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Expression of nk2.1a during early development of the thyroid gland in zebrafish.

We show here that a zebrafish orthologue of the Thyroid Transcription Factor-1 (TTF-1), nk2.1a, is expressed in the developing thyroid gland. Using a fate mapping approach we found that an early nk2.1a expression domain in the endoderm adjacent to the heart follows morphogenetic movements of the lower jaw, ending up in the region in which the mature thyroid gland is located. We therefore suggest that nk2.1a labels the thyroid precursor cells from somitogenesis stages onwards.

Animals↗

The early development of Kraepelin's ideas on classification: a conceptual history.

Psychiatry still lives in a Kraepelinian world and scholarship on the work of Kraepelin has consequently been unhistorical. Selective exegesis of the various editions of his textbook has led to a rigid view of his contribution. However, Kraepelin lived and wrote during an important period of European intellectual history and his work can only be understood against this background. This paper analyses the development of his views in terms of the 'Research Programme' he planned early in his life and whose objective was the creation of a stable description and classification of the psychoses. This Kraepelin eventually achieved by longitudinally studying patient cohorts in terms of methodological criteria such as illness course and incurability. In the event, this methodology allowed him to identify by correlation 'clinical pictures' that both represented the 'essence' of the disease and provided a taxonomic criterion. Although avowedly atheoretical, Kraepelin thus managed to construct (influenced by Kahlbaum and Wundt) an empirical support for his Kantian categorization of the psychoses. A discussion of the cultural variables that moulded these ideas is included.

Germany↗

Early development of delinquency within divorced families: evaluating a randomized preventive intervention trial.

This paper reports on an experimental test of coercion theory early onset model of delinquency. Results are from the Oregon Divorce Study-II, a randomized preventive intervention trial with a sample of 238 recently separated mothers and their sons in early elementary school. The objective was to experimentally manipulate parenting variables hypothesized to influence development of delinquent behaviors. Multiple-method assessment spanned 36 months. Because the intervention focused on parent training, we expected that any intervention effects on changes in child outcomes would be mediated by hypothesized intervening mechanisms. Linear growth models showed significantly greater reduction in boys' delinquency and deviant peer affiliation in the experimental group relative to the controls. Subsequent models using no method overlap in constructs demonstrated that the intervention effect on delinquency operated through growth in parenting and reduction in deviant peer affiliation.

Adult↗

Effects of 5 azacytidine on DNA methylation and early development of sea urchins and ascidia.

5-azacytidine (5-azaCR), an analogue of cytidine, inhibits nuclear DNA methylation in early sea urchin embryos. This inhibition is specific and dose-dependent. Exposure of sea urchin embryos at any stage between one-cell and blastula, to micromolar quantities of 5-azaCR invariably inhibits development beyond the blastula stage. In a substantial number of embryos arrested at the blastula stage, spicule formation proceeds although other morphological differentiation is lacking. No significant effect on development is seen if sea urchin embryos are exposed to 5-azaCR at post-blastula stages. 5-azaCR also inhibits the development of a mosaic egg such as the ascidian Phallusia mammilata at the blastula stage, indicating that both regulative (sea urchin) and mosaic (ascidian) embryos respond more or less similarly to 5-azaCR treatment.

Animals↗

Temporal expression of three mouse lens fiber cell membrane protein genes during early development.

PURPOSE: Three different lens fiber cell intrinsic membrane proteins, MIP (Major Intrinsic Protein), MP19, and connexin50 (Cx50), have separately been implicated as causative candidates for congenital cataracts. The aim of this study was to determine gene transcript expression of these three proteins during successive stages of mouse embryonic development. METHODS: Total RNA was prepared from mouse embryos taken at days 9-10 (E9-E10) of gestation, heads of day 11-13 (E11-E13) embryos, and lenses of adult mice. Coupled reverse transcriptase-polymerase chain reaction (RT-PCR) was used to determine gene transcript expression of MIP, Cx50, and MP19 during embryonic development. The products of RT-PCR were further cloned into the TOPO(TM) TA vector, and further analyzed by double strand nucleotide sequencing. RESULTS: Cx50 gene expression was observed throughout the developmental period observed (E9-E13). MIP transcripts were first observed at mouse embryonic day 11.5 (E11.5) and synthesis continued throughout the developmental period observed. The gene for MP19 (Lim2) begins to express at mouse embryo day 12 (E12) and synthesis continued throughout the developmental period observed. mRNA levels for all three proteins appear to remain steady from these early embryonic stages through adulthood. CONCLUSIONS: The identified early expression of Cx50, MIP, and Lim2 transcripts in mouse embryonic stages suggests that all three proteins play very important, probably quite different, roles in lens fiber cell differentiation. Variation in the temporal expression of these three genes during the course of development suggests a critical gene-coordinated regulation throughout lens fiber cell development. These three genes clearly play important roles in early normal lens development since it is known that mutations in the sequence of each membrane protein results in cataractogenesis.

