Combination of in silico and in situ hybridisation approaches to identify potential Dll1 associated miRNAs during mouse embryogenesis.
MicroRNAs (miRNAs) have regulatory functions during vertebrate embryogenesis. They are short approximately 21bp long endogenously expressed single-stranded RNAs, which preferentially bind to complementary sequences in the 3' untranslated regions (UTR) of mRNAs and typically down-regulate the respective target mRNAs by translational repression or enhanced mRNA degradation. The Notch ligand Delta-like 1 (Dll1) is expressed in a highly dynamic pattern and has pleiotropic functions during embryogenesis and in adult tissues. Here, we report an interspecies in silico analysis to identify 16 miRNAs, which potentially bind to the mouse, human and chicken Dll1 3'UTRs. To analyze whether these miRNAs could regulate Dll1 gene expression during somitogenesis and neurogenesis, we performed a systematic whole mount in situ hybridisation screen, followed by radioactive in situ hybridisation on sections, using LNA modified DNA probes in mouse embryos. We find that 7 miRNAs (miR-34a, miR-103, miR-107, miR-130a, miR-130b, miR-449a and miR-449c) are expressed in developing somites, limbs, restricted regions of the brain and neural tube between 9.5 dpc and 12.5 dpc. This suggests that these miRNAs could possibly target the Dll1 3'UTR in these regions. The other miRNAs are not expressed or below the detection limit and thus are unlikely to regulate Dll1 at the analyzed embryonic stages.
Hoesel
Bastian
B
Institute of Experimental Genetics, Helmholtz Zentrum München (GmbH), Neuherberg, Germany.
Bhujabal
Zambarlal
Z
Przemeck
Gerhard K H
GK
Kurz-Drexler
Annerose
A
Weisenhorn
Daniela M Vogt
DM
Angelis
Martin Hrabé de
MH
Beckers
Johannes
J
D020413:3' Untranslated Regions / Q000235:genetics; D000818:Animals; D002135:Calcium-Binding Proteins; D002642:Chick Embryo; D002645:Chickens; D019295:Computational Biology / Q000379:methods; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000302:isolation & purification* / Q000378:metabolism*; D011485:Protein Binding
D020413:3' Untranslated Regions; D002135:Calcium-Binding Proteins; C512925:Dlk1 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D035683:MicroRNAs