STUDIES ON THE MECHANISM OF INSECTICIDAL ACTION OF ORGANO-PHOSPHORUS COMPOUNDS WITH PARTICULAR REFERENCE TO THEIR ANTIESTERASE ACTIVITY

The preparation of an extract of the mealworm larvae, Tenebrio molitor L. which hydrolyses ethyl butyrate and o -nitrophenyl acetate, but not acetylcholine, is described. The inhibition of this esterase by TEPP-containing materials and parathion was determined.
An enzyme that hydrolysed o -nitrophenyl acetate and was inhibited by a TEPP-containing material was demonstrated in the five other insect species used.
The relative toxicities as contact insecticides to adult Tribolium castaneum Hbst. of ten samples of TEPP-containing materials were compared with their relative activities as esterase inhibitors. There was not an exact quantitative correlation between TEPP content estimated chemically, insecticidal activity and anti-esterase activity; but the correlation was sufficient to suggest interdependence of these factors.
Eggs of Diataraxia oleracea L. and Ephestia kühniella Zell, were shown to contain an enzyme that hydrolysed o -nitrophenyl acetate and was inhibited by the TEPP-containing materials. This enzyme was present in eggs less than 24 hr. old, i.e. before there was any visible signs of development. The TEPP was shown to be toxic to these eggs and in high concentrations kills at an early stage of development before differentiation of the nervous system. This, in conjunction with the other evidence, suggests that esterases other than the choline-esterase of the nervous system are important when considering the toxic action of these compounds.
Comparison of the anti-esterase activity and toxicity of parathion and TEPP-containing materials as insecticides showed that although the TEPP materials were the more potent enzyme inhibitors, parathion was the more potent contact insecticide to five species of insects. This appears to be due to the relative instability of TEPP. The study of the rates of action of the two poisons applied at different concentrations supports this view.     

Effect of a juvenoid on embryonic development of Earias fabia (Stoll) (Lepidoptera: Noctuidae)

Eggs of Earias fabia were treated with three different quantities of a juvenoid, 6–7 epoxy-3-ethyl-1-(p-ethyl phenoxy)-7 methylnonane at 5 distinct stages. These were: (a) before blastoderm formation; (b) before gastrulation; (c) before segmentation; (d) before blastokinesis; and (e) just after blastokinesis. The effect on embryonic development was studied. If sufficient hormone is administered early enough, development may be arrested at stages as early as blastoderm formation. With moderate and smaller doses, developmental arrest mostly occurred at certain well-defined stages, such as germ-band formation; before blastokinesis; post blastokinesis; when the embryo assumes its larval form; at the time of chitin formation and during embryonic moulting. Observations also bring in focus the importance of the quantity of hormone and its relationship to the speed of its action. Toxic action is apparent when eggs are treated with relatively larger quantities of the compound: application of juvenoid to younger stages causes fewer deaths.     

STUDIES OF THE TOXICITY TO WORKER HONEY-BEES (APIS MELLIFERA L.) OF CERTAIN CHEMICALS USED IN PLANT PROTECTION

Laboratory techniques are described for the estimation of the stomach poison, direct and residual film contact poison and fumigant poison effects of chemicals to adult worker honey-bees.
The toxicity of eleven chemicals used in plant protection has been investigated by these methods. The order of effectiveness as stomach and contact poisons was: parathion, TEPP, γ-BHC, dieldrin, aldrin, chlordane, o , o -diethyl- o -ethylmercaptoethyl thiophosphate (constituent of Systox), bisdimethylamino fluorophosphine oxide, toxaphene and the sodium salts of 2:4-D and MCPA: as residual films, dieldrin, aldrin, γ-BHC, parathion, chlordane, and o , o -diethyl- o -ethylmercapto-ethyl thiophosphate (constituent of Systox); toxaphene, TEPP and bisdimethylamino fluorophosphine oxide had no measurable effect; as fumigants, dieldrin, γ-BHC, aldrin, parathion, and chlordane; the remainder had no measurable effect.     

