HP1-beta is required for development of the cerebral neocortex and neuromuscular junctions.

HP1 proteins are thought to be modulators of chromatin organization in all mammals, yet their exact physiological function remains unknown. In a first attempt to elucidate the function of these proteins in vivo, we disrupted the murine Cbx1 gene, which encodes the HP1-beta isotype, and show that the Cbx1(-/-) -null mutation leads to perinatal lethality. The newborn mice succumbed to acute respiratory failure, whose likely cause is the defective development of neuromuscular junctions within the endplate of the diaphragm.

The sex determination in Ellobius lutescens remains bizarre.

Mammalian sex determination and gonad differentiation are the result of a complex interaction of fine-tuned spatial and temporal gene expression with threshold levels of individual genes. The male pathway is initiated by SRY. Some exceptional mammals determine male sex without the SRY gene and even without a Y chromosome.

Sex chromosome genes directly affect brain sexual differentiation.

Sex differences in the brain are caused by differences in gonadal secretions: higher levels of testosterone during fetal and neonatal life cause the male brain to develop differently than the female brain. In contrast, genes encoded on the sex chromosomes are not thought to contribute directly to sex differences in brain development, even though male (XY) cells express Y-chromosome genes that are not present in female (XX) cells, and XX cells may have a higher dose of some X-chromosome genes. Using mice in which the genetic sex of the brain (XX versus XY) was independent of gonadal phenotype (testes versus ovaries), we found that XY and XX brain cells differed in phenotype, indicating that a brain cell's complement of sex chromosomes may contribute to its sexual differentiation.

Sry does not fully sex-reverse female into male behavior towards pups.

Investigations in mice suggest that the Y-chromosomal genes affect certain behaviors. Here, we studied whether a part of the Y chromosome, the Sxr locus, has an effect on induction of motivation for parental care (pup retrieval) or of parental aggression towards pups (infanticide). XX females, XX males with the Sxr locus on the X chromosome, and XY males of the C57BL/6J strain were tested.

Protein kinase C isoenzyme: selective expression pattern of protein kinase C-θ during mouse development.

Protein kinase C (PKC)-θ, a serine/threonine protein kinase and novel PKC subfamily member, has been recently identified as an essential component of the T cell synapse which activates the NF-kB signaling cascade leading to expression of the IL-2 gene during T cell activation. By RNA in situ hybridization to whole-body embryo sections it is shown that the murine PKCθ is specifically expressed in tissues with hematopoietic and lymphopoietic activity. Expression is also evident in skeletal muscle.

Exclusion of SOX9 as the testis determining factor in Ellobius lutescens: evidence for another testis determining gene besides SRY and SOX9.

In mammals the initiation of testis determination usually depends on the Y-chromosomal gene SRY. A few species, however, escape from this rule with a testis determination that is independent of SRY. The mole vole Ellobius lutescens is one of these species.

Developmental profile of Sry transcripts in mouse brain.

Transient activation of the gene Sry in the gonadal ridge during a brief period of embryonic development is believed to function as a key signal for sex determination. However, a number of reports suggest that Sry expression is not as restricted in space and time as one would expect if its role was confined to directing male-specific differentiation in the early gonadal anlage. We have previously reported the occurrence of Sry/SRY transcripts in adult murine and human brain.

The Y-chromosomal genes SRY and ZFY are transcribed in adult human brain.

Sexual differentiation of the brain is thought to be regulated by hormonal signals from the developing male gonad. However, more-recent experimental and clinical data throw some doubt on the general validity of the "classical" steroid hypothesis and suggest that additional intervening factors or mechanisms need to be considered. In particular, it is now envisaged that neurons are capable of acquiring sex-specific properties independently of their hormonal environment.

Ontogenetic expression and splicing of estrogen receptor-alpha and beta mRNA in the rat midbrain.

Several studies have shown that estrogen is important for the differentiation of midbrain dopaminergic neurons. This is supported by the previous demonstration of estrogen synthesis in the perinatal ventral midbrain. The present study attempts to characterize the expression pattern of nuclear estrogen receptors (ER-alpha/beta) mRNAs in the ventral rat midbrain during development.

