Second Day Of Incubation Research Articles

Embryonic development of chicken (Gallus Gallus Domesticus) From 1st to 19th Day-ectodermal structures

Birds occupy a prominent place in the Brazilian economy not only in the poultry industry but also as an animal model in many areas of scientific research. Thus the aim of this study was to provide a description of macro and microscopic aspects of the ectoderm-derived structures in chicken embryos / fetuses poultry (Gallus gallus domesticus) from 1st to 19th day of incubation. 40 fertilized eggs, from a strain of domestic chickens, with an incubation period of 2-19 days were subjected to macroscopic description, biometrics, light, and scanning microscopy. All changes observed during the development were described. The nervous system, skin and appendages and organs related to vision and hearing began to be identified, both macro and microscopically, from the second day of incubation. The vesicles from the primitive central nervous system-forebrain, midbrain, and hindbrain-were identified on the third day of incubation. On the sixth day of incubation, there was a clear vascularization of the skin. The optic vesicle was first observed fourth day of development and on the fifth day there was the beginning of the lens formation. Although embryonic development is influenced by animal line as well as external factors such as incubation temperature, this paper provides a chronological description for chicken (Gallus gallus domesticus) during its embryonic development.

Answer to February 2013 Photo Quiz

Answer: Leptotrichia buccalis bacteremia. On the second day of incubation, two FN anaerobic-blood-culture bottles yielded an anaerobic Gram-negative bacillus that could not be identified by the routine identification methods used in our laboratory. The isolate was sent to Mayo Medical Laboratories

Factors in ventricular and atrioventricular valve growth: An embryologist's perspective

1058-9813/$ – see front matter © 2010 Elsevier Ireland doi:10.1016/j.ppedcard.2010.02.010 Growth of the prenatal heart is predominantly based on adding new cells (hyperplasia), in contrast to the postnatal heart, where further increase of ventricular mass is due almost entirely to increase in myocyte size (hypertrophy) soon after birth. The key epigenetic factors regulating cardiac growth are the hemodynamics (volume/ preload and pressure/afterload), whose gradual increase reflects the changing demands of the growing embryo and fetus.While genesmay play a significant part in the etiology of congenital heart disease, hemodynamic perturbations also can lead to predictable changes in morphogenesis and produce cardiac lesions. At present, developmental perturbations seem to produce many of the common congenital defects, while genetic abnormalities are clearly linked to the many chromosomal syndromes that have been well described. The aim of this article is to provide an overview of our knowledge of the growth and development of ventricles and valves gained from experimental fetal animal models. Most hemodynamic perturbation procedures by this author have been performed in the chick model. The incubation period of the chick embryo is 21 days and the tubular heart starts to beat on the second day of incubation. The chambers start to differentiate by the third day, which includes the development of the atrioventricular and outflow tract cushions. Ventricular and outflow tract septation is completed by day 8, i.e., after one third of the incubation period. Consequently, the developmental events proceed over a relatively short period of time, allowing the effects of hemodynamic perturbations to be directly studied.

Effect of Potassium Dihydrogen Phosphate and Bovine Manure Compost on the Degradation of Chlorothalonil in Soil

