Activation of Pulmonary and Cardiac Cell Populations in Response to Environmental Hypoxia or Lung Injury Using Transgenic Mouse Strains Expressing Phenotype-Sensitive Fluorescent Reporter Proteins

Variation in immune homeostasis, immune system stability, in organ systems such as the lungs is likely to shape the host response to infection at these exposed tissues. We evaluated immune homeostasis in immune cell populations in the lungs of the Collaborative Cross (CC) mouse genetic reference population. We found vast heritable variation in leukocyte populations with the frequency of many of these cell types showing distinct patterns relative to classic inbred strains C57BL/6J and BALB/cJ. We identified 28 quantitative trait loci (QTL) associated with variation in baseline lung immune cell populations, including several loci that broadly regulate the abundance of immune populations from distinct developmental lineages, and found that many of these loci have predictive value for influenza disease outcomes, demonstrating that genetic determinants of homeostatic immunity in the lungs regulate susceptibility to virus-induced disease. All told, we highlight the need to assess diverse mouse strains in understanding immune homeostasis and resulting immune responses.

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Live imaging of apoptosis in a novel transgenic mouse highlights its role in neural tube closure

The Journal of Cell Biology ◽

10.1083/jcb.201104057 ◽

2011 ◽

Vol 195 (6) ◽

pp. 1047-1060 ◽

Cited By ~ 125

Author(s):

Yoshifumi Yamaguchi ◽

Naomi Shinotsuka ◽

Keiko Nonomura ◽

Kiwamu Takemoto ◽

Keisuke Kuida ◽

...

Keyword(s):

Transgenic Mouse ◽

Neural Tube ◽

Resonance Energy Transfer ◽

Resonance Energy ◽

Time Lapse ◽

Neural Tube Closure ◽

Caspase Activation ◽

Fluorescent Reporter ◽

Time Lapse Imaging ◽

Tube Closure

Many cells die during development, tissue homeostasis, and disease. Dysregulation of apoptosis leads to cranial neural tube closure (NTC) defects like exencephaly, although the mechanism is unclear. Observing cells undergoing apoptosis in a living context could help elucidate their origin, behavior, and influence on surrounding tissues, but few tools are available for this purpose, especially in mammals. In this paper, we used insulator sequences to generate a transgenic mouse that stably expressed a genetically encoded fluorescence resonance energy transfer (FRET)–based fluorescent reporter for caspase activation and performed simultaneous time-lapse imaging of apoptosis and morphogenesis in living embryos. Live FRET imaging with a fast-scanning confocal microscope revealed that cells containing activated caspases showed typical and nontypical apoptotic behavior in a region-specific manner during NTC. Inhibiting caspase activation perturbed and delayed the smooth progression of cranial NTC, which might increase the risk of exencephaly. Our results suggest that caspase-mediated cell removal facilitates NTC completion within a limited developmental window.

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Abstract 341: Transplantation of Cardiac Tissue Sheets Including Defined Cardiovascular Cell Populations Differentiated from Human-Induced Pluripotent Stem Cells Ameliorates Cardiac Dysfunction After Subacute Myocardial Infarction

Circulation Research ◽

10.1161/res.111.suppl_1.a341 ◽

2012 ◽

Vol 111 (suppl_1) ◽

Author(s):

Hidetoshi Masumoto ◽

Tadashi Ikeda ◽

Tatsuya Shimizu ◽

Teruo Okano ◽

Ryuzo Sakata ◽

...

Keyword(s):

Myocardial Infarction ◽

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Cardiac Dysfunction ◽

Cardiac Tissue ◽

Vascular Endothelial Cell ◽

Cell Populations ◽

Cardiac Cell ◽

Induced Pluripotent

BACKGROUNDS: To realize cardiac regeneration with human induced pluripotent stem cells (hiPSCs), efficient differentiation from hiPSCs to defined cardiac cell populations (cardiomyocytes [CMs]/ endothelial cells [ECs]/ vascular mural cells [MCs]), and transplantation technique for fair engraftment are required. Recently, we reported that mouse ES cell-derived cardiac tissue sheet transplantation to rat myocardial infarction (MI) model ameliorated cardiac function after MI (Stem Cells, in press). Here we tried to extend this technique to hiPSCs. METHODS & RESULTS: We have reported an efficient cardiomyocyte differentiation protocol based on a monolayer culture (PLoS One, 2011), in which cardiac troponin-T (cTnT)-positive CMs robustly appeared with 50-80% efficiency. In this study, we further modified the protocol to induce vascular cells (ECs/MCs) together with CMs by vascular endothelial cell growth factor supplementation, resulted in proportional differentiation of cTnT+-CMs (62.7±11.7% of total cells), VE-cadherin+-ECs (7.8±4.9%) and PDGFRb+-MCs (18.2±11.0%) at differentiation day 15 (n=12). Then, these cells were transferred onto temperature-responsive culture dishes (UpCell dishes; CellSeed, Tokyo, Japan) to form cardiac tissue sheets including defined cardiac populations. After 4 days of culture, we successfully collected self-pulsating cardiac tissue sheets with 7.0×10 5 ±2.3 (n=12) of cells consisted of CMs (46.9±15.9% of total cells), ECs (4.1±3.7%), and MCs (22.5±15.7%). Three-layered hiPSC-derived cardiac sheets were transplanted to a MI model of athymic rat heart one week after MI. In transplantation group, echocardiogram showed a significant improvement of systolic function of left ventricle (fractional shortening: 22.6±5.0 vs 36.5±8.0%, p<0.001, n=20) and a decrease in akinetic length (20.8±9.7 vs 2.5±7.7%, p<0.001, n=20) (pre-treatment vs 4weeks after transplantation). We also succeeded in generation of larger sheets (1.6 inch diameter) with the same method. CONCLUTIONS: Transplantation of hiPSC-derived cardiac tissue sheets significantly ameliorates cardiac dysfunction after MI. Thus, we developed a valuable technological basis for hiPSC-based cardiac cell therapy.