Avian Egg Odour Encodes Information on Embryo Sex,Fertility and Development

Avian chemical communication is a rapidly emerging field, but has been hampered by a critical lack of information on volatile chemicals that communicate ecologically relevant information (semiochemicals). A possible, but as yet unexplored, function of olfaction and chemical communication in birds is in parent-embryo and embryo-embryo communication. Communication between parents and developing embryos may act to mediate parental behaviour, while communication between embryos can control the synchronicity of hatching. Embryonic vocalisations and vibrations have been implicated as a means of communication during the later stages of development but in the early stages, before embryos are capable of independent movement and vocalisation, this is not possible. Here we show that volatiles emitted from developing eggs of Japanese quail (Coturnix japonica) convey information on egg fertility, along with the sex and developmental status of the embryo. Specifically, egg volatiles changed over the course of incubation, differed between fertile and infertile eggs, and were predictive of embryo sex as early as day 1 of incubation. Egg odours therefore have the potential to facilitate parent-embryo and embryo-embryo interactions by allowing the assessment of key measures of embryonic development long before this is possible through other modalities. It also opens up the intriguing possibility that parents may be able to glean further relevant information from egg volatiles, such as the health, viability and heritage of embryos. By determining information conveyed by egg-derived volatiles, we hope to stimulate further investigation into the ecological role of egg odours.     

STUDIES IN THE MODE OF ACTION OF INSECTICIDES

Acetylcholine, choline chloride, acetyl-β-methylcholine, benzoylcholine, carbamyl choline, adrenaline and d -tubocurarine are non-toxic when injected into the locust. Prostigmine is also non-toxic, and eserine considerably less toxic to the locust than to man.
The toxic effect of tetraethyl pyrophosphate (TEPP) cannot be antagonized by injection of atropine or enhanced by d -tubocurarine.
The injection of acetylcholine chloride following injection of TEPP does not affect subsequent mortality.
These findings are discussed, and it is suggested that the physiology of the nervous system of the insect is unlike that of the mammal, neither cholinesters nor adrenaline being concerned in it. Phosphorus insecticides are thought to inhibit a general esterase not specifically connected with cholinesters.     

Embryo ecology: Developmental synchrony and asynchrony in the embryonic development of wild annual fish populations

Embryo–environment interactions are of paramount importance during the development of all organisms, and impacts during this period can echo far into later stages of ontogeny. African annual fish of the genus Nothobranchius live in temporary pools and their eggs survive the dry season in the dry bottom substrate of the pools by entering a facultative developmental arrest termed diapause. Uniquely among animals, the embryos (encased in eggs) may enter diapause at three different developmental stages. Such a system allows for the potential to employ different regulation mechanisms for each diapause. We sampled multiple Nothobranchius embryo banks across the progressing season, species, and populations. We present important baseline field data and examine the role of environmental regulation in the embryonic development of this unique system. We describe the course of embryo development in the wild and find it to be very different from the typical development under laboratory conditions. Development across the embryo banks was synchronized within and across the sampled populations with all embryos entering diapause I during the rainy season and diapause II during the dry season. Asynchrony occurred at transient phases of the habitat, during the process of habitat desiccation, and at the end of the dry season. Our findings reveal the significance of environmental conditions in the serial character of the annual fish diapauses.     

Consistency of passerine embryo development and the use of embryonic staging in studies of hatching failure

Hatching failure is widespread in birds, and is usually the result of embryo death rather than infertility. Embryo death can result from both intrinsic and extrinsic factors, some of which may vary across the developmental period. Determining the point at which an embryo died during development may therefore help us to understand the underlying cause of death. Here we describe simple criteria that can be used by field ornithologists to establish the developmental stage of dead embryos found in unhatched passerine eggs, and explain how this can be used to estimate the date of embryo death. We compared the pattern of embryo development over the incubation period for three species, the Zebra Finch Taeniopygia guttata, Blue Tit Cyanistes caeruleus and Great Tit Parus major. We also compared rates of Zebra Finch embryo development under artificial and standard (parental) incubation. Embryo development rates were remarkably similar across the three species and between Zebra Finch embryos under artificial and natural incubation conditions. We therefore suggest that the pattern of embryo development in the Zebra Finch may provide a model for other small passerines with similar incubation periods, but acknowledge that further interspecific comparisons are required before this model is considered more widely applicable. By estimating embryo death dates using our approach, ornithologists will be able to determine temporal patterns of embryo mortality in relation to extrinsic environmental conditions. This approach may shed light on how extrinsic factors such as climate and parental behaviour influence embryo survival in wild birds.     

Structure, localization and potential role of a novel molluscan trypsin inhibitor in Lymnaea.