The effects of potassium dihydrogen phosphate (KH2PO4) and bovine manure compost (BMC) on the degradation and metabolism of chlorothalonil were examined in a sandy loam soil under laboratory conditions. In non-sterilized, non-amended soil, chlorothalonil degradation half-life was 8.8 days. However, it was up to 19.0 days in sterilized non-amended soil, suggesting that the degradation rate was about 2-fold slower than that in non-sterilized non-amended soil. Biological mechanisms accounted for 54.7% of chlorothalonil degradation in non-sterilized soil, indicating that the indigenous microorganisms in soil play an important role in the degradation process. In non-sterilized soil, the more acutely toxic metabolite, 4-OH-chlorothalonil (HTI), of chlorothalonil started forming after the second day of incubation. The concentration of HTI reached its maximum level (2.9 μ g g− 1) at 10 days after treatment, and no further degradation of HTI was observed in the following span of 20 days (10–30 days of incubation). However, the degradation rate of chlorothalonil was decreased substantially by the addition of KH2PO4 with a half-life of 17.5 days. No formation of HTI was observed before 10 days of incubation and no significant difference of the metabolite concentration at the end of experiment was observed between KH2PO4-amended and non-amended treatments, which denoted that the addition of KH2PO4 did not reduce the formation of HTI in the longer incubation course. In BMC-amended soil, the degradation rate was about 1.8 times faster than in non-amended soil. At 30 days of incubation, both chlorothalonil and HTI were degraded to a lower level in BMC-amended soil than in non-amended soil. The application of farm litter is a common fertilizing practice in vegetable fields in China, and thus this practice could not only improve soil fertility but also promote the removal of chlorothalonil and its metabolite HTI to further increase safety in crop rotations.

A New Central venous Catheter Cap: Decreased Microbial Growth and Risk for Catheter-Related Bloodstream Infection

Catheter-related blood stream infection (CRBI) is a major cause of morbidity and mortality, and is a source of significant healthcare expenditures in patients that require central venous catheters for intravenous nutrition, chemotherapy, and other products. The source of many catheter-related infections is contamination of the catheter hub. Herein an antimicrobial catheter cap, the AB Cap is described. The AB Cap device is a catheter cleaning device designed to keep needleless luer valves clean by encapsulating them in a cleaning solution. This device was evaluated using an in vitro model of hub contamination with Staphylococcus aureus, Staphylococcus epidermidis (S. epidermidis), Klebsiella pneumonia (K. pneumonia), Pseudomonas aeruginosa, Escherichia coli and Candida albicans (C. albicans). Following hub contamination on days 1, 3, 5 and 7, saline was infused through the AB Cap and effluent collected from the efferent end. The effluent fluid was cultured for the index organisms, and allowed to incubate in culture for up to 7 days. Negative control caps were not contaminated and positive controls lacked cleaning solution and were contaminated. Microbial growth developed for all index organisms, and generally within 1 day of culture growth following the first day of contamination (day 1) in effluent from all positive controls, while no growth occurred in effluent from negative controls. No growth of any organism occurred in any of the test items after the first day of contamination. Growth of three organisms was detected in two of the three test AB Caps following contamination day 3, after 1-4 days of incubation. All organisms could be cultured in the effluent from two of the three test items at contamination day 5, generally by the second day of incubation. One test item remained free of growth for the entire test period except for one organism. By day 7, this particular test item grew an additional organism and the testing was concluded. All positive growth test items remained positive on subsequent inoculations during culture of newly obtained effluent with the exception of test item A, from which effluent following inoculation on day 3 showed growth of S. epidermidis and K. pneumonia, but no growth for these organisms from effluent obtained on inoculation day 5. In addition, effluent from test item C showed growth of C. albicans from inoculation day 5, but no growth from effluent obtained on inoculation day 7. The growth of S. epidermidis from effluent of test item A from the day 3 inoculation, and C. albicans from effluent of test items B and C did not occur until day 4 of incubation, suggesting a very small amount of contamination. An antimicrobial catheter cap is not a complete substitute for a proper catheter cleaning technique and other anti-infection precautions. However, we describe a unique catheter cap that significantly decreased the likelihood of a catheter-related infection from a non-cleaned cap in an in vitro model.

