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2 results on '"MESH : Embryo"'

1. Widespread lipoplex-mediated gene transfer to vascular endothelial cells and hemangioblasts in the vertebrate embryo

Author

Daniel Scherman, Daisuke Sugiyama, Rodolphe Gautier, Karine Bollerot, Samuel Tozer, Thierry Jaffredo, Virginie Escriou, Laboratoire de Biologie du Développement (LBD), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Biologie Paris Seine (IBPS), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Unité de Pharmacologie Chimique et Génétique (UPCG - UMR_S 640/UMR 8151), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC), Laboratoire de Biologie du Développement ( LBD ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Centre National de la Recherche Scientifique ( CNRS ), Unité de Pharmacologie Chimique et Génétique ( UPCG - UMR_S 640/UMR 8151 ), Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL ( ENSCP ) -Institut des sciences du Médicament -Toxicologie - Chimie - Environnement ( IFR71 ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL ( ENSCP ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Génétique, Pharmacologie Chimique Et

Subjects
Vascular Endothelial Growth Factor A, MESH : Va, MESH : Animals, Genetically Modified, MESH: Va, MESH : RNA, Small Interfering, MESH: Hematopoietic Stem Cells, Animals, Genetically Modified, Mice, 0302 clinical medicine, MESH: Genetic Vectors, RNA interference, MESH: RNA, Small Interfering, MESH: Animals, MESH: Endothelial Cells, RNA, Small Interfering, 0303 health sciences, MESH : Neovascularization, Physiologic, Gene Transfer Techniques, Embryo, Transfection, [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences, MESH : Mice, Transgenic, MESH : Genetic Vectors, Cell biology, [SDV.SP] Life Sciences [q-bio]/Pharmaceutical sciences, Haematopoiesis, medicine.anatomical_structure, Hemangioblast, MESH: Endothelium, Vascular, Stem cell, Functional genomics, MESH: Neovascularization, Physiologic, Endothelium, MESH: Mice, Transgenic, Genetic Vectors, Neovascularization, Physiologic, Mice, Transgenic, MESH: Gene Transfer Techniques, Biology, MESH: Animals, Genetically Modified, 03 medical and health sciences, MESH : Mice, medicine, Animals, MESH : Gene Transfer Techniques, MESH: Mice, 030304 developmental biology, MESH : Embryo, MESH : Endothelial Cells, MESH : Hematopoietic Stem Cells, MESH: Embryo, Endothelial Cells, Embryo, Mammalian, Hematopoietic Stem Cells, Molecular biology, [ SDV.SP ] Life Sciences [q-bio]/Pharmaceutical sciences, MESH : Endothelium, Vascular, MESH : Liposomes, Liposomes, MESH: Liposomes, Endothelium, Vascular, MESH : Animals, 030217 neurology & neurosurgery, Developmental Biology
Abstract

We report here a method that allows fast, efficient, and low-cost screening for gene function in the vascular system of the vertebrate embryo. Through intracardiac delivery of nucleic acids optimally compacted by a specific cationic lipid, we are able to induce in vivo endothelial cell-specific gain-of-function during development of the vascular network in the chick embryo. When the nucleic acids are delivered during the period of intraembryonic hematopoiesis, aortic hemangioblasts, the forerunners of the hematopoietic stem cells known to derive from the aortic endothelium, are also labeled. Similarly, we show that siRNA could be used to induce loss-of-function in vascular endothelial cells. This gene transfer technique was also applied to the mouse embryo with a high efficiency. The present method allows large-scale analysis and may represent a new and versatile tool for functional genomics.

Published
2006

2. Expression analysis of murine genes using in situ hybridization with radioactive and nonradioactively labeled RNA probes

Author

Anne, Chotteau-Lelièvre, Pascal, Dollé, Françoise, Gofflot, Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Louis Pasteur - Strasbourg I, Toussaint, Jean-Luc, Institut de génétique et biologie moléculaire et cellulaire ( IGBMC ), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), and Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
MESH: Gene Expression, MESH: Paraffin Embedding, Gene Expression, MESH : RNA Probes, [ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, MESH: Histological Techniques, In Vitro Techniques, [SDV.BBM.BM] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Sensitivity and Specificity, MESH : Frozen Sections, Mice, MESH: Pregnancy, MESH: In Situ Hybridization, Pregnancy, MESH : Mice, Animals, Frozen Sections, MESH : Female, MESH: Animals, MESH: Mice, In Situ Hybridization, Paraffin Embedding, MESH : Embryo, MESH: Frozen Sections, MESH : Paraffin Embedding, Histological Techniques, MESH: Digoxigenin, MESH: Embryo, MESH : Digoxigenin, MESH : In Situ Hybridization, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, RNA Probes, Embryo, Mammalian, MESH: Sensitivity and Specificity, MESH : Gene Expression, MESH : Pregnancy, Female, MESH : Histological Techniques, MESH: RNA Probes, MESH : Animals, MESH : Sensitivity and Specificity, Digoxigenin, MESH: Female
Abstract

International audience; The term in situ hybridization (ISH) refers to all methods allowing the detection of specific DNA (gene loci) or RNA (gene expression products) sequences, using molecular hybridization (base pairing) of labeled nucleic acid probes to target molecules within "intact" cell populations in tissue sections or whole organisms, cultured cells, or chromosomal spreads. For more than two decades, ISH has been one of the main approaches used to characterize gene expression patterns in all laboratory animal models, especially in the context of embryonic development, as well as in human tissue or cell samples for both research and diagnostic purposes. Here, we describe several ISH protocols applied to the analysis of mouse embryos and tissues; this organism has become a reference for mammalian experimental genetics. These protocols use in vitro transcribed RNAs as probes for detection. Radiolabeled probes (using 35S as a radioisotope) allow sensitive ISH on sections of paraffin-embedded material, whereas nonradioactively (digoxigenin) labeled probes can be used both for hybridization of whole embryos (whole-mount ISH) and frozen tissue sections.

Published
2006

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