Quail Embryos Research Articles

FGF and TGFβ signaling link form and function during jaw development and evolution

How does form arise during development and change during evolution? How does form relate to function, and what enables embryonic structures to presage their later use in adults? To address these questions, we leverage the distinct functional morphology of the jaw in duck, chick, and quail. In connection with their specialized mode of feeding, duck develop a secondary cartilage at the tendon insertion of their jaw adductor muscle on the mandible. An equivalent cartilage is absent in chick and quail. We hypothesize that species-specific jaw architecture and mechanical forces promote secondary cartilage in duck through the differential regulation of FGF and TGFβ signaling. First, we perform transplants between chick and duck embryos and demonstrate that the ability of neural crest mesenchyme (NCM) to direct the species-specific insertion of muscle and the formation of secondary cartilage depends upon the amount and spatial distribution of NCM-derived connective tissues. Second, we quantify motility and build finite element models of the jaw complex in duck and quail, which reveals a link between species-specific jaw architecture and the predicted mechanical force environment. Third, we investigate the extent to which mechanical load mediates FGF and TGFβ signaling in the duck jaw adductor insertion, and discover that both pathways are mechano-responsive and required for secondary cartilage formation. Additionally, we find that FGF and TGFβ signaling can also induce secondary cartilage in the absence of mechanical force or in the adductor insertion of quail embryos. Thus, our results provide novel insights on molecular, cellular, and biomechanical mechanisms that couple musculoskeletal form and function during development and evolution.

Potency of polycyclic aromatic hydrocarbons in chicken and Japanese quail embryos.

Birds are receptors of concern for polycyclic aromatic hydrocarbons (PAHs), yet limited data describing the relative potency of PAH congeners are available for avian species. In the present study, we determined embryonic median lethal dose (LD50) values for 5 PAH congeners in chicken (Gallus gallus) and one PAH congener in Japanese quail (Coturnix japonica). Graded concentrations of each test compound were injected into the air cell of chicken or quail eggs before incubation. Embryos were monitored through development (quail) or hatching (chicken). All PAHs tested caused dose-dependent increases in embryo mortality, but few other effects (e.g., weight changes, deformities) were observed. In chicken, windows of developmental sensitivity were identified between embryonic days 4 and 9 and between embryonic days 20 and 22. The rank order potency of benzo[k]fluoranthene (BkF; 76 μg/kg) ≈ dibenz[ah]anthracene (83 μg/kg) > indeno[1,2,3-cd]pyrene (325 μg/kg) > benzo[a]pyrene (461 μg/kg) > benz[a]anthracene (529 μg/kg) corresponded well with previous in vitro estimates in birds. Previously published ethoxyresorufin-O-deethylase median effect concentrations from cultured chicken embryo hepatocytes were highly predictive of our LD50s (p < 0.001, r2 = 0.99). To explore differences in sensitivity between species, Japanese quail eggs were injected with BkF, the most potent PAH. We found that chicken and quail were nearly equally sensitive to BkF. The present results contribute to our developing understanding of variability in responses to PAHs among congeners and species. Environ Toxicol Chem 2018;37:1556-1564. © 2018 SETAC.

Hematology from embryo to adult in the bobwhite quail (Colinus virginianus): Differential effects in the adult of clutch, sex and hypoxic incubation