Birthdates of the tyrosine hydroxylase immunoreactive neurons in the hypothalamus of male and female rats.

This study determined the birthdates of the tyrosine hydroxylase-(TH) immunoreactive (IR) neurons in the zona incerta (ZI), periventricular nucleus (PeVN) and arcuate nucleus (AN) of male and female rats. 'Long-survival' [3H]thymidine autoradiography combined with TH immunocytochemistry, the first enzyme of catecholamine synthesis, was used. In males, TH-IR neurons originate in the ZI between embryonic days (E) 12 and 13, while in the PeVN and AN this process is prolonged until E16.

Activation of dopaminergic D1 receptors promotes morphogenesis of developing striatal neurons.

The early dopaminergic input from the midbrain may play an important role in the development of the basal ganglia. We therefore investigated whether and how dopamine affects the morphogenesis of striatal target neurons. Dissociated cell cultures of embryonic day 17 rat striatum were raised for seven days.

Sex differentiation of rat hippocampal GABAergic neurons.

In order to analyse mechanisms of sex differentiation of the hippocampus at the cellular level, the differentiation of hippocampal GABAergic neurons was studied in vitro. Serum-supplemented and serum-free dissociated cell cultures were raised from the hippocampus of embryonic day 17 male and female rat embryos for up to 14 days in vitro. This time period roughly corresponds to the critical phase for sex differentiation of the rat brain as determined in vivo.

Pre- and postnatal development of dopaminergic neuron numbers in the male and female mouse midbrain.

Quantitative information about dopaminergic neuron numbers in the mesencephalon is needed to assess the significance of physiological cell death in the regulation of the development of this neural system. Therefore, stereological techniques were applied to determine absolute numbers of mesencephalic neurons immunoreactive to tyrosine hydroxylase during the ontogenetic period between embryonic day (E) 13 and postnatal day (P) 90. Male and female CBA/J mice were examined separately.

Genotype-dependent sex differentiation of dopaminergic neurons in primary cultures of embryonic mouse brain.

In order to investigate genetic factors that interfere with hormone-mediated sex differentiation of dopaminergic neurons, we raised sex-specific primary cultures from embryonic day 13 diencephalon (D) or mesencephalon (M) of three different strains of mice, NMRI, CBA/J, and BALBc/J. Part of the cultures were maintained for 6 or 13 days in vitro (DIV) in medium containing 17 beta-estradiol or testosterone. The cultures were analyzed for sex differences in numbers of tyrosine hydroxylase-immunoreactive neurons, endogenous dopamine (DA) levels, and specific uptake of [3H]DA.

Ontogeny of aromatase messenger ribonucleic acid and aromatase activity in the rat midbrain.

Estrogen formation catalyzed by neural aromatase is crucial for the sexual differentiation of the brain. Ontogenic expression of aromatase mRNA and aromatase activity were studied in male and female rat midbrains. Aromatase mRNA was transiently expressed in both sexes showing maximum levels on postnatal day (P)2 and being absent on P20 and in adults.

Transcription of the Y chromosomal gene, Sry, in adult mouse brain.

The Y chromosomal gene Sry encodes a putative transcription factor which appears to serve as a master switch initiating testicular development. Here we show that this gene is transcribed in hypothalamus, midbrain, and testis of adult male but not adult female mice. In contrast to its circular transcripts in adult testis, those in brain are linear and may be translated.

Effects of sex and estrogen on tyrosine hydroxylase mRNA in cultured embryonic rat mesencephalon.

In order to elucidate cellular events responsible for sex differentiation of the nigro-striatal system, we studied the influence of estrogen on the expression of tyrosine hydroxylase (TH) in sex-specific dissociated cell cultures of embryonic day 14 rat mesencephalon. Cultures were raised in the absence or presence of 17 beta-estradiol (10(-12) M) and hybridized with a [35S]oligonucleotide specific to TH. Cultured cells and tissues were probed for estrogen receptor (ER) transcripts by hemi-nested PCR.

Neurotoxicity of dopamine and protective effects of the NMDA receptor antagonist AP-5 differ between male and female dopaminergic neurons.

