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PubMed1990

Scintigraphic evaluation of digital circulation during the developmental and acute phases of equine laminitis.

Scintigraphy was used to evaluate digital circulation at 24 h intervals in 11 control horses and in nine horses affected with acute laminitis created by administration of a high-starch ration. Following intra-arterial injection of technetium-99m macroaggregated albumin into the brachiocephalic trunk, static images were acquired of the right front foot. Dynamic radionuclide angiograms and static blood pool images were also obtained after jugular vein injection of technetium-99m diethylenetriamine pentaacetic acid. These procedures were performed on standing horses, using either minimal or no tranquillisation. Images were analysed quantitatively for parameters indicative of circulation to the whole foot and to specific regions within the foot. There was no evidence of reduced total blood flow to the lamellae during either the developmental or acute phases of laminitis. Total flow tended to increase throughout the peripheral/external regions of the foot, but statistically significant elevations were consistently present only within the lamellae. The increased total blood flow to the lamellae may have been due to elevated capillary flow and/or elevated arteriovenous shunt flow. This study did not support lamellar ischaemia as the primary cause of acute equine laminitis.

Acute Disease↗
PubMed2000

A chemical flow system mimics waves of gene expression during segmentation.

The early vertebrate developmental process of somitogenesis involves bands of gene expression that form periodically at the posterior end of the presomitic mesoderm (PSM) and traverse it with decreasing width and velocity. We have constructed a chemical flow system that, based on the novel flow-distributed oscillator (FDO) mechanism of wave pattern formation, reproduces key physical features of the PSM and observe concentration waves having similar spatio-temporal behavior. This suggests that the gene expression waves can be understood qualitatively in terms of phase dynamics in an open flow of a self-oscillating medium and that chemical flow systems can be used to mimic and model biological pattern formation during axial growth. In fact, expressions for wavelength and wave velocity derived from phase dynamics are found to be in quantitative agreement with measurements from both the biological and the chemical systems. This indicates that they, despite their significant differences, have common dynamics.

Animals↗
PubMed2006

Transcriptome profiling of the tubular porcine conceptus identifies the differential regulation of growth and developmentally associated genes.

Gastrulation and trophectoderm elongation of the porcine conceptus coincide with peak conceptus estrogen secretion from gestational day 11 to day 12. The current study aim was to identify genes required for elongation by defining the transcriptome profile of this dynamic tubular stage. The gastrulation and proliferative status of ovoid, tubular, and filamentous conceptuses were also examined. Polarization of the embryonic disc and growth throughout the conceptus were evident. An unamplified and two distinct amplified serial analysis of gene expression (SAGE) libraries were generated from tubular conceptus mRNA. Comparing the three libraries at 12,000 tags/library indicated small-amplified RNA-SAGE was a reliable amplification procedure. The unamplified library was increased to 42,415 tags and statistical analyses of tag frequencies with previously generated ovoid and filamentous libraries revealed the differential expression (P < 0.05) of 483 and 364 tags between ovoid:tubular or tubular:filamentous libraries, respectively. Annotated transcripts known to be involved in development and also potentially regulated by estrogen (cytokeratins 8 and 18, stratifin, midkine, and glycolytic enzymes) were further analyzed by real-time PCR. The majority of glycolytic enzyme transcripts were constitutively expressed or downregulated at the filamentous stage. Likewise, cytokeratin mRNAs were less abundant in filamentous conceptuses, whereas stratifin and midkine were more abundant in tubular conceptuses. Analysis of protein revealed distinct expression patterns for cytokeratin 18, stratifin, and midkine. The function(s) of these factors and potential modulation by estrogen clearly needs to be elucidated to understand their physiological role in normal conceptus development.

Animals↗
PubMed2003

Dynamics of infection in tick vectors and at the tick-host interface.

Tick-borne flaviviruses are common, widespread, and successfully adapted to their mode of transmission. Most tick vectors of flaviviruses are ixodid species. These ticks are characterized by a comparatively long life cycle, lasting several years, during which the infecting virus may be maintained from one developmental stage of the tick to the next. Hence ticks act as highly efficient reservoirs of flaviviruses. Many tick-borne flaviviruses are transmitted vertically, from adult to offspring, although the frequency is too low to maintain the viruses solely in the tick population. Instead, the survival of tick-borne flaviviruses is dependent on horizontal transmission, both from an infected tick to a susceptible vertebrate host and from an infected vertebrate to uninfected ticks feeding on the animal. The dynamics of transmission and infection have traditionally been considered in isolation: in the tick, following virus uptake in the infected blood meal, infection of the midgut, passage through the hemocoel to the salivary glands, and transmission via the saliva; and in the vertebrate host, virus delivery into the skin at the site of tick feeding, infection of the draining lymph nodes, and dissemination to target organs. However, there is now compelling evidence of a complex interaction between the tick vector and its vertebrate host that affects virus transmission profoundly. The feeding site in the skin is a battleground in which the hemostatic, inflammatory, and immune responses of the host are countered by antihemostatic, anti-inflammatory, and immunomodulatory molecules (mostly proteins and peptides) secreted in tick saliva. Here we speculate that exploitation of the tick pharmacopeia, rather than development of viremia, is the key step in successful tick-borne flavivirus transmission.

Animals↗
PubMed2004

Functional genomics of cell elongation in developing cotton fibers.

Cotton fibers are single-celled seed trichomes of major economic importance. Factors that regulate the rate and duration of cell expansion control fiber morphology and important agronomic traits. For genetic characterization of rapid cell elongation in cotton fibers, approximately 14,000 unique genes were assembled from 46,603 expressed sequence tags (ESTs) from developmentally staged fiber cDNAs of a cultivated diploid species ( Gossypium arboreum L.). Conservatively, the fiber transcriptome represents 35-40% of the genes in the cotton genome. In silico expression analysis revealed that rapidly elongating fiber cells exhibit significant metabolic activity, with the bulk of gene transcripts, represented by three major functional groups - cell wall structure and biogenesis, the cytoskeleton and energy/carbohydrate metabolism. Oligonucleotide microarrays revealed dynamic changes in gene expression between primary and secondary cell wall biogenesis showing that fiber genes in the dbEST are highly stage-specific for cell expansion - a conclusion supported by the absence of known secondary cell wall-specific genes from our fiber dbEST. During the developmental switch from primary to secondary cell wall syntheses, 2553 "expansion-associated" fiber genes are significantly down regulated. Genes (81) significantly up-regulated during secondary cell wall synthesis are involved in cell wall biogenesis and energy/carbohydrate metabolism, which is consistent with the stage of cellulose synthesis during secondary cell wall modification in developing fibers. This work provides the first in-depth view of the genetic complexity of the transcriptome of an expanding cell, and lays the groundwork for studying fundamental biological processes in plant biology with applications in agricultural biotechnology.

Cell Growth Processes↗
PubMed2005

Differentiating plant cells switched to proliferation remodel the functional organization of nuclear domains.

The immature pollen grain, the microspore, under stress conditions can switch its developmental program towards proliferation and embryogenesis. The comparison between the gametophytic and sporophytic pathways followed by the microspore permitted us to analyse the nuclear changes in plant differentiating cells when switched to proliferation. The nucleus is highly dynamic, the architecture of its well organised functional domains--condensed chromatin, interchromatin region, nuclear bodies and nucleolus--changing in response to DNA replication, RNA transcription, processing and transport. In the present work, the rearrangements of the nuclear domains during the switch to proliferation have been determined by in situ molecular identification methods for the subcellular localization of chromatin at different functional states, rDNA, elements of the nuclear machinery (PCNA, splicing factors), signalling and stress proteins. The study of the changes in the nuclear domains was determined by a correlative approach at confocal and electron microscopy levels. The results showed that the switch of the developmental program and the activation of the proliferative activity affected the functional organization of the nuclear domains, which accordingly changed their architecture and functional state. A redistribution of components, among them various signalling molecules which targeted structures within the interchromatin region upon translocation from the cytoplasm, was also observed.

Cell Cycle↗
PubMed2004

Taste bud cell dynamics during normal and sodium-restricted development.

