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PubMed2005

Moment-to-moment dynamics of ADHD behaviour.

BACKGROUND: The behaviour of children with Attention-Deficit/Hyperactivity Disorder is often described as highly variable, in addition to being hyperactive, impulsive and inattentive. One reason might be that they do not acquire complete and functional sequences of behaviour. The dynamic developmental theory of ADHD proposes that reinforcement and extinction processes are inefficient because of hypofunctioning dopamine systems, resulting in a narrower time window for associating antecedent stimuli and behaviour with its consequences. One effect of this may be that the learning of behavioural sequences is delayed, and that only short behavioural sequences are acquired in ADHD. The present study investigated acquisition of response sequences in the behaviour of children with ADHD. METHODS: Fifteen boys with ADHD and thirteen boys without, all aged between 6-9 yr, completed a computerized task presented as a game with two squares on the screen. One square was associated with reinforcement. The task required responses by the computer mouse under reinforcement contingencies of variable interval schedules. Reinforcers were cartoon pictures and small trinkets. Measures related to response location (spatial dimension) and to response timing (temporal dimension) were analyzed by autocorrelations of consecutive responses across five lags. Acquired response sequences were defined as predictable responding shown by high explained variance. RESULTS: Children with ADHD acquired shorter response sequences than comparison children on the measures related to response location. None of the groups showed any predictability in response timing. Response sequencing on the measure related to the discriminative stimulus was highly related to parent scores on a rating scale for ADHD symptoms. CONCLUSION: The findings suggest that children with ADHD have problems with learning long sequences of behaviour, particularly related to response location. Problems with learning long behavioural sequences may ultimately lead to deficient development of verbally governed behaviour and self control. The study represents a new approach to analyzing the moment-to-moment dynamics of behaviour, and provides support for the theory that reinforcement processes are altered in ADHD.

Journal Article↗
PubMed2006

[Distribution characteristic of NDRG2 expression in human fetal tissues.].

NDRG2, one of the new N-Myc downstream-regulated gene (NDRG) gene families, is believed to be involved in cell growth event. However, the exact function is still unknown. Identification of the tissue or cell types expressing this gene in vivo will provide clues in clarifying its physiological roles. Using RT-PCR and Western blot, we analyzed the expression level of NDRG2 mRNA and protein in human fetal tissues from different gestational ages. The anti-NDRG2 monoclonal antibody, which has been proved to react specifically with NDRG2 protein, was further used to analyze the cellular location of NDRG2 protein in various human fetal tissues by immunohistochemistry. We found that NDRG2 expression was developmentally dynamic, being generally lower in the early stages of development and markedly increasing during the later stages. NDRG2 mRNA and protein distribution were generally consistent in heart and lung. One of the differences was that NDRG2 protein appeared later than mRNA in kidney. Another unmatched expression was found in liver. NDRG2 mRNA appeared later than protein in liver. In human fetal tissues at sixteen and twenty-eight weeks of gestation, NDRG2 protein immunoreactions could be seen in epithelium of small intestine, epithelium of large intestine, superficial layer of epidermis, whisker follicles, epithelium of small bronchus, hepatocytes, cardiac myocytes, thymus corpuscles and epithelium of renal tubule, and the immunoreactions in those tissues from twenty-eight weeks of gestation was stronger than that from sixteen weeks of gestation. In the present study, we demonstrate the expression pattern and cellular location of NDRG2 protein in a large set of human fetal tissues. This is the first demonstration of NDRG2 protein expression in human fetal tissues. Taken together, the results suggest that NDRG2 protein found in a variety of tissues is not a tissue-specific protein, and may play important roles in histogenesis and organogenesis.

Fetus↗
PubMed1996

Structure, development, and maintenance of the nerve net of the body column in Hydra.

