Prenatal uterine environment and sexual differentiation of rats

prenatal factors relevant to hormonal environment on the sexual differentiation of behavior, morphology and central nervous system in rats. The effects of such factors as prenatal sex composition of the litter and position in utero on the sexual differentiation of normally developed (i.e. untreated) male and female rats was examined. In addition, the effects of experimentally induced changes in the perinatal hormonal milieu on the central nervous system and behavior of male rats were assessed. This general introduction provides an overview of the effects of hormones on reproductive morphology and behavior, and function and morphology of the central nervous system of mammals, with emphasis on rats. Current questions and hypotheses that led to the experiments presented in this thesis will be outlined

Plasma testosterone in fetal rats and their mothers on day 19 of gestation

Plasma testosterone levels were higher in pooled samples from male fetuses than from female fetuses on day 19 of pregnancy. Plasma testosterone from female fetuses with males located caudally in the uterus was higher than from females that lacked such males. Testosterone level of both male and female fetuses was correlated with maternal testosterone. No correlation was found between maternal testosterone and number of males in the litter, male-to-female ratio, or litter size. These results corroborate earlier findings of a sex difference in plasma testosterone levels on fetal day 19 in rats, and provide support for the hypothesis that female rats receive androgens from males located caudally in the uterus. No evidence was found that testosterone of pregnant females is affected by the sex ratio or size of her litter

SDN-POA volume, sexual behavior, and partner preference of male rats affected by perinatal treatment with ATD

The present study investigated 1) the importance of the aromatization process during the perinatal period for the development of the sexually dimorphic nucleus in the preoptic area of the hypothalamus (SDN-POA) of male rats, and 2) the relationship between SDN-POA volume and parameters of masculinization in male rats that were treated perinatally with the aromatase-inhibitor ATD. Males were treated with ATD either prenatally or pre- and neonatally, or with the vehicle. Masculine sexual behavior and partner preference were investigated in adulthood. Thereafter, animals were sacrificed and SDN-POA volume was measured. The SDN-POA volume was reduced in both the prenatally and the pre- and neonatally treated group, with a larger reduction in the latter than in the former group. Combined pre- and neonatal ATD treatment resulted in reduced frequency of mounts, intromissions, and ejaculations, as well as a reduced preference for a female over a male. The SDN-POA size was significantly and positively correlated with frequency of masculine sexual behavior, as well as preference for a female over a male

Chromatin interaction of TATA-binding protein is dynamically regulated in human cells

International audienceGene transcription in mammalian cells is a dynamic process involving regulated assembly of transcription complexes on chromatin in which the TATA-binding protein (TBP) plays a central role. Here, we investigate the dynamic behaviour of TBP by a combination of fluorescence recovery after photobleaching (FRAP) and biochemical assays using human cell lines of different origin. The majority of nucleoplasmic TBP and other TFIID subunits associate with chromatin in a highly dynamic manner. TBP dynamics are regulated by the joint action of the SNF2-related BTAF1 protein and the NC2 complex. Strikingly, both BTAF1 and NC2 predominantly affect TBP dissociation rates, leaving the association rate unchanged. Chromatin immunoprecipitation shows that BTAF1 negatively regulates TBP and NC2 binding to active promoters. Our results support a model for a BTAF1-mediated release of TBP-NC2 complexes from chromatin

Dissection of cell cycle-dependent dynamics of Dnmt1 by FRAP and diffusion-coupled modeling

DNA methyltransferase 1 (Dnmt1) reestablishes methylation of hemimethylated CpG sites generated during DNA replication in mammalian cells. Two subdomains, the proliferating cell nuclear antigen (PCNA)-binding domain (PBD) and the targeting sequence (TS) domain, target Dnmt1 to the replication sites in S phase. We aimed to dissect the details of the cell cycle–dependent coordinated activity of both domains. To that end, we combined super-resolution 3D-structured illumination microscopy and fluorescence recovery after photobleaching (FRAP) experiments of GFP-Dnmt1 wild type and mutant constructs in somatic mouse cells. To interpret the differences in FRAP kinetics, we refined existing data analysis and modeling approaches to (i) account for the heterogeneous and variable distribution of Dnmt1-binding sites in different cell cycle stages; (ii) allow diffusion-coupled dynamics; (iii) accommodate multiple binding classes. We find that transient PBD-dependent interaction directly at replication sites is the predominant specific interaction in early S phase (residence time Tres ≤10 s). In late S phase, this binding class is taken over by a substantially stronger (Tres ∼22 s) TS domain-dependent interaction at PCNA-enriched replication sites and at nearby pericentromeric heterochromatin subregions. We propose a two-loading-platform-model of additional PCNA-independent loading at postreplicative, heterochromatic Dnmt1 target sites to ensure faithful maintenance of densely methylated genomic regions