Hematology and its regulation in developing birds have been primarily investigated in response to relatively short-term environmental challenges in the embryo. Yet, whether any changes induced in the embryo persist into adulthood as a hematological form of “fetal programming” is unknown. We hypothesized that: 1) chronic as opposed to acute hypoxic incubation will alter hematological respiratory variables in embryos of bobwhite quail (Colinus virginianus), and 2) alterations first appearing in the embryo will persist into hatchlings through into adulthood. To test these hypotheses, we first developed an embryo-to-adult profile of normal hematological development by measuring hematocrit (Hct), red blood cell concentration ([RBC]), hemoglobin concentration ([Hb]), mean corpuscular volume, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration, as well plasma osmolality. Hct, [RBC] and [Hb] in normoxic-incubated birds (controls) steadily increased from ~22%, ~1.6 × 106 μL−1 and ~7 g% in day 12 embryos to almost double the values at maturity in adult birds. Both cohort and sex affected hematology of normoxic-incubated birds. A second population, incubated from day 0 (d0) in 15% O2, surprisingly revealed little or no significant difference from controls in hematology in embryos. In hatchlings and adults, hypoxic incubation caused no significant modification to any variables. Compared to major hematological effects caused by hypoxic incubation in chickens, the hematology of the bobwhite quail embryo appears to be minimally affected by hypoxic incubation, with very few effects induced during hypoxic incubation actually persisting into adulthood.

Stochastic clonal expansion of “superstars” enhances the reserve capacity of enteric nervous system precursor cells

We quantified cell population increase in the quail embryo enteric nervous system (ENS) from E2.5 (about 1500 cells) to E12 (about 8 million cells). We then probed ENS proliferative capacity by grafting to the chorio-allantoic membrane large (600 cells) and small (40 cells) populations of enteric neural crest (ENC) cells with aneural gut. This demonstrated that ENC cells show an extremely high capacity to regulate their proliferation while forming the ENS. Previous mathematical models and clonal label experiments revealed that a few dominant ENS “superstar” cell clones emerge but most clones are small. The model implied that “superstars” arise stochastically, but the same outcome could arise if “superstars” were pre-determined. We investigated these two modes mathematically and by grafting experiments with large and small numbers of ENCs, each including one EGFP-labelled ENC cell. The stochastic model predicts that the frequency of “superstar” detection increases as the ENC population decreases, the pre-determined model does not. Experimentally, as predicted by the stochastic model, the frequency of “superstar” detection increased with small ENC cell number. We conclude that ENS “superstar” clones achieve this status stochastically. Clonal dominance implies that clonal diversity is greatly reduced and in this case, somatic mutations may affect the phenotype. We suggest that somatic mutations coupled with loss of clonal diversity may contribute to variable penetrance and expressivity in individuals with genetically identical ENS pathologies.

Chick muscle development.

Striated muscle is the most abundant tissue in the body of vertebrates and it forms, together with the skeleton, the locomotory system required both for movement and the creation of the specific body shape of a species. Research on the embryonic development of muscles has a long tradition both in classical embryology and in molecular developmental biology. While the gene networks regulating muscle development have been discovered mostly in the mouse through genetics, our knowledge on cell lineages, muscle morphogenesis and tissue interactions regulating their formation is to a large extent based on the use of the avian model. This review highlights present knowledge of the development of skeletal muscle in vertebrate embryos. Special focus will be placed on the contributions from chicken and quail embryo model systems.

Blood Lipid Profile of Coturnix coturnix japonica Fed whith Organic Feed and Supplement Curcuma longa

&lt;p class="IsiAbstrakIndo"&gt;&lt;span lang="EN-GB"&gt;People’s concern about quail egg consumption has triggered the efforts to produce quail eggs with low cholesterol content by using organic feed. Organic feed is one type of feed rations made of mixing organic material. In this study, standard organic feed (rice bran, yellow corn, soybean, and fishmeal) and organic feed containing mackerel, cassava leaves, and turmeric powder was administere to female japanese quail. Organic feed containing mackerel, cassava leaves, and turmeric powder can affect the Japanese quail (&lt;/span&gt;&lt;em&gt;&lt;span lang="EN-GB"&gt;Coturnix coturnix japonica L.&lt;/span&gt;&lt;/em&gt;&lt;span lang="EN-GB"&gt;) lipid profile. This research used experimental methods. Twenty-seven female Japanese quails were divided into 3 treatment groups (PO: commercial feed, P1: standard organic feed, P2: feed containing organic turmeric powder, cassava leaves, and mackerel) for 145 days with 9 repeat. Data obtained were in the form of average daily feed intake and blood lipid profiles of each treatment group. Further data analysis was performed by analysis of variance (ANOVA) followed by Duncan test. The results showed that administration of organic feed containing mackerel, cassava leaves, and turmeric powder can reduce the levels of cholesterol, triglycerides, and LDL but increase the HDL levels in Japanese quail. Supplementation of turmeric powder improve the chemical quality of Japanese quail eggs, so it is good for the development of quail embryos as well as for consumption. It can be concluded that organic feed and turmeric powder supplementation stabilize blood lipid profile and improve the reproduction of Japanese quail egg low cholesterol, so good for the development of embryo quail and for consumption.&lt;/span&gt;&lt;/p&gt;

