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PubMed2005

Oligodendrocyte development and myelination in GFP-transgenic zebrafish.

Green fluorescent protein (GFP) transgenic zebrafish technology has been employed to directly visualize and analyze dynamic developmental processes, such as cell migration and morphogenesis. Stable transgenic zebrafish that express GFP in oligodendrocytes can be a valuable tool to visualize complex myelination processes in vivo, as well as to conduct rapid mutagenesis screens for defective myelination mutants. We investigated whether two myelin gene promoters, the zebrafish P0 promoter and the mouse proteolipid protein (PLP) promoter, drive GFP expression in zebrafish oligodendrocytes. Transiently, both promoters drive enhanced GFP (EGFP) expression in morphologically identifiable oligodendrocytes, premyelinating oligodendrocytes, and possible oligodendrocyte precursors. We have established a stable transgenic zebrafish line, tg(plp:EGFP) zebrafish, at the F1 generation, which expresses enhanced GFP (EGFP) driven by the mouse PLP promoter. In this transgenic line, EGFP-expressing cells are visually detectable around 24-hr postfertilization (hpf), and later at 54 hpf, these cells start exhibiting the clear morphologic characteristics of oligodendrocytes. Shortly afterward, EGFP-expressing oligodendrocytes establish a ventral dominant distribution pattern throughout the central nervous system. This transgenic zebrafish line is likely to serve as a useful tool, in which normal myelination as well as abnormal myelination can be recorded under time-lapse confocal microscopy. Furthermore, it has the potential to greatly facilitate mutagenesis screening for novel dysmyelinating mutants.

Animals↗
PubMed1993

A comparison of the development of neuropeptide and MAP2 immunocytochemical labeling in the macaque visual cortex during pre- and postnatal development.

The appearance of Substance P (SP) and Neuropeptide Y (NPY) has been studied using light microscopic immunocytochemical labeling throughout the complete developmental span of Macaca nemestrina monkey striate cortex. In the adult, 80% of the NPY+ neurons occur in the white matter (WM) and most of the remainder are medium to large multipolar neurons in layer 2. Fibers occur in all layers except 4C and are very numerous, given the relatively small number of NPY+ cell bodies. NPY+ neurons first were seen at embryonic day (E) 75. Most neurons were in the intermediate zone (IZ), but a few were in the immature cortical plate (CP). An adult-like distribution was present by E125 for neurons and by birth for fibers, but fiber staining intensity and number increased to postnatal year 1 (P1yr). In adult cortex, numerous SP+ nonpyramidal neurons were present in layers 2-6 and WM, but SP+ fibers were surprisingly infrequent. During development, significant numbers of SP+ neurons were not seen in the CP until E113-125. Later prenatal ages had a prominent plexus of SP+ cell bodies and fibers at the layer 5/6 border. This plexus disappeared by P12wk due to either down-regulation of SP or cell death. SP+ neurons in IZ/WM were very sparse until birth after which they increased in number and staining intensity up to P1yr, suggesting a postnatal up-regulation of SP in a preexisting WM subpopulation. Cell densities were determined for SP, NPY, and the neuron-specific marker microtubule-associated protein 2 (MAP2) to clarify the developmental dynamics of IZ/WM neurons. MAP2+ cell densities in WM peaked around birth and then declined 20% in the outer half and 77% in the inner half of WM. SP+ cell density rose 57% from birth to P20wk and then declined 20% into adulthood. NPY+ cell density was fairly constant prenatally and then rose 300% by adulthood. Neuropeptide cell density changes took place predominantly in the outer WM. These data indicate that cell death does occur in the general population of monkey striate cortical WM neurons. In contrast, both SP+ and NPY+ cells are characterized by minimal cell death and a late expression of neuropeptides which causes an increase in neuropeptide+ cell density in postnatal WM.

Animals↗
PubMed1996

Comparison of the growth and fate of fetal spinal iso- and allografts in the adult rat injured spinal cord.