The anatomy and developmental dynamics of the nerve net in the body column of Hydra viridissima were examined immunocytochemically with a monoclonal antibody (CC04) that recognizes an antigen in nerve cells and with an antiserum against vasopressin. CC04+ neuron cell bodies, their neurites, and vasopressin-like-immunoreactive (VLI+) neurites could be clearly visualized on whole-mount preparations. All neurites of the CC04+ neurons in the body column were VLI+. However, only half of the VLI+ neurites in the body column were CC04+. Immunocytochemical analysis of macerated preparations showed that half of the neurons in the gastric region of the body column were CC04+. These results suggest that most of the neurons in the gastric region are VLI+. The density of the VLI+ neurites was uniform along the entire length of the body column. The CC04+ neuron density in the gastric region remained constant at all stages of asexual development and during foot regeneration. After pulse-labeling with 5-bromo-2'-deoxyuridine (BrdU), CC04+ neurons with labeled nuclei appeared in the body column. We conclude that neuron density in the gastric region is maintained at a constant value by insertion of new neurons in parallel with continuous epithelial cell division.

Animals↗
PubMed1996

Application of immunocytochemistry for detection of proliferating cell populations during tooth development.

BACKGROUND: The production of monoclonal antibodies to cell cycle-related molecules provides the basis for immunochemical studies on cell kinetics. METHODS: Immunocytochemistry permits the tissue localization of replicating cells, whereas flow cytometry defines the exact position of immunoreactive cells in the cell cycle and ensures a quantitative analysis of the growth fraction. Bromo-deoxyuridine-antibody can be used to reveal S phase-traversing cells, whereas the immunoreactivity for the Proliferating Cell Nuclear Antigen defines the G1, S, and G2-M subpopulations of the cell cycle. RESULTS: Odontogenic cells produce secretory products (e.g., enamel and dentine matrix proteins and growth factors) and express receptors and oncogenes during specific stages of their differentiation. CONCLUSIONS: The simultaneous detection of cell cycle-related antigens and differentiation markers using double immunochemical staining may be useful to clarify the role of putative regulatory molecules in the control of cell growth during odontogenesis, thus unveiling molecular mechanisms that regulate developmental dynamics.

Animals↗
PubMed1993

Accelerating language acquisition.

How much can the development of language and other skills be accelerated in the general population? High correlations between early verbal and mental competencies and parent and teacher language socialization practices suggest enormous potential for widespread improvement. Here we report follow-up research in progress in studies of late adolescent children from diverse ethnic and educational backgrounds who participated in a language enrichment programme during infancy in the home or day-care. In 39 of 44 home-stimulated children located to date (nearly all from college-educated families) 62-93% were: in gifted or advanced programmes, obtaining high grades, avid readers and skilled in writing (over half read before school and wrote creative material independently) and generally highly skilled in verbal, mathematical and other academic domains. They also excelled socially and in sports, and showed intellectual independence. Additional subjects and data (on competence, later experiences and Scholastic Aptitude Test [SAT] scores) are currently being collected. Preliminary data analyses suggest that although early language enrichment can in the short term easily increase competence in all groups well beyond norms generated by current socialization practices, long-term outcomes are a complex function of developmental dynamics between the early, complex, foundation of high skills and motivation for learning, and the interaction with facilitative parental resources.

Adolescent↗
PubMed1992

Planar induction of convergence and extension of the neural plate by the organizer of Xenopus.

This paper demonstrates that convergence and extension within the neural plate of Xenopus laevis are regulated by planar inductive interactions with the adjacent Spemann organizer. The companion article (Keller et al.: Developmental Dynamics 193:199-217, 1992) showed that the prospective hindbrain and spinal cord occupy a very short and very wide area just above the Spemann organizer in the early gastrula and that these regions converge and extend greatly during gastrulation and neurulation, using a sequence of radial and mediolateral cell intercalations. In this article, we show that "planar" contact of these regions with the organizer at their vegetal edge until stage 11 is sufficient to induce convergence and extension, after which their convergence and extension become autonomous. Grafts of the organizer in planar contact with uninduced ectodermal tissues induce these ectodermal tissues to converge and extend by a planar inductive signal from the organizer. Labeling of the inducing or responding tissues confirms that only planar interactions occur. Neural convergence and extension are actually hindered in explants deliberately constructed so that vertical interactions occur. These results show unambiguously that the Spemann organizer induces the extraordinary and precocious convergence and extension movements of the Xenopus neural plate by planar interactions acting over short distances.