Effects of liming on the microbial biomass and its activities in soils long-term contaminated by toxic elements

The effects of liming by CaO and CaCO<sub>3</sub> on soil microbial characteristics were studied during laboratory incubation of long-term contaminated arable and grassland soils from the vicinity of lead smelter near Příbram (Czech Republic). The CaO treatment showed significant negative effects on soil microbial biomass C and its respiratory activity in both studied soils, despite the fact that microbial biomass C in the grassland soil increased sharply during the first day of incubation. The metabolic quotient (qCO<sub>2</sub>) in soils amended by CaO showed greater values than the control from the second day of incubation, indicating a possible stress of soil microbial pool. The vulnerability of organic matter to CaO could be indicated by the availability of K<sub>2</sub>SO<sub>4</sub>-extractable carbon that increased sharply, particularly at the beginning of the experiment. The amendment of soils by CaCO<sub>3 </sub>moderately increased the soil microbial biomass. The respiratory activity and qCO<sub>2</sub> increased sharply during the first day of incubation, however it is not possible to ascribe them only to microbial activities, but also to CaCO<sub>3</sub> decomposition in hydrogen carbonates, water and CO<sub>2</sub>. The pH values increased more sharply under CaO treatment in comparison to CaCO<sub>3</sub> treatment. The improvement of soil pH by CaCO<sub>3</sub> could be therefore more convenient for soil microbial communities.

The changing cellular environments of hematopoiesis in human development in utero

The hematopoietic system is indispensable from the earliest stages of development and adapts to the rapidly changing anatomy of the embryo and fetus; this takes place in such different anatomic locations as the yolk sac blood island, hepatic parenchyme, aorta-gonads-mesonephros paravascular mesenchyme, and bone marrow primary logette. We herein summarize our investigation of these serial blood-forming events in the human embryo and fetus. The access to early stages of human development, availability of a large panoply of molecular markers for human blood cell lineages, and recent development of robust assays for the earliest human hematopoietic stem cells have allowed us to gain relatively clear insight into the developmental sequence that underlies the ontogeny of human blood cells. Conversely, the control exerted by these diverse cellular environments on the emergence of human hematopoietic cells remains elusive, as is the case in animal models. We nonetheless present preliminary attempts to decipher the structure and molecular characteristics of the distinct cellular "niches" in which blood cells are produced during human gestation.

Effects of Small-Scale Turbulence on Bacteria: A Matter of Size

We examined the influence of small-scale turbulence and its associated shear on bacterioplankton abundance and cell size. We incubated natural microbial assemblages and bacteria-only fractions and subjected them to treatments with turbulence and additions of mineral nutrients and/or organic carbon. Bacterial abundance was not affected directly by turbulence in bacteria-only incubations. In natural microbial assemblage incubations, bacterial concentrations were higher under turbulence than in still-water controls when nutrients were added. In general, in the turbulence treatments bacteria increased significantly in size, mainly due to elongation of cells. The addition of inorganic nutrients had a negative effect on bacterial size, but a significantly positive effect on abundance independently of other factors such as turbulence and the presence of predators. Flagellate grazing did not trigger an increase in bacterial size as a grazing resistance response in unmixed containers. With the addition of organic carbon, bacteria elongated and partly settled to the bottom of the containers, in both the turbulent and still treatment, but bacterial abundance did not further increase. Furthermore, bacteria aggregated in the turbulence treatments after the second day of incubation even in the absence of other components of the microbial community. We found that turbulence and the associated shear increase bacterial size and change bacterial morphology, at least under certain nutrient conditions. This might be due to a physiological response (enhanced growth rate and/or unbalanced growth) or due to the selection of opportunistic strains when organic carbon is in excess compared to mineral nutrients. We suggest that shear associated with turbulent flow enhances the DOM flux to bacteria directly as well as indirectly through enhanced grazing activity and photosynthetic release. The formation of bacterial aggregates and filaments under turbulence might give selective advantage to bacteria in terms of nutrient uptake and grazing resistance.

