title: A whole-mount in situ hybridization method for microRNA detection in Caenorhabditis elegans.
abstract: Whole-mount in situ hybridization (WISH) is an outstanding method to decipher the spatiotemporal expression patterns of microRNAs (miRNAs) and provides important clues for elucidating their functions. The first WISH method for miRNA detection was developed in zebrafish. Although this method was quickly adapted for other vertebrates and fruit flies, WISH analysis has not been successfully used to detect miRNAs in Caenorhabditis elegans Here, we show a novel WISH method for miRNA detection in C. elegans Using this method, mir-1 miRNA was detected in the body-wall muscle where the expression and roles of mir-1 miRNA have been previously elucidated. Application of the method to let-7 family miRNAs, let-7, mir-48, mir-84, and mir-241, revealed their distinct but partially overlapping expression patterns, indicating that miRNAs sharing a short common sequence were distinguishably detected. In pash-1 mutants that were depleted of mature miRNAs, signals of mir-48 miRNA were greatly reduced, suggesting that mature miRNAs were detected by the method. These results demonstrate the validity of WISH to detect mature miRNAs in C. elegans.
authors.affiliation: Genetic Strains Research Center, National Institute of Genetics, Research Organization of Information and Systems, Mishima 411-8540, Japan Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies), Mishima 411-8540, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Andachi
authors.affiliation: Genetic Strains Research Center, National Institute of Genetics, Research Organization of Information and Systems, Mishima 411-8540, Japan Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies), Mishima 411-8540, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Kohara
mesh_terms: D000818:Animals; D017173:Caenorhabditis elegans / Q000196:embryology / Q000235:genetics* / Q000254:growth & development; D017403:In Situ Hybridization; D007814:Larva / Q000378:metabolism; D035683:MicroRNAs / Q000378:metabolism*; D009154:Mutation
chemical_list: D035683:MicroRNAs
keywords: C. elegans; in situ hybridization; let-7; miRNA; mir-1