Eggs and egg masses of the freshwater gastropod mollusc Lymnaea provide a microenvironment for developing embryos. Secretions of the exocrine albumen gland of Lymnaea are packaged in the eggs of an egg mass before the eggs are laid externally. The perivitelline fluid that directly surrounds individual oocytes is the main source of nutrition for developing embryos. During early stages of development, the perivitelline fluid is initially internalized by pinocytosis and degraded by lysosomes; in later stages, the embryo ingests the fluid. We previously found that the albumen gland produces large amounts of Lymnaea epidermal growth factor. The albumen gland also appears to produce significant amounts of a novel Lymnaea trypsin inhibitor (LTI), a second peptide that was purified and characterized from Lymnaea albumen gland extracts. The primary structure was determined by microsequence analysis, mass spectrometry, and C-terminal sequence analysis, and showed that LTI is a 57-residue glycosylated peptide. Comparison of the LTI sequence with other known serine protease inhibitors indicates that LTI is a member of the bovine pancreatic trypsin inhibitor family. Reverse phase-high performance liquid chromatography, microsequence analysis, mass spectrometry, and immunocytochemistry demonstrated that abundant amounts of intact LTI are packaged in egg masses. The presence of a trypsin inhibitor in the perivitelline fluid compartment of the egg mass may minimize digestion of peptides and proteins in the perivitelline fluid that are important for the development of the embryo, for example, Lymnaea epidermal growth factor.     

Observations on the female reproductive organs and the poison apparatus of Caraphractus ductus Walker (Hymenoptera:Mymaridae)

Caraphractus cinctus is an arrhenotokous mymarid parasitizing the eggs of Dytiscidae under water. In the newly emerged female only fully formed eggs are present in the ovaries and the earlier stages of ovarian development have been studied in the pupa. The two ovaries each contain from 10 to 20 ovarioles depending upon the size of the female. The two lateral oviducts unite to form the vagina which is bent upon itself when laying is not in progress. The eggs are stored in the ovarioles and the female has remarkable control over the deposition of the eggs, since in most cases she rejects host eggs already parasitized, after probing them with her ovipositor. The spermatheca is a rigid capsule and the spermathecal duct at its base has a deep U-shaped bend. There is a large spermathecal gland opening by its own duct into the spermathecal duct after the bend. The poison apparatus is well developed though the female does not kill the egg or paralyse the embryo host. The poison gland is of unusual shape being compact and rounded distally instead of tubular. Dufour's gland is large and buoyant. The ducts of both glands lead to the base of the ovipositor. The possible effect of their secretions in rendering a once parasitized Agabus egg generally unacceptable for further laying is discussed.     

CARBOHYDRATE METABOLISM IN THE EGGS OF THE SILKWORM, BOMBYX MORI. I. ABSENCE OF PHOSPHOFRUCTOKINASE IN THE DIAPAUSING EGG

In the diapausing eggs of the silkworm, Bombyx mori , glycogen is rapidly converted to sorbitol and glycerol, and this conversion is reversed at termination of the diapause (C hino , 1958). To elucidate the pathway leading to this polyol formation and its regulatory mechanisms, enzymes concerning carbohydrate metabolism were surveyed in diapausing as well as in developing eggs of the silkworm.
Most of the enzyme activities concerning citric acid cycle are low at the beginning of the embryogenesis and during diapause, but increase at the later stages of the development. Making an exception, reduction rate of malate and fumarate was rather high from the onset of the embryonic development. Several glycolytic enzymes were also studied. Most remarkable fact is that phosphofructokinase activity could not be demonstrated in the diapausing and also in the early stages of the developing eggs. Other enzymes, viz. α-glycerophosphate dehydrogenase, aldolase, glyceraldehyde-3-phosphate dehydrogenase were detected from the beginning of the embryogenesis.
Absence of phosphofructokinase, together with the high activity in glucose-6-phosphate dehydrogenase, suggests that predominant pathway in carbohydrate metabolism in the early stages of embryogenesis and in the diapause period is by way of pentose phosphate pathway. This supposition is confirmed by the experiments using labeled glucose. Incorporation of the label into glycerol of the diapausing eggs was three to four fold when G-6-¹⁴C was injected into pupae as compared with the case of G-1-¹⁴C injection. The above experiments provide evidence supporting the theory that glycogen is converted into sorbitol and glycerol mostly by way of the pentose phosphate pathway in the diapausing eggs.     

