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Abstract
MicroRNAs play important roles in regulating development
at both transcriptional and post-transcriptional levels.
Here we report twenty nine microRNAs from mouse testis
that are differentially expressed as the prepuberal testis
differentiates to the adult testis. Using computational
analyses to identify potential microRNA target mRNAs, we
identify several possible male germ cell target mRNAs. One
highly conserved sequence in the 3'UTR of transition
protein 2 (Tnp2) mRNA, a testis-specific and
post-transcriptionally regulated mRNA in post-meiotic germ
cells, is complementary to Mirn122a.
Mirn122a is enriched in
late stage male germ cells and is predominantly on
polysomes. Mirn122a, but not another
non-complementary
miRNA, inhibits the activity of a luciferase reporter
construct containing the 3'UTR of Tnp2. Site
directed
mutations of Mirn122a indicate that base pairing of
the
5'-region of Mirn122a to its complementary site in
the 3'
UTR of Tnp2 mRNA is essential for the
down-regulation of
luciferase activity. Real Time RT-PCR and ribonuclease
protection assays reveal that the Mirn122a directed
decrease of the Tnp2 reporter gene activity results
from
mRNA cleavage. We propose that specific microRNAs such as
Mirn122a could be involved in the
post-transcriptional
regulation of mRNAs such as Tnp2 in the mammalian
testis.
Key words:
Testis •
Gametogenesis •
Gene regulation •
Spermatid •
Spermatogenesis
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