Animals↗

Perinatal supply and metabolism of long-chain polyunsaturated fatty acids: importance for the early development of the nervous system.

The long-chain polyunsaturated fatty acids, arachidonic (AA) and docosahexaenoic acid (DHA), are essential structural lipid components of biomembranes. During pregnancy, long-chain polyunsaturated fatty acids (LC-PUFA) are preferentially transferred from mother to fetus across the placenta. This placental transfer is mediated by specific fatty acid binding and transfer proteins. After birth, preterm and full-term babies are capable of converting linoleic and alpha-linolenic acids into AA and DHA, respectively, as demonstrated by studies using stable isotopes, but the activity of this endogenous LC-PUFA synthesis is very low. Breast milk provides preformed LC-PUFA, and breast-fed infants have higher LC-PUFA levels in plasma and tissue phospholipids than infants fed conventional formulas. Supplementation of formulas with different sources of LC-PUFA can normalize LC-PUFA status in the recipient infants relative to reference groups fed human milk. Some, but not all, randomized, double-masked placebo-controlled clinical trials in preterm and healthy full-term infants demonstrated benefits of formula supplementation with DHA and AA for development of visual acuity up to 1 year of age and of complex neural and cognitive functions. From the available data, we conclude that LC-PUFA are conditionally essential substrates during early life that are related to the quality of growth and development. Therefore, a dietary supply during pregnancy, lactation, and early childhood that avoids the occurrence of LC-PUFA depletion is desirable, as was recently recommended by an expert consensus workshop of the Child Health Foundation.

Arachidonic Acid↗

Early development of protein metabolic perturbations in the liver and skeletal muscle of tumour-bearing rats. A model system for cancer cachexia.

In rats into which a fast-growing ascites hepatoma (Yoshida AH-130) had been transplanted, tumour growth elicited a marked loss of body weight until the animal's death in about 2 weeks. Overall tissue protein metabolism was simultaneously studied in vivo in the gastrocnemius muscle and liver after labelling with [14C]bicarbonate. Early and progressive atrophy developed in the gastrocnemius muscle, the underlying metabolic imbalance being expressed by an elevation in the apparent protein-degradation rate, with no changes in the apparent synthesis rate. A transient hyperplastic response preceded waste in the liver, both states being associated with alterations in protein-degradation rate: an initial decrease during liver growth, then an acceleration as liver regressed. Protein-synthesis rates, virtually unchanged during liver growth, were elevated in the subsequent phase, although not sufficient to balance the enhanced breakdown. Thus, in the tumour host tissues examined, altered states of protein turnover appeared to result mostly from changes in rates of protein breakdown. In sharp contrast with the negative protein balance in the host, the ascites hepatoma cells had the ability to grow or at least, in advanced stages, to maintain a stationary state.

Animals↗

[A preliminary study on early development of goat (Capra hircus) reconstituted embryos].

Goat reconstituted embryos (REs) have been produced by electrofusion-mediated nuclear transplantation method. Single cell derived from normal embryos or REs developed to 8-cell morula stage or the inner cell mass (ICM) of early blastocyst stage was used to fuse with enucleated mature egg (26-28 hrs after injection of LRH). According to the results summarized in table 1 and 2, we decided to adopt the method to embed REs in agarose and then transfer into goat oviduct lumen of host mother for 4-6 days in vivo culture. Normal fertilized eggs seem to develop synchronously, but that of REs are not. Tables 4 and 5 reveal that REs and embryos reconstituted successively can develop normally, no significant difference was found among their development rates. All these experimental results indicate that nuclei of some blastomeres from normal embryos or REs (derived from eight cells to morula or ICM) retained their totipotency for further development. These nuclei can be reprogrammed in host ooplasm, and developed to term.

Animals↗

Transport and steady-state concentration of plasma proteins in the vitreous humor of the chicken embryo: implications for the mechanism of eye growth during early development.