Taste bud volume increases over the postnatal period to match the number of neurons providing innervation. To clarify age-related changes in fungiform taste bud volume, the current study investigated developmental changes in taste bud cell number, proliferation rate, and life span. Taste bud growth can largely be accounted for by addition of cytokeratin-19-positive taste bud cells. Examination of taste bud cell kinetics with 3H-thymidine autoradiography revealed that cell life span and turnover periods were not altered during normal development but that cells were produced more rapidly in young rats, a prominent modification that could lead to increased taste bud size. By comparison, dietary sodium restriction instituted during pre- and postnatal development results in small taste buds at adulthood as a result of fewer cytokeratin-19-positive cells. The dietary manipulation also had profound influences on taste bud growth kinetics, including an increased latency for cells to enter the taste bud and longer life span and turnover periods. These studies provide fundamental, new information about taste bud development under normal conditions and after environmental manipulations that impact nerve/target matching.

Animals↗
PubMed2000

Anatomy of commitment: an in vivo study.

A self-evaluation of one small but innovative agency providing supported living and other community supports to individuals with developmental disabilities is presented. Key elements that appear to perpetuate a culture of commitment among direct support and supervisory staff are described. Agency staff people have achieved the capacity to listen and respond to individuals and honor their choices. In order to assure that individuals get what they need, staff members take appropriate initiative, work outside of their assigned hours, and support one another. Whereas most human service organizations function within the parameters of well-defined policies and procedures, the agency described here operates more flexibly. The observed pattern of organizational dynamism and reflexivity is compared to that of a living organism.

Adult↗
PubMed2001

Developmental regulation and complex organization of the promoter of the non-coding hsr(omega) gene of Drosophila melanogaster.

The nucleus-limited large non-coding hsr(omega)-n RNA product of the 93D or the hsr(omega) gene of Drosophila melanogaster binds to a variety of RNA-binding proteins involved in nuclear RNA processing. We examined the developmental and heat shock induced expression of this gene by in situ hybridization of nonradioactively labelled riboprobe to cellular transcripts in intact embryos, larval and adult somatic tissues of wild type and an enhancer-trap line carrying the hsr(omega) 05241 allele due to insertion of a P-LacZ-rosy+ transposon at -130 bp position of the hsr(omega) promoter. We also examined LacZ expression in the enhancer-trap line and in two transgenic lines carrying different lengths of the hsr(omega) promoter upstream of the LacZ reporter. The hsr(omega) gene is expressed widely at all developmental stages; in later embryonic stages, its expression in the developing central nervous system was prominent. In spite of insertion of a big transposon in the promoter, expression of the hsr(omega) 05241 allele in the enhancer-trap line, as revealed by in situ hybridization to hsr(omega) transcripts in cells, was similar to that of the wild type allele in all the embryonic, larval and adult somatic tissues examined. Expression of the LacZ gene in this enhancer-trap line was similar to that of the hsr(omega) RNA in all diploid cell types in embryos and larvae but in the polytene cells, the LacZ gene did not express at all, neither during normal development nor after heat shock. Comparison of the expression patterns of hsr(omega) gene and those of the LacZ reporter gene under its various promoter regions in the enhancer-trap and transgenic lines revealed a complex pattern of regulation, which seems to be essential for its dynamically varying expression in diverse cell types.

Animals↗
PubMed2005

Cancer in xeroderma pigmentosum and related disorders of DNA repair.

Nucleotide-excision repair diseases exhibit cancer, complex developmental disorders and neurodegeneration. Cancer is the hallmark of xeroderma pigmentosum (XP), and neurodegeneration and developmental disorders are the hallmarks of Cockayne syndrome and trichothiodystrophy. A distinguishing feature is that the DNA-repair or DNA-replication deficiencies of XP involve most of the genome, whereas the defects in CS are confined to actively transcribed genes. Many of the proteins involved in repair are also components of dynamic multiprotein complexes, transcription factors, ubiquitylation cofactors and signal-transduction networks. Complex clinical phenotypes might therefore result from unanticipated effects on other genes and proteins.

Child↗
PubMed2005

Novel splice variants associated with one of the zebrafish dnmt3 genes.

