Detection of mRNA in Whole Mounts of Mouse Embryos Using Digoxigenin Riboprobes

AbstractSeveral different transgenic tamoxifen-inducible cre reporter lines have been used to analyse the contribution of embryonic precursors to the development of the mononuclear phagocyte system in mice. Here we show that tamoxifen treatment of the mother at 8.5dpc with doses commonly-used in lineage trace studies produces a 4-5-fold expansion of the embryonic leukocyte populations by 10.5dpc, detected in whole mounts of embryos using aCsf1rreporter gene or separately by expression ofCsf1r, Itgam(CD11b),Adgre1(F4/80) orPtprc(CD45) mRNA. These findings indicate that tamoxifen cannot be considered a neutral agonist in macrophage lineage trace studies.Summary sentenceTreatment of pregnant mice with tamoxifen in early gestation produces a large expansion of the embryonic macrophage population.

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Three-Dimensional Reconstruction Using Stereo Pairs from Serial Thick Sections

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100080249 ◽

1977 ◽

Vol 35 ◽

pp. 562-563

Author(s):

Robert Glaeser ◽

Thomas Bauer ◽

David Grano

Keyword(s):

Three Dimensional ◽

Dimensional Structure ◽

Serial Sections ◽

Thin Sections ◽

3 Dimensional ◽

Transmission Electron ◽

Freeze Dried ◽

Dimensional Reconstruction ◽

Stereo Imagery ◽

Whole Mounts

In transmission electron microscopy, the 3-dimensional structure of an object is usually obtained in one of two ways. For objects which can be included in one specimen, as for example with elements included in freeze- dried whole mounts and examined with a high voltage microscope, stereo pairs can be obtained which exhibit the 3-D structure of the element. For objects which can not be included in one specimen, the 3-D shape is obtained by reconstruction from serial sections. However, without stereo imagery, only detail which remains constant within the thickness of the section can be used in the reconstruction; consequently, the choice is between a low resolution reconstruction using a few thick sections and a better resolution reconstruction using many thin sections, generally a tedious chore. This paper describes an approach to 3-D reconstruction which uses stereo images of serial thick sections to reconstruct an object including detail which changes within the depth of an individual thick section.

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Human Adenovirus Uptake by Preirnplantation Mouse Embryos

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100094607 ◽

1972 ◽

Vol 30 ◽

pp. 268-269

Author(s):

D. G. Chase ◽

W. Winters ◽

L. Piko

Keyword(s):

Hela Cells ◽

Human Adenovirus ◽

Mouse Embryos ◽

Equilibrium Density ◽

Cell Stage ◽

Virus Attachment ◽

Preimplantation Mouse Embryos ◽

Embryo Culture Medium ◽

Preimplantation Mouse ◽

Mouse Cells

Although the outlines of human adenovirus entry and uncoating in HeLa cells has been clarified in recent electron microscope studies, several details remain unclear or controversial. Furthermore, morphological features of early interactions of human adenovirus with non-permissive mouse cells have not been extensively documented. In the course of studies on the effects of human adenoviruses type 5 (AD-5) and type 12 on cultured preimplantation mouse embryos we have examined virus attachment, entry and uncoating. Here we present the ultrastructural findings for AD-5.AD-5 was grown in HeLa cells and purified by successive velocity gradient and equilibrium density gradient centrifugations in CsCl. After dialysis against PBS, virus was sedimented and resuspended in embryo culture medium. Embryos were placed in culture at the 2-cell stage in Brinster's medium.

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Preplate neurons in embryonic mouse cortex: Ultrastructural observations using photoconversion of the fluorescent lipophilic dye dil

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100124938 ◽

1992 ◽

Vol 50 (1) ◽

pp. 906-907

Author(s):

Linda C. Hassinger ◽

James E. Crandall

Keyword(s):

Mouse Embryos ◽

Embryonic Mouse ◽

Cortical Afferents ◽

Mouse Cortex ◽

Carbocyanine Dye ◽

Increased Sensitivity

We have begun to look directly at small numbers of afferent axons to early generated neurons that form the preplate in the developing mouse cortex. The carbocyanine dye Dil (1’1, dioctadecyl-3,3,3’3’-tetramethyl-indocarbocyanine) has proved especially useful for this goal. DiI labels axons and their terminals with greater sensitivity and without some of the disadvantages of axon filling with HRP. The increased sensitivity provided by labeling embryonic axons with DiI has given us new insights into the development of cortical afferents. For instance, we reported originally that afferents from the thalamus were present below the cortex as early as embryonic day 15 (E15) based on HRP injections into mouse embryos. By using DiI placements into the thalamus in aldehyde-fixed brains, we now know that thalamic fibers reach the cortex 24 hrs earlier.

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Localization of Intracisternal a Particle Antigens in Neoplastic Cells and Preimplantation Mouse Embryos

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s1431927600003044 ◽

1980 ◽

Vol 38 ◽

pp. 696-697

Author(s):

Thomas T.F. Huang ◽

Patricia G. Calarco

Keyword(s):

Endoplasmic Reticulum ◽

Normal Development ◽

Mouse Embryos ◽

Neoplastic Cells ◽

Large Numbers ◽

Preimplantation Mouse Embryos ◽

Preimplantation Mouse ◽

Intracisternal A Particle ◽

Normal Feature

The stage specific appearance of a retravirus, termed the Intracisternal A particle (IAP) is a normal feature of early preimplantation development. To date, all feral and laboratory strains of Mus musculus and even Asian species such as Mus cervicolor and Mus pahari express the particles during the 2-8 cell stages. IAP form by budding into the endoplasmic reticulum and appear singly or as groups of donut-shaped particles within the cisternae (fig. 1). IAP are also produced in large numbers in several neoplastic cells such as certain plasmacytomas and rhabdomyosarcomas. The role of IAP, either in normal development or in neoplastic behavior, is unknown.

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