Strontium Transport, Distribution, and Toxic Effects on Maize Seedling Growth

Two-day-old seedlings of maize (Zea mays L.) were incubated on 3 mM and 35 μM solutions of Sr(NO3)2, and the toxic effects of strontium were assessed by measuring, in the course of four days of incubation, the daily increments of the primary root length and also the root and shoot length by day 7 of incubation, and the length of the fully elongated cells. Sodium rhodizonate, a reagent developing the colored complex with Sr, was used to follow Sr distribution in maize tissues and organs following 2, 24, 48, and 168 h of incubation. Sr was found in all root tissues as soon as after 24 h of incubation; it accumulated mostly in the cell apoplast, whereas its content in the protoplasts was considerably lower. Strontium readily crossed the endodermal barrier via the symplast and was immobilized predominantly in the pericycle cell walls; therefore, it did not hamper root branching. Strontium did not affect the final cell length and hindered root growth (at the concentration of 3 mM) by inhibiting cell division. In the shoots, Sr was found in the xylem cell walls in the vascular bundles of coleoptile, mesocotyl, and leaves on the second day of incubation, an evidence for high Sr mobility. We conclude that the transport of Sr differs from the transport of such heavy metals, as Cd, Pb, and Ni, and is similar in many aspects to the distribution of calcium, another alkaline earth metal, probably due to similar physical and chemical properties of their ions.

The availability of DTPA extracted heavy metals during laboratory incubation of contaminated soils with glucose amendments

The laboratory incubation with glucose treatment was carried out in order to estimate the possible effects of increased microbial activity on heavy metal availability. The soils from vicinity of a lead smelter operating for more than 200 years were used for the experiment. The DTPA-extractable heavy metal contents increased after glucose addition and mostly reached the highest values the second day of the incubation. The comparative study, where the chloroform fumigation was used prior to the incubation in order to decrease the microbial activities, showed especially from second day of incubation significantly lower DTPA-extractable metal contents compared to non-fumigated treatments. The interactions among the maximum possible availability of DTPA-extractable heavy metal fractions and native soil microbial characteristics were studied in differently contaminated arable and grassland soils. Irrespective of different heavy metal contents in soils, significant correlations were found among the maximum percentage increase of DTPA-extractable Pb and Cd and the ratio Bc/TOC and metabolic quotient (qCO<sub>2</sub>) which may be a result of the important role of organic matter and microbial characteristics in soils on the heavy metal availability.

Influence of media nutrients on synthesis of lignin peroxidase from Aspergillus sp.

The effect of carbon and nitrogen sources, lignocellulosic substrates, and metal ions on lignin peroxidase (LiP) activity of Aspergillus sp., which was isolated from a mangrove area, was studied. Glucose (1%) was found to be the best carbon source. Among the various lignocellulosic substrates used, coir pith at 3% concentration increased LiP activity twofold on the second day of incubation. Peptone and KNO3 completely inhibited the enzyme synthesis while (NH4)2SO4 at 12.5 mM produced maximum activity. Since seawater contained all the requisite metal ions, any added ions had a negative effect on activity. Cu2+ had the most inhibiting effect while K+ the least. When all the optimized conditions were provided, in nitrogen- and carbon-sufficient medium, a maximum LiP activity of 345 U/mL was obtained on the second day of incubation.

Characterization of the B12- and iron-sulfur-containing reductive dehalogenase from Desulfitobacterium chlororespirans.

The United Nations and the U.S. Environmental Protection Agency have identified a variety of chlorinated aromatics that constitute a significant health and environmental risk as "priority organic pollutants," the so-called "dirty dozen." Microbes have evolved the ability to utilize chlorinated aromatics as terminal electron acceptors in an energy-generating process called dehalorespiration. In this process, a reductive dehalogenase (CprA), couples the oxidation of an electron donor to the reductive elimination of chloride. We have characterized the B12 and iron-sulfur cluster-containing 3-chloro-4-hydroxybenzoate reductive dehalogenase from Desulfitobacterium chlororespirans. By defining the substrate and inhibitor specificity for the dehalogenase, the enzyme was found to require an hydroxyl group ortho to the halide. Inhibition studies indicate that the hydroxyl group is required for substrate binding. The carboxyl group can be replaced by other functionalities, e.g. acetyl or halide groups, ortho or meta to the chloride to be eliminated. The purified D. chlororespirans enzyme could dechlorinate an hydroxylated PCB (3,3',5,5'-tetrachloro-4,4'-biphenyldiol) at a rate about 1% of that with 3-chloro-4-hydroxybenzoate. Solvent deuterium isotope effect studies indicate that transfer of a single proton is partially rate-limiting in the dehalogenation reaction.