BACKGROUND: DNA methylation and the methyltransferases are known to be important in vertebrate development and this may be particularly true for the Dnmt3 family of enzymes because they are thought to be the de novo methyltransferases. Mammals have three Dnmt3 genes; Dnmt3a, Dnmt3b, and Dnmt3L, two of which encode active enzymes and one of which produces an inactive but necessary cofactor. However, due to multiple promoter use and alternative splicing there are actually a number of dnmt3 isoforms present. Six different dnmt3 genes have recently been identified in zebrafish. RESULTS: We have examined two of the dnmt3 genes in zebrafish that are located in close proximity in the same linkage group and we find that the two genes are more similar to each other than they are to the other zebrafish dnmt3 genes. We have found evidence for the existence of several different splice variants and alternative splice sites associated with one of the two genes and have examined the relative expression of these genes/variants in a number of zebrafish developmental stages and tissues. CONCLUSION: The similarity of the dnmt3-1 and dnmt3-2 genes suggests that they arose due to a relatively recent gene duplication event. The presence of alternative splice and start sites, reminiscent of what is seen with the human DNMT3s, demonstrates strong parallels between the control/function of these genes across vertebrate species. The dynamic expression levels of these genes/variants suggest that they may well play a role in early development and this is particularly true for dnmt3-2-1 and dnmt3-1. dnmt3-2-1 is the predominantly expressed form prior to zygotic gene activation whereas dnmt3-1 predominates post zygotic gene activation suggesting a distinct developmental role for each.

Alternative Splicing↗
PubMed2006

Foot differentiation and genomic plasticity in Hydra: lessons from the PPOD gene family.

In Hydra, developmental processes are permanently active to maintain a simple body plan consisting of a two-layered, radially symmetrical tube with two differentiated structures, head and foot. Foot formation is a dynamic process and includes terminal differentiation of gastric epithelial cells into mucous secreting basal disc cells. A well-established marker for this highly specialized cell type is a locally expressed peroxidase (Hoffmeister et al. 1985). Based on the foot-specific peroxidase activity, the gene PPOD1 has been identified (Hoffmeister-Ullerich et al. 2002). Unexpectedly, this approach led to the identification of a second gene, PPOD2, with high sequence similarity to PPOD1 but a strikingly different expression pattern. Here, we characterize PPOD2 in more detail and show that both genes, PPOD1 and PPOD2, are members of a gene family with differential complexity and expression patterns in different Hydra species. At the genomic level, differences in gene number and structure within the PPOD gene family, even among closely related species, support a recently proposed phylogeny of the genus Hydra and point to unexpected genomic plasticity within closely related species of this ancient metazoan taxon.

Animals↗
PubMed1990

Pattern regulation in the ventral histoblasts of the housefly: induction of sternal pattern abnormalities by mechanical wounding of larval epidermal cells.

In higher Diptera, two nests of diploid cells called the ventral histoblasts, located one on either side of each abdominal segment among the polytene larval epidermal cells, give rise to the sternite and its surrounding pleura. During metamorphosis of the insect, these two groups of cells migrate and meet with each other in the midventral region of the developing adult. The cuticular pattern elements and pigmentation in the fifth sternite of the male housefly, when compared to those of other segments as well as the tergites of both sexes, are quite distinct. The above-mentioned features, coupled with the smaller number and predictable occurrence of one of the pattern elements in this sternite, viz, the primary forceps, help one to determine the developmental potential of the histoblast nest and the regulation of its potential which occur at the time of fusion of the two contralateral nests of this segment. A simple operation of slitting the larval epidermal cells (LEC) in a hemisegment in the vicinity of the histoblast nest or extirpation or rotation of a small rectangular piece of LEC between the ventral nest and the midventral line produced pattern abnormalities including mirror image duplication in the hemisternite. An analysis of these pattern abnormalities in the different segments and, in particular, in the fifth segment provides a dynamic picture of the formation of the median sternite. Further, these abnormalities indicate the significance of the presence of the intervening pleural cells between the confronting hemisternites under experimental conditions. Thus, each of the fifth ventral nests has the developmental potential to form more than half of the final sternite pattern. Possible mechanisms for the formation of the normal median sternite during metamorphosis and for the formation of duplicated hemisternites and their fusion products under experimental conditions are discussed in light of current models of pattern regulation.

Animals↗
PubMed1999

Developmental expression of the murine Prl-1 protein tyrosine phosphatase gene.

