NOTCH1 Gain of Function in Germ Cells Causes Failure of Spermatogenesis in Male Mice

NOTCH1 is a member of the NOTCH receptor family, a group of single-pass trans-membrane receptors. NOTCH signaling is highly conserved in evolution and mediates communication between adjacent cells. NOTCH receptors have been implicated in cell fate determination, as well as maintenance and differentiation of stem cells. In the mammalian testis expression of NOTCH1 in somatic and germ cells has been demonstrated, however its role in spermatogenesis was not clear. To study the significance of NOTCH1 in germ cells, we applied a cre/loxP approach in mice to induce NOTCH1 gain- or loss-of function specifically in male germ cells. Using a Stra8-icretransgene we produced mice with conditional activation of the NOTCH1 intracellular domain (NICD) in germ cells. Spermatogenesis in these mutants was progressively affected with age, resulting in decreased testis weight and sperm count. Analysis of downstream target genes of NOTCH1 signaling showed an increased expression of Hes5, with a reduction of the spermatogonial differentiation marker, Neurog3 expression in the mutant testis. Apoptosis was significantly increased in mouse germ cells with the corresponding elevation of pro-apoptotic Trp53 and Trp63genes' expression. We also showed that the conditional germ cell-specific ablation of Notch1 had no effect on spermatogenesis or male fertility. Our data suggest the importance of NOTCH signaling regulation in male germ cells for their survival and differentiation.

Without women: Masculinities, gay male sexual culture and sexual behaviors in Miami, Florida

Recent studies have reported alarmingly high rates of HIV infection and risky sexual behaviors among gay men in Miami, Florida. Previous research has suggested that the risky sexual behaviors of many gay men reflect the pursuit of intimacy and love, and that barriers to intimate relationships among gay men may stem from traditional masculinity norms. This dissertation examines the meanings which gay men ascribe to their sexual behaviors, as well as the intersections of those meanings with both traditional masculinity constructions and Miami's gay male sexual culture. ^ The study is based upon participant observation, print media content analysis, surveys and ethnographic interviews of a purposive snowball sample of 30 Cuban American, Puerto Rican, African American and Anglo gay men who reside in Miami-Dade County, Florida. Analysis of research questions was accomplished through grounded theory methods and descriptive and non-parametric statistics, including Pearson chi-square, Fisher's Exact and Mann-Whitney U tests. ^ The study shows that culturally-specified masculinity norms vary in the relative importance ascribed to heterosexual prowess, economic providership and competitiveness. These cultural differences appear important not only to the timing of sexual awareness and to the strength of homosexual stereotyping as effeminacy, but also to men's strategies in coming out as gay. The meanings men attributed to their sexual behaviors were, however, constructed in response to both inherited masculinity norms and the hypermasculine structure of Miami's gay male sexual culture. In addition to providing an ethnographic account of this subculture, the study elaborates men's issues relative to casual sex and committed relationships. Unprotected anal intercourse with casual partners during the previous twelve months was associated with growing up without one's father in the home, having been teased for effeminacy during childhood, being defensive about one's masculinity, not trusting men, having been cheated on by boyfriends, and believing that long-term gay male relationships are problematic. ^ It is concluded that the continuing epidemic of HIV infections among local gay men, as well as the hypermasculine form of the gay sexual subculture itself, are nihilistic symptoms embedded in the masculinist gender structure of the larger society. ^

Longitudinal effects of family factors on alcohol use and alcohol-related problems among males from pre-adolescence to adulthood

The goal of this study was to examine the longitudinal effects of five family factors on alcohol use among adolescent males. The family factors included familism (family pride, loyalty, and cohesion), parent derogation (being put down by parents), parent/child communication, family alcohol problems and family drug problems. The study focused on the effects of the family factors reported by a sample of 451 White-non-Hispanic and African American males during early and mid-adolescence on (1) the intensity of alcohol use in mid-adolescence, and (2) the number of problems associated with alcohol use during the transition to young adulthood. The study also explored racial differences in the effects of the family factors. The data for this study were derived from a two-phase longitudinal epidemiologic cohort study of male and female adolescents enrolled in middle schools in Miami, FL. Data were collected at four points between 1990 and 2001. Linear and logistical regressions were used to analyze the effects of the family variables on the dependent variables. ^ The results of the analyses indicated that all of the family variables except family drug problems were statistically significant predictors of the level of alcohol use and alcohol-related problems. Familism had a moderate influence on both of the dependent variables at all data points, while parent derogation, parent/child communication and family alcohol problems were weak predictors. While the family factors varied by race, their impact on the dependent variables did not vary substantially. ^ This study had methodological shortcomings related to measurement and design that may have contributed to the weak influence of the variables. Future studies should explore possible mediating effects of these variables, and should employ more sensitive measures that are culturally appropriate. The results suggest that, since early family factors have long-term effects on children's substance-using behaviors, the family environment should be addressed in prevention and intervention efforts. ^