The nature and origin of the proteins of the vitreous humor were examined in chickens during embryonic and early posthatching stages. The major proteins of the vitreous humor were similar in electrophoretic mobility to plasma proteins at all ages examined. Earlier studies from our laboratory and experiments described below showed that plasma proteins continuously entered and left the eye throughout its development. From these data it was concluded that the majority of vitreous-humor proteins were derived from the blood. The protein concentration of the vitreous humor was 13% of that of the plasma from embryonic Days 6 through 15 (E6 through E15). After E15, the relative protein concentration in the vitreous humor declined with respect to the plasma and reached 4% of the plasma protein concentration at hatching. Several possibilities were considered to account for how proteins can rapidly enter and leave the eye, yet maintain a steady-state concentration in the vitreous humor that is less than one-seventh of that in the blood.

Animals↗

Bcl-2 expression during T-cell development: early loss and late return occur at specific stages of commitment to differentiation and survival.

During T-cell development CD3-CD4-CD8- (double-negative) thymocytes proliferative and produce an enormous number of CD3loCD4+CD8+ (double-positive) thymocytes which are destined to die intrathymically unless rescued by positive selection. Those which survive become mature CD3hiCD4/8+ (single-positive) cells and are the precursor of peripheral blood lymphocytes. The product of the bcl-2 protooncogene has been implicated in preventing programmed cell death and is required for prolonged lymphocyte survival following maturation. Previously we and others have reported that Bcl-2 protein expression is biphasic, being high in proliferating double-negative stem cells, low in all double-positive thymocytes except for 1-5% of these cells, and restored in mature, single-positive thymocytes. However, it remained unclear which signaling and selection events regulate Bcl-2 during T-cell maturation. Now we have utilized four-color flow cytometry in normal and genetically altered mice for a detailed analysis of Bcl-2 expression as it relates to T-cell receptor (TCR) expression and positive selection. These studies show that (i) expression of a transgenic TCR in double-negative thymocytes does not lead to premature loss of Bcl-2; thus, Bcl-2 downregulation is not solely due to TCR expression; (ii) Bcl-2 expression is lost at the early transitional CD3-/loCD4-CD8+ stage, prior to expression of CD4; (iii) the Bcl-2+ double-positive thymocytes are those which have undergone positive selection; and (iv) upregulation of Bcl-2 during positive selection requires participation of the CD4 or CD8 co-receptor. These results demonstrate that Bcl-2 and TCR expression are regulated independently during T-cell development, and suggest a role for the CD4 or CD8 co-receptor in Bcl-2 induction during positive selection.

Animals↗

Phenotypic expression of monoamines and GABA in the early development of human telencephalon, transient or not transient.

We review the phenotypic expression of molecules involved in monoamine and GABA neurotransmission in the developing human brain. Recent experimental reports have analyzed neurotransmitter signaling before the onset of synaptogenesis, which could act to influence early developmental events such as proliferation, migration, and differentiation of animal brain development. Such signaling may also occur in human development. The expression of molecules involved in neurotransmission in precocious human brain may reflect either the differentiation of a permanent neurotransmitter system of the adult brain or transient expression to serve specific developmental functions different from those in the adult brain. We review the changes observed in the expression of various catecholamine markers such as tyrosine-hydroxylase (TH) immunoreactivity in various neuronal populations of the developing human telencephalon. The specific transporter for serotonin, serotonin transporter (SERT) has been detected in fibers of the internal capsule (IC) during the restricted time period of 12-14 gestational weeks in humans. These serotonin-containing fibers do not correspond to serotoninergic ascending axons from the raphe nuclei. They may be the human counterpart of the thalamo-cortical axons that have been shown to uptake serotonin during the critical period of development of the sensory systems in rodents. GABA phenotypes are expressed in numerous cells of the human ganglionic eminence (GE) and cerebral wall at the end of the embryonic period proper. These results are similar to that described at comparable developmental stages in the mouse and support the hypothesis of an early migration from ganglionic progenitors in humans. But one cannot exclude a transient expression of GABA within the post-mitotic neurons, which could influence early developmental events. In conclusion, data showing the phenotypic expression of molecules in discrete areas of the brain at various points in the protracted human development require careful interpretation.

Animals↗

Early development of retinal ganglion cell dendrites in the marsupial Setonix brachyurus, quokka.

