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PubMed1986

Selective expression of rat pancreatic genes during embryonic development.

We present the developmental profiles of the mRNAs of 10 selectively expressed pancreatic exocrine genes and of insulin. The mRNA profiles fall into three related classes, but each profile is in some respect unique. The data on gene expression suggest there are four developmental states of the exocrine pancreas: early morphogenesis and low-level gene expression (the protodifferentiated state), the embryonic differentiated state, a modulated state in neonatal animals, and the adult differentiated state. Each state is characterized by distinct ratios of the exocrine mRNAs and presumably involves a distinct regulatory transition. This complex differentiative program must involve multiple regulatory molecules.

Amylases↗

PubMed1997

The CRE-binding protein dCREB-A is required for Drosophila embryonic development.

We have previously described the cloning of a cyclic AMP response-element (CRE)-binding protein, dCREB-A, in Drosophila melanogaster that is similar to the mammalian CRE-binding protein CREB. dCREB-A is a member of the bZIP family of transcription factors, shows specific binding to the (CRE), and can activate transcription in cell culture. In this report, we describe the gene structure for dCREB-A, protein expression patterns throughout development and the necessary role for this gene in embryogenesis. The 4.5-kb transcript is encoded in six exons that are distributed over 21 kb of DNA. There are seven start sites and no TATA consensus sequences upstream. The dCREB-A protein is expressed in the nuclei of the embryonic salivary gland, proventriculus and stomadeum. Late in embryogenesis, tracheal cell nuclei and specific nuclei within the segments show staining with anti-dCREB-A antibodies. In adult female ovaries, dCREB-A is expressed in the stage 9 through stage 11 follicle cell nuclei. Null mutations of the dCREB-A gene give rise to animals that no longer express dCREB-A protein and die late in embryogenesis before or at hatching. The absolute requirement of dCREB-A for embryogenesis demonstrates a nonredundant function for a CRE-binding protein that will be useful in studying the role of specific signal transduction cascades in development.

Animals↗

PubMed1998

Expression of non-classical islet hormone-like peptides during the embryonic development of the pancreas.

Understanding of islet embryogenesis may prove to be key in the design of future therapies for diabetes directed at re-initiating islet growth, with the goal to replace and/or replenish the impaired beta-cell mass in the disease. In this context, studies of islet neurohormonal peptides, known to play a role in the local regulation of islet function, and their expression during islet embryogenesis are important. Here we review our studies on the embryonic islet expression of islet amyloid polypeptide (IAPP) and the PP-fold peptides pancreatic polypeptide (PP), peptide YY (PYY) and neuropeptide Y (NPY). IAPP, which is constitutively expressed in beta- and delta-cells in the adult rat, was found to occur in the assumed pluripotent islet progenitor cell, together with PYY, glucagon, and to a lesser extent with insulin. As development proceeds, the insulin/IAPP phenotype is segregated from that of PYY/glucagon; with the formation of islet-like structures, insulin/IAPP-expressing cells primarily occupy their central portions, while PYY/glucagon-expressing cells are found in their periphery. At the time of formation of islet-like structures, expression of NPY is induced in the insulin/IAPP-containing cells. Whereas NPY-expression ceases at birth, PYY is constitutively expressed in non-beta-cells in the mature rat. Expression of PP is induced just prior to birth in a separate population of islet cells, occasionally co-expressed with PYY. Although a clear role for these peptides during embryogenesis has not been identified, they conceivably could play a role in the control of insulin secretion, islet growth and islet blood flow.

Amyloid↗

PubMed1986

Chick embryonic development following exposure to caffeine and nicotine.

Because cigarette smoking is considered to be deleterious to the fetus and caffeine is held in suspect as a potential human teratogen, the combined effects of caffeine and nicotine on early chick embryos were investigated. Treatment of the embryos at 48 h incubation with both caffeine (1 mg) and nicotine (1 mg) resulted in a high incidence of embryonic death and developmental defects. At 72 h incubation teratogenicity was potentiated following the same treatment. Embryonic growth was not affected. Embryotoxic interactions of this nature might account for congenital anomalies of doubtful etiology.