Neurodevelopmental and neurodegenerative disorders related to dopaminergic transmission typically exhibit a sex-specific prevalence. In order to investigate the underlying cellular mechanisms, primary cultures of dissociated embryonic rat midbrain were subject to a 24 h treatment with dopamine in concentrations between 1 and 1000 microM. Dopamine caused a dose-dependent loss of neurons and reduction of neurites immunoreactive to tyrosine hydroxylase with a LD50 of about 100 microM.

Activation of cultured rat hypothalamic dopaminergic neurons by long-term but not short-term treatment with prolactin.

Pituitary prolactin (PRL) secretion is inhibited by hypothalamic GABAergic and dopaminergic (DAergic) systems. PRL, in turn, appears to be capable of activating these neurons, thus, providing for a negative feedback regulation. We have recently shown that cultured hypothalamic GABAergic- but not DAergic neurons respond to PRL with a rapid increase in intracellular free calcium.

Differentiation of hypothalamic GABAergic neurons in vitro: absence of effects of sex and gonadal steroids.

The inhibitory neurotransmitter gamma-aminobutyric acid (GABA) is involved in the control of sexually dimorphic brain functions, such as pituitary secretion and reproductive behavior. Hypothalamic GABAergic systems in vivo exhibit sexually dimorphic functional properties. Sexual dimorphisms in the rat brain are currently thought to be brought about by the organizational influence of gonadal steroids during the perinatal developmental period.

Ontogeny of synaptophysin and synaptoporin in the central nervous system: differential expression in striatal neurons and their afferents during development.

The expression of the synaptic vesicle antigens synaptophysin (SY) and synaptoporin (SO) was studied in the rat striatum, which contains a nearly homogeneous population of GABAergic neurons. In situ hybridization revealed high levels of SY transcripts in the striatal anlage from embryonic day (E) 14 until birth. In contrast, SO hydridization signals were low, and no immunoreactive cell bodies were detected at these stages of development.

Uptake and subcellular distribution of azithromycin in human phagocytic cells. Demonstration of the antibiotic in neutrophil polymorphonuclear leucocytes and monocytes by autoradiography and electron microscopy.

Azithromycin (CAS 83905-01-5), a new azalide antibiotic, reached high concentrations in alveolar macrophages obtained from patients. Uptake of [14C] azithromycin by macrophages resulted in an intracellular/extracellular concentration ratio of approximately 634 after 90 min incubation. Electron microscopic autoradiography provided visual evidence for the marked uptake and localization of [3H] azithromycin in human neutrophils and monocytes isolated from healthy volunteers.

Sex steroids do not alter sex differences in tyrosine hydroxylase activity of dopaminergic neurons in vitro.

In order to distinguish the effects of genetic sex from those of sex hormones on the sexual differentiation of dopaminergic neurons, catecholamine synthesis was studied in gender-specific cultures of embryonic day-14 rat diencephalon. In addition to embryos from normal dams, embryos were used whose mothers had been treated with the estrogen antagonist tamoxifen or the testosterone antagonist cyproterone acetate on days 12 and 13 of gestation. Cultures from embryos of untreated dams were fed daily with a medium containing 17 beta-estradiol or testosterone.

Sex differences of hypothalamic prolactin cells develop independently of the presence of sex steroids.

There is evidence for a hypothalamic prolactin (PRL) system that expresses sexually dimorphic traits. The aim of this in vitro study is to gain an insight into the process of sexual differentiation of hypothalamic PRL cells. In particular, we wanted to determine whether sexual differentiation of these cells can occur independently of the surge of gonadal testosterone which, in the male rat embryo, takes place at embryonic day (E) 18 and is commonly believed to start the critical period of sexual differentiation of the brain.

Sex differences in densities of dopaminergic fibers and GABAergic neurons in the prenatal rat striatum.

On the basis of observations on dopaminergic neurons developing in gender-specific cultures of embryonic rat mesencephalon, we have hypothesized that as yet unknown sexual dimorphisms might be found in projection areas of dopaminergic neurons. Therefore we searched for possible sex differences in the striatum during the period when massive ingrowth of mesencephalic afferents occurs and the striatal gamma-aminobutyric acid (GABA)ergic neurons differentiate. Male and female rats of embryonic days (E) 16, 18, 20, and 21 were fixed by perfusion through the heart.