The dendritic morphology of retinal ganglion cells was studied in flat-mounted retinae of the marsupial Setonix brachyurus, quokka. In the adults, horseradish peroxidase (HRP) was applied to the vitread surface of flattened retinae. Wide-, large-, medium-, and small-field classes appeared to correspond to gamma, alpha, delta, and beta cells, respectively, in the cat (Boycott and Wässle, J. Physiol. 249:397-419, 1974). To reveal the early stages of dendritic development, HRP was placed on the optic nerve of isolated eye cups from the day of birth to postnatal day (P) 63 when the area centralis is beginning to form (Dunlop and Beazley, Dev. Brain Res. 23:81-90, 1985). Youngest cells lacked dendrites and had an elongate soma in the cytoblastic layer with an endfoot contacting the ventricular surface. Once in the ganglion cell layer, the soma was rounded and dendrites appeared as short, unbranched processes. Most cells were asymmetric or "polarised" with the axon arising from the side nearest the optic disk and dendrites from the opposite side. Polarity was maintained in cells with longer, branched dendrites. A small proportion of cells exhibited a reversed polarity in which the axon arose from the side nearest the retinal edge and dendrites towards the disk. Cells appeared to acquire an approximately symmetric, adult-like tree by the addition of new primary dendrites between the existing ones and the axon hillock. Wide-, large-, medium-, and small-field cells were evident from P6, P25, P31, and P40, respectively. Spines were observed on dendrites and axons during development but were rare in the adult. Some dendro-axons were seen at all ages examined. The existence of an initial axodendritic polarity in retinal ganglion cells supports the hypothesis that the axon hillock is the determinant of dendritic geometry (Maffei and Perry, Dev. Brain Res. 41:185-194, 1988). Polarity may also contribute to the establishment of "radial orientation" in which the long axis of the elliptical dendritic tree of cells outside the area centralis points towards central retina and the weighted centre is displaced towards the retinal periphery (Leventhal and Schall, J. Comp. Neurol. 220:465-475, 1983).

Animals↗

Astrocyte metabolism is disturbed in the early development of experimental hydrocephalus.

The proper diagnosis of the arrested or the progressive form of hydrocephalus has a critical impact on treatment, but remains difficult. The assessment of early changes in cerebral metabolism might help in the development of adequate non-invasive diagnostic tools. This study examined the alterations in label incorporation in neurotransmitter amino acids and other compounds in kaolin-induced progressive hydrocephalus in rats by means of magnetic resonance spectroscopy (MRS) combined with the administration of [1-13C]glucose and [1,2-13C]acetate. Some 2, 4 and 6 weeks after kaolin injection into the cisterna magna, cerebrum, brainstem and cerebellum were dissected. Interestingly, labelling of most amino acids derived from [1-13C]glucose showed no alterations, whereas labelling from [1,2-13C]acetate was affected. Two weeks after induction of hydrocephalus the taurine concentration was decreased, whereas the concentration of [1,2-13C]lactate was increased in the cerebrum and that of [1,2-13C]GABA in the brainstem. Furthermore, labelling from [1,2-13C]acetate was significantly decreased in [4,5-13C]glutamate, [1,2-13C]glutamate and [1,2-13C]GABA in cerebrum from 4 weeks after hydrocephalus induction. The concentration of N-acetylaspartate, a neuronal marker, was unchanged. However, labelling of the acetyl group from [1-13C]glucose was decreased in cerebellum and brainstem at 6 weeks after the induction of hydrocephalus. As glucose is metabolized predominately by neurones, whereas acetate is exclusively taken up by astrocytes, these results indicate that mostly astrocytic, and only later neuronal, metabolism is disturbed in the kaolin model of hydrocephalus. If verified in patients using in vivo MRS, impaired astrocyte metabolism might serve as an early indication for operative treatment.

Acetates↗

Activation of a beta-tubulin gene during early development of the plasmodium in Physarum polycephalum.

Uninucleate amoebae of Physarum polycephalum strain CL undergo apogamic development to form multinucleate plasmodia via an intermediate stage of large, uninucleate cells irreversibly committed to plasmodial development. This amoebal-plasmodial transition involves major changes in tubulin gene expression and the organization of microtubular structures. We analysed the expression of the betC locus, which encodes the plasmodial-specific beta 2-tubulin, during plasmodial development. A key question addressed was the timing of expression of betC in relation to the last open mitosis of the amoeba and the first closed mitosis of the plasmodium during the transition. Culture conditions were improved to yield partly synchronous differentiating cultures containing 50-60% committed cells, in order to facilitate biochemical analysis of development. Northern blotting indicated that betC RNA was virtually absent from amoebae and from early differentiating cultures. However, betC transcripts could already be detected in differentiating cultures containing only 0.1% of committed cells; the relative amount of betC transcripts increased as the percentage of committed cells in differentiating cultures increased. In fully developed plasmodia, there was at least a 330-fold increase in the betC transcript level compared to that in amoebae. We conclude that betC is activated during the amoebal-plasmodial transition immediately before or during the commitment event. Small amounts of beta 2-tubulin polypeptide could first be detected by Western blotting around the stage of the first closed mitosis. Thus beta 2-tubulin may participate in the first closed mitosis that committed cells undergo during their development into plasmodia.