Live tissue antibody injection: A novel method for imaging ECM in limb buds and other tissues.

Understanding the morphogenesis and differentiation of tissues and organs from progenitor fields requires methods to visualize this process. Despite an ever-growing recognition that ECM plays an important role in tissue development, studies of ECM movement, and patterns in live tissue are scarce. Here, we describe a method in which a living limb bud is immunolabeled prior to fixation using fluorescent antibodies that recognize two ECM constituents, fibronectin and fibrillin 2. The results show that undifferentiated mesenchyme in quail embryos can be distinguished from prechondrogenic cellular condensations, in situ, via ECM antibodies-indicating the developmental transition from naïve mesenchyme to committed skeletal tissue. We conclude that our live tissue injection method is a general approach that allows visualization of the structural characteristics and the distribution pattern of ECM scaffolds, in situ. With slight modifications, the method will produce robust fluorescence images of ECM scaffolds in any suitable tissue mass and allow multiple kinds of optical analyses including virtual 3D reconstructions.

Uptake of radiolabeled 3,3',4,4'-tetrachlorobiphenyl into Japanese quail egg compartments and embryo following air cell and albumen injection.

The avian embryo is an excellent model for testing adverse developmental effects of environmental chemicals as well as uptake and movement of xenobiotics within the egg compartments. Before incubation at embryonic day 0, 14 C 3,3',4,4'-tetrachlorobiphenyl (14 C PCB 77) was injected into Japanese quail eggs either onto the air cell or into the albumen. All egg components were collected on embryonic day 1, 5, or 10, and concentrations of 14 C PCB 77 were measured in various egg components (shell, membrane, yolk, albumen, and embryo). The results showed measurable 14 C PCB 77 in all egg components, with changing concentrations in each egg component over the course of embryonic development. Specifically, concentrations in the shell content decreased between embryonic days 1 and 10, increased in albumen from embryonic days 1 to 5 and then decreased at embryonic day 10, and increased in both yolk and embryo from embryonic days 1 to 10. Vehicle and injection site both influenced 14 C PCB 77 allantoic fluid concentrations, with little effect on other egg components except for the inner shell membrane. The fatty acid vehicle injected into the albumen yielded the highest 14 C PCB 77 recovery. These findings demonstrate dynamic movement of toxicants throughout the egg components during avian embryonic development and a steady increase of relatively low levels of 14 C PCB 77 in the embryo compared with the yolk, albumen, and shell, suggesting that embryonic uptake (i.e., exposure) mirrors utilization of egg components for nutrition and growth during development. Environ Toxicol Chem 2018;37:126-135. © 2017 SETAC.

Critical developmental windows for morphology and hematology revealed by intermittent and continuous hypoxic incubation in embryos of quail (Coturnix coturnix).

Hypoxia during embryonic growth in embryos is frequently a powerful determinant of development, but at least in avian embryos the effects appear to show considerable intra- and inter-specific variation. We hypothesized that some of this variation may arise from different protocols that may or may not result in exposure during the embryo’s critical window for hypoxic effects. To test this hypothesis, quail embryos (Coturnix coturnix) in the intact egg were exposed to hypoxia (~15% O2) during “early” (Day 0 through Day 5, abbreviated as D0-D5), “middle” (D6-D10) or “late” (D11-D15) incubation or for their entire 16–18 day incubation (“continuous hypoxia”) to determine critical windows for viability and growth. Viability, body mass, beak and toe length, heart mass, and hematology (hematocrit and hemoglobin concentration) were measured on D5, D10, D15 and at hatching typically between D16 and D18 Viability rate was ~50–70% immediately following the exposure period in the early, middle and late hypoxic groups, but viability improved in the early and late groups once normoxia was restored. Middle hypoxia groups showed continuing low viability, suggesting a critical period from D6-D10 for embryo viability. The continuous hypoxia group experienced viability reaching <10% after D15. Hypoxia, especially during late and continuous hypoxia, also inhibited growth of body, beak and toe when measured at D15. Full recovery to normal body mass upon hatching occurred in all other groups except for continuous hypoxia. Contrary to previous avian studies, heart mass, hematocrit and hemoglobin concentration were not altered by any hypoxic incubation pattern. Although hypoxia can inhibit embryo viability and organ growth during most incubation periods, the greatest effects result from continuous or middle incubation hypoxic exposure. Hypoxic inhibition of growth can subsequently be “repaired” by catch-up growth if a final period of normoxic development is available. Collectively, these data indicate a critical developmental window for hypoxia susceptibility during the mid-embryonic period of development.

Histologis Testis pada Keturunan F1 dari Induk Puyuh (Coturnix coturnix japonica L.) yang diberi Suplemen Serbuk Kunyit (Curcuma longa L.) dalam Pakan

Quail child has a tremendous potential to continue further descent because quails have almost all the potential that can be exploited by humans, either for consumption or economic. Children quail superior quality can be produced from superior quality parent, so the parent quail quality must be considered. Additional food in the form of powder turmeric supplements can increase the phytoestrogen which stimulates the liver to produce vitellogenin as the material forming the yolk. Egg yolk is a source of nutrients for the development of quail embryos. This study aimed to analyze the differences in the quality of the parent male quail chicks were given supplements of turmeric powder to the testis weight, testis size, number of spermatogonia and spermatids amount contained in the seminiferous tubules. The study design used was completely randomized design (CRD) that is 3 treatments and 5 replications. By using quail male child as much as 15 tails as test animals, divided into three groups, namely K0: F1 generation of the female quail are not given turmeric powder supplement, K1: F1 generation of the female quail turmeric powder supplemented with a dose of 54 mg/tail/day and K2: kuturunan F1 of the parent quail turmeric powder supplemented with a dose of 108 mg/tail/day. The variables measured were testis weight, testis size, number of spermatogonia and spermatids number. Data were analyzed using Analysis of Variance (ANOVA) followed by a further test Duncan Multiple Range Test (DMRT) at the 95% significance level. The results showed that the quality of parent child quail turmeric powder supplemented with a dose of 108 mg/tail/day for better views of the process of spermatogenesis. Keywords: Child quail (Coturnix coturnix japonica. L), Turmeric, Testis, spermatogonia, spermatids.

The intersensory redundancy hypothesis: Extending the principle of unimodal facilitation to prenatal development.

Selective attention to different properties of stimulation provides the foundation for perception, learning, and memory. The Intersensory Redundancy Hypothesis (IRH) proposes that early in development information presented redundantly across two or more modalities (multimodal) selectively recruits attention to and enhances perceptual learning of amodal properties, whereas information presented to a single sense modality (unimodal) enhances perceptual learning of modality-specific properties. The present study is the first to assess this principle of unimodal facilitation in non-human animals in prenatal development. We assessed bobwhite quail embryos' prenatal detection of pitch, a modality-specific property, under conditions of unimodal and bimodal (synchronous or asynchronous) exposure. Chicks exposed to prenatal unimodal auditory stimulation or asynchronous bimodal (audiovisual) stimulation preferred the familiarized maternal call over a novel pitch-modified maternal call following hatching, whereas chicks exposed to redundant (synchronous) audiovisual stimulation failed to prefer the familiar call over the pitch-modified call. These results provide further evidence that selective attention is recruited to specific stimulus properties of events in early development and that these biases are evident even during the prenatal period.

