Dicer is a key player in oocyte maturation.
OBJECTIVE
Apply Dicer siRNA to study functions of Dicer and miRNA during oogenesis.


METHODS
Mouse oocytes were injected with Dicer siRNA and negative control siRNA and then matured in vitro. After IVM, oocytes were examined for maturation rates, spindle and chromosomal organization, and various gene expressions.


RESULTS
Dicer siRNA significantly reduced maturation rates, increased abnormal spindle and chromosomal organization, and reduced the transcripts of Dicer miRNAs, spindle formation proteins (plk1 and AURKA) and spindle check points (Bub1, Bublb). Depletion of bulb16 markedly prohibited the first polar body extrusion and increased the incidence of misaligned chromosomes and abnormal meiotic spindle assembly.


CONCLUSIONS
Dicer siRNA triggered a cascade reduction for gene expressions starting from Dicer to miRNAs than to spindle assembly proteins and checkpoints which led to abnormal spindle and chromosomal organization. Thus, Dicer and miRNA appeared to play an important role during oogenesis and were essential for meiotic completion.
Liu
Hung-Ching
HC
Department of Reproductive Medicine, Weill Medical College of Cornell University, New York, NY 10021, USA. hcliu@med.cornell.edu
Tang
YaXu
Y
He
Zhiying
Z
Rosenwaks
Zev
Z
D000818:Animals; D016009:Chi-Square Distribution; D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D004722:Endoribonucleases / Q000378:metabolism*; D005260:Female; D005455:Fluorescent Antibody Technique; D008540:Meiosis / Q000502:physiology*; D051379:Mice; D008845:Microinjections; D018613:Microscopy, Confocal; D009866:Oogenesis / Q000502:physiology*; D034741:RNA, Small Interfering; D020133:Reverse Transcriptase Polymerase Chain Reaction; D043244:Ribonuclease III
D034741:RNA, Small Interfering; D004722:Endoribonucleases; C509781:Dicer1 protein, mouse; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases
Science. 2001 Sep 21;293(5538):2269-71
11486053
Nature. 2001 May 24;411(6836):494-8
11373684
Zygote. 2000 Feb;8(1):87-96
10840878
Cell. 2000 Mar 31;101(1):25-33
10778853
EMBO J. 2000 Oct 2;19(19):5194-201
11013221
Nat Genet. 2003 Nov;35(3):215-7
14528307
Dev Biol. 2007 Feb 1;302(1):104-17
17022963
Curr Opin Cell Biol. 2004 Jun;16(3):223-9
15145345
Genes Dev. 2001 Jan 15;15(2):188-200
11157775
Biol Reprod. 2006 Dec;75(6):877-84
16943360
Cell Cycle. 2006 Sep;5(18):2130-7
16969117
Genes Dev. 2007 Mar 15;21(6):644-8
17369397
Mol Reprod Dev. 2003 Mar;64(3):292-301
12548662
EMBO J. 2002 Sep 2;21(17):4663-70
12198168
Nat Cell Biol. 2000 Feb;2(2):70-5
10655585
PLoS One. 2009 Nov 02;4(11):e7701
19888327
Development. 2000 Oct;127(19):4147-56
10976047
Genes Dev. 2007 Mar 15;21(6):682-93
17369401
Trends Genet. 1999 Sep;15(9):358-63
10461204
Biol Reprod. 2009 Apr;80(4):795-803
19129509
Cell. 1999 Oct 15;99(2):133-41
10535732
Cell. 1999 Oct 15;99(2):123-32
10535731
Proc Natl Acad Sci U S A. 2002 Dec 10;99 Suppl 4:16499-506
12169664
Mol Reprod Dev. 2005 Feb;70(2):111-21
15570624
Reproduction. 2006 May;131(5):861-74
16672351
Nature. 2002 Jul 11;418(6894):244-51
12110901
Cell. 2003 Jun 13;113(6):673-6
12809598
Nat Rev Genet. 2004 Jul;5(7):522-31
15211354
Mol Reprod Dev. 2002 Dec;63(4):413-21
12412042
Nat Cell Biol. 2006 Oct;8(10):1114-23
16980957
Curr Top Microbiol Immunol. 2008;320:225-56
18268847
Nature. 2003 Sep 25;425(6956):415-9
14508493
J Biol Chem. 2005 Mar 11;280(10):9330-5
15613470
Genesis. 2002 Apr;32(4):269-76
11948914
Nat Genet. 2003 Nov;35(3):217-8
14528306
Cell. 2004 Jan 23;116(2):281-97
14744438
Cell. 2005 Aug 26;122(4):553-63
16122423
Science. 2009 Nov 13;326(5955):991-4
19965510
Curr Biol. 2003 Dec 2;13(23):R925-36
14654021
Curr Biol. 2002 Sep 3;12(17):1484-95
12225663
Genes Dev. 2006 Jul 1;20(13):1732-43
16766679
J Reprod Immunol. 2002 Jan;53(1-2):171-80
11730914
Methods Mol Biol. 2009;518:157-73
19085139
Science. 2001 Aug 3;293(5531):834-8
11452083
Cancer Res. 2005 Aug 1;65(15):6719-25
16061653