Altered Gravity and Early Heart Development in Culture

The macromolecules comprising the cytoskeleton and extracellular matrix of cells may be sensitive to gravitation. Since early development of organs depends on dynamic interactions across cell surfaces, altered gravity may disturb development. We investigated this possibility for heart development. Previous studies showed that the extracellular matrix glycoprotein fibronectin (Fn) is necessary for normal heart development. We cultured precardiac tissue explants in a high aspect ratio bioreactor vessel (HARV) to simulate microgravity. We observed tissue morphology, contraction, and Fn distribution by immunolocalization in HARV rotated and control (lxg) explants, cultured 18 hr. We also measured Fn amount by immunoassay. Explants in HARV were rotated at 6 rpm to achieve continuous freefall. Thirty-five of 37 control, but only 1 of 37 matched rotated explants exhibited contractions. Tissue architecture was identical. Immunolocalization of Fn showed remarkable differences which may be related to the development of contractions. The Fn staining in the HARV explants was less intense in all areas. Areas of linear staining along epithelia were present but shorter, and there was less intercellular staining in both mesenchymal tissue and myocardium. Initial immunoassay results of 5 matched pairs of explants showed a 22% reduction in total tissue Fn in the HARV rotated samples. Our results indicate that altered gravity in the HARV reduced the amount and distribution of Fn, as assessed by two independent criteria. This was correlated with a reduction in the development of contractile activity

Homocysteine and Cardiac Neural Crest Cell Cytoskeletal Proteins in the Chick Embryo

Elevated serum homocysteine (Hcys) is correlated with cardiovascular disease and with embryonic malformations related to neural crest cells (NCCs). We predicted Hcys may alter the balance of actin networks, stress fibers and focal adhesions, altering migration. We cultured neural tube explants in control and Hcys-treated medium and visualized actin, α-actinin, vinculin, filamin, and LIM3 protein in NCCs migrating outward. In Hcys, phalloidin-stained actin in stress fibers was brighter, and vinculin was more abundant in focal adhesions and lamellipodia. α-actinin and LIM3 were also enhanced around nuclei and in focal adhesions, and α-actinin also in filopodia. Filamin was unchanged. Hcys caused more spreading and migration of NCCs, but not more cell-cell adhesions. The findings support our hypothesis that Hcys adjusts NCCs for greater adhesion and migration. Its effect on LIM3 suggests it may modulate signaling that adjusts the cytoskeleton for enhanced migration, leading to mistimed and defective development of target tissues

Is High Folic Acid Intake a Risk Factor for Autism?—A Review

Folate is required for metabolic processes and neural development. Insuring its adequate levels for pregnant women through supplementation of grain-based foods with synthetic folic acid (FA) in order to prevent neural tube defects has been an ongoing public health initiative. However, because women are advised to take multivitamins containing FA before and throughout pregnancy, the supplementation together with natural dietary folates has led to a demographic with high and rising serum levels of unmetabolized FA. This raises concerns about the detrimental effects of high serum synthetic FA, including a rise in risk for autism spectrum disorder (ASD). Some recent studies have reported a protective effect of FA fortification against ASD, but others have concluded there is an increased risk for ASD and other negative neurocognitive development outcomes. These issues are accompanied by further health questions concerning high, unmetabolized FA levels in serum. In this review, we outline the reasons excess FA supplementation is a concern and review the history and effects of supplementation. We then examine the effects of FA on neuronal development from tissue culture experiments, review recent advances in understanding of metabolic functional blocks in causing ASD and treatment for these with alternative forms such as folinic acid, and finally summarize the conflicting epidemiological findings regarding ASD. Based on the evidence evaluated, we conclude that caution regarding over supplementing is warrante

Expression of Cadherin-11 during Organogenesis in the Chick Embryo

Cadherin-11 (cad-11) is primarily a mesenchymal cadherin that appears in delaminating neural crest cells. Its expression correlates with morphogenetic events and the pattern has been studied in mouse, rat and Xenopus embryos, but not during avian organogenesis. Our purpose was to investigate this pattern in the chick embryo during organogenesis using immunolocalization and in situ hybridization. Cad-11 was expressed in mesenchyme around the pharynx and aortic arches, eyes, auditory vesicles, lung buds, stomach, and nasal placodes. Neural expression included some cranial ganglia and also new neuroepithelium within the tail bud region undergoing secondary neurulation. We also found expression in epithelia of the developing circulatory and digestive organs. The limb buds, pineal rudiment and mesonephros were also positive. Cad-11 expression became more widespread with development. Our findings support the role of cad-11 as a mesenchymal cadherin, but provide evidence for a wider role that includes epithelial morphogenesis and secondary neurulation

Xenopus Development from Late Gastrulation to Feeding Tadpole in Simulated Microgravity

Microgravity (microG) is known to influence cytoskeletal structure, but its effects on cell migration are not well understood. To examine the effects of altered gravity on neural crest cell (NCC) migration, we inserted Xenopus laevis embryos into two separate microG-simulating slow turning lateral vessels (STLVs) just before neurulation (stage 11-12), and exposed them until feeding stage (stage 45), when the jaws and branchial apparatus are fully functional. To evaluate apparatus-related artifacts, we used two different STLVs and a vibration control as well as a stationary control vessel. Larval growth, pattern of NCC-derived cartilage formation, and incidence of malformations were analyzed using immunolocalization and wholemount staining of cartilage with Alcian blue. Interestingly, the two STLVs often yielded different or conflicting results. Many differences, such as increased cartilage size, attenuated Hoxa2 expression, and increased cell division, may be attributed mainly to vibration of the rotating vessels. However, tadpoles that developed in simulated microgravity (both STLVs, but not the vibration control) showed significantly more skeletal abnormalities, with stronger effects on cartilages derived from NCCs than those derived mainly from mesoderm. We conclude that migrating NCCs of Xenopus are sensitive to the altered gravitational environment of STLVs, and that studies relying on bioreactors to simulate microgravity also need to take variation in apparatus into account

Isotope ratios of H, C, and O in CO2 and H2O of the Martian atmosphere

Stable isotope ratios of H, C, and O are powerful indicators of a wide variety of planetary geophysical processes, and for Mars they reveal the record of loss of its atmosphere and subsequent interactions with its surface such as carbonate formation. We report in situ measurements of the isotopic ratios of D/H and O-18/O-16 in water and C-13/C-12, O-18/O-16, O-17/O-16, and (CO)-C-13-O-18/(CO)-C-12-O-16 in carbon dioxide, made in the martian atmosphere at Gale Crater from the Curiosity rover using the Sample Analysis at Mars (SAM)'s tunable laser spectrometer (TLS). Comparison between our measurements in the modern atmosphere and those of martian meteorites such as ALH 84001 implies that the martian reservoirs of CO2 and H2O were largely established similar to 4 billion years ago, but that atmospheric loss or surface interaction may be still ongoing