Most studies investigating early fetal CNS graft-host interactions and host immune responses have been performed using intracerebral transplantation paradigms. The purpose of this study was to establish the early developmental dynamics of fetal graft integration with the injured host spinal cord and to determine whether fetal allografts in this environment are subject to rejection. ACI rat fetal spinal cord (FSC) tissue was grafted into acute lesion cavities of adult WF rat spinal cords. Graft development and/or rejection was followed from 1 to 45 days posttransplantation with morphometric, histological, and immunocytochemical methods. We determined that all FSC grafts in acute resection lesions of the adult rat spinal cord undergo an early substantial cellular attrition, but following favorable attachment to healthy host tissue margins, they rebound and grow to fill the lesion cavity by approximately 45 days. We also determined that FSC allografts into nonimmunosuppressed adult recipients are consistently rejected, but only after an early period of growth and maturation. The onset of rejection is characterized by extensive cellular infiltration coincidental with graft and host MHC antigen expression. The implications of delayed graft development and graft-host integration are discussed relative to interconnectivity and long-term potential for graft-derived benefits. The observed rejection response was characteristic of first-order allograft rejection and underscores a lack of immunological privilege in the microenvironment of the injured spinal cord.

Age Factors↗
PubMed1990

Audiovocal interactions during development? Vocalisation in deafened young horseshoe bats vs. audition in vocalisation-impaired bats.

Horseshoe bats (Rhinolophus rouxi) were deafened in their 3rd-5th postnatal week. Subsequently their vocalisations were monitored to evaluate the impact of audition on the development of echolocation pulses. Hearing impairment affected the echolocation pulses as follows: the frequency of the constant frequency (CF) component was altered by between +4 kHz and -14 kHz, and the dominance of the second harmonic of the pulses was neutralised by a relative increase in intensity of the first and third harmonics. A second experiment focused on possible influences of acoustical self-stimulation with echolocation pulses on the establishment of auditory fovea representation in the inferior colliculus (IC). Frequency control of echolocation pulses was disrupted by larynx denervation. Thereafter, the bats produced multiharmonic echolocation signals (4-11 harmonics) varying in frequency. IC tonotopy, however, as monitored by stereotaxic electrophysiology, showed the same developmental dynamics as seen in control specimens (Fig. 10). Both experiments indicate that throughout postnatal development echolocation pulses are under auditory feedback control, whereas maturation of the auditory fovea and shifts in its frequency tuning represent an innate process. The significance of this postnatal development might be the adjustment of the vocal motor system of each bat to the frequency of its 'personal' auditory fovea.

Animals↗
PubMed1989

Resistance and countertransference in child and adolescent psychotherapy.

Countertransference brings to light the influence of both anxiety and family processes on the child's or adolescent's maturation and character development. The significance of developmental, dynamic, and family-interpersonal factors have all been stressed as part of the latent communication inherent in resistance. Family influences on resistance and countertransference have to be the subject of analytic scrutiny. The engagement of children and adolescents in the therapeutic process seriously threatens parents' defensive operations both as couples and as individuals. Countertransference responses afford the opportunity not only to clarify the meaning of resistances but also to pinpoint anxieties that have been reexperienced by the child or adolescent with the analyst at points of threat and disorganization. The twofold therapeutic task with children and adolescents consists of delineating and counteracting the family's negative impact on the child's development and self, in addition to addressing the children's contribution to their own pathological traits and immaturities. Countertransference anxiety confirms the dynamic and family implications of the analyst's participation with the patient. Countertransference resistance suggests the analyst's lack of openness in investigating how the treatment has evoked the child's anxiety, the parent's anxiety, and the analyst's anxiety. Resistance-transference-countertransference exchanges reveal the child's or adolescent's efforts to move towards health while emerging from the internalized family.

Adolescent↗
PubMed1990

Primitive megalencephaly in children: natural history, medium term prognosis with special reference to external hydrocephalus.

We studied 74 children with primitive megalencephaly retrospectively with attention directed to familial megalencephaly, birth history, enlarged pericerebral subarachnoid space (SAS) (idiopathic external hydrocephalus), head and statural growth dynamics, developmental and school prognosis, morphological findings and development of subdural haematoma. In the megalencephalic children, no significant differences were found between those with normal or those with enlarged pericerebral SAS. Out of 62, 31 children (50%) were already megalencephalic at birth. Of 74, 37 children (50%) showed variable degrees of developmental delay which in 18 was transient. Eight out of 74 were mentally retarded. Of 52 children at school age, 42 attend normal schools and 10, of whom 7 are mentally retarded, attend special schools. Three children showed subdural haematoma resulting from apparently minor trauma or occurring spontaneously. We suggest that idiopathic external hydrocephalus represents a variant of primitive megalencephaly with transient increase of intracranial pressure and that it could predispose to the development of idiopathic (spontaneous or non-traumatic) subdural haematoma.