Abnormalities, Drug-Induced↗

PubMed2005

The effects of dimethylated and alkylated polycyclic aromatic hydrocarbons on the embryonic development of the Japanese medaka.

The Japanese medaka (Oryzias latipes) early-life stage assay was used to investigate the effects of a number of commercially available dimethylated polycyclic aromatic hydrocarbons (PAHs) (3,6-dimethylphenanthrene, 7,12-dimethylbenz[a]anthracene, and 4,6-dimethyldibenzothiophene) and their unsubstituted congeners, dimethylated and unsubstituted tertiary mixtures, and a complex environmental mixture (with elevated C2-substituted dibenzothiophene) on embryo larval development. Unsubstituted PAHs showed trends of increased blue sac disease (BSD) relative to dimethylated PAHs, although the severity of BSD induction varied. Results demonstrated that the dibenzothiophene congeners were the strongest inducers of BSD of the commercial PAHs tested. These compounds reduced the hatching success of embryonic medaka, an effect that was enhanced in the mixture. The base neutral extract significantly increased the frequency and severity of BSD abnormalities, while significantly reducing larval hatch length. Based on these results, a sublethal maximum allowable toxicant concentration (MATC) of 13.91 microg PAHs/L was calculated.

Animals↗

PubMed1995

wingless and DWnt4, 2 Drosophila Wnt genes, have related expression, regulation and function during the embryonic development.

The multigenic Wnt family encode secreted signalling molecules with important regulatory functions in various developmental processes. This paper reports an analysis of the relationships, in terms of structure, expression and function, that exist between the Drosophila genes wingless, the orthologue of the mammalian Wnt1 proto-oncogene, and DWnt4, a new member of the Wnt family. The 2 genes are physically clustered, are transcribed in overlapping embryonic territories under the control of the same regulatory molecules. Co-expression and co-regulation suggest first, that the close physical linkage results from the sharing of cis-control elements and second, that the 2 Wnt signals cooperate in developmental patterning events. Antisense RNA experiments revealed that signalling by DWnt4 is essential for cells from the anterior compartment of each parasegment to adopt a denticled fate. We propose that wingless and DWnt4 achieve opposite, but complementary functions in intrasegmental cell patterning of the embryonic ectoderm.

Animals↗

PubMed1997

Both apolipoprotein E and A-I genes are present in a nonmammalian vertebrate and are highly expressed during embryonic development.

Apolipoprotein E (apoE) is associated with several classes of plasma lipoproteins and mediates uptake of lipoproteins through its ability to interact with specific cell surface receptors. Besides its role in cardiovascular diseases, accumulating evidence has suggested that apoE could play a role in neurodegenerative diseases, such as Alzheimer disease. In vertebrates, apoA-I is the major protein of high-density lipoprotein. ApoA-I may play an important role in regulating the cholesterol content of peripheral tissues through the reverse cholesterol transport pathway. We have isolated cDNA clones that code for apoE and apoA-I from a zebrafish embryo library. Analysis of the deduced amino acid sequences showed the presence of a region enriched in basic amino acids in zebrafish apoE similar to the lipoprotein receptor-binding region of human apoE. We demonstrated by whole-mount in situ hybridization that apoE and apoA-I genes are highly expressed in the yolk syncytial layer, an extraembryonic structure implicated in embryonic and larval nutrition. ApoE transcripts were also observed in the deep cell layer during blastula stage, in numerous ectodermal derivatives after gastrulation, and after 3 days of development in a limited number of cells both in brain and in the eyes. Our data indicate that apoE can be found in a nonmammalian vertebrate and that the duplication events, from which apoE and apoA-I genes arose, occurred before the divergence of the tetrapod and teleost ancestors. Zebrafish can be used as a simple and useful model for studying the role of apolipoproteins in embryonic and larval nutrition and of apoE in brain morphogenesis and regeneration.

Amino Acid Sequence↗

PubMed1995

Primary structure of the long and short splice variants of mouse collagen XII and their tissue-specific expression during embryonic development.

