Primordial germ cells (PGCs) in mice have been recognized histologically as alkaline phosphatase (AP) activity-positive cells at 7.2 days post coitum (dpc) in the extra-embryonic mesoderm. However, mechanisms regulating PGC formation are unknown, and an appropriate in vitro system to study the mechanisms has not been established. Therefore, we have developed a primary culture of explanted embryos at pre- and early-streak stages, and have studied roles of cell and/or tissue interactions in PGC formation. The emergence of PGCs from 5.5 dpc epiblasts was observed only when they were co-cultured with extra-embryonic ectoderm, which may induce the conditions required for PGC formation within epiblasts. From 6.0 dpc onwards, PGCs emerged from whole epiblasts as did a fragment of proximal epiblast that corresponds to the area containing presumptive PGC precursors without neighboring extra-embryonic ectoderm and visceral endoderm. Dissociated epiblasts at these stages, however, did not give rise to PGCs, indicating that interactions among a cluster of a specific number of proximal epiblast cells is needed for PGC differentiation. In contrast, we observed that dissociated epiblast cells from a 6.5-b (6.5+15-16 hours) to 6.75 dpc embryo that had undergone gastrulation gave rise to PGCs. Our results demonstrate that stage-dependent tissue and cell interactions play key roles in PGC determination.
Reference
Ang
S.-L.
, Rossant
J.
(1993
) Anterior mesendoderm induces mouse Engrailed genes in explant cultures.
Development
118
, 139
–149
Chiquoine
A. D.
(1954
) The identification, origin and migration of the primordial germ cells in the mouse embryo.
Anat. Rec
118
, 135
–146
Ciruna
B. G.
, Schwartz
L.
, Harpal
K.
, Yamaguchi
T. P.
, Rossant
J.
(1997
) Chimeric analysis of fibroblast growth factor receptor-1 (Fgfr1) function: a role for FGFR1 in morphogenetic movement through the primitive streak.
Development
124
, 2829
–2841
Copp
A. J.
, Roberts
H. M.
, Polani
P. E.
(1986
) Chimaerism of primordial germ cells in the early postimplantation mouse embryo following microsurgical grafting of posterior primitive streak cells in vitro.
J. Embryol. Exp. Morph
95
, 95
–115
Dolci
S.
, Williams
D. E.
, Ernst
M. K.
, Resnick
J. L.
, Brannan
C. I.
, Lock
L. F.
, Lyman
S. D.
, Scott Boswell
H.
, Donovan
P. J.
(1991
) Requirement for mast cell growth factor for primordial germ cell survival in culture.
Nature
352
, 809
–811
Donovan
P. J.
, Scott
D.
, Cairns
L. A.
, Heasman
J.
, Wylie
C. C.
(1986
) Migratory and postmigratory mouse primordial germ cells behave differently in culture.
Cell
44
, 831
–838
Downs
K. M.
, Davies
T.
(1993
) Staging of gastrulating mouse embryos by morphological landmarks in the dissecting microscope.
Development
118
, 1255
–1266
Eddy
E. M.
(1975
) Germ plasm and the differentiation of the derm cell line.
Int. Rev. Cytol
43
, 229
–280
Faust
C.
, Schumacher
A.
, Holdener
B.
, Magnuson
T.
(1995
) The eed mutation disrupts anterior mesoderm production in mice.
Development
121
, 273
–285
Gardner
R. L.
, Rossant
J.
(1979
). Investigation of the fate of 4.5 day post-coitum mouse inner cell mass cells by blastocyst injection.
J. Embryol. Exp. Morph
53
, 141
–152
Ginsburg
M.
, Snow
M. H. L.
, McLaren
A.
(1990
) Primordial germcells in the mouse embryo during gastrulation.
Development
110
, 521
–528
Godin
I.
, Deed
R.
, Cooke
J.
, Zsebo
K.
, Dexter
M.
, Wylie
C. C.
(1991
) Effects of the steel gene product on mouse primordial germ cells in culture.
Nature
352
, 807
–809
Herrmann
B. G.
(1991
) Expression pattern of the Brachyury gene in whole-mount Twis/Twis mutant embryos.
Development
113
, 913
–917
Kataoka
H.
, Takakura
N.
, Nishikawa
S.
, Tsuchida
K.
, Kodama
H.
, Kunisada
T.
, Risau
W.
, Kita
T.
, Nishikawa
S.-I.
(1997
) Expressions of PDGF receptor alpha, c-Kit and Flk1 genes clustering in mouse chromosome 5 define distinct subsets of nascent mesodermal cells.
Dev. Growth Differ
39
, 729
–740
Kelly
S. J.
(1977
) Studies of the developmental potential of 4-and 8-ccell stage mouse blastomeres.