Onset of microglial entry into developing quail retina coincides with increased expression of active caspase-3 and is mediated by extracellular ATP and UDP.

Microglial cell precursors located in the area of the base of the pecten and the optic nerve head (BP/ONH) start to enter the retina of quail embryos at the 7th day of incubation (E7), subsequently colonizing the entire retina by central-to-peripheral tangential migration, as previously shown by our group. The present study demonstrates a precise chronological coincidence of the onset of microglial cell entry into the retina with a striking increase in death of retinal cells, as revealed by their active caspase-3 expression and TUNEL staining, in regions dorsal to the BP/ONH area, suggesting that dying retinal cells would contribute to the microglial cell inflow into the retina. However, the molecular mechanisms involved in this inflow are currently unclear. Extracellular nucleotides, such as ATP and UDP, have previously been shown to favor migration of microglia towards brain injuries because they are released by apoptotic cells and stimulate both chemotaxis and chemokinesis in microglial cells via signaling through purinergic receptors. Hence, we tested here the hypothesis that ATP and UDP play a role in the entry and migration of microglial precursors into the developing retina. For this purpose, we used an experimental model system based on organotypic cultures of E6.5 quail embryo retina explants, which mimics the entry and migration of microglial precursors in the in situ developing retina. Inhibition of purinergic signaling by treating retina explants with either apyrase, a nucleotide-hydrolyzing enzyme, or suramin, a broad spectrum antagonist of purinergic receptors, significantly prevents the entry of microglial cells into the retina. In addition, treatment of retina explants with either exogenous ATP or UDP results in significantly increased numbers of microglial cells entering the retina. In light of these findings, we conclude that purinergic signaling by extracellular ATP and UDP is necessary for the entry and migration of microglial cells into the embryonic retina by inducing chemokinesis in these cells.

High-resolution Episcopic Microscopy (HREM) - Simple and Robust Protocols for Processing and Visualizing Organic Materials

We provide simple protocols for generating digital volume data with the high-resolution episcopic microscopy (HREM) method. HREM is capable of imaging organic materials with volumes up to 5 x 5 x 7 mm3 in typical numeric resolutions between 1 x 1 x 1 and 5 x 5 x 5 µm3. Specimens are embedded in methacrylate resin and sectioned on a microtome. After each section an image of the block surface is captured with a digital video camera that sits on the phototube connected to the compound microscope head. The optical axis passes through a green fluorescent protein (GFP) filter cube and is aligned with a position, at which the bock holder arm comes to rest after each section. In this way, a series of inherently aligned digital images, displaying subsequent block surfaces are produced. Loading such an image series in three-dimensional (3D) visualization software facilitates the immediate conversion to digital volume data, which permit virtual sectioning in various orthogonal and oblique planes and the creation of volume and surface rendered computer models. We present three simple, tissue specific protocols for processing various groups of organic specimens, including mouse, chick, quail, frog and zebra fish embryos, human biopsy material, uncoated paper and skin replacement material.

High-resolution Episcopic Microscopy (HREM) - Simple and Robust Protocols for Processing and Visualizing Organic Materials.

We provide simple protocols for generating digital volume data with the high-resolution episcopic microscopy (HREM) method. HREM is capable of imaging organic materials with volumes up to 5 x 5 x 7 mm3 in typical numeric resolutions between 1 x 1 x 1 and 5 x 5 x 5 µm3. Specimens are embedded in methacrylate resin and sectioned on a microtome. After each section an image of the block surface is captured with a digital video camera that sits on the phototube connected to the compound microscope head. The optical axis passes through a green fluorescent protein (GFP) filter cube and is aligned with a position, at which the bock holder arm comes to rest after each section. In this way, a series of inherently aligned digital images, displaying subsequent block surfaces are produced. Loading such an image series in three-dimensional (3D) visualization software facilitates the immediate conversion to digital volume data, which permit virtual sectioning in various orthogonal and oblique planes and the creation of volume and surface rendered computer models. We present three simple, tissue specific protocols for processing various groups of organic specimens, including mouse, chick, quail, frog and zebra fish embryos, human biopsy material, uncoated paper and skin replacement material.

