MicroRNA biogenesis is required for mouse primordial germ cell development and spermatogenesis.
BACKGROUND
MicroRNAs (miRNAs) are critical regulators of transcriptional and post-transcriptional gene silencing, which are involved in multiple developmental processes in many organisms. Apart from miRNAs, mouse germ cells express another type of small RNA, piwi-interacting RNAs (piRNAs). Although it has been clear that piRNAs play a role in repression of retrotransposons during spermatogenesis, the function of miRNA in mouse germ cells has been unclear.


RESULTS
In this study, we first revealed the expression pattern of miRNAs by using a real-time PCR-based 220-plex miRNA expression profiling method. During development of germ cells, miR-17-92 cluster, which is thought to promote cell cycling, and the ES cell-specific cluster encoding miR-290 to -295 (miR-290-295 cluster) were highly expressed in primordial germ cells (PGCs) and spermatogonia. A set of miRNAs was developmentally regulated. We next analysed function of miRNA biogenesis in germ cell development by using conditional Dicer-knockout mice in which Dicer gene was deleted specifically in the germ cells. Dicer-deleted PGCs and spermatogonia exhibited poor proliferation. Retrotransposon activity was unexpectedly suppressed in Dicer-deleted PGCs, but not affected in the spermatogonia. In Dicer-deleted testis, spermatogenesis was retarded at an early stage when proliferation and/or early differentiation. Additionally, we analysed spermatogenesis in conditional Argonaute2-deficient mice. In contrast to Dicer-deficient testis, spermatogenesis in Argonaute2-deficient testis was indistinguishable from that in wild type.


CONCLUSIONS
These results illustrate that miRNAs are important for the proliferation of PGCs and spermatogonia, but dispensable for the repression of retrotransposons in developing germ cells. Consistently, miRNAs promoting cell cycling are highly expressed in PGCs and spermatogonia. Furthermore, based on normal spermatogenesis in Argonaute2-deficient testis, the critical function of Dicer in spermatogenesis is independent of Argonaute2.
Hayashi
Katsuhiko
K
Wellcome Trust/Cancer Research United Kingdom Gurdon Institute, University of Cambridge, Cambridge, United Kingdom.
Chuva de Sousa Lopes
Susana M
SM
Kaneda
Masahiro
M
Tang
Fuchou
F
Hajkova
Petra
P
Lao
Kaiqin
K
O'Carroll
Donal
D
Das
Partha P
PP
Tarakhovsky
Alexander
A
Miska
Eric A
EA
Surani
M Azim
MA
D000818:Animals; D060565:Argonaute Proteins; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D053487:DEAD-box RNA Helicases / Q000502:physiology; D019175:DNA Methylation; D004722:Endoribonucleases / Q000502:physiology; D015852:Eukaryotic Initiation Factor-2 / Q000502:physiology; D005260:Female; D005786:Gene Expression Regulation; D005854:Germ Cells / Q000254:growth & development* / Q000378:metabolism; D051034:Inhibitor of Apoptosis Proteins / Q000378:metabolism; D019426:Integrases; D020084:Long Interspersed Nucleotide Elements / Q000502:physiology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics*; D034741:RNA, Small Interfering / Q000502:physiology; D043244:Ribonuclease III; D013091:Spermatogenesis / Q000502:physiology*; D013737:Testis / Q000166:cytology / Q000502:physiology; D044767:Ubiquitin-Protein Ligases
C526735:Ago2 protein, mouse; D060565:Argonaute Proteins; D015852:Eukaryotic Initiation Factor-2; D051034:Inhibitor of Apoptosis Proteins; D035683:MicroRNAs; D034741:RNA, Small Interfering; C494834:Birc2 protein, mouse; D044767:Ubiquitin-Protein Ligases; C045073:Cre recombinase; D019426:Integrases; D004722:Endoribonucleases; C509781:Dicer1 protein, mouse; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases