Using a Regression Approach to Study the Influence of Male Fetuses on the Genital Morphology of Neonatal Female Rats

Among newborn female rats considerable variability is found in genital morphology (e.g., anogenital distance, AGD). Presumably, such differences are related to prenatal androgen exposure, with greater exposure resulting in larger AGD's and thus in a trend toward masculinization. The source of prenatal androgen in female fetuses is unclear, but a role for male uterine mates has been implicated. The present study investigated the effect of a number of prenatal factors related to number and position of males in utero on female AGD in two strains of rats. Because such prenatal factors often show systematic covariance, a methodology was used that enabled statistical control over variables that could not be /experimentally controlled. Results confirmed the importance of caudal males to Female AGD,and identified two additional intra-uterine variables salient to female genital masculinization, namely the distance of the female fetus from the nearest caudal male, and the overall number of males sharing the same uterine horn. An increase in number of adjacent males was, contrary to previous reports, associated with a decrease in AGD, but this effect was limited to one strain. There was considerable variation in AGD across the two strains, and, more importantly, across litters, suggesting the importance of factors impacting the litter as a whole rather than specific individuals within the litter.