Animals↗
PubMed1984

Effects of genic substitution at the brown locus on the differentiation of epidermal melanocytes in newborn mouse skin.

In the dorsal epidermis of both C57BL/10J (black, BB) and C57BR/cdJ (brown, bb) mice, the number of melanocytes positive to the dopa reaction (melanocyte population) increases from birth to day 3 or 4, and then gradually decreases. However, the number of melanoblasts plus melanocytes positive to the combined dopa-premelanin reaction (melanoblast-melanocyte population) remains constant until day 3 or 4 and then decreases in the two strains. Despite the similarity of the developmental dynamics in both black and brown mice, there is a significant difference in the number of differentiated melanocytes. Melanocytes are more numerous and more dopa-reactive in brown mice than in black. The maximal density of the melanoblast-melanocyte population on day 3 or 4 does not differ in brown and black mice. Moreover, the maximal density of the melanocyte population in brown epidermis does not differ from that of the melanoblast-melanocyte population of both brown and black. These results indicate that b allele, when homozygous, enhances the differentiation of epidermal melanoblasts by inducing high tyrosinase activity.

Alleles↗
PubMedDate not supplied

Visualization of molecular and cellular events with green fluorescent proteins in developing embryos: a review.

During the past 5 years, green fluorescent protein (GFP) has become one of the most widely used in vivo protein markers for studying a number of different molecular processes during development, such as promoter activation, gene expression, protein trafficking and cell lineage determination. GFP fluorescence allows observation of dynamic developmental processes in real time, in both transiently and stably transformed cells, as well as in live embryos. In this review, we include the most up-to-date use of GFP during embryonic development and point out the unique contribution of GFP visualization, which resulted in novel discoveries.

Animals↗
PubMed2001

Postnatal development of catecholamine inputs to the paraventricular nucleus of the hypothalamus in rats.

Adrenergic and noradrenergic neural projections to the paraventricular nucleus of the hypothalamus (PVN) contribute importantly to viscerosensory modulation of pituitary hormone secretion. Immaturity of ascending catecholamine pathways may partially underlie the documented hyporesponsiveness of PVN neurosecretory cells to certain interoceptive stimuli in rats during the first few weeks of postnatal development. To explore this possibility, the present study compared the distribution and number of dopamine-beta-hydroxylase (DBH)- and phenylethanolamine-N-methyltransferase (PNMT)-positive neurons projecting to the PVN in newborn and adult rats. In addition, a quantitative analysis of DBH- and PNMT-immunoreactive fibers in the medial parvocellular subnucleus, dorsal division (PVNmpd) and posterior magnocellular subnucleus, lateral division (PVNpml) was performed in adult rats and in developing rats on postnatal day (P)1, P7, P14, and P21. The numbers of PVN-projecting neurons in the A1, C1, A2/C2, C3, or A6 catecholamine cell groups were similar in newborn and adult rats, as were the proportions of PVN-projecting neurons in each region that were PNMT-positive. However, fewer PVN-projecting neurons in the C1 and C3 regions expressed DBH immunolabeling in newborn rats compared to adults. DBH immunolabeling increased progressively in the PVNmpd and PVNpml between postnatal days P1 and P21, when adult-like levels were achieved. Conversely, PNMT immunolabeling in the same PVN subdivisions was most dense at P1, gradually decreasing to adult-like levels by P21. These dynamic developmental changes in catecholamine synthetic enzyme immunolabeling densities in the PVN may reflect functional changes in noradrenergic and adrenergic signaling capacity in rats during the first few weeks of postnatal development.

Aging↗
PubMed2005

Development of Hsp25 expression compartments is not constrained by Purkinje cell defects in the Lurcher mouse mutant.