Accumulation and consumption of odorous compounds in feedlot soils under aerobic, fermentative, and anaerobic respiratory conditions.

Agricultural odors present an increasingly difficult challenge to livestock producers, yet very little information is available on the microbiology of odor production or microbial factors that regulate the emission of odors. This study examined the microbial potential for odor production and odor consumption in two soils from a cattle production facility in central Nebraska. The two soils tested were collected from a feedlot pen and a runoff ditch below the pen and contained high- and low-fecal matter content, respectively. These soils were tested for their ability to produce and consume a mixture of VFA and aromatic compounds (phenols and indoles) under aerobic, fermentative, and anaerobic respiratory conditions, with NO3-, Fe(III), Mn(IV), and SO4(2-) serving as anaerobic terminal electron acceptors, over a 6-wk incubation. The pen soil had greater (P < 0.05) initial total VFA content (40 micromol/g soil) and produced more VFA during incubation than the feedlot ditch soil, whereas total aromatic compound concentrations were not significantly different between soils. The general pattern of odor compound accumulation and consumption did not differ between soils. Oxygen and nitrate treatments produced very little VFA and consumed acetate more rapidly than the other treatments, which produced large quantities of short-chain VFA and consumed acetate only after all other VFA were consumed. When VFA and aromatic compound consumption was compared across all the treatments, aerobic incubation proved most effective, and all compounds were rapidly consumed by the second day of incubation. Of the anaerobic treatments examined, nitrate proved most effective, followed by Fe, with VFA consumed by d 5 and 21, respectively. Anaerobic incubation with sulfate produced more VFA than the fermentative incubation, and anaerobic incubation with oxidized Mn produced the largest quantities of VFA, which remained high throughout the six-wk incubation. Aromatic compounds were more easily consumed aerobically and were only slowly consumed in the anaerobic treatments. We conclude from this study that cattle feedlot soils possessed a varying, potentially exploitable capacity for odor consumption when alternate electron acceptors were available.

Noninvasive assessment of glucose and pyruvate uptake by human embryos after intracytoplasmic sperm injection and during the formation of pronuclei

Objective: To improve in vitro culture conditions and human embryo selection before transfer after IVF with intracytoplasmic sperm injection (ICSI). Design: A controlled, randomized, prospective study. Setting: University hospital–based IVF-ET program. Patient(s): Couples undergoing ICSI. Intervention(s): Culture of human embryos in the presence of 1 mM or 5.56 mM glucose and metabolic measurements with the use of noninvasive microfluorescence assays immediately after ICSI to the time of transfer. Main Outcome Measure(s): Embryo development, implantation rate, and glucose and pyruvate uptake. Result(s): Fertilization rates, early embryo development, and implantation rates were not significantly different between 1 mM and 5.56 mM glucose. Pyruvate uptake was significantly higher during the formation of the pronuclei, at 15 ± 0.7 and 11.4 ± 1.3 pmol/embryo/h for fertilized and unfertilized oocytes, respectively. Pyruvate uptake did not correlate with cleavage stage or embryo morphology. However, during the second day of incubation, pyruvate uptake was significantly higher for the untransferred embryos of pregnant women compared with nonpregnant women, at 17.9 ± 1.5 and 10.8 ± 1.0 pmol/embryo/h, respectively. Conclusion(s): The increased level of pyruvate uptake during fertilization reflects the increased demand for energy necessary for the formation of the pronuclei. However, the metabolic measurements could not improve the selection of embryos with the best implantation potential. Finally, the reduction of glucose concentration in the culture medium failed to improve embryo viability.