J. Exp. Zool
200
, 365
–376
Lawson
K. A.
, Dunn
N. R.
, Roelen
B. A. J.
, Zeinstra
L. M.
, Davis
A. M.
, Wright
C. V. E.
, Korving
J. P. W. F. M.
, Hogan
B. L. M.
(1999
) Bmp4 is required for the generation of primordial germ cells in the mouse embryo.
Genes Dev
13
, 424
–436
Matsui
Y.
, Toksoz
D.
, Nishikawa
S.
, Nishikawa
S.
, Williams
D.
, Zsebo
K.
, Hogan
B. L. M.
(1991
) Effect of Steel factor and leukemia inhibitory factor on murine primordial germ cells in culture.
Nature
353
, 750
–752
Matsui
Y.
, Zsebo
K.
, Hogan
B. L. M.
(1992
) Derivation of pluripotential embryonic stem cells from murine primordial germ cells in culture.
Cell
70
, 841
–847
Mintz
B.
, Russell
E. S.
(1957
) Gene-induced embryological modifications of primordial germ cells in the mouse.
J. Exp. Zool
134
, 207
–237
Okamoto
K.
, Okazawa
H.
, Okuda
A.
, Sakai
M.
, Muramatsu
M.
, Hamada
H.
(1990
) A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells.
Cell
60
, 461
–472
Quinn
P.
, Barros
C.
, Whittingham
D. G.
(1982
) Preservation of hamster oocytes to assay the fertilizing capacity of human spermatozoa.
J. Reprod. Fertil
66
, 161
–168
Sasaki
H.
, Hogan
B.
(1993
) Differential expression of multiple fork head related genes during gastrulation and axial pattern formation in the mouse embryo.
Development
118
, 47
–59
Schöler
H. R.
, Dressler
G. R.
, Balling
R.
, Rohdewohld
H.
, Gruss
P.
(1990
) Oct-4: a germline-specific transcription factor mapping to the mouse t-complex.
EMBO J
9
, 2185
–2195
Shimazaki
T.
, Okazawa
H.
, Fujii
H.
, Ikeda
M.
, Tamai
K.
, McKay
R. D. G.
, Muramatsu
M.
, Hamada
H.
(1993
) Hybrid cell extinction andre-expression of Oct-3 function correlates with differentiation potential.
EMBO J
12
, 4489
–4498
Snow
M. H. L.
(1981
) Autonomous development of parts isolated from primitive-streak-stage mouse embryos. Is development clonal?.
J. Embryol. Exp. Morphol
65
, 269
–287
Soriano
P.
, Jaenisch
R.
(1986
) Retroviruses as probes for mammalian development: Allocation of cells to the somatic and germ cell lineages.
Cell
46
, 19
–29
Sun
X.
, Meyers
E. N.
, Lewandoski
M.
, Martin
G. R.
(1999
) Taegeted disruption of Fgf8 causes failure of cell migration in the gastrulating mouse embryo.
Genes Dev
13
, 834
–1846
Tam
P. L. P.
, Zhou
S. X.
(1996
) The allocation of epiblast cells to ectodermal and germ-line lineages is influenced by the position of the cells in the gastrulating mouse embryo.
Dev. Biol
178
, 124
–132
Williams
D. E.
, de Vries
P.
, Namen
A. E.
, Widmer
M. B.
, Lyman
S. D.
(1992
) The Steel factor.
Dev. Biol
151
, 368
–376
Wilson
V.
, Manson
L.
, Skarnes
W. C.
, Beddington
R. S.
(1995
) The T gene is necessary for normal mesodermal morphogenetic cell movements during gastrulation.
Development
121
, 877
–886
Wilson
V.
, Rashbass
P.
, Beddington
R. S.
(1993
) Chimeric analysis of T (Brachyury) gene function.
Development
117
, 1321
–1331
Yamaguchi
T. P.
, Dumont
D. J.
, Conlon
R. A.
, Breitman
M. L.
, Rossant
J.
(1993
) flk-1, an flt -related receptor tyrosine kinase is an early marker for endothelial call precursors.
Development
118
, 489
–498
Yasui
K.
, Sasaki
H.
, Arakaki
R.
, Uemura
M.
(1997
) Distribution pattern of HNF-3proteins in developing embryos of two mammalian species, the house shrew and the mouse.
Dev. Growth Differ
39
, 667
–676
Yoshinaga
K.
, Muramatsu
H.
, Muramatsu
T.
(1991
) Immunohistochemical localization of the carbohydrate antigen 4C9 in the mouse embryo: a reliable marker of mouse primordial germ cells.
Differentiation
48
, 75
–82
This content is only available via PDF.
© 2001 by Company of Biologists
2001
-JournalMeeting.png?versionId=3737)
(update)-ImmuneSI.png?versionId=3737)
-GSWorkshop.png?versionId=3737)
(update)-InPreprints.png?versionId=3737)
-FocalPlaneNetworkLaunch.png?versionId=3737)