Effects of selective and combined activation of estrogen receptor α and β on reproductive organ development and sexual behaviour in Japanese quail (Coturnix japonica).

Excess estrogen exposure of avian embryos perturbs reproductive organ development in both sexes and demasculinizes the reproductive behaviors of adult males. We have previously shown that these characteristic effects on the reproductive organs also can be induced by exposure of Japanese quail (Coturnix japonica) embryos to selective agonists of estrogen receptor alpha (ERα). In contrast, the male copulatory behavior is only weakly affected by developmental exposure to an ERα agonist. To further elucidate the respective roles of ERα and ERβ in estrogen-induced disruption of sexual differentiation, we exposed Japanese quail embryos in ovo to the selective ERα agonist 16α-lactone-estradiol (16αLE2), the selective ERβ agonist WAY-200070, or both substances in combination. The ERα agonist feminized the testes in male embryos and reduced cloacal gland size in adult males. Furthermore, anomalous retention and malformations of the Müllerian ducts/oviducts were seen in embryos and juveniles of both sexes. The ERβ agonist did not induce any of these effects and did not influence the action of the ERα agonist. Male copulatory behavior was not affected by embryonic exposure to either the ERα- or the ERβ-selective agonist but was slightly suppressed by treatment with the two compounds combined. Our results suggest that the reproductive organs become sexually differentiated consequent to activation of ERα by endogenous estrogens; excessive activation of ERα, but not ERβ, during embryonic development may disrupt this process. Our results also suggest that the demasculinizing effect of estrogens on male copulatory behavior is only partly mediated by ERα and ERβ, and may rather involve other estrogen-responsive pathways.

Incubation Temperature Alters Temperature-Dependent Oxygen Consumption in Northern Bobwhite Quail Hatchlings (Colinus virginianus).

This study investigated the effect of mismatching incubation and posthatch temperatures in northern bobwhite quail hatchlings. Quail embryos were incubated at 35.5° or 37.5°C. Metabolic rates were then measured in hatchlings acclimated to either the same or the opposite temperature treatment. While hatchlings expressed higher oxygen consumption when posthatch temperature did not match incubation temperature, the effect of mismatching temperatures was significant only when posthatch temperature was higher than incubation temperature. Our data suggest that bobwhite quail hatchlings may express increased metabolism due to mismatches between incubation and posthatch temperatures. More specifically, the nature or direction of the mismatch can determine the magnitude of the metabolic effect. These findings highlight the importance of considering the context of specific conditions experienced throughout ontogeny when observing phenotypic outcomes.

Goose embryonic development from oviposition through 16 hours of incubation

Normal tables provide an objective step-wise description of the morphological development of an embryo. Such tables have been described for the chicken, turkey, quail, and duck embryos, but there is no such staging table for goose embryos. As the goose has one of the longest incubation periods of all the poultry species and embryo mortality during incubation is relatively high, a normal table of goose embryo development would be useful in assessing the morpho-genetic status of the goose embryo before and during incubation. In this study, embryos were isolated from commercial White Koluda goose eggs stored no longer than four days in a cool room (18°C) prior to incubation and after 4, 8, 12, and 16 h of incubation. Embryo staging was based on the normal tables described for the chicken by Eyal-Giladi and Kochav (EGK) and Hamburger and Hamilton (HH). Goose embryos from unincubated eggs were at Stage X and XI EGK and after 16 h of incubation the majority of embryos were between Stages 2 and 4 HH. Our results suggest that while the stage of development of the embryo in the unincubated goose egg is similar to that reported for the chicken, although the diameter of goose embryo is slighter larger. Following incubation, a goose embryo advances more slowly than a chicken embryo up to 16 h of incubation.

The differential expression of MC1R regulators in dorsal and ventral quail plumages during embryogenesis: Implications for plumage pattern formation.