Cholesterol-degrading bacteria : isolation, characterization and bioconversion

Agrobacterium sp. M4, a gram negative, motile, oxidase- and catalase-positive bacterium isolated from 410 colonies from soil was found to degrade cholesterol with high efficiency (within 9 days). The first and most probably the main metabolite of cholesterol degradation was detectable on TLC from the second day of incubation, and it was identified as 4-cholestene-3-one. No substances with cyclopentenoperhydrophenanthrene structure was found under U.V. radiation at 256 and 336 nm, or by staining with sulphuric acid after 9 days of incubation. Morphological, cultural and biochemical characteristics of the isolate placed it in the genus ‘Agrobacterium’ although its urease activity was negative. Further investigation on this newly isolated strain is under way.

Effect of rice plants on methane production and rhizospheric metabolism in paddy soil

In order to elucidate the effects of rice plants on CH4 production, we conducted experiments with soil slurries and planted rice microcosms. Methane production in anoxic paddy soil slurries was stimulated by the addition of rice straw, of unsterile or autoclaved rice roots, and of the culture fluid in which rice plants had axenically been cultivated. The addition of these compounds also increased the concentrations of acetate and H2, precursors of CH4 production, in the soil. Planted compared to unplanted paddy soil microcosms exhibited lower porewater CH4 concentrations but higher CH4 emission rates. They also exhibited higher sulfate concentrations but similar nitrate concentrations. Concentrations of acetate, lactate and H2 were not much different between planted and unplanted microcosms. Pulse labeling of rice plants with14CO2 resulted during the next 5 days in transient accumulation of radioactive lactate, propionate and acetate, and after the second day of incubation in the emission of14CH4. Most of the radioactivity (40–70%) was incorporated into the above-ground biomass of rice plants. However, during a total incubation of 16 days about 3–6% of the applied radioactivity was emitted as14CH4, demonstrating that plant-derived carbon was metabolized and significantly contributed to CH4 production. The sequence of the appearance of radioactive products and their specific radioactivities indicate that CH4 was produced from root exudates by a microbial community consisting of fermenting and methanogenic bacteria.

Accumulation of free amino acids and humic substances in a freshwater modular system and their ecological significance

1. Methods for measuring concentrations of total and individual free amino acids (TFAA, FAA), other amino compounds and humic substances (HS) in media containing the following three treatments and the control are described: (i) the non‐axenic aquatic macrophyte Ceratophyllum demersum alone; (ii) the pulmonate snail, Biomphalaria glabrata alone; (iii) the snail, plant and the epiphytic bacteria and algae together as a four‐component modular system; (iv) control without organisms.2. TFAA accumulated to give asymptotic values of 2.5 μm, 10 μm and 15 μm in treatments (i), (ii) and (iii), respectively, by the end of the 30‐day incubation period. The mass‐specific accumulation rate in the treatment with the snail alone [89.4 nmol g–1 (wet weight minus shell) day–1] was approximately ten times that with the plant alone [8.4 nmol g–1 (wet mass) day–1]. No FAA or HS could be detected at any time in (iv).3. On the second day of incubation the concentrations of TFAA and of some individual amino acids were significantly higher in the treatment containing the snail and plant together than the sum of the concentrations in the treatments with the plant and snail alone, presumably due to an increase in the snails’ metabolic rate and ‘sloppy feeding’.4. The differences in the relative abundance of amino compounds accumulating in media conditioned by snails alone (e.g. much larger proportions of ammonia, ethanolamine and phosphoserine than in plant‐conditioned media) and plants alone (e.g. larger proportions of asparagine and glutamine than in snail‐conditioned media) suggest that snails and plants may derive mutual benefits by exchanging amino compounds.5. The accumulation patterns of the more recalcitrant HS differ from those of the amino acids in two respects. First, the HS concentrations continued to increase throughout the 30‐day incubation period in all three treatments. Second, most of the HS in the module originates from the plant as both the concentrations and mass‐specific accumulation rates were significantly higher in the treatment with the plant alone [2.6 mg l–1, 9.09 μg g–1 (wet mass) day–1, respectively] than in the treatment with the snail alone [0.75 mg l–1 and 7.7 μg g–1 (wet mass) day–1, respectively].6. The possible reasons for the differences in the accumulation patterns of FAA and HS in the three treatments are discussed. Evidence is also given in support of the testable hypothesis that the four components of the module derive several mutual benefits, including those arising from the release of dissolved organic matter, such as FAA and HS, by living organisms.

