It is well-documented that circadian rhythms are controlled by the circadian master clock of the mammalian brain, located in the suprachiasmatic nucleus (SCN) of the hypothalamus. The SCN clockwork is a cell autonomous mechanism consisting of a series of interlocked transcriptional/post-translational feedback loops. In turn, the SCN controls the seasonal rhythmicity of various biological processes, in particular the secretion pattern of hormones. Although the effects of gonadal hormones on circadian rhythmicity are clearly established, how the SCN integrates and regulates these hormonal stimuli remains unknown. We have previously found that clock genes are expressed in the choroid plexus (CP). Therefore, we compared the circadian expression of these genes in female and male rat CP. We show that there is a 24-h rhythm in the expression of Per2 and Cry2 in males and females. Bmal1 and Per1 expression also varied along the day, but only in females. Bmal1, Clock and Per1 mRNA did not show any significant differences in the CP of males. Moreover, data from cultured CP cells collected at different timepoints revealed significant circadian rhythms in mRNA abundance of Bmal1, Clock and Per2. In conclusion, our data show that the rat CP expresses all canonical clock genes and that their circadian expression differs between genders suggesting that hormones can regulate circadian rhythmicity in CP.

The choroid plexus (CP) are highly vascularized branched structures that protrude into the ventricles of the brain, and form a unique interface between the blood and the cerebrospinal fluid (CSF), the blood-CSF barrier, that are the main site of production and secretion of CSF. Sex hormones are widely recognized as neuroprotective agents against several neurodegenerative diseases, and the presence of sex hormones cognate receptors suggest that it may be a target for these hormones. In an effort to provide further insight into the neuroprotective mechanisms triggered by sex hormones we analyzed gene expression differences in the CP of female and male rats subjected to gonadectomy, using microarray technology. In gonadectomized female and male animals, 3045 genes were differentially expressed by 1.5-fold change, compared to sham controls. Analysis of the CP transcriptome showed that the top-five pathways significantly regulated by the sex hormone background are olfactory transduction, taste transduction, metabolism, steroid hormone biosynthesis and circadian rhythm pathways. These results represent the first overview of global expression changes in CP of female and male rats induced by gonadectomy and suggest that sex hormones are implicated in pathways with central roles in CP functions and CSF homeostasis.

Congenital hypogonadotropic hypogonadism (CHH) is characterized by lack of normal pubertal development due to deficient gonadotropin-releasing hormone (GnRH) secretion or action, and is caused by genetic defects in several genes. Mutations in the CHD7 gene cause CHARGE syndrome (Coloboma of the eye, Heart defects, Atresia of the choanae, Retardation of growth and development, Genital hypoplasia and Ear abnormalities), but have also been found in patients with isolated CHH. The aim of this study was to identify CHD7 mutations in patients with CHH. Fifty Portuguese patients with CHH were screened for mutations in the CHD7 gene by DNA sequencing. Eight (16%) patients had CHD7 rare sequence variants that consisted of six missense (p.Gly388Glu, p.His903Pro, p.Thr1082Ile, p.Val1452Leu, p.Asp1854Gly, and p.Arg2065His) and two synonymous (p.Ser559Ser, and p.Ala2785Ala) mutations. Five of these mutations have never been reported before. Three CHD7 mutations occurred in patients that had mutations in additional CHH-genes. This study uncovered novel genetic variants that expand the known spectrum of mutations associated with CHH. The frequency of CHD7 mutations in this cohort was higher than that of other major CHH-genes and confirms the importance of including CHD7 in the genetic testing of CHH, even in the absence of additional CHARGE features.

Evidence from the literature suggests that testosterone (T) plays an important role in the neural structure, physiology, and function of the hippocampus (HP). However, many of the genes involved and underlying mechanisms remain to be elucidated. To shed light on this issue, we explored the transcriptome of HP in orchidectomized (OOX) rats to identify T-dependent gene expression in rat HP. RNA from OOX and sham HP animals were processed and measured by the Applied Biosystems microarray platform. The results showed a total of 271 genes differentially expressed between OOX vs. sham animals. Overall, T depletion resulted in the upregulation of 98 genes, including genes associated with neurogenesis and behavior. Of particular interest was the downregulation of 173 genes, with known functions, including signal transduction or neurological system processes. Our data shows that T depletion results in significantly altered hippocampal gene expression profiles and constitutes a starting tool to elucidating the molecular mechanisms involved in the action of androgens in the physiology of the HP.