Brain↗
PubMed2001

Juvenile-hormone-dependent interaction of actin and spectrin is crucial for polymorphic differentiation of the larval honey bee ovary.

Programmed cell death in the worker ovary of Apis mellifera reduces the number of ovarioles during metamorphosis from 150-200 primordia to less than 10. In contrast, practically all ovarioles in the ovary of queens survive to the adult stage. The correct formation and persistence of polyfusomes has been suggested as a critical factor for ovariole survival. We have analyzed the developmental dynamics of F-actin and alpha-spectrin in fusomes of queen and worker larvae, and in juvenile-hormone-treated worker larvae. Small fusomes containing actin and spectrin can be detected in the ovaries of fourth instar larvae in both castes. After molting to the fifth instar, the actin-spectrin association persists in the enlarged fusomes of queen ovarioles. In workers, actin dissociates from the fusomal and cortical alpha-spectrin. Coinciding with the appearance of apoptosis markers, large agglomerates of actin are detectable in worker ovarioles. Treatment of fourth-instar worker larvae with juvenile hormone rescues ovarioles from apoptosis and maintains the actin-spectrin association. Juvenile-hormone-dependent actin-spectrin interaction is thus one of the earliest steps in the differentiation of a polymorphic ovary. Plasticity in ovariole numbers as a result of hormone-dependent fusome formation may be a more widespread phenomenon in insects, extending beyond caste polymorphism in highly eusocial Hymenoptera.

Actins↗
PubMed2000

Developmental cognitive neuropsychology of number processing and calculation: varieties of developmental dyscalculia.

This article provides a brief overview about the current state of cognitive developmental neuropsychology of developmental dyscalculia (DD) as well as results from a Zurich study that investigates different subtypes of DD according to various aspects of numerical abilities that are impaired or preserved. The differential effects of impairments of one particular numerical area on the development of other numerical abilities are highlighted in the case of a 17 year old boy with severe DD and Developmental Gerstmann Syndrome. A comprehensive model of developmental dynamics of number processing and calculation abilities will be proposed in the last section with respect to the development of intelligence theory.

Adolescent↗
PubMed2007

Lamins A and C are present in the nuclei of early porcine embryos, with lamin A being distributed in large intranuclear foci.

Gametogenesis and embryogenesis are dynamic developmental stages marked by extensive modifications in the organization of the genome and nuclear architecture. In the literature it is conveyed that only B-type lamins are required in these early stages of development and that A-type lamins are not present or required until differentiation of specific cell types associated with specialized tissue is initiated. To assess the presence of nuclear structures that are putatively involved in genome regulation, we investigated the distribution of lamin proteins throughout the early stages of porcine embryonic development, using testes tissue sections, oocytes and in-vitro fertilized (IVF) porcine embryos and employing anti-lamin antibodies. We have shown that anti-lamin A staining is present at the one-cell, two-cell, four-cell, and six- to eight-cell stages of early porcine embryo development, but diminishes at the morulae and blastocyst stages. Large intranuclear anti-lamin A foci are prominent in the early preimplantation stages. Both anti-lamin A/C and anti-lamin B staining were clearly present in all embryonic stages. Immature porcine oocytes revealed lamin rings using the monoclonal anti-lamin A/C antibody and many immature oocytes exhibited a pale rim staining pattern with anti-lamin A antibody. A-type lamins were not observed in sperm precursor cells. Thus, we have shown that A-type lamins and B-type lamins are present at the nuclear envelope in very early porcine embryos and that lamin A is also found in large intranuclear aggregates in two-cell to eight-cell embryos but is lacking from later embryonic stages.

Animals↗
PubMed1995

Differentiation of photoreceptors, glia, and neurons in the retina of the cichlid fish Aequidens pulcher; an immunocytochemical study.

