Abstract

For prevention and cure of AIDS, it is important to know the mechanism of how cell lineage for T lymphocyte is determined during embryonic hematopoiesis. BMPs specify the fate of mesodermal tissues more ventral. Hematopoietic cells are derived from the most ventral mesoderm, therefore, BMPs are hypothesized to play important roles during specification of T lymphocyte lineage in the hematopoietic tissues. Our basic strategy to prove this hypothesis is to culture embryos that have floxed alleles for BMPs or BMP receptors using whole embryo culture system, and then to microinject Cre recombinase expressing plasmids in the interested region of embryos. Through this method, we could achieve region specific gene disruption. For this purpose, first, we established the culture method for the embryos as early as day 5.5 embryos. We found that DMEM supplemented with 75% rat serum worked better than 100 % rat serum for E5.5 embryos whereas 100 % rat serum works better for E5.75 embryos. We are underway to optimize the condition to achieve the Cre-dependent gene disruption by Cre plasmids. The knowledge that will be acquired through this research may give us better understanding for the primitive and definitive hematopoiesis.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Intramural Research (Z01)
Project #
1Z01ES071004-05
Application #
6838566
Study Section
(LRDT)
Project Start
Project End
Budget Start
Budget End
Support Year
5
Fiscal Year
2003
Total Cost
Indirect Cost

  Institution

Name
U.S. National Inst of Environ Hlth Scis
Department
Type
DUNS #
City
State
Country
United States
Zip Code

  Publications

Miura, Shigeto; Mishina, Yuji (2007) The DVE changes distal epiblast fate from definitive endoderm to neurectoderm by antagonizing nodal signaling. Dev Dyn 236:1602-10
Di-Gregorio, Aida; Sancho, Margarida; Stuckey, Daniel W et al. (2007) BMP signalling inhibits premature neural differentiation in the mouse embryo. Development 134:3359-69
Song, Lanying; Fassler, Reinhard; Mishina, Yuji et al. (2007) Essential functions of Alk3 during AV cushion morphogenesis in mouse embryonic hearts. Dev Biol 301:276-86
Miura, Shigeto; Davis, Shannon; Klingensmith, John et al. (2006) BMP signaling in the epiblast is required for proper recruitment of the prospective paraxial mesoderm and development of the somites. Development 133:3767-75
Park, Changwon; Lavine, Kory; Mishina, Yuji et al. (2006) Bone morphogenetic protein receptor 1A signaling is dispensable for hematopoietic development but essential for vessel and atrioventricular endocardial cushion formation. Development 133:3473-84
Kishigami, Satoshi; Komatsu, Yoshihiro; Takeda, Haruko et al. (2006) Optimized beta-galactosidase staining method for simultaneous detection of endogenous gene expression in early mouse embryos. Genesis 44:57-65
Fukuda, Tomokazu; Scott, Gregory; Komatsu, Yoshihiro et al. (2006) Generation of a mouse with conditionally activated signaling through the BMP receptor, ALK2. Genesis 44:159-67
Kishigami, Satoshi; Mishina, Yuji (2005) BMP signaling and early embryonic patterning. Cytokine Growth Factor Rev 16:265-78
Gaussin, Vinciane; Morley, Gregory E; Cox, Luk et al. (2005) Alk3/Bmpr1a receptor is required for development of the atrioventricular canal into valves and annulus fibrosus. Circ Res 97:219-26
Oh, Hidemasa; Chi, Xuan; Bradfute, Steven B et al. (2004) Cardiac muscle plasticity in adult and embryo by heart-derived progenitor cells. Ann N Y Acad Sci 1015:182-9

Showing the most recent 10 out of 16 publications