Type XII collagen, a member of the FACIT group of extracellular matrix proteins, consists of molecules that are trimers of alpha 1(XII) chains. The three chains in each molecule form a cross-shaped structure with a central globule from which a triple-helical tail and three finger-like regions (containing von Willebrand factor A-like regions (containing von Willebrand factor A-like domains and fibronectin type III repeats) extend. cDNA cloning/sequencing of chicken alpha 1(XII) collagen and protein studies with mouse, bovine, and human material suggest that the alpha 1(XII) collagen gene gives rise to two molecular variants, differing in the length of the finger-like regions, by alternative splicing of the primary transcript. To provide a basis for studies of the function of the two variants in an organism that can be genetically manipulated, we have isolated and sequenced mouse cDNAs encoding both splice variants. The sequence provides the first complete nucleotide and amino acid sequence of mammalian type XII collagen. From these cDNAs we have generated digoxigenin-labeled RNA probes for in situ hybridization of developing mouse embryos to find out whether the splicing mechanism responsible for generation of the two forms is developmentally regulated. The results, combined with Northern blot and RT-PCR analysis of RNA from embryos at various developmental stages, demonstrate that the long form of collagen XII, XIIA, is the predominant form at early stages (ED7 and 11); at later stages of development (ED15 and 17) the short form, XIIB, becomes the major form. As the short form becomes the major product, the long splice variant continues to be expressed in several tissues, even after birth. An exception is dermis, which is positive for the long form up to embryonic day 15, but negative at day 18, when only the short form RNA can be detected.

Alternative Splicing↗

PubMed1998

High expression of the HMG box factor sox-13 in arterial walls during embryonic development.

Members of the Sox gene family of transcription factors are defined by the presence of an 80 amino acid homology domain, the High Mobility Group (HMG) box. Here we report the cloning and initial analysis of murine Sox-13 . The 984 amino acids Sox-13 protein contains a single HMG box, a leucine zipper motif and a glutamine-rich stretch. These characteristics are shared with another member of the Sox gene family, Sox-6. High level embryonic expression of Sox-13 occurs uniquely in the arterial walls of 13.5 days post coitum (dpc) mice and later. Low level expression was observed in the inner ear of 13.5 dpc mice and in a limited number of cells in the thymus of 16.5 dpc mice, from which Sox-13 was originally cloned. At 18.5 dpc, Sox-13 is expressed in the tracheal epithelium below the vocal cord and in the hair follicles. The Sox-13 protein binds to the consensus HMG box motif, AACAAAG, but does not transactivate transcription through a concatamer of this motif. Sox-13, like other members of the Sox family likely plays an important role in development.

Amino Acid Sequence↗

PubMed1985

Embryonic development of the rat cerebellum. I. Delineation of the cerebellar primordium and early cell movements.

Short-survival and long-survival thymidine radiograms, and methacrylate-embedded tissue from normal and X-irradiated rat embryos were used to delineate the neuroepithelial source of the cerebellum and trace the earliest cell movements. The cerebellar anlage, crescent shaped, is demarcated by two ventricular landmarks, the anterior extension of the tela choroidea of the fourth ventricle and the embryonic cerebellar fissure. The cerebellar tela choroidea extends from the medullary fourth ventricle posteromedially to the lateral recess of the pontine fourth ventricle anterolaterally. The embryonic cerebellar fissure begins caudally as a single midline incision beneath the fused posterior cerebellar primordium, then splits to follow the unfused cerebellar halves, first separating each from the isthmus then from the pons. The cerebellar primordium is divided into three parts. The lateral cerebellar primordium caps the lateral recess of the fourth ventricle; it is contiguous with the pons medially and separated ventrally from the anlage of the cochlear nuclei by the tela choroidea. The subisthmal cerebellar primordium is situated beneath the isthmus, medially lining the isthmus canal. Laterally and posteriorly, it is continuous with the lateral and postisthmal primordia. The postisthmal cerebellar primordium caps the postisthmal recess of the fourth ventricle and extends to the medullary fourth ventricle. As we shall describe later, each of these primordia is a source of different components of the developing cerebellum. Most cells of the superficially located nuclear transitory zone are labeled with 3H-thymidine administered on day E14 but not thereafter. A high proportion of the cells of the deeper cortical transitory zone could still be labeled on day E15. This supports the assumption made earlier that the first is composed of differentiating deep neurons, the second of Purkinje cells. The cells of the nuclear transitory zone originate in the lateral cerebellar primordium near the junction with the tela choroidea prior to the formation of the germinal trigone and migrate in a superficial position medially. Beginning on day E16, the nuclear transitory zone splits into two components. One has transversely oriented cells that seem to be the source of a decussating fiber tract, presumably the hook bundle of Russell. The other is composed of longitudinally oriented cells that apparently contribute fibers to the ipsilateral superior cerebellar peduncle. The translocation of the cells of the nuclear transitory zone from the cerebellar surface to its depth, to form the deep nuclei, and the radial migration of the cells of the cortical transitory zone to the surfa

Animals↗

PubMed2005

Prediction of a coding sequence for a novel type II keratin from N-terminal sequences of mouse epidermal proteins site-specifically deiminated in embryonic development.

BACKGROUND: Epidermal keratinization involves various post-translational modifications including the deimination of arginine residues. Major deiminated proteins are derived from keratin K1. Two preferred deimination sites were identified in the V subdomain of mouse K1. An antibody against one of the deiminated peptide sequences (ACP) recognized deiminated mouse and human K1, and stained the cornified layers of human and infant mouse epidermis. ACP also stained the outermost layer of mouse embryonic epidermis. Western blotting revealed minor proteins showing strong ACP-positive signals in the mouse embryonic epidermal extract in which deiminated K1 derivatives were hardly detected. OBJECTIVE: To characterize ACP-positive proteins expressed in mouse embryonic epidermis. METHODS: ACP-positive proteins were isolated by preparative gel electrophoresis for N-terminal sequencing followed by blast searches for matching sequences in the protein and nucleotide database. RESULTS: We obtained N-terminal sequences of two ACP-positive proteins. A cDNA clone in the est_mouse database has an open reading frame for 202 amino acid residues containing both sequenced peptides. The deduced sequence shows typical features of the N-terminal portion of type II keratins. A virtually identical sequence to this reading frame is present in a genomic contig of chromosome 15 on which keratin type II genes are clustered. Sequential searches for overlapping cDNA clones in the est_mouse database along with similar searches in the genomic contig formulated a hypothetical cDNA sequence encoding a putative protein of 572 amino acid residues tentatively called K1-emb. CONCLUSION: We predicted a sequence of novel type II keratin site-specifically deiminated in embryonic mouse epidermis.

Amino Acid Sequence↗

PubMed1997

Otoconial formation in the chick: changing patterns of tetracycline incorporation during embryonic development and after hatching.

The antibiotic tetracycline (TC) is incorporated into calcifying tissues and serves as a fluorescent marker for identifying calcifying sites in bone and otoconia. Fluorescent labeling was performed at different stages in chick embryos and newly hatched chicks. The stagewise changes in the intensity, location and time course of fluorescent labeling were assessed. TC/egg (1-2 mg) was injected into the yolk sacs of embryos on the 4th, 8th and 11th embryonic days (ED), and the embryos were then killed at specified times after injection. In newly hatched chicks, TC was injected daily intraperitoneally with 0.06 mg TC/g body weight for 7 days and the chicks were killed on the 8th day after hatching. Embryos injected on the 4th ED and killed on the 6th ED showed intense fluorescence in the saccular otoconial layer. All maculae from embryos killed after injection on the 4th ED showed uniformly intense fluorescence throughout the otoconial layer. By the 9th ED, otoconia in all three maculae (saccular, utricular, and lagenar) fluoresced. Maculae from embryos killed after injection on the 8th ED showed uniformly intense fluorescence throughout the otoconial layer or intense localized fluorescence mainly in the upper half of the otoconial layer. All maculae from embryos killed after injection on the 11th ED showed moderate to weak fluorescence primarily in the lower half of the otoconial layer. All maculae from posthatched chicks showed very weak fluorescence throughout the otoconial layer. Otoconia at the periphery of the maculae generally showed weak fluorescence in embryos that had been injected on the 4th and 8th ED, but not in embryos injected on the 11th ED. TC, which competes for calcium binding sites, may inhibit the formation of some otoconia. The formation of giant otoconia may reflect subtle changes in the crystallization microenvironment on these occasions. In brief, the results suggest that: (a) otoconial formation in the saccule precedes that of the utricle and lagena; (b) otoconial formation occurs during the early period (beginning the 6th ED); (c) otoconial formation is stratified, with those in the upper layer forming first and those in the lower layer forming last.