Light-microscopic immunocytochemistry was carried out to investigate the developmental dynamics of several neurochemical markers in the retina of blue acara (Aequidens pulcher). As a rule, double-label experiments were performed in order to determine the absolute and relative timing of the appearance of these markers. The diameter of eye-ball (from 0.6 to 1.2 mm) and the body length (from 4.6 to 9.4 mm) enlarged in parallel during the observation period of 2 to 9 days after spawning (day 2-9); hatching took place usually on day 2. Immunoreactive proliferating cell nuclear antigen (ir-PCNA) was present in all neuroblasts (the embryonic homogeneous cell stage; day 1.0-2.0), but was lost progressively in a center-to-periphery and apparent proximal-to-distal sequence as the cells and layers differentiated. In late larvae and juveniles, ir-PCNA was confined to a ring of dividing neuroblasts at the retinal margin and to a population of scattered rod precursors in the outer nuclear layer. Immunoreactive structures of representative antigens progressively appeared after ir-PCNA had decayed. Around hatching, at the synaptic separation stage (day 2.0-2.5), luteinizing hormone-releasing hormone-ir centrifugal fibers, visinin-ir cones, glial fibrillary acidic protein-ir structures and gamma-aminobutyric acid-ir cell bodies appeared, which were followed by the emergence of rhodopsin-ir rods and tyrosine hydroxylase-ir interplexiform cells (on day 2.5-3.0) and serotonin-, neuropeptide Y- and substance P-ir amacrine cells (on day 3.0-4.0). The results indicate that photoreceptor cells, and especially rods start to differentiate at an earlier stage of retinogenesis than has previously been proposed. In addition, an extraretinal tissue in the brain identified as the prospective pineal organ was found to be visinin- and rhodopsin-immunoreactive on day 1.5-2.0 before these photoreceptor-specific antigens became positive in the retina.

Animals↗
PubMed1996

Exposure of postnatal rats to glucocorticoids suppresses the development of choline acetyltransferase-immunoreactive neurons: role of adrenal steroids in the development of forebrain cholinergic neurons.

Rat forebrain cholinergic neurons undergo dynamic developmental changes, showing a continuous increase in choline acetyl-transferase (ChAT) activity, during the early postnatal period. In adult rats, increases in circulating glucocorticoids result in decreases in activity of forebrain neuronal ChAT, thus raising the possibility that postnatal development of forebrain cholinergic neurons results from low levels of these hormones. In the rat, the first 2 weeks postnatally are characterized by very low levels of adrenal steroids. To understand the role of endogenous glucocorticoids in the development of forebrain cholinergic neurons, we studied the changes in ChAT immunoreactivity in forebrain cholinergic neurons of postnatal rats which had received daily subcutaneous injection of the synthetic glucocorticoid dexamethasone for 8 days. Immunohistochemical analysis of the rat pup forebrain revealed nearly complete obliteration of ChAT-immunoreactive neurons in the caudate-putamen, especially in the dorsolateral region of the rostral part. At the same stage, treatment with dexamethasone induced significant decreases in both number and length of dendritic branches of ChAT-immunoreactive neurons in the substantia innominata and the diagonal band. Despite the marked alterations in the caudate-putamen and diagonal band, the ChAT-immunoreactive neurons in other forebrain structures such as globus pallidus and medial septal nucleus showed little change. In the caudate-putamen, Nissl staining and specific labeling for nuclear DNA fragmentation exhibited no increase in number of dying cells following dexamethasone treatment, therefore indicating that the loss of ChAT immunoreactivity is not due to glucocorticoid-induced cholinergic cell death. These observations demonstrated that the development of cholinergic neurons in rat pups was inhibited by prolonged glucocorticoid exposure, suggesting that low levels of adrenal steroids may promote the postnatal development of these neurons.

Adrenal Cortex Hormones↗
PubMed2004

Premature adolescent autonomy: parent disengagement and deviant peer process in the amplification of problem behaviour.

Premature autonomy describes a developmental dynamic where parents of high-risk adolescents reduce their involvement and guidance when confronted with challenges of problem behaviour and the influence of deviant friendships. This dynamic was tested on the sample of Oregon Youth Study boys (N=206), whose family management practices and friendships were observed on videotaped interaction tasks. Latent growth curve models were used to examine longitudinal trends between deviant friendship interactions and family management. Direct observations of deviant friendship process at age 14 were associated with degradation in family management during adolescence. A comparison of antisocial and well-adjusted boys clarified that parents of antisocial boys (started early and persisted) decreased family management around puberty, in comparison to parents of well-adjusted boys who maintained high levels of family management through adolescence. In predicting late adolescent problem behaviour, there was a statistically reliable interaction between family management degradation and deviant peer involvement in adolescence in support of the premature autonomy hypothesis. Adolescent males involved in deviant friendships, and whose parents decreased their family management, were most likely to use marijuana and commit antisocial acts at age 18. The implications for interventions that target adolescents are discussed.

Adolescent↗
PubMed2004

Origin and developmental fate of vestigial tooth primordia in the upper diastema of the field vole (Microtus agrestis, Rodentia).

OBJECTIVE: Odontogenesis in voles is a convenient model to test hypotheses on tooth development generated from investigations in the mouse. Similar to other rodents, the functional dentition of the vole includes a toothless diastema. At its mesial end, a vestigial tooth bud has been found in the upper jaw of vole embryos. The aim of this study was to analyse the developmental dynamics of vestigial tooth structures in the upper diastema of the field vole and to compare it with the situation in the mouse. DESIGN: The development of odontogenic structures in the upper diastema of the field vole was investigated using serial histological sections and three-dimensional (3D) computer-aided reconstruction. RESULTS: A transient continuous dental lamina in the upper diastema of the field vole extended mesially to the first molar primordium, but was not continuous with the dental lamina in the incisor region. At its mesial limit, a large vestigial tooth primordium was regularly present. A further distinct vestigial bud was located mesially to the first molar primordium. The segmentation of the dental lamina suggested a potential to give rise to further vestiges in the upper diastema of the vole. CONCLUSIONS: In the prospective diastema of the vole exists as in the mouse a continuous dental lamina. Beside the prominent vestigial tooth bud in the mesial diastema, a further large bud was transiently located in front of the molars. The incorporation of dental epithelium into the first upper molar (M(1)) primordium in the vole differs from that in the mouse.

Animals↗
PubMed2004

Retinoic acid modulates the retinoblastoma protein during adipocyte terminal differentiation.

Terminal differentiation is characterized by a permanent withdrawal of cells from the cell cycle. Retinoblastoma protein (RB) has been involved in cell cycle progression. Accumulating evidence also implicates RB in the promotion of differentiation of many cell types. We present new insights into the role of RB and other cell cycle regulatory proteins in adipocyte differentiation and on the role of retinoic acid (RA) in the regulation of the latter process. It is shown that RA reduces RB expression and enhances RB phosphorylation by a mechanism that involves down-regulation of the cyclin-dependent kinase inhibitor (CKI) p21(Cip1), having this fact as important consequences for both the cell cycle progression and the adipocyte differentiation process. The effects of RA result in the blockage of adipogenesis, but may also favor the retention of a pool of adipose cells able to re-enter the cell cycle, which may be important for the developmental dynamics of adipose tissue in vivo. In addition, these results reinforce the idea that there is a cross-talk between the cell cycle machinery and the adipocyte differentiation machinery that can be modulated by external signals, including nutrients.

3T3 Cells↗
PubMed2006

A quantitative approach to the study of cell shapes and interactions during early chordate embryogenesis.

BACKGROUND: The prospects of deciphering the genetic program underlying embryonic development were recently boosted by the generation of large sets of precisely organized quantitative molecular data. In contrast, although the precise arrangement, interactions, and shapes of cells are crucial for the fulfilment of this program, their description remains coarse and qualitative. To bridge this gap, we developed a generic software, 3D Virtual Embryo, to quantify the geometry and interactions of cells in interactive three-dimensional embryo models. We applied this approach to early ascidian embryos, chosen because of their simplicity and their phylogenetic proximity to vertebrates. RESULTS: We generated a collection of 19 interactive ascidian embryos between the 2- and 44-cell stages. We characterized the evolution with time, and in different cell lineages, of the volume of cells and of eight mathematical descriptors of their geometry, and we measured the surface of contact between neighboring blastomeres. These analyses first revealed that early embryonic blastomeres adopt a surprising variety of shapes, which appeared to be under strict and dynamic developmental control. Second, we found novel asymmetric cell divisions in the posterior vegetal lineages, which gave birth to sister cells with different fates. Third, during neural induction, differences in the area of contact between individual competent animal cells and inducing vegetal blastomeres appeared important to select the induced cells. CONCLUSIONS: In addition to novel insight into both cell-autonomous and inductive processes controlling early ascidian development, we establish a generic conceptual framework for the quantitative analysis of embryo geometry that can be applied to other model organisms.