The development of a species difference in the local distribution of brain estrogen receptive cells

Estrogen receptors (ER) are crucial for estrogen-dependent brain differentiation and for estrogen-dependent neural functions of vertebrates. ER are expressed in a area-specific pattern in the vertebrate brain. The mechanisms that lead to the area-specific expression of ER are unknown. Here we use a species difference in the ER distribution between quail and chick and isochronic and isotopic quail to chick transplants to investigate mechanisms underlying the development of area-specific expression of ER. The entire neural tube rostral to the 6th somite ( n = 2) or rostral to the otic capsules ( n = 4) were transplanted at the second day of incubation (E2). In immunocytochemical stainings with the ER antibody H222Spγ, there is a defined cluster of intensely immunostained cells in the ventrolateral preoptic area of adult quails (the QERN). At the time of grafting, the entire brain primordium lacks ER. The QERN expresses ER as early as E13. The homologous area of the chicken brain does not differentiate cells that contain ER at any stage after ER are first detectable in the chicken brain at E11. In contrast, quail-chick chimeras develop the QERN phenotype in the ventrolateral preoptic area similar to quails. This implies that some signal which commits cells to the QERN phenotype (ER expression) is present in the quail brain primordium rostral to the otic capsules by embryonic day 2, and that this signal is unaffected by subsequent exposure to the global chick environment.

Excess Iodide and Thyroid Function in the Developing Chick Embryo

The effects of adding excess iodide (I -) to egg yolks were studied on the quantity of iodide transferred to the chick embryo and on thyroid function from Days 8 to 18 of incubation. Following injection of either 0.5 or 1 mg of iodide into the yolk on the second day of incubation, embryonic plasma iodine levels increased significantly over levels of controls at the same age. These increases were proportional to the amount of excess iodide in the yolk. Moreover, a constant ratio between the iodine levels of the allantoic fluid and the plasma was maintained for any given day of incubation. Up to Day 10, the iodine content of the embryonic thyroid was closely correlated with the increased plasma iodine concentrations. After Day 10, concomitant with the establishment of the hypothalamo-adenohypophyseal-thyroid axis, the increase in thyroid iodine no longer paralleled that of plasma iodine, but was only twice that of controls. As thyroid weight doubled in these treated embryos, thyroid iodine concentrations of control and treated embryos were similar. Iodide excess did not block thyroid iodine organification, nor was there any modification of iodine distribution among the different iodo-amino acids. Moreover, despite the doubling of thyroid hormone content in the goiters of treated embryos, circulating levels of thyroid hormones in control and treated embryos were not different.

Pathfinding by cranial nerve VII (facial) motorneurons in the chick hindbrain.

Cranial nerve VII (facial) motorneurons begin extending axons through rhombomeres 4 and 5 (R4 and R5) in the chick hindbrain on the second day of incubation. Without crossing the midline, facial motorneuron axons extend laterally from a ventromedial cell body location. All facial motorneuron axons leave the hindbrain through a discrete exit site in R4. To examine the importance of the exit site in R4 on motorneuron pathfinding, we ablated R4 before motorneuron axonogenesis. We find that mechanisms intrinsic to R5 direct the initial lateral orientation of R5 motorneuron axons. Upon reaching a particular lateral position, all R5 motorneuron axons must turn. In normal embryos the axons all turn rostrally to reach the nerve exit in R4. In embryos with R4 ablated, sometimes the axons turn rostrally and sometimes they turn caudally. A model combining permissive fields and chemotropic cues is presented to account for our observations.