The expression of the murine Prl-1 protein tyrosine phosphatase gene was examined in normal embryos from E10.5 through E18.5. Prl-1 mRNA was detected in the brain, neural tube, and dorsal root ganglia, and in several non-neuronal tissues, including the skeletal system. Heart and skeletal muscle were consistently negative. At E13.5, Prl-1 was expressed in the condensing prechondrogenic cells of the vertebrae, whereas at E18.5, Prl-1 mRNA was localized to the hypertrophic chondrocytes. The dynamic expression of Prl-1 during cartilage differentiation may suggest a functional role in skeletal development.

Amino Acid Sequence↗
PubMed2002

Analysis of spatial and temporal gene expression patterns in blastula and gastrula stage chick embryos.

Studies on the genetic basis of rostral-caudal specification, neural induction, and head development require knowledge of the relevant gene expression patterns. Gaps in our understanding of gene expression have led us to examine the detailed spatiotemporal expression patterns of 19 genes implicated in early development, to learn more about their potential role in specifying and patterning early developmental processes leading to head formation. Here, we report the expression patterns of these markers in blastula- and gastrula-stage chick embryos, using whole-mount in situ hybridisation. Nodal, Fgf8, Bmp7, Chordin, Lim1, Hnf3beta, Otx2, Goosecoid, Cerberus, Hex, Dickkopf1, and Crescent are all already expressed by the time the egg is laid. When the primitive streak has reached its full length, a later group of genes, including Ganf, Six3, Bmp2, Bmp4, Noggin, Follistatin, and Qin (BF1), begins to be expressed. We reassess current models of early rostral patterning based on the analysis of these dynamic spatiotemporal expression patterns.

Animals↗
PubMed2005

Gene expression profile analysis of mouse colon embryonic development.

During late embryogenesis, the mouse colon develops from a pseudostratified, undifferentiated endoderm to a single-layered columnar epithelium with accompanying mesenchymal maturation. To identify regulatory genetic programs underlying these morphological changes, we profiled gene expression of the developing mouse colon by microarray from embryonic day (E)13.5 to E18.5. Unbiased cluster analysis of 13,484 cDNA elements revealed two distinct groups of genes whose expression changes reflect the dynamic morphological events of the epithelium and mesenchyme during this period. Additional analyses revealed two subsets of genes whose expression is either upregulated or downregulated over the same developmental period. Of those genes whose expression increases from E13.5 to E18.5 (n = 158), known functions include acquisition and/or maintenance of colonic differentiation. Genes whose transcription is downregulated over this period (n = 49) have demonstrated roles in nuclear organization, transcriptional regulation, and cell proliferation. These results provide the basis for a molecular portrait of colonic development during late embryogenesis and should be a valuable resource for investigators interested in colonic development and neoplasia, as well as comparative organogenesis.

Animals↗
PubMed1987

Expression of fasciclin I and II glycoproteins on subsets of axon pathways during neuronal development in the grasshopper.

The "labeled pathways" hypothesis predicts that axon fascicles in the embryonic neuropil are differentially labeled by surface recognition molecules used for growth cone guidance. To identify candidates for such recognition molecules, we generated monoclonal antibodies (MAbs) that recognize surface antigens expressed on subsets of axon fascicles in the grasshopper embryo. The 3B11 and 8C6 MAbs immunoprecipitate 70- and 95-kd membrane glycoproteins called fasciclin I and II, respectively, which are expressed on different subsets of axon fascicles during development. These two glycoproteins are expressed regionally on particular portions of embryonic axons in correlation with their patterns of fasciculation, dynamically during the period of axon outgrowth in a manner consistent with a role in growth cone guidance, and at other times and places during embryogenesis, suggesting multiple developmental roles.

Animals↗
PubMed2004

Gastrulation dynamics: cells move into focus.

During vertebrate gastrulation, a relatively limited number of blastodermal cells undergoes a stereotypical set of cellular movements that leads to formation of the three germ layers: ectoderm, mesoderm and endoderm. Gastrulation, therefore, provides a unique developmental system in which to study cell movements in vivo in a fairly simple cellular context. Recent advances have been made in elucidating the cellular and molecular mechanisms that underlie cell movements during zebrafish gastrulation. These findings can be compared with observations made in other model systems to identify potential general mechanisms of cell migration during development.

Animals↗