Melanin pigmentation patterns are ubiquitous in animals and function in crypsis, physical protection, thermoregulation and signalling. In vertebrates, pigmentation patterns formed over large body regions as well as within appendages (hair/feathers) may be due to the differential distribution of pigment producing cells (melanocytes) and/or regulation of the melanin synthesis pathway. We took advantage of the pigmentation patterns of Japanese quail embryos (pale ventrum and patterned feathers dorsally) to explore the role of genes and their transcripts in regulating the function of the melanocortin-1-receptor (MC1R) via 1. activation: pro-opiomelanocortin (POMC), endoproteases prohormone convertase 1 (PC1) and 2 (PC2), and 2. inhibition—agouti signaling and agouti-related protein (ASIP and AGRP, respectively). Melanocytes are present in all feather follicles at both 8 and 12 days post-fertilisation (E8/E12), so differential deposition of melanocytes is not responsible for pigmentation patterns in embryonic quail. POMC transcripts expressed were a subset of those found in chicken and POMC expression within feather follicles was strong. PC1 was not expressed in feather follicles. PC2 was strongly expressed in all feather follicles at E12. ASIP transcript expression was variable and we report four novel ASIP transcripts. ASIP is strongly expressed in ventral feather follicles, but not dorsally. AGRP expression within feather follicles was weak. These results demonstrate that the pale-bellied quail phenotype probably involves inhibition of MC1R, as found previously. However, quail may require MC1R activation for eumelanogenesis in dorsal feathers which may have important implications for an understanding of colour pattern formation in vertebrates.

Coordinated movement is influenced by prenatal light experience in bobwhite quail chicks (Colinus virginianus)

Sensory-motor development begins early during embryogenesis and is influenced by sensory experience. Little is known about the prenatal factors that influence the development of motor coordination. Here we investigated whether and to what extent prenatal light experience can influence the development of motor coordination in bobwhite quail hatchlings. Quail embryos were incubated under four light conditions: no light (dark), 2h of total light (2HR), 6h of total light (6HR), and diffused sunlight (controls). Hatchlings were video recording walking down a runway at three developmental ages (12, 24, and 48h). Videos were assessed for forward locomotion, a measurement of motor coordination, falls, a measurement of motor instability, and motivation to complete the task. We anticipated a linear decline of coordination with a reduction in prenatal light experience and improved coordination with age. Furthermore, as motor coordination becomes more laborious we anticipated motivation to complete the task would decline. However, our findings revealed hatchlings did not uniformly improve with age as expected, nor did the reduction of light result in a linear reduction in motor coordination. Instead, we found a more complex relationship with 6HR and 2HR hatchlings showing distinct patterns of stability and instability. Similarly, we found a reduction in motivation within the 6HR light condition. It appears that prenatal light exposure influences the development of postnatal motor coordination and we discuss these finding in light of neurodevelopmental processes influenced by light experience.

Basal filopodia and vascular mechanical stress organize fibronectin into pillars bridging the mesoderm-endoderm gap

Cells may exchange information with other cells and tissues by exerting forces on the extracellular matrix (ECM). Fibronectin (FN) is an important ECM component that forms fibrils through cell contacts and creates directionally biased geometry. Here, we demonstrate that FN is deposited as pillars between widely separated germ layers, namely the somitic mesoderm and the endoderm, in quail embryos. Alongside the FN pillars, long filopodia protrude from the basal surfaces of somite epithelial cells. Loss-of-function of Ena/VASP, α5β1-integrins or talin in the somitic cells abolished the FN pillars, indicating that FN pillar formation is dependent on the basal filopodia through these molecules. The basal filopodia and FN pillars are also necessary for proper somite morphogenesis. We identified a new mechanism contributing to FN pillar formation by focusing on cyclic expansion of adjacent dorsal aorta. Maintenance of the directional alignment of the FN pillars depends on pulsatile blood flow through the dorsal aortae. These results suggest that the FN pillars are specifically established through filopodia-mediated and pulsating force-related mechanisms.