THE EFFECT OF BODY WEIGHT ON THE RESISTANCE TO INSECTICIDES OF THE LAST-INSTAR LARVA OF DIATARAXIA OLERACEA L., THE TOMATO MOTH

At a constant temperature of 24° C. the final larval instar of Diataraxia oleracea lasts about 10 days, during which its resistance to DDT and γ-BHC as contact insecticides progressively increases up to the 5th or 6th day. It then suddenly decreases, this coinciding with cessation of feeding and the beginning of prepupal formation.
Between the 2nd and the 6th days the gross body weight of the last-instar larva increases from about 0.27 to 0.65g. Under the conditions of the experiments, the LD50 of parathion, as a stomach poison, was linearly related to body weight; on the same basis TEPP was slightly less, and lead arsenate, slightly more, toxic to the larger than to the smaller larvae. However, DDT as a stomach or contact insecticide, and γ-BHC as a stomach poison were notably less toxic to the larger larvae. For example, the increase in LD50 for an increase in larval body weight of × 2 was about × 11 for DDT as a stomach poison and about × 12 as a contact insecticide.
The order of effectiveness of the above insecticides as stomach poisons for the last-instar larva of D. oleracea was parathion > DDT > γ-BHC > TEPP = lead arsenate. Zinc fluoarsenate and rotenone were relatively non-toxic. Larvae of D. oleracea were repelled by food leaf treated with an extract of natural pyrethrins.     

Initiation of embryo implantation and maintenance of early pregnancy in the rat by chlordecone (Kepone).

The effect of chlordecone (Kepone), an insecticide/fungicide with reproductive toxicity, on the early stages of pregnancy in the rat was studied. Intraperitoneal injection of chlordecone into adult virgin female Holtzman strain rats before mating, in doses as high as 80 mg/kg, did not prevent fertilization, early development of the embryo to the blastocyst stage, transport of the embryo through the oviduct, or its implantation into the uterus. However, a single dose of 60 or 80 mg/kg, but not 20 or 40 mg/kg, before mating significantly reduced the concentration of progesterone in the serum of rats undergoing normal embryo implantation 5 days later. A dose of 80 mg/kg of chlordecone reduced progesterone levels in the serum by more than 50% within 48 hr in ovariectomized rats with Silastic tubing implants containing crystalline progesterone. This dose of chlordecone induced deciduomata formation in progesterone-primed ovariectomized rats to the same extent as 1 microgram of estradiol benzoate. The minimal effective single dose of chlordecone to initiate implantation of blastocysts in the uteri of hypophysectomized progesterone-primed rats, and to maintain embryo development for at least 5 days, was 50 mg/kg. Daily doses of 20 mg/kg for 3 or 5 days were effective at initiating implantation but did not maintain pregnancy. The latter treatment, however, did not prevent initiation of implantation or embryo development induced by subsequent administration of estrone. The results are consistent with the view that chlordecone is a weak estrogen that has both nongenomic and genomic estrogenic actions.     

Evolution of germ cell development in tetrapods: comparison of urodeles and amniotes

SUMMARY. The embryonic development of germ cells in tetrapods is described, focusing on groups with the inductive mode of germ cell specification. In mammals PGCs are induced early in the gastrulation process, they are internalized with future extraembryonic mesoderm in the early posterior primitive streak, and specified soon thereafter. Strong evidence indicates that a similar process occurs in turtles and some other reptiles. In amniotes, the PGCs appear well before formation of the gonad in the posterior trunk, resulting in a period in which they are located outside the embryo before their migration to the gonad. In contrast, in urodeles the PGCs appear relatively late, and throughout development maintain a position close to precursors of the somatic cells of the gonad so that migration is not required. In lampreys early development of germ cells is strikingly similar to that in urodeles, suggesting this is the primitive process. As amniotes evolved large yolky eggs and better access to nutrition, development of the posterior half of the trunk became more dependent on cell proliferation; this was followed or accompanied by a shift of early germ cell development to the equivalent of the early primitive streak. A similar process may have occurred as some basal vertebrates developed large yolky eggs.     

Vibration analysis on incubating eggs and its relation to embryonic development

Coucke (1998) was the first to use acoustic resonance analysis to monitor embryo development in chicken eggs. He remarked that at around 100 hours of incubation, the course of the resonant frequency and damping changed abruptly in the case of fertile eggs. He also showed that these changes were related to a physiologic event during early embryonic development. The objective of our study is to monitor the course of the vibration parameters during the early incubation of chicken eggs and to relate these changes to egg and embryo characteristics. A total of 72 Hybro eggs were incubated vertically in a small incubator at standard conditions. Several egg parameters were measured before incubation. During the early stages of incubation the vibration behavior of these eggs was monitored. The time at which the damping of the vibration suddenly changed, the diameter of the eggs and their interaction were found to be significant explanatory variables in order to predict hatching time. A correlation coefficient r of 0.72 was obtained.     

