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Blood, Vol. 95 No. 4 (February 15), 2000:
pp. 1274-1282
Hemopoietic lineage commitment decisions: in vivo evidence from a
transgenic mouse model harboring µLCR- pro-LacZ as a
transgene
Thalia Papayannopoulou,
Gregory V. Priestley,
Alex Rohde,
Kenneth R. Peterson, and
Betty Nakamoto
From the Department of Medicine, Division of Hematology, University
of Washington, Seattle, WA and the Department of Biochemistry and
Molecular Biology, University of Kansas, Kansas City,
KS.
A substantial body of published data suggests activation of
lineage-specific genes in multipotential hemopoietic cells before their
unilineage commitment. Because the behavior and plasticity of cells
isolated in vitro away from microenvironmental constraints exercised in
vivo may be altered, one wonders whether similar findings can be
observed in a physiologic setting in vivo. We used a transgenic mouse
model harboring human micro LCR together with  promoter sequences as
a transgene to examine activation of lineage-specific programs in vivo.
By using LacZ as a reporter, we had the ability to detect,
quantitate, and select live cells with different levels of LacZ
activation. We found strong expression of LacZ by X-gal
staining in 2 lineages erythroid and megakaryocytic. Activation in the
latter was a novel finding not previously observed when similar
transgenes were used. We also found activation of µLCR-pro at low
levels in progenitor cells of granulocytic-macrophagic, erythroid, or
megakaryocytic lineage detected by in vitro assays, suggesting
activation before commitment to a specific lineage pathway. In
particular, the expression of LacZ was graded among progenitors, so that in a proportion of them activation occurred only
after commitment to erythroid or megakaryocytic lineage. In addition,
we found quantitative reduction in LacZ expression between
fetal liver and bone marrow-derived cells, the basis of which is
unclear. Collectively our data provide in vivo evidence supporting the
view that lineage-specific genes are expressed in a graded fashion in
pluripotential cells before their irreversible unilineage commitment.
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