scholarly journals Tie2-Cre Transgenic Mice: A New Model for Endothelial Cell-Lineage Analysis in Vivo

2001 ◽  
Vol 230 (2) ◽  
pp. 230-242 ◽  
Author(s):  
Yaz Y Kisanuki ◽  
Robert E Hammer ◽  
Jun-ichi Miyazaki ◽  
S.Clay Williams ◽  
James A Richardson ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Transgenic Mice ◽  
Cell Lineage ◽  
New Model ◽  
Lineage Analysis

Highly efficient lentiviral-mediated human cytokine transgenesis on the NOD/scid background

Blood ◽  
2004 ◽  
Vol 103 (2) ◽  
pp. 580-582 ◽  
Author(s):  
Isabel Punzon ◽  
Luis M. Criado ◽  
Alfredo Serrano ◽  
Fernando Serrano ◽  
Antonio Bernad
Keyword(s):  
Transgenic Mice ◽  
Lentiviral Vector ◽  
Cell Lineage ◽  
Transgenic Animals ◽  
Hematopoietic Tissue ◽  
Nonobese Diabetic ◽  
Human Blood Cell ◽  
Macrophage Colony ◽  
Human Cytokine

Abstract Human neo-organ formation from stem cells can only be assayed by in vivo xenotransplantation. The human nonobese diabetic–severe combined immunodeficient (HuNOD/scid) CD34+ cell transplantation is a model that allows examination of hematopoietic tissue formation, although human hematopoietic cell maturation is abortive. Conventional humanization of the cytokine microenvironment has depended on generation of human cytokine-transgenic mice in strains appropriate for conventional plasmid microinjection, followed by backcrossing, a costly and time-consuming approach. Lentiviral vector infection of single-cell embryos was recently reported to produce transgenic animals. Using this approach, we have generated direct human granulocyte-macrophage colony-stimulating factor (hGM-CSF) transgenic mice from lentivirus-microinjected NOD/scid embryos, with 68% efficiency and 100% penetrance; this allowed us to obtain NOD/scid transgenic mice with considerable savings of resources. This powerful technique should assist in producing novel mouse models for the study of human blood cell lineage development and other human neo-organs from stem cell xenotransplantation for which a similar “humanization” rationale may be required.

scholarly journals Expression of c-MYC under the control of GATA-1 regulatory sequences causes erythroleukemia in transgenic mice.

1995 ◽  
Vol 181 (5) ◽  
pp. 1603-1613 ◽  
Author(s):  
R C Skoda ◽  
S F Tsai ◽  
S H Orkin ◽  
P Leder
Keyword(s):  
Transgenic Mice ◽  
Late Onset ◽  
Cell Lineage ◽  
Erythropoietin Receptor ◽  
Leukemic Cells ◽  
Regulatory Sequences ◽  
Beta Globin ◽  
Promoter Construct ◽  
Transgenic Lines

To study oncogenesis in the erythroid lineage, we have generated transgenic mice carrying the human c-MYC proto-oncogene under the control of mouse GATA-1 regulatory sequences. Six transgenic lines expressed the transgene and displayed a clear oncogenic phenotype. Of these, five developed an early onset, rapidly progressive erythroleukemia that resulted in death of the founder animals 30-50 d after birth. Transgenic progeny of the sixth founder, while also expressing the transgene, remained asymptomatic for more than 8 mo, whereupon members of this line began to develop late onset erythroleukemia. The primary leukemic cells were transplantable into nude mice and syngeneic hosts. Cell lines were established from five of the six leukemic animals and these lines, designated erythroleukemia/c-MYC (EMY), displayed proerythroblast morphology and expressed markers characteristic of the erythroid lineage, including the erythropoietin receptor and beta-globin. Moreover, they also manifested a limited potential to differentiate in response to erythropoietin. Studies in the surviving transgenic line indicated that, contrary to our expectations, the transgene was not expressed in the mast cell lineage. That, coupled with the exclusive occurrence of erythroleukemia in all the transgenic lines, suggests that the GATA-1 promoter construct we have used includes regulatory sequences necessary for in vivo erythroid expression only. Additional sequences would appear to be required for expression in mast cells. Further, our results show that c-MYC can efficiently transform erythroid precursors if expressed at a vulnerable stage of their development.

scholarly journals Visualizing calcium responses to acetylcholine convection along endothelium of arteriolar networks in Cx40BAC-GCaMP2 transgenic mice

2011 ◽  
Vol 301 (3) ◽  
pp. H794-H802 ◽  
Author(s):  
Pooneh Bagher ◽  
Michael J. Davis ◽  
Steven S. Segal
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Transgenic Mice ◽  
Calcium Signaling ◽  
Muscarinic Receptors ◽  
Side Branch ◽  
Widespread Application ◽  
Cell Calcium ◽  
Flow Stream

