The role of miRNA-200a in the early stage of the mandibular development

Ahmed Salah Yassin, Kenji Hoshi, Fumie Terao, Mariko Umeda, Ichiro Takahashi

Research output: Contribution to journalArticle

Abstract

Background MicroRNAs are non-coding small RNA molecules that regulate various cellular functions. In this study, we analyzed the function of microRNA-200a (miR-200a) during the development of primary and secondary cartilages in the mandible during the embryonic stage. Materials and methods Mandibular processes were excised from ICR mouse embryos. Meckel's cartilage and the complex of condylar and angular cartilages were examined as primary and secondary cartilages, respectively. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and in situ hybridization were performed to examine the expression of miR-200a during normal mandibular development. Transfection of a miR-200a-mimic and -inhibitor were performed to analyze the effect of miR-200a on mandibular cartilage formation. Prior to transfection, DiI labeling was performed to detect the transfection sites and examine the effect of miR-200a on Meckel's and condylar cartilage formation. Nine days after transfection, Alcian blue staining and quantification were performed to analyze the formation of the cartilage. Results The expression levels of miR-200a gradually increased from embryonic day 9–14 in mandibular processes. However, the expression levels of miR-200a in Meckel's cartilage were nearly identical from embryonic day 12–16. The positive hybridization signals were observed in Meckel's cartilage and mesenchymal condensation of mandibular condylar cartilage. Alcian blue analysis showed a significant decrease in the complex of condylar and angular cartilage formation in the lateral-anterior-miR-200a-mimic transfected samples compared to miR-200a-inhibitor-transfected and control samples. Conclusion miR-200a negatively regulates the formation of condylar cartilage in early mandibular development.

Original languageEnglish
Pages (from-to)197-206
Number of pages10
JournalOrthodontic Waves
Volume76
Issue number4
DOIs
Publication statusPublished - Dec 1 2017

Fingerprint

MicroRNAs
Cartilage
Transfection
Alcian Blue
Small Untranslated RNA
Inbred ICR Mouse
Mandible
Reverse Transcription
In Situ Hybridization
Embryonic Structures
Staining and Labeling
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Orthodontics

Cite this

The role of miRNA-200a in the early stage of the mandibular development. / Yassin, Ahmed Salah; Hoshi, Kenji; Terao, Fumie; Umeda, Mariko; Takahashi, Ichiro.

In: Orthodontic Waves, Vol. 76, No. 4, 01.12.2017, p. 197-206.

Research output: Contribution to journalArticle

@article{d26a9f12b5164914a73dae321a5c2233,
title = "The role of miRNA-200a in the early stage of the mandibular development",
abstract = "Background MicroRNAs are non-coding small RNA molecules that regulate various cellular functions. In this study, we analyzed the function of microRNA-200a (miR-200a) during the development of primary and secondary cartilages in the mandible during the embryonic stage. Materials and methods Mandibular processes were excised from ICR mouse embryos. Meckel's cartilage and the complex of condylar and angular cartilages were examined as primary and secondary cartilages, respectively. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and in situ hybridization were performed to examine the expression of miR-200a during normal mandibular development. Transfection of a miR-200a-mimic and -inhibitor were performed to analyze the effect of miR-200a on mandibular cartilage formation. Prior to transfection, DiI labeling was performed to detect the transfection sites and examine the effect of miR-200a on Meckel's and condylar cartilage formation. Nine days after transfection, Alcian blue staining and quantification were performed to analyze the formation of the cartilage. Results The expression levels of miR-200a gradually increased from embryonic day 9–14 in mandibular processes. However, the expression levels of miR-200a in Meckel's cartilage were nearly identical from embryonic day 12–16. The positive hybridization signals were observed in Meckel's cartilage and mesenchymal condensation of mandibular condylar cartilage. Alcian blue analysis showed a significant decrease in the complex of condylar and angular cartilage formation in the lateral-anterior-miR-200a-mimic transfected samples compared to miR-200a-inhibitor-transfected and control samples. Conclusion miR-200a negatively regulates the formation of condylar cartilage in early mandibular development.",
author = "Yassin, {Ahmed Salah} and Kenji Hoshi and Fumie Terao and Mariko Umeda and Ichiro Takahashi",
year = "2017",
month = "12",
day = "1",
doi = "10.1016/j.odw.2017.06.001",
language = "English",
volume = "76",
pages = "197--206",
journal = "Orthodontic Waves",
issn = "1344-0241",
publisher = "Elsevier BV",
number = "4",

}

TY - JOUR

T1 - The role of miRNA-200a in the early stage of the mandibular development

AU - Yassin, Ahmed Salah

AU - Hoshi, Kenji

AU - Terao, Fumie

AU - Umeda, Mariko

AU - Takahashi, Ichiro

PY - 2017/12/1

Y1 - 2017/12/1

N2 - Background MicroRNAs are non-coding small RNA molecules that regulate various cellular functions. In this study, we analyzed the function of microRNA-200a (miR-200a) during the development of primary and secondary cartilages in the mandible during the embryonic stage. Materials and methods Mandibular processes were excised from ICR mouse embryos. Meckel's cartilage and the complex of condylar and angular cartilages were examined as primary and secondary cartilages, respectively. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and in situ hybridization were performed to examine the expression of miR-200a during normal mandibular development. Transfection of a miR-200a-mimic and -inhibitor were performed to analyze the effect of miR-200a on mandibular cartilage formation. Prior to transfection, DiI labeling was performed to detect the transfection sites and examine the effect of miR-200a on Meckel's and condylar cartilage formation. Nine days after transfection, Alcian blue staining and quantification were performed to analyze the formation of the cartilage. Results The expression levels of miR-200a gradually increased from embryonic day 9–14 in mandibular processes. However, the expression levels of miR-200a in Meckel's cartilage were nearly identical from embryonic day 12–16. The positive hybridization signals were observed in Meckel's cartilage and mesenchymