scholarly journals A reporter mouse for in vivo detection of DNA damage in embryonic germ cells

genesis ◽  
2020 ◽  
Vol 58 (8) ◽  
Author(s):  
Jordana C. Bloom ◽  
John C. Schimenti
Keyword(s):  
Dna Damage ◽  
Germ Cells ◽  
Reporter Mouse ◽  
Embryonic Germ Cells ◽  
In Vivo Detection

A Reporter Mouse for In Vivo Detection of DNA Damage in Embryonic Germ Cells

2020 ◽  
Author(s):  
Jordana C. Bloom ◽  
John C. Schimenti
Keyword(s):  
Dna Damage ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Strand Break ◽  
Dependent Manner ◽  
Dna Double Strand Breaks ◽  
Specific Expression ◽  
Exogenous Dna ◽  
Embryonic Germ Cells

AbstractMaintaining genome integrity in the germline is essential for survival and propagation of a species. In both mouse and human, germ cells originate during fetal development and are hypersensitive to both endogenous and exogenous DNA damaging agents. Currently, mechanistic understanding of how primordial germ cells respond to DNA damage is limited in part by the tools available to study these cells. We developed a mouse transgenic reporter strain expressing a 53BP1-mCherry fusion protein under the control of the Oct4ΔPE embryonic germ cell-specific promoter. This reporter binds sites of DNA double strand breaks (DSBs) on chromatin, forming foci. Using ionizing radiation as a DNA double strand break-inducing agent, we show that the transgenic reporter expresses specifically in the embryonic germ cells of both sexes and forms DNA damage induced foci in both a dose- and time-dependent manner. The dynamic time-sensitive and dose-sensitive DNA damage detection ability of this transgenic reporter, in combination with its specific expression in embryonic germ cells, makes it a versatile and valuable tool for increasing our understanding of DNA damage responses in these unique cells.

Developmental fate of embryonic germ cells (EGCs), in vivo and in vitro

2003 ◽  
Vol 71 (2) ◽  
pp. 135-141 ◽  
Author(s):  
Gabriela Durcova-Hills ◽  
Florence Wianny ◽  
Julie Merriman ◽  
Magdalena Zernicka-Goetz ◽  
Anne McLaren
Keyword(s):  
Germ Cells ◽  
Embryonic Germ Cells ◽  
Developmental Fate

scholarly journals Novel TALEN-generated mCitrine-FANCD2 fusion reporter mouse model for in vivo research of DNA damage response

DNA Repair ◽  
2020 ◽  
Vol 94 ◽  
pp. 102936
Author(s):  
Maja Sabol ◽  
M. Aydın Akbudak ◽  
Dominika Fricova ◽  
Inken Beck ◽  
Radislav Sedlacek
Keyword(s):  
Dna Damage ◽  
Mouse Model ◽  
Dna Damage Response ◽  
Reporter Mouse ◽  
Damage Response

11:20 AM: Human Embryonic Germ Cells Generate Adipose Tissue In Vivo

2006 ◽  
Vol 135 (2_suppl) ◽  
pp. P103-P103
Author(s):  
Alexander T Hillel ◽  
Michael Shamblott ◽  
Jennifer Elisseeff
Keyword(s):  
Adipose Tissue ◽  
Germ Cells ◽  
Embryonic Germ Cells

scholarly journals Association of culture of mouse urogenital complexes in media containing rodent sera with the appearance of primordial germ cell-like cells

Reproduction ◽  
2003 ◽  
pp. 519-526 ◽  
Author(s):  
T Mayanagi ◽  
K Ito ◽  
J Takahashi
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Primordial Germ Cell ◽  
Calf Serum ◽  
Culture Method ◽  
Embryonic Germ Cells ◽  
Alkaline Phosphatase Staining

Primordial germ cells differentiate into germ cells and have the ability to reacquire totipotency. Mouse primordial germ cells are identified by alkaline phosphatase staining of the extraembryonic mesoderm, and they proliferate and migrate to reach the genital ridges. Mouse primordial germ cells have never been maintained in culture exclusively for longer than a week without differentiation or dedifferentiation. Moreover, primordial germ cells have not been proliferated with urogenital complexes in vitro, because gonad culture has never been successful. It was thought that primordial germ cells could proliferate in a culture of urogenital complex under modified medium conditions resembling those in vivo; however, organ culture of mouse gonad has been performed with fetal calf serum or equine serum, and those sera produce conditions different from those in vivo. Therefore, mouse urogenital complexes were cultured in media containing rodent sera. As a result, it was possible to proliferate primordial germ cell-like cells outside gonads, and these cells very closely resembled primordial germ cells. In addition, motile primordial germ cell-like cells could be obtained. The ability to maintain primordial germ cell-like cells in culture by this intra-species culture method is important in the study of gametogenesis. Furthermore, this method is useful as a source of stem cells such as embryonic germ cells.

Effects of a Monoclonal Antibody to Glycoprotein IIb/IIIa (P256) and of Enzymically Derived Fragments of P256 on Human Platelets

1991 ◽  
Vol 65 (04) ◽  
pp. 432-437 ◽  
Author(s):  
A W J Stuttle ◽  
M J Powling ◽  
J M Ritter ◽  
R M Hardisty
Keyword(s):  
Monoclonal Antibody ◽  
Flow Cytometry ◽  
Platelet Aggregation ◽  
Calcium Ions ◽  
Human Platelet ◽  
Human Platelets ◽  
In Vivo Detection ◽  
Fibrinogen Binding ◽  
Specific Antagonist

SummaryThe anti-platelet monoclonal antibody P256 is currently undergoing development for in vivo detection of thrombus. We have examined the actions of P256 and two fragments on human platelet function. P256, and its divalent fragment, caused aggregation at concentrations of 10−9−3 × 10−8 M. A monovalent fragment of P256 did not cause aggregation at concentrations up to 10−7 M. P256–induced platelet aggregation was dependent upon extracellular calcium ions as assessed by quin2 fluorescence. Indomethacin partially inhibited platelet aggregation and completely inhibited intracellular calcium mobilisation. Apyrase caused partial inhibition of aggregation. Aggregation induced by the divalent fragment was dependent upon fibrinogen and was inhibited by prostacyclin. Aggregation induced by the whole antibody was only partially dependent upon fibrinogen, but was also inhibited by prostacyclin. P256 whole antibody was shown, by flow cytometry, to induce fibrinogen binding to indomethacin treated platelets. Monovalent P256 was shown to be a specific antagonist for aggregation induced by the divalent forms. In–111–labelled monovalent fragment bound to gel-filtered platelets in a saturable and displaceable manner. Monovalent P256 represents a safer form for in vivo applications

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