Four transverse zones can be distinguished in the adult mouse cerebellar cortex based on differential expression of cell-specific antigens, termination patterns of mossy fiber afferents, and phenotypes of mouse mutants with cerebellar defects: the anterior zone (AZ), central zone (CZ), posterior zone (PZ), and nodular zone (NZ). In the heterozygous Lurcher (Lc/+) mouse a zonally restricted abnormality in Purkinje cell development is seen. The Purkinje cell-specific antigen zebrin II is normally differentially expressed in all four zones of the adult cerebellum, but in the Lc/+ mutant is confined to the PZ and NZ, caudal to a transverse boundary in the dorsal aspect of lobule VIII. In this study we wanted to understand why zebrin II expression is arrested at this boundary and whether the Lc mutation affects the differentiation of additional Purkinje cell antigens in a similar manner. To determine this, we took advantage of the dynamic developmental timetable of another Purkinje cell antigen, the small heat shock protein Hsp25. Using immunohistochemistry we demonstrate that cerebellar maturation anterior to the CZ/PZ transverse boundary appears to be unaffected by the Lc allele, in that initial progression of Hsp25 expression in the Lc/+ cerebellum was similar to controls. Double-labeling experiments with anti-Hsp25 and anti-calbindin suggest that characteristic banding patterns of Hsp25 in Lc/+ cerebellum develop and are preserved despite cell loss. Thus, since simple temporal or spatial models cannot account for the zonal restriction seen during Lc/+ cerebellar development, the abnormality may be zebrin II-specific.

Animals↗
PubMed1992

Nerve ring of the hypostome in hydra. I. Its structure, development, and maintenance.

The anatomy and developmental dynamics of the nerve ring in the hypostome of Hydra oligactis were examined immunocytochemically with an antiserum against a neuropeptide and with neuron-specific monoclonal antibodies. The nerve ring is unique in the mesh-like nerve net of hydra. It is a distinct neuronal complex consisting of a thick nerve bundle running circumferentially at the border between the hypostome and tentacle zone. Immunostaining showed that the nerve ring was heterogeneous and contained at least four different subsets of neurons. During head regeneration and budding, the nerve ring appeared only after the nerve net of ganglion and sensory cells had formed. Every epithelial cell is continuously displaced with neurons toward either head or foot in an adult hydra. However, the ectoderm in the immediate vicinity of, and including, the nerve ring constitutes a stationary zone that is not displaced. Tissue immediately above this zone is displaced toward the tip of the hypostome, while tissue below is displaced along the tentacles. Correspondingly, the production of new neurons in the ring as measured by their differentiation kinetics is much slower than in surrounding areas. Thus, the nerve ring is static and stable in contrast to the dynamic features of the nerve net of hydra.

Animals↗
PubMed2004

Significance of the cranial neural crest.

The cranial neural crest has long been viewed as being of particular significance. First, it has been held that the cranial neural crest has a morphogenetic role, acting to coordinate the development of the pharyngeal arches. By contrast, the trunk crest seems to play a more subservient role in terms of embryonic patterning. Second, the cranial crest not only generates neurons, glia, and melanocytes, but additionally forms skeletal derivatives (bones, cartilage, and teeth, as well as smooth muscle and connective tissue), and this potential was thought to be a unique feature of the cranial crest. Recently, however, several studies have suggested that the cranial neural crest may not be so influential in terms of patterning, nor so exceptional in the derivatives that it makes. It is now becoming clear that the morphogenesis of the pharyngeal arches is largely driven by the pharyngeal endoderm. Furthermore, it is now apparent that trunk neural crest cells have skeletal potential. However, it has now been demonstrated that a key role for the cranial neural crest streams is to organise the innervation of the hindbrain by the cranial sensory ganglia. Thus, in the past few years, our views of the significance of the cranial neural crest for head development have been altered. Developmental Dynamics 229:5-13, 2004.

Animals↗
PubMed2004

Complementation of melanocyte development in SOX10 mutant neural crest using lineage-directed gene transfer.