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Generation of Novel Plasmodium falciparum NF135 and NF54 Lines Expressing Fluorescent Reporter Proteins Under the Control of Strong and Constitutive Promoters

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2020.00270 ◽

2020 ◽

Vol 10 ◽

Cited By ~ 1

Author(s):

Shinya Miyazaki ◽

Annie S. P. Yang ◽

Fiona J. A. Geurten ◽

Catherin Marin-Mogollon ◽

Yukiko Miyazaki ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Fluorescent Reporter ◽

Reporter Proteins

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Characterization of Learning, Motivation, and Visual Perception in Five Transgenic Mouse Lines Expressing GCaMP in Distinct Cell Populations

Frontiers in Behavioral Neuroscience ◽

10.3389/fnbeh.2020.00104 ◽

2020 ◽

Vol 14 ◽

Author(s):

Peter A. Groblewski ◽

Douglas R. Ollerenshaw ◽

Justin T. Kiggins ◽

Marina E. Garrett ◽

Chris Mochizuki ◽

...

Keyword(s):

Visual Perception ◽

Transgenic Mouse ◽

Learning Motivation ◽

Cell Populations ◽

Distinct Cell ◽

Mouse Lines

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Tetrodotoxin-insensitive sodium channels in a cardiac cell line from a transgenic mouse

AJP Cell Physiology ◽

10.1152/ajpcell.1992.262.3.c724 ◽

1992 ◽

Vol 262 (3) ◽

pp. C724-C730 ◽

Cited By ~ 11

Author(s):

A. Sculptoreanu ◽

M. Morton ◽

C. L. Gartside ◽

S. D. Hauschka ◽

W. A. Catterall ◽

...

Keyword(s):

Skeletal Muscle ◽

Cell Line ◽

Transgenic Mouse ◽

Sodium Channels ◽

Sodium Current ◽

Cardiac Cell ◽

Dependent Protein Kinase ◽

Electrophysiological Properties ◽

Maximal Activation ◽

Dependent Protein

The electrophysiological properties of a cardiac cell line (MCM1) originating from a transgenic mouse were characterized. The dominant current in these cells is a sodium current that is insensitive to concentrations of tetrodotoxin (TTX) up to 100 microM. It activates and inactivates rapidly with half-maximal activation at -40 mV and half-maximal inactivation at -79 mV. This sodium current is reduced by agents that increase intracellular adenosine 3',5'-cyclic monophosphate (cAMP) and activate cAMP-dependent protein kinase including isoproterenol, 8-bromo-cAMP, and isobutylmethylxanthine. The phenylalkylamine desmethoxyverapamil blocks the TTX-insensitive sodium current in MCM1 cells in both tonic and use-dependent fashion. Membrane depolarization enhances this block. It is proposed that the TTX-insensitive sodium current in these cells may be similar in origin to the embryonic type of TTX-insensitive sodium current described in other cardiac and skeletal muscle preparations.

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Sharing of Ia antigens between species. I. Detection of Ia specificities shared by rats and mice.

Journal of Experimental Medicine ◽

10.1084/jem.146.2.381 ◽

1977 ◽

Vol 146 (2) ◽

pp. 381-393 ◽

Cited By ~ 26

Author(s):

D H Sachs ◽

G W Humphrey ◽

J K Lunney

Keyword(s):

Lymphoid Cells ◽

Mouse Strains ◽

Cell Populations ◽

Complex Chemical ◽

Cross Reactions ◽

Ia Antigens ◽

Mouse Cells ◽

The Cross ◽

Distribution Studies ◽

Rats And Mice

A mouse anti-rat xenogeneic antiserum, B10.D2 anti-BN, has been found to react with a subpopulation of lymphoid cells of certain mouse strains. The corresponding alloantiserum, B10.D2 anti-B10.BR, reacted in analogous fashion with lymphoid cells of BN rats. In the case of the cross-reaction on mouse cells, mapping studies indicated that at least part of the reactivity was with the product of gene(s) determined by the I-A subregion of the H-2 complex. Chemical isolation studies with radiolabeled cell surface preparations indicated that the antigens detected in both mouse and rat had mol wt characteristic of Ia antigens (35,000 and 28,000 dalton molecules). Testing of fractionated spleen cell populations revealed that the cross-reactive antigens were expressed predominatly on B cells, but that a subpopulation of T cells were also reactive. Wider strain and species distribution studies are in progress to determine the extent of such Ia cross-reactions between species and to further assess the practical and theoretical importance of such cross-reactions.

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