Effects of Thyroxine Exposure on Osteogenesis in Mouse Calvarial Pre-Osteoblasts

The incidence of craniosynostosis is one in every 1,800-2500 births. The gene-environment model proposes that if a genetic predisposition is coupled with environmental exposures, the effects can be multiplicative resulting in severely abnormal phenotypes. At present, very little is known about the role of gene-environment interactions in modulating craniosynostosis phenotypes, but prior evidence suggests a role for endocrine factors. Here we provide a report of the effects of thyroid hormone exposure on murine calvaria cells. Murine derived calvaria cells were exposed to critical doses of pharmaceutical thyroxine and analyzed after 3 and 7 days of treatment. Endpoint assays were designed to determine the effects of the hormone exposure on markers of osteogenesis and included, proliferation assay, quantitative ALP activity assay, targeted qPCR for mRNA expression of Runx2, Alp, Ocn, and Twist1, genechip array for 28,853 targets, and targeted osteogenic microarray with qPCR confirmations. Exposure to thyroxine stimulated the cells to express ALP in a dose dependent manner. There were no patterns of difference observed for proliferation. Targeted RNA expression data confirmed expression increases for Alp and Ocn at 7 days in culture. The genechip array suggests substantive expression differences for 46 gene targets and the targeted osteogenesis microarray indicated 23 targets with substantive differences. 11 gene targets were chosen for qPCR confirmation because of their known association with bone or craniosynostosis (Col2a1, Dmp1, Fgf1, 2, Igf1, Mmp9, Phex, Tnf, Htra1, Por, and Dcn). We confirmed substantive increases in mRNA for Phex, FGF1, 2, Tnf, Dmp1, Htra1, Por, Igf1 and Mmp9, and substantive decreases for Dcn. It appears thyroid hormone may exert its effects through increasing osteogenesis. Targets isolated suggest a possible interaction for those gene products associated with calvarial suture growth and homeostasis as well as craniosynostosis. © 2013 Cray et al

Elongation, proliferation & migration differentiate endothelial cell phenotypes and determine capillary sprouting

<p>Abstract</p> <p>Background</p> <p>Angiogenesis, the growth of capillaries from preexisting blood vessels, has been extensively studied experimentally over the past thirty years. Molecular insights from these studies have lead to therapies for cancer, macular degeneration and ischemia. In parallel, mathematical models of angiogenesis have helped characterize a broader view of capillary network formation and have suggested new directions for experimental pursuit. We developed a computational model that bridges the gap between these two perspectives, and addresses a remaining question in angiogenic sprouting: how do the processes of endothelial cell elongation, migration and proliferation contribute to vessel formation?</p> <p>Results</p> <p>We present a multiscale systems model that closely simulates the mechanisms underlying sprouting at the onset of angiogenesis. Designed by agent-based programming, the model uses logical rules to guide the behavior of individual endothelial cells and segments of cells. The activation, proliferation, and movement of these cells lead to capillary growth in three dimensions. By this means, a novel capillary network emerges out of combinatorially complex interactions of single cells. Rules and parameter ranges are based on literature data on endothelial cell behavior in vitro. The model is designed generally, and will subsequently be applied to represent species-specific, tissue-specific in vitro and in vivo conditions.</p> <p>Initial results predict tip cell activation, stalk cell development and sprout formation as a function of local vascular endothelial growth factor concentrations and the Delta-like 4 Notch ligand, as it might occur in a three-dimensional in vitro setting. Results demonstrate the differential effects of ligand concentrations, cell movement and proliferation on sprouting and directional persistence.</p> <p>Conclusion</p> <p>This systems biology model offers a paradigm closely related to biological phenomena and highlights previously unexplored interactions of cell elongation, migration and proliferation as a function of ligand concentration, giving insight into key cellular mechanisms driving angiogenesis.</p

Gene expression profiling in human craniosynostoses: a tool to investigate the molecular basis of suture ossification

Non-sy ndromic craniosy nostoses (NSC) occur as isolated skull malf ormations due to the premature ossif ication of one (single suture f orms) or more (complex f orms) calv arial sutures and represent the most f requent f orm of craniosy nostosis worldwide. The etiology of NSC is still largely unknown, as a genetic basis can be rarely demonstrated especially in single-suture f orms. In these cases, during the prenatal/perinatal dev elopment of af f ected patients, only one suture undergoes a premature direct ossif ication within an otherwise phy siologically grown skull. This could suggest that def inite somatic alterations, possibly due to unclear env ironmental agents, occur locally at the site of premature suture f usion during skull dev elopment. A promising tool to inv estigate the molecular mechanisms that may orchestrate this ev ent is the comparativ e analy sis of suture- and sy nostosis-deriv ed tissues and cells. Particularly , this rev iew will f ocus on the dif f erent studies that attempted to clarif y this issue using genome-wide microarray -based technologies f or the comparativ e analy sis of gene expression prof iles. All relev ant results will be comprehensiv ely rev iewed, possibly compared and critically discussed

To eat or not to eat? debris selectivity by marine turtles

Marine debris is a growing problem for wildlife, and has been documented to affect more than 267 species worldwide. We investigated the prevalence of marine debris ingestion in 115 sea turtles stranded in Queensland between 2006-2011, and assessed how the ingestion rates differ between species (Eretmochelys imbricata vs. Chelonia mydas) and by turtle size class (smaller oceanic feeders vs. larger benthic feeders). Concurrently, we conducted 25 beach surveys to estimate the composition of the debris present in the marine environment. Based on this proxy measurement of debris availability, we modeled turtles' debris preferences (color and type) using a resource selection function, a method traditionally used for habitat and food selection. We found no significant difference in the overall probability of ingesting debris between the two species studied, both of which have similar life histories. Curved carapace length, however, was inversely correlated with the probability of ingesting debris; 54.5% of pelagic sized turtles had ingested debris, whereas only 25% of benthic feeding turtles were found with debris in their gastrointestinal system. Benthic and pelagic sized turtles also exhibited different selectivity ratios for debris ingestion. Benthic phase turtles had a strong selectivity for soft, clear plastic, lending support to the hypothesis that sea turtles ingest debris because it resembles natural prey items such as jellyfish. Pelagic turtles were much less selective in their feeding, though they showed a trend towards selectivity for rubber items such as balloons. Most ingested items were plastic and were positively buoyant. This study highlights the need to address increasing amounts of plastic in the marine environment, and provides evidence for the disproportionate ingestion of balloons by marine turtles