Acetylcholine evokes endothelium-dependent vasodilation subsequent to a rise in intracellular calcium. Despite widespread application in human and animal studies, calcium responses to intravascular ACh have not been visualized in vivo. Microiontophoresis of ACh in tissue adjacent to an arteriole activates abluminal muscarinic receptors on endothelial cells within a “local” region of diffusion, but it is unknown whether ACh released in such fashion gains access to the flow stream resulting in further actions downstream. To test this hypothesis and provide new insight into calcium signaling in vivo, we studied the cremaster muscle microcirculation of anesthetized male Cx40BAC-GCaMP2 transgenic mice ( n = 22; 5–9 mo; 33 ± 1 g) expressing the fluorescent calcium sensor GCaMP2 selectively in arteriolar endothelial cells. Submaximal ACh stimuli were delivered using microiontophoresis (1-μm pipette tip, 500 nA). With stimulus duration <500 ms or with the micropipette positioned within one vessel diameter (∼30 μm) away from an arteriole, endothelial cell calcium fluorescence was restricted to the region of ACh diffusion (<200 μm). In contrast, with the micropipette tip positioned immediately adjacent to an arteriole or within its lumen, calcium fluorescence encompassed entire networks downstream. The velocity of downstream calcium signaling in response to ACh increased with centerline velocity of fluorescent tracer microbeads ( r2 > 0.99; range: <1 mm/s to >10 mm/s). Diverting arteriolar blood flow into a side branch redirected downstream fluorescence responses to ACh; occluding flow abolished responses. Blocking luminal muscarinic receptors (intravascular glycopyrrolate; 6 μg/kg) inhibited downstream responses reversibly. Through visualizing the actions of a “local” ACh stimulus on endothelial cell calcium fluorescence in vivo, the present findings illustrate that transmural diffusion and convection of an agonist can activate entire networks of arteriolar endothelial cells concomitant with its delivery in the flow stream.

scholarly journals Endogenous CRISPR arrays for scalable whole organism lineage tracing

2018 ◽  
Keyword(s):  
Genetic Engineering ◽  
Dna Sequences ◽  
Single Cell Analysis ◽  
Tumor Biology ◽  
Cell Lineage ◽  
Lineage Tracing ◽  
Cell Analysis ◽  
Central Importance ◽  
Lineage Analysis

AbstractThe last decade has seen a renewed appreciation of the central importance of cellular lineages to many questions in biology (especially organogenesis, stem cells and tumor biology). This has been driven in part by a renaissance in genetic clonal-labeling techniques. Recent approaches are based on accelerated mutation of DNA sequences, which can then be sequenced from individual cells to re-create a “phylogenetic” tree of cell lineage. However, current approaches depend on making transgenic alterations to the genome in question, which limit their application. Here, we introduce a new method which completely avoids the need for prior genetic engineering, by identifying endogenous CRISPR target arrays suitable for lineage analysis. In both mouse and zebrafish we identify the highest quality compact arrays as judged by equal base composition, 5’ G sequence, minimal likelihood of residing in the functional genome, minimal off targets and ease of amplification. We validate multiple high quality endogenous CRISPR arrays, demonstrating their utility for lineage tracing. Our technique thus can produce deep and broad lineages in vivo, while removing the dependence on genetic engineering, and also avoiding the need for single-cell analysis.

Vascular endothelial cell lineage-specific promoter in transgenic mice

Development ◽  
1995 ◽  
Vol 121 (4) ◽  
pp. 1089-1098 ◽  
Author(s):  
T.M. Schlaeger ◽  
Y. Qin ◽  
Y. Fujiwara ◽  
J. Magram ◽  
T.N. Sato
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Vascular Endothelial Cells ◽  
Es Cells ◽  
Vascular Endothelial Cell ◽  
Cell Lineage ◽  
Vascular Endothelial ◽  
Lacz Reporter

Vascular endothelial cells play essential roles in the function and development of the cardiovascular system. However, due to the lack of lineage-specific markers suitable for molecular and biochemical analyses, very little is known about the molecular mechanisms that regulate endothelial cell differentiation. We report the first vascular endothelial cell lineage-specific (including angioblastic precursor cells) 1.2 kb promoter in transgenic mice. Moreover, deletion analysis of this promoter region in transgenic embryos revealed multiple elements that are required for the maximum endothelial cell lineage-specific expression. This is a powerful molecular tool that will enable us to identify factors and cellular signals essential for the establishment of vascular endothelial cell lineage. It will also allow us to deliver genes specifically into this cell type in vivo to test specifically molecules that have been implicated in cardiovascular development. Furthermore, we have established embryonic stem (ES) cells from the blastocysts of the transgenic mouse that carry the 1.2 kb promoter-LacZ reporter transgene. These ES cells were able to differentiate in vitro to form cystic embryoid bodies (CEB) that contain endothelial cells determined by PECAM immunohistochemistry. However, these in vitro differentiated endothelial cells did not express the LacZ reporter gene. This indicates the lack of factors and/or cellular interactions which are required to induce the expression of the reporter gene mediated by this 1.2 kb promoter in this in vitro differentiation system. Thus this system will allow us to screen for the putative inducers that exist in vivo but not in vitro. These putative inducers are presumably important for in vivo differentiation of vascular endothelial cells.

scholarly journals in vivo Cell Lineage Analysis During Chemical Hepatocarcinogenesis in Rats Using Retroviral-Mediated Gene Transfer: Evidence for Dedifferentiation of Mature Hepatocytes

2002 ◽  
Vol 82 (6) ◽  
pp. 781-788 ◽  
Author(s):  
Jérôme Gournay ◽  
Isabelle Auvigne ◽  
Virginie Pichard ◽  
Catherine Ligeza ◽  
Marie-Pierre Bralet ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Cell Lineage ◽  
Lineage Analysis
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