胡子鲶的胚胎发育

用无膜卵的培养方法,比较详细地观察了胡子鲶胚胎发育的过程,并从早期个体发育的本质着眼提出了卵裂、细胞分化和器官分化三个主要胚胎发育阶段。分析了胚胎发育与环境条件的适应性。根据胡子鲶胚胎发育的特点,初步讨论了苗种生产中应该注意的几个问题,同时提出胡子鲶可以作为鱼类细胞工程研究的一个比较理想的实验鱼材料。       

千年笛鲷早期发育及视蛋白基因表达规律分析

初步观察千年笛鲷早期发育各个时期的形态特征, 同时使用实时荧光定量PCR方法对4种视蛋白基因在早期发育中的表达规律进行分析。研究观察到千年笛鲷卵为圆球形, 属浮性卵, 中心有一明显的油球。在水温(24.50.5)℃的条件下, 千年笛鲷胚胎发育共经历6个发育阶段18个时期, 从受精卵到孵化一共经历24h。仔鱼经历1215d发育为稚鱼, 30d35d发育为幼鱼。同时对7个胚胎发育时期和2个仔鱼发育时期4种视蛋白(LWS、SWS1、SWS2、RH)基因的表达情况进行检测, 在下包1/2、胚孔封闭、视囊这3个时期有显著性表达(P0.05), 尤其在胚孔封闭时期, 表达量达到最高。其余时期4种基因的表达水平显著下降, 但在2个仔鱼时期表达量比孵化期略有增加。结果表明千年笛鲷4种视蛋白基因在早期表达过程中与神经胚的形成有密切的联系。       

WHOLE-EMBRYO CULTURE AND THE STUDY OF MAMMALIAN EMBRYOS DURING ORGANOGENESIS

1. Simple and reliable methods are now available for growing rat and mouse embryos in culture at all stages of organogenesis. Primitive-streak embryos can be maintained for up to 5 days in culture while they develop to early foetal stages. Older embryos are maintained for progressively shorter periods and the most advanced stage that can be supported is equivalent to the rat foetus of 15 days' gestation. 2. The rates of protein synthesis and differentiation of the younger embryos in vitro are similar, and of head-fold embryos identical, to those in vivo. After the formation of the limb buds growth is slower, with protein synthesis more retarded than differentiation, resulting in embryos or foetuses that are well formed but smaller than in vivo. This slowing of growth of the older embryos in culture is probably caused by the lack of a functional allantoic placenta. 3. The embryos of some other species, including the guinea-pig, hamster, rabbit and opossum have also been maintained in culture during organogenesis but the results are not yet as good as those for rats and mice. 4. Maximum growth of rat embryos explanted with the visceral yolk sac intact is obtained in undiluted homologous serum, though adequate growth for many studies can be maintained in mixtures of serum with chemically defined tissue-culture media. The best results are obtained in serum prepared from blood centrifuged before clotting has occurred (I.C. serum) and heat-inactivated. The importance of a high concentration of serum in the culture medium may be related to the mechanisms for uptake, transport and digestion of macromolecules by the rodent yolk sac. 5. There is no convincing evidence for a changing rate of oxygen consumption during organogenesis but there is strong evidence for changes in energy metabolism. At the beginning of organogenesis, the embryo shows a high rate of anaerobic glycolysis and of pentose-shunt activity. During the following days these decline while activity of the Krebs' cycle and electron-transport system increases. Anoxia, or exposure of the embryo to carbon monoxide, increases glycolysis and reduces growth rate. 6. The earliest stages of the formation of the heart and blood circulation can be closely observed in culture. The heart rate of the 111/2-day rat embryo is about 160 beats per minute at 38°C, and falls by about 7% per degree for lower temperatures. Several drugs that are cardioactive in the adult also affect the frequency of the heartbeat in the embryo, and the pattern of response suggests that the adrenergic receptors in the embryo develop before the cholinergic receptors. Experiments in which embryo and yolk sac were cultured separately, as well as together, have indicated that haemopoiesis can occur in the embryo only after a migration of stem cells from the yolk sac. 7. Microsurgery has been successfully applied to embryos in culture in studies on morphogenetic movements, heart development, axial rotation, limb-bud regeneration and placenta formation. Biochemical studies of normal morphogenesis have been few, but one has shown a high rate of hyaluronate synthesis by the embryo which may be related to the maintenance and expansion of extracellular spaces and the formation of the neural folds. 8. Embryos are particularly sensitive to teratogenic agents during organogenesis. Teratogens that have been studied on whole embryos in culture include trypan blue, antisera, hyperthermia, anaesthetics, and abnormal concentrations of vitamins, oxygen and glucose. Many of the malformations induced have been similar to those obtained after administration of the same agents in vivo and have demonstrated a direct teratogenic effect on the embryo independent of the maternal metabolism. It is suggested that culture methods may provide a valuable additional screening procedure for new drugs and other potentially embryopathic agents.     