Animals↗
PubMed2003

Cyclin E expression during development in Caenorhabditis elegans.

Our interest in the coordination of cell cycle control and differentiation has led us to investigate the Caenorhabditis elegans cye-1 gene encoding the G(1) cell cycle regulator cyclin E. We have studied the expression and function of cye-1 by using monoclonal antibodies directed against CYE-1 protein, cye-1::GFP reporter genes, and a cye-1 chromosomal deletion mutation. We show that a ubiquitous embryonic pattern of expression becomes restricted and dynamic during postembryonic development. Promoter analysis reveals a relatively small region of cis-acting sequences that are necessary for the complex pattern of expression of this gene. Our studies demonstrate that two other G(1) cell cycle genes, encoding cyclin D and CDK4/6, have similarly compact promoter requirements. This suggests that a relatively simple mechanism of regulation may underlie the dynamic developmental patterns of expression exhibited by these three G(1) cell cycle genes. Our analysis of a new cye-1 deletion allele confirms and extends previous studies of two point mutations in the gene.

Amino Acid Sequence↗
PubMed2000

Lithium treatment in ovo: effects on embryonic heart rate, natural death of ciliary ganglion neurons, and brain expression of a highly conserved chicken homolog of human MTG8/ETO.

Understanding the action of the mood stabilizer lithium is dependent on availability of experimental models where lithium treatment at clinically relevant concentrations induces marked phenotypic and genotypic changes. Here we report on such changes in the chicken embryo. Lithium chloride (0.6 mM), applied in ovo 60 h after incubation, markedly delayed the heart rate increase observed from ED2.5 to ED5, and induced the brain expression of a new chicken gene cETO from ED7 to ED15. At the same time the overall developmental dynamics and embryo survival, or the expression of chicken gephyrin were not significantly affected. Furthermore, lithium treatment (0.3 mM, 48 h after incubation) abolished the difference in neuronal number between ED12 ciliary ganglia developing in the presence or absence of postganglionic target muscles. We show that cETO is a close homologue of the human transcription factor MTG8/ETO; named after its location on chromosome 8, and participation in chromosomal translocation 8;21 in myeloid leukemia. The mRNA and protein levels of ETO and gephyrin had a parallel course in chicken brain development suggesting that the expression of both genes is regulated mainly at the level of gene transcription. However, the patterns of expression were markedly different. ETO peaked at ED7 and decreased five-fold at ED15. In contrast, gephyrin levels increased five-fold from ED7 to ED15. We propose that the induction of ETO expression, in concert with lithium-induced upregulation of other genes, such as PEBP2beta and bcl-2, is participating in the neuroprotective effect of chronic lithium treatment.

Amino Acid Sequence↗
PubMed2002

Development of the catecholaminergic system in the early zebrafish brain: an immunohistochemical study.

Tyrosine hydroxylase-containing cells (TH cells) were investigated immunohistochemically in early and late postembryonic zebrafish brain sections (at 2 and 5 days postfertilization [dpf]) yielding an improved neuroanatomical resolution of spatiotemporal developmental dynamics of the catecholaminergic system. Additionally, double-immunolabel preparations for visualizing TH cells and cells containing the proliferating cell nuclear antigen (PCNA cells) were carried out allowing for a prosomeric interpretation of early forebrain TH cell clusters. Many TH cell populations recently described in the adult zebrafish brain could be identified in the present study by location and cell type already in the 5 dpf (e.g. eight of 12 adult diencephalic TH cell populations) and 2 dpf (e.g. five of 12 adult TH cell populations) zebrafish brain. Early and adult diencephalic TH cells are restricted to the pretectum (P1) and ventral thalamus (P3) in the alar plate, and to various TH groups in the basal plate posterior tuberculum (P3), as well as to various populations in the hypothalamus (secondary prosencephalon). The alar plate ventral thalamic and most anterodorsal posterior tubercular TH cell populations range among the earliest detectable ones. There was no indication of migration of TH cells from the midbrain-hindbrain boundary or anterior neural ridge into the diencephalon.

Animals↗