An in vitro gene complementation approach has been developed to dissect gene function and regulation in neural crest (NC) development and disease. The approach uses the avian RCAS virus to express genes in NC cells derived from transgenic mice expressing the RCAS receptor TVA, under the control of defined promoter elements. Constructs for creating TVA transgenic mice were developed using site-specific recombination GATEWAY (GW), compatible vectors that can also be used to facilitate analysis of genomic fragments for transcriptional regulatory elements. By using these GW vectors to facilitate cloning, transgenic mouse lines were generated that express TVA in SOX10-expressing NC stem cells under the control of the Pax3 promoter. The Pax3-tv-a transgene was bred onto a Sox10-deficient background, and the feasibility of complementing genetic NC defects was demonstrated by infecting the Pax3-tv-a cells with an RCAS-Sox10 expression virus, thereby rescuing melanocyte development of Sox10-deficient NC cells. This system will be useful for assessing genetic hierarchies in NC development. Developmental Dynamics 229:54-62, 2004.

Animals↗
PubMed2004

Homocysteine inhibits cardiac neural crest cell formation and morphogenesis in vivo.

Elevated homocysteine increases the risk of neurocristopathies. Here, we determined whether elevating homocysteine altered the proliferation or number of chick neural crest cells that form between the midotic and third somite in vivo. Homocysteine increased the number of neural tube cells but decreased neural crest cell number. However, the sum total of cells was not different from controls. In controls, the 5-bromo-2'-deoxyuridine-labeling index was higher in newly formed neural crest cells than in their progenitors, paralleling reports showing these progenitors must pass the restriction point before undergoing epithelial-mesenchymal transition. Homocysteine decreased the labeling index of newly formed neural crest cells, suggesting that it inhibited cell cycle progression of neural crest progenitors or the S-phase entry of newly formed neural crest cells. Homocysteine also inhibited neural crest dispersal and decreased the distance they migrated from the neural tube. These results show neural crest morphogenesis is directly altered by elevated homocysteine in vivo. Developmental Dynamics 229:63-73, 2004.

Animals↗
PubMed2004

Neural crest and the origin of ectomesenchyme: neural fold heterogeneity suggests an alternative hypothesis.

The striking similarity between mesodermally derived fibroblasts and ectomesenchyme cells, which are thought to be derivatives of the neural crest, has long been a source of interest and controversy. In mice, the gene encoding the alpha subunit of the platelet-derived growth factor receptor (PDGFRalpha) is expressed both by mesodermally derived mesenchymal cells and by ectomesenchyme. Whole-mount immunostaining previously revealed that PDGFRalpha is present in the cephalic neural fold epithelium of early murine embryos (Takakura et al. [1997] J Histochem Cytochem 45:883-893). We now show that, within the neural fold, a sharp boundary exists between E-cadherin-expressing non-neural epithelium and the neural epithelium of the dorsal ridge. In addition, we found that cells coexpressing E-cadherin and PDGFRalpha are present in the non-neural epithelium of the neural folds. These observations raise the possibility that at least some PDGFRalpha(+) ectomesenchyme originates from the lateral non-neural domain of neural fold epithelium. This inference is consistent with previous reports (Nichols [ 1981] J Embryol Exp Morphol 64:105-120; Nichols [ 1986] Am J Anat 176:221-231) that mesenchymal cells emerge precociously from an epithelial neural fold domain resembling the primitive streak in the early embryonic epiblast. Therefore, we propose the name "metablast" for this non-neural epithelial domain to indicate that it is the site of a delayed local delamination of mesenchyme similar to involution of mesoderm during gastrulation. We further propose the testable hypothesis that neural crest and ectomesenchyme are developmentally distinct progenitor populations and that at least some ectomesenchyme is metablast-derived rather than neural crest-derived tissue. Developmental Dynamics 229:118-130, 2004.

Animals↗
PubMed2004

Slow degeneration of zebrafish Rohon-Beard neurons during programmed cell death.

