ABSTRACT
Sex steroid hormones act on developing neural circuits regulating the hypothalamic-pituitary-gonadal axis and are involved in hormone-sensitive behaviours. These hormones act mainly via nuclear receptors, such as oestrogen receptor (ER)-α and androgen receptor (AR). By using immunohistochemistry, we analysed the expression level of ERα and AR throughout perinatal life [at embryonic (E) day 19 and postnatal (P) days 5, 15 and 25] and in adulthood in several hypothalamic nuclei controlling reproduction in both wild-type and aromatase knockout (ArKO) (i.e. which cannot convert testosterone into oestradiol) mice to determine whether there are sex differences in hypothalamic ERα and AR expression and, if so, whether these are established by the action of oestradiol. As early as E19, ERα immunoreactivity (-IR) was observed at same expression levels in both sexes in the anteroventral periventricular nucleus (AVPv), the medial preoptic area (MPOA), the bed nucleus of the stria terminalis (BnST), the ventrolateral part of the ventromedial hypothalamic nucleus and the arcuate nucleus (ARC). Sex differences (female > male) in ERα-IR were observed not only during the prepubertal period in the BnST (P5 to P25) and the MPOA (P15), but also in adulthood in these two brain regions. Sex differences in AR-IR (male > female) were observed at P5 in the AVPv and ARC, and at P25 in the MPOA and ARC, as well as in adulthood in all hypothalamic regions analysed. In adulthood, gonadectomy and hormonal treatment (oestradiol or dihydrotestosterone) also strongly modulated ERα-IR and AR, respectively. Taken together, sex differences in ERα-IR and AR-IR were observed in all hypothalamic regions analysed, although they most likely do not reflect the action of oestradiol because ArKO mice of both sexes showed expression levels very similar to wild-type mice throughout perinatal development.
Subject(s)
Aging/metabolism , Estrogen Receptor alpha/metabolism , Hypothalamus/growth & development , Hypothalamus/metabolism , Receptors, Androgen/metabolism , Animals , Embryo, Mammalian , Female , Hypothalamus/embryology , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Sex Characteristics , Tissue DistributionABSTRACT
The SP/KLF transcription factor family contains over 25 members sharing a DNA-binding domain composed of three zinc fingers of the C(2)H(2) type. We previously identified the sixth member of the SP subfamily (Sp6). The 5' end of the Sp6 transcript was not cloned and was predicted bioinformatically. A mouse molar tooth cDNA was then isolated differing from the Sp6 sequence by its 5' end, and was named epiprofin. Sp6 and epiprofin are currently used as synonyms. Here, we show that the Sp6 transcript possesses a first exon distinct from the epiprofin one: the Sp6 gene thus uses two promoters, generating two transcript variants which differ in their first exon. Furthermore, we identified an Sp6 opposite strand transcript (Sp6os) and examined, by quantitative RT-PCR experiments, the presence and the abundance of these two transcripts in mouse tissues. We also mapped the mouse locus by FISH to chromosome 11D.
Subject(s)
Kruppel-Like Transcription Factors/genetics , Promoter Regions, Genetic/genetics , RNA, Antisense/genetics , Animals , Base Sequence , Gene Expression , Gene Expression Profiling , In Situ Hybridization, Fluorescence , Male , Mice , Mice, Inbred Strains , Molecular Sequence Data , Physical Chromosome Mapping , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Transcription Factors/genetics , Transcription Initiation SiteABSTRACT
The neural mechanisms controlling mate recognition and heterosexual partner preference are sexually differentiated by perinatal actions of sex steroid hormones. We previously showed that the most important action of oestrogen during prenatal development is to defeminise and, to some extent, masculinise brain and behaviour in mice. Female mice deficient in alpha-foetoprotein (AFP) due to a targeted mutation in the Afp gene (AFP-KO) do not show any female sexual behaviour when paired with an active male because they lack the protective action of AFP against maternal oestrogens. In the present study, we investigated whether odour preferences, another sexually differentiated trait in mice, are also defeminised and/or masculinised in AFP-KO females due to their prenatal exposure to oestrogens. AFP-KO females of two background strains (CD1 and C57Bl/6j) preferred to investigate male over female odours when given the choice between these two odour stimuli in a Y-maze, and thus remained very female-like in this regard. Thus, the absence of lordosis behaviour in these females cannot be explained by a reduced motivation of AFP-KO females to investigate male-derived odours. Furthermore, the presence of a strong male-directed odour preference in AFP-KO females suggests a postnatal contribution of oestrogens to the development of preferences to investigate opposite-sex odours.
Subject(s)
Estrogens/physiology , Mating Preference, Animal/physiology , Prenatal Exposure Delayed Effects , Smell/physiology , alpha-Fetoproteins/physiology , Analysis of Variance , Animals , Choice Behavior , Female , Mice , Mice, Inbred C57BL , Mice, Knockout , Pregnancy , Sex Differentiation/physiology , alpha-Fetoproteins/geneticsABSTRACT
This work shows that the proximal promoter of the mouse Afp gene contains a Ku binding site and that Ku binding is associated with down-regulation of the transcriptional activity of the Afp promoter. The Ku binding site is located in a segment able to adopt a peculiar structured form, probably a hairpin structure. Interestingly, the structured form eliminates the binding sites of the positive transcription factor HNF1. Furthermore, a DNAse hypersensitive site is detected in footprinting experiments done with extracts of AFP non-expressing hepatoma cells. These observations suggest that the structured form is stabilised by Ku and is associated with extinction of the gene in AFP non-expressing hepatic cells.
