS Contrary to predictions, the heterosexual twins expressed more symptoms of specific distress-hostility (r = .30), paranoid ideation (r = .26), and psychoticism (r = .24)-than their gay cotwins. As predicted, heterosexual men were less favorable to seeking help (r = .25) and expressed greater emphasis on masculine norms (r = .26) than their cotwins. Within each group of men, unique aspects of masculine norms were significantly related to attitudes toward psychological help-seeking behavior. CONCLUSION The findings lend credence to the hypothesis that social environments influence attitudes and behaviors that are stereotypically masculine and potentially detrimental to mens health.

The Biology of Sexual Orientation and Gender Identity

The two largest sex differences in behavior between men and women are sexual orientation and gender identity. Sexual orientation refers to the direction of sexual attraction to the same or other sex/gender. Gender identity refers to peoples inner sense of maleness or femaleness. Although these phenomena are a fundamental part of human life, their underlying mechanisms remain poorly understood. The dominant theory is that prenatal hormones of gonadal origin play a major role in both sexual orientation and gender identity. However, a review of the evidence shows that hormones fail to explain either of these traits in the vast majority of humans. Furthermore, correlations with several parameters, including digit length ratio, handedness, fraternal birth order, and brain anatomy, do not provide clear causal etiology to masculine or feminine sexual behavior. Emerging studies on twins and the application of linkage analysis provide arguments for a genetic influence on sexual orientation and is reviewed in this chapter.