Cell Differentiation↗

Early development of the serotonergic and dopaminergic nervous system in Spisula solidissima (surf clam) larvae.

We have defined the development of the serotonergic and dopaminergic components of the central nervous system in the early Spisula solidissima (surf clam) embryo using HPLC and immunocytochemistry. HPLC analysis reveals norepinephrine, dopamine, and serotonin are present at 24 h post-fertilization. Immunocytochemistry shows that the serotonergic nervous system emerges during the late trochophore stage with the development of a single serotonergic cell, C/A1, in the cerebral/apical ganglion. After 48 h, a second serotonergic cell forms, C/A2, which is connected to C/A1 by two serotonergic processes, and a single serotonergic cell emerges in the visceral ganglion, V1. At 72 h, a new serotonergic cell body develops in the cerebral/apical ganglion, C/A3. After 96 h, the cerebral/apical ganglion and visceral ganglion are connected by a serotonergic process. Expression of the dopamine receptor, D2, begins by 24 h with a generalized expression in the region of the developing gut. D2 expression in the gut ceases by 48 h. At 48 h, a network of fibers forms dorsolateral to the mouth. By 72 h, D2 expressing projections emerge from this network.

Animals↗

Cytoskeletal changes during oogenesis and early development of Xenopus laevis.

The frog oocyte is well known for studies on the control of gene expression, but has been used much less in studies on the cytoskeleton. However, frog oocytes are very large single cells, whose cytoplasmic movements and asymmetries are fundamental to the correct development of the subsequent embryo. One particular example of asymmetrically distributed cytoplasm is germ plasm, thought to be important in the formation of the germ line. Data are presented that show that germ plasm is highly concentrated mass of cytoskeletal elements, which include tubulin, and an intermediate filament protein of molecular weight 55 X 10(3). The distribution of these molecules has been studied during oogenesis and during early post-fertilization development. The implications of these findings are discussed.

Animals↗

Synthesis of the foetal protein fetuin by early developing neurons in the immature neocortex.

The presence of the foetal protein fetuin has previously been demonstrated by immunocytochemistry to be specifically confined to the primordial plexiform layer, the early cortical plate and subplate zone cells in the developing neocortex of a number of species. In order to investigate its origin there, we have applied in situ hybridization in paraffin sections of Bouin's fixed foetal sheep brain, using a short anti-sense oligonucleotide probe. The distribution of fetuin mRNA has been compared with that of the protein by using anti-fetuin antibodies and immunocytochemistry. This allowed us to confirm that fetuin is synthesised initially in cells of the primordial plexiform layer and subsequently cortical plate and subplate cells. On the other hand, cells in the ventricular zone that are fetuin (protein) positive do not contain detectable fetuin mRNA. The time course of fetuin mRNA expression in the developing neocortex follows closely the previously described pattern of fetuin (protein) distribution in the sheep brain, apart from its absence from the ventricular zone where its origin is probably by uptake from cerebrospinal fluid.

Animals↗

Fibrillarin is essential for early development and required for accumulation of an intron-encoded small nucleolar RNA in the mouse.

Fibrillarin, a protein component of C/D box small nucleolar ribonucleoproteins (snoRNPs), directs 2'-O-methylation of rRNA and is also involved in other aspects of rRNA processing. A gene trap screen in embryonic stem (ES) cells resulted in an insertion mutation in the fibrillarin gene. This insertion generated a fusion protein that contained the N-terminal 132 amino acids of fibrillarin fused to a beta-galactosidase-neomycin phosphotransferase reporter. As a result, the N-terminal GAR domain was present in the fusion protein but the methyltransferase-like domain was missing. The ES cell line with the targeted fibrillarin allele was transmitted through the mouse germ line, creating heterozygous animals. Western blot analyses showed a reduction in fibrillarin protein levels in the heterozygous knockout animals. Animals homozygous for the mutation were inviable, and massive apoptosis was observed in early Fibrillarin(-/-) embryos, showing that fibrillarin is essential for development. Fibrillarin(+/-) live-born mice displayed no obvious growth defect, but heterozygous intercrosses revealed a reduced ratio of +/- to +/+ mice, showing that some of the Fibrillarin heterozygous embryos die in utero. Analyses of tissue samples and cultured embryonic fibroblasts showed no discernible alteration in pre-rRNA processing or the level of the U3 snoRNA. However, the level of the intron-encoded box C/D snoRNA U76 was clearly reduced. This suggests a high requirement for snoRNA synthesis during an early stage in development.

Amino Acid Sequence↗