Animals↗

PubMed2003

Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor.

The biochemical properties of beclin 1 suggest a role in two fundamentally important cell biological pathways: autophagy and apoptosis. We show here that beclin 1-/- mutant mice die early in embryogenesis and beclin 1+/- mutant mice suffer from a high incidence of spontaneous tumors. These tumors continue to express wild-type beclin 1 mRNA and protein, establishing that beclin 1 is a haploinsufficient tumor suppressor gene. Beclin 1-/- embryonic stem cells have a severely altered autophagic response, whereas their apoptotic response to serum withdrawal or UV light is normal. These results demonstrate that beclin 1 is a critical component of mammalian autophagy and establish a role for autophagy in tumor suppression. They both provide a biological explanation for recent evidence implicating beclin 1 in human cancer and suggest that mutations in other genes operating in this pathway may contribute to tumor formation through deregulation of autophagy.

Animals↗

PubMed1993

Local sources of retinoic acid coincide with retinoid-mediated transgene activity during embryonic development.

We have assessed whether retinoic acid (RA) comes from local sources or is available widely to activate gene expression in embryos. We used an RA-responsive indicator cell line, L-C2A5, to localize RA sources. In these cells, an RA-sensitive promoter/lacZ reporter construct used previously by us to produce indicator transgenic mice is induced globally by RA in medium or locally by RA released at physiological concentrations (1 nM) from AG-1X2 resin beads. Furthermore, the cells are differentially responsive to the 9-cis and all-trans isomers of RA at low concentrations. Indicator transgenic mice with the same promoter/reporter construct were used to identify regions of RA-mediated gene activation. There are distinct domains of lacZ expression in the cervical and lumbar spinal cords of embryonic indicator mice. This pattern might reflect localized RA sources or restricted spatial and temporal expression of RA receptors, binding proteins, or other factors. To resolve this issue we compared the pattern of transgene activation in indicator cell monolayers cocultured with normal embryonic spinal cords with that in transgenic spinal cords. The explants induced reporter gene expression in L-C2A5 monolayers in a pattern identical to that in transgenic mice: alar regions of the cervical and lumbar cord were positive whereas those in the thoracic and sacral regions were not. We conclude that restricted sources of RA in the developing spinal cord mediate the local activation of RA-inducible genes. Thus, region-specific gene activation in embryos can be mediated by precisely localized sources of inductive molecules like RA.

Animals↗

PubMed1987

Early embryonic development and preimplantation changes in the uterus of the bat Rhinopoma hardwickei hardwickei (Gray) (Rhinopomatidae).

Rhinopoma hardwickei hardwickei has an annual reproductive cycle. Although many of the females become inseminated from the latter half of February until about the middle of April, ovulation has not been recorded until the 11th of March. A single follicle reached full development and released one ovum from either of the ovaries with nearly equal frequency, and a single conceptus was carried in the ipsilateral uterine cornu during each cycle. The embryo descended into the uterus as an early morula and attained the bilaminar blastocyst stage before undergoing implantation. As the morula advanced in age, the embryonic surface of the zona became progressively more basophilic. Hence in advanced morulae, the inner surface of the zona pellucida took a dark stain with hematoxylin and appeared like a distinct thin membrane, while the rest of the thickness of the zona was eosinophilic. Although progestational changes commenced in both uterine cornua, they became augmented in the uterine cornu on the side of ovulation and blastocyst attachment. After blastocyst attachment, the contralateral cornu reverted to an anestrus condition. The progestational changes became less conspicuous from the cranial to the caudal end of the uterus. Evidently, there was a linear gradient in the progestational response of the uterus with the cranial end being most responsive and the caudal end least responsive. The precise mechanism which brings this about is not known.