Selfed embryo death in Pinus taeda: a phenotypic profile

Selective elimination of selfed embryos, or inbreeding depression, is shared among many members of the Pinaceae but it has not been fully characterized at the phenotypic level. Here, two death pattern model hypotheses are tested using 10 621 Pinus taeda embryos sampled in two cohorts. Cones from a single pedigree based on selfed, outbred, parent-offspring and offspring-parent matings were destructively sampled weekly before, during and after fertilization. Selfed embryo deaths adhered to two patterns over the course of development: death was linear with respect to days from fertilization; and a stage-specific death peak occurred during the early embryogeny stage. This death peak occurred from 23 to 36 d after fertilization in the 2004 cohort and from 27 to 34 d after fertilization in the 2006 cohort. Of those selfed embryos that died, 64-83% died at stages where a single dominant embryo was elongating inside the female gametophyte. Additional genetic models are needed to account for the stage-specific death component of selfed P. taeda embryos.     

FINE STRUCTURE OF THE ZYGOTE AND EARLY EMBRYO IN QUERCUS GAMBELII

This investigation begins with the late zygote and traces ultrastructural development to the late globular stage of the embryo. Two nucleoli and satellite nucleoli sometimes occur in the zygote nucleus. Mitochondria, dictyosomes, cytoplasmic ribosomes, rough ER, and lipid bodies are numerous in the zygote. Microbodies are occasionally seen. The cell wall becomes well developed before the first division. No plasmodesmata occur in the zygote wall. The basal cell of the proembryo and the suspensor cells of the later embryo have very dense cytoplasm with a high concentration of cytoplasmic ribosomes. The nuclei are very electron opaque. The terminal cell and the cells of the embryo proper have a fine structure similar to that of the zygote. Plastids increase in number, size, starch content, and amount of thylakoid lamellae as the embryo develops. Mitochondria are numerous and appear active at all stages. Dictyosome activity, ribosomal aggregation, and the amount of ER are highest during the late globular stage. Lipid bodies are present up to the early globular stage, then disappear. The inner cell walls of the embryo are thin and have many plasmodesmata. These walls begin to thicken at the late globular stage, and at this time the size of the embryo begins to show an increase over that of the zygote. The results show a corresponding increase in the amount and activity of the metabolic machinery as the development of the embryo progresses. Lipids are probably more important as a nutrient source in the zygote and early embryo; starch becomes more important in the late stages. Absorption of nutrient material into the embryo sac and developing embryo appears to be from the chalazal end.     

In vivo electroporation: a new frontier for gene delivery and embryology

One of the key techniques in developmental biology is introducing transgenes into tissues and analyzing their subsequent effects on morphogenesis and organogenesis. In mammals, the transgenic approach is a way to misexpress foreign genes in various tissues and organs. However, targeting expression to certain tissues is totally dependent on the availability of specific promoters. Hence, it is not an easy task to control transgene expression temporally and spatially during embryogenesis. Further, if the transgene is toxic, embryonic development can be disrupted, resulting in premature death before the desired stages of development. As alternative systems, Xenopus and zebrafish are used frequently. In these vertebrate models, overexpression of genes can be carried out by injecting synthetic RNAs into eggs. However, genetic techniques in these systems are limited only to early development, prohibiting the precise analysis of gene effects on organogenesis in later stages. In contrast, the chick embryo has long served as a powerful and useful model system, holding a unique position in the field of developmental biology. Although trials of transgenic chicks have never been successful, easy accessibility to the developing embryo through a window opened in an eggshell enables performance of a variety of techniques, such as time-lapse cinephotomatography, microsurgical manipulations (including chick/quail chimeras), transplantation of cells and tissues, New's in vitro culture, etc. (Bortier et al., 1996; Douarin et al., 1996; Selleck, 1996). In addition to these experimental advantages, retrovirus-mediated gene delivery, and recently, adenovirus-mediated misexpression have been employed routinely in chick embryos (Leber et al., 1996; Morgan and Fekete, 1996).