Direct effects of retinoic acid on entry of fetal male germ cells into meiosis in mice.
In both male and female germ cells of mice, retinoic acid (RA) is a meiosis-inducing factor. In the present study, we used a germ cell culture system to examine the direct effects of RA on meiotic initiation in male germ cells at the stage when they normally enter mitotic arrest to determine the extent to which fetal male germ cells can respond to exogenous RA to alter their sex-specific pathway. Male germ cells between 13.5 and 15.5 days postcoitum (dpc) were isolated from Pou5fl-green fluorescent protein transgenic fetuses and cultured with or without RA for up to 6 days. In the absence of RA, male germ cells did not undergo DNA replication and did not enter meiosis in culture. However, in the presence of RA, male germ cells isolated at 13.5 dpc expressed Stra8 and initiated the meiotic process. The ratio of cells entering meiosis gradually decreased as cells were isolated progressively at later stages. By 15.5 dpc, isolated male germ cells lost their ability to respond to RA signaling. These cells remained dispersed as single cells and progressed along the male differentiation pathway, as evidenced by the establishment of male-specific methylation imprints regardless of the presence or absence of RA. We conclude that male germ cells maintain sexual bipotency until 14.5 dpc that can be reversed by the addition of RA. Once male germ cells enter mitotic arrest, however, they appear to be committed irreversibly to the male-specific differentiation pathway even in the presence of exogenously added RA.
Ohta
Kohei
K
Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii 96813, USA.
Lin
Yanling
Y
Hogg
Nathanael
N
Yamamoto
Miyuki
M
Yamazaki
Yukiko
Y
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D018797:Cell Cycle Proteins; D002478:Cells, Cultured; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D004261:DNA Replication; D004268:DNA-Binding Proteins; D047109:Fetal Development; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D018392:Genomic Imprinting; D008297:Male; D008540:Meiosis; D051379:Mice; D008822:Mice, Transgenic; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D062085:RNA, Long Noncoding; D012333:RNA, Messenger / Q000378:metabolism; D022661:RNA, Untranslated / Q000235:genetics / Q000378:metabolism; D019849:Sex Determination Processes; D013094:Spermatozoa / Q000166:cytology / Q000378:metabolism; D014212:Tretinoin / Q000502:physiology*
D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C094335:H19 long non-coding RNA; D009687:Nuclear Proteins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D011506:Proteins; D062085:RNA, Long Noncoding; D012333:RNA, Messenger; D022661:RNA, Untranslated; C490060:SYCP3 protein, human; C102019:Stra8 protein, mouse; D014212:Tretinoin; C498796:Dnmt3l protein, mouse; D004248:DNA (Cytosine-5-)-Methyltransferases
Development. 2003 Aug;130(15):3579-86
12810604
Nat Rev Mol Cell Biol. 2004 Dec;5(12):983-97
15573136
J Cell Sci. 2008 Oct 1;121(Pt 19):3233-42
18799790
Science. 2006 Apr 28;312(5773):596-600
16574820
Hum Mol Genet. 2000 Nov 22;9(19):2885-94
11092765
Biol Reprod. 2010 Feb;82(2):433-43
19864318
Philos Trans R Soc Lond B Biol Sci. 1995 Nov 29;350(1333):229-33
8570686
Cell Cycle. 2008 Mar 1;7(5):656-64
18256537
Biol Reprod. 2008 Jul;79(1):35-42
18322276
Science. 2001 Aug 10;293(5532):1089-93
11498579
Dev Biol. 1995 Apr;168(2):683-5
7729599
Stem Cells. 2008 Feb;26(2):339-47
18024419
Genomics. 2002 Apr;79(4):530-8
11944985
Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16916-21
12481029
Biol Reprod. 2008 Mar;78(3):537-45
18032419
Development. 1990 Jul;109(3):635-46
2401216
Proc Natl Acad Sci U S A. 2008 Sep 30;105(39):14976-80
18799751
Chromosoma. 1982;85(3):427-37
6180868
Development. 2002 Apr;129(8):1983-93
11934864
Biol Reprod. 2004 Mar;70(3):687-93
14613895
Mech Dev. 2002 Jul;115(1-2):157-60
12049782
Nature. 1990 Nov 29;348(6300):450-2
2247150
Development. 2008 Apr;135(8):1415-25
18321981
Proc R Soc Lond B Biol Sci. 1975 Jul 1;190(1099):187-97
238208
Nature. 2004 Jun 24;429(6994):900-3
15215868
Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2474-9
16461896
Symp Soc Exp Biol. 1984;38:7-23
6400220
Dev Biol. 2000 Oct 15;226(2):209-19
11023681
Nat Genet. 2006 Dec;38(12):1430-4
17115059
Genes Dev. 2008 Feb 15;22(4):430-5
18281459
Dev Biol. 1997 Jul 1;187(1):107-13
9224678
Development. 1991 May;112(1):265-8
1769333
Biol Reprod. 2007 Oct;77(4):697-706
17615405
Proc Natl Acad Sci U S A. 2005 Aug 9;102(32):11361-6
16055553
Biol Cell. 2009 Jul 31;101(10):587-98
19419345
Development. 2002 Mar;129(5):1155-64
11874911
J Reprod Fertil. 1999 Jul;116(2):379-84
10615264
Science. 2003 Aug 29;301(5637):1239-41
12947200
Histochem Cell Biol. 2003 May;119(5):355-62
12736726