Animals↗

PubMed1998

Cloning and expression pattern of Xenopus prx-1 (Xprx-1) during embryonic development.

Homeobox genes are expressed both temporally and spatially during vertebrate development, and regulate the tissue-specific expression of other genes. A Xenopus paired-related homeobox- 1 (Xprx-1) cDNA was cloned. Xprx-1 had a paired-related homeodomain, but did not contain a paired-box. The sequence of Xprx-1 had a high level of homology with K-2(mouse) and Prx-1 (chicken), thus Xprx-1 is assumed to be the Xenopus homolog of these genes. Xprx-1 transcripts were maternally restricted, in Xenopus embryos, and a decrease in the late blastula stage was followed by an increase in zygotic transcripts after gastrulation. The transcripts were localized to the animal hemisphere of the late blastula and were concentrated in the branchial arches of the tail-bud stage embryo. In animal cap experiments, Activin A dose-dependently induced Xprx-1 gene expression. These results suggest that Xprx-1 plays a role in early Xenopus development similar to other species.

Activins↗

PubMed1997

Establishement of the dorso-ventral pattern during embryonic development of drosophila melanogasater: a logical analysis

This report focuses on dorso-ventral patterning in the segmented region of the Drosophila melanogaster embryo. According to the concept of positional information, this pattern results from the different response of cells to the Dorsal-protein morphogen. This protein shows a distribution gradient along the dorso-ventral axis, with the highest concentration on the ventral side. Using the generalized logical formalism developed by R. Thomas and co-workers, the different cellular responses were analysed in terms of the intracellular loops between the regulatory genes. Two positive loops were found to be involved, each constituting a switch which can be acted upon by the Dorsal morphogen to determine the different cell types that make up the embryonic dorso-ventral pattern. The novelty in this use of generalized logical formalism is the employment of a multilevel variable to represent a morphogen gradient. The proposed model accounts for the essential qualitative effects of the Dorsal gradient in the dorso-ventral determination process. Three main conclusions may be drawn. Firstly, the gene twist needs to have two functional threshold concentrations, one for autoactivation and the other for activation of the gene snail. Secondly, the autoactivation threshold must be smaller than that which activates snail. Thirdly, the action of the gene snail on the maintenance function of the gene twist is crucial for cells to be able to choose between the mesoderm or neuroectoderm developmental pathways. Furthermore, it is predicted that if the gene snail shows autoregulation, this will not be crucial for the determination of the embryonic D-V pattern. Copyright 1997 Academic Press Limited Copyright 1997 Academic Press Limited

Journal Article↗

PubMedDate not supplied

Transient expression of a surface antigen on a small subset of neurones during embryonic development.

During development, neurones find and interconnect with their targets in a remarkably precise way. The unfolding of neuronal specificity involves a series of highly specific recognition events which are likely to be coordinated by the spatial and temporal expression of many different surface molecules. At early stages of development, neuronal recognition occurs most dramatically at the tips of growing axons, at growth cones and their filopodia. Previous studies on the grasshopper embryo suggest that specific filopodial contacts lead to the stereotyped patterns of selective axonal fasciculation; these results support the 'labelled pathways' hypothesis which predicts that the different neighbouring axon fascicles in the embryonic neuropil within filopodial grasp are differentially labelled. To uncover the molecular labels on fasciculating embryonic axons, we screened 2,000 monoclonal antibodies generated against the embryonic neuroepithelium. Here we describe three antibodies which reveal surface antigens whose temporal and spatial expression during embryogenesis correlate with the predictions of the model. In particular, the Mes-2 antibody recognizes an antigen which is transiently expressed on the surface of only 4 out of approximately 1,000 neurones in each metathoracic hemisegment during a short period of embryogenesis. The growth cones of two of these neurones fasciculate in the periphery and innervate the same target. Moreover, they transiently express the Mes-2 surface antigen while doing so.

Animals↗