Rohon-Beard cells are large, mechanosensory neurons located in the dorsal spinal cord of anamniote vertebrates. In most species studied to date, these cells die during development. We followed labeled Rohon-Beard cells in living zebrafish embryos and found that they degenerate slowly, over many days. During degeneration, the soma shrinks and finally disappears, and the processes become beady in appearance and finally break apart, but they do not retract. Zebrafish Rohon-Beard cells apparently fragment their DNA, as revealed by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) labeling, before undergoing degenerative morphologic changes. We also followed the development of labeled dorsal root ganglion neurons, as they are developing at the same stages that Rohon-Beard cells are degenerating. We found that, although axons of both cell types extend into similar regions, Rohon-Beard cells degenerate normally in mutants lacking dorsal root ganglia, providing evidence that interactions between the two cell types are not responsible for Rohon-Beard cell degeneration. Developmental Dynamics 229:30-41,2004.

Animals↗
PubMed2004

Long-distance cue from emerging dermis stimulates neural crest melanoblast migration.

Neural crest melanoblasts display unique navigational abilities enabling them to colonize the dorsal path between ectoderm and somite. One signal shown here to elicit melanoblast migration is a chemotactic cue supplied by the emerging dermis. Until dermis emerges, melanoblasts fail to enter the dorsal path. The dermis emerges from a site that is too distant to stimulate migration by cell contact. Instead, surgeries show that dermis elicits migration from a distance. When dermis is grafted distally, neural crest cells enter the path precociously. Moreover, large grafts recruit melanoblasts from the control sides (without increasing crest cell numbers) as well as a few crest cells from ventral somite. Because other grafted tissues fail to stimulate migration, the dermis stimulus is specific. This report is the first documentation that trunk neural crest cells can be guided chemotactically. It also extends evidence that migration is exquisitely sensitive to temporal-spatial patterns of somite morphogenesis. Developmental Dynamics 229:99-108, 2004.

Animals↗
PubMed2005

Experimental analyses of the function of the proepicardium using a new microsurgical procedure to induce loss-of-proepicardial-function in chick embryos.

The proepicardium (PE) is a primarily extracardiac progenitor cell population that colonizes the embryonic heart and delivers the epicardium, the subepicardial and intramyocardial fibroblasts, and the coronary vessels. Recent data show that PE-derived cells additionally play important regulatory roles in myocardial development and possibly in the normal morphogenesis of the heart. Developmental Dynamics 233, 2005. Research on the latter topics profits from the fact that loss-of-PE-function can be experimentally induced in chick embryos. So far, two microsurgical techniques were used to produce such embryos: (1) blocking of PE cell transfer with pieces of the eggshell membrane, and (2) mechanical excision of PE. Both of these techniques, however, have their shortcomings. We have searched, therefore, for new techniques to eliminate the PE. Here, we show that loss-of-PE-function can be induced by photoablation of the PE. Chick embryos were treated in ovo by means of a window in the eggshell at Hamburger and Hamilton (HH) stage 16 (iday 3). The pericardial coelom was opened, and the PE was externally stained with a 1% solution of Rose Bengal by means of a micropipette. Photoactivation of the dye was accomplished by illumination of the operation field with visible light. Examination on postoperative day 1 (iday 4, HH stages 19/20) disclosed complete removal of PE in every experimental embryo. On iday 9 (HH stages 33/34), the survival rate of experimental embryos was 35.7% (15 of 42). Development of the PE-derivatives was compromised in the heart of every survivor. The abnormalities encompassed hydro- or hemopericardium, epicardium-free areas with aneurysmatic outward bulging of the ventricular wall, thin myocardium, defects of the coronary vasculature, and abnormal tissue bridges between the ventricles and the pericardial wall. Our results show that photoablation of the PE is a powerful technique to induce long-lasting loss-of-PE-function in chick embryos. We have additionally obtained new data that suggest that the embryonic epicardium may make important contributions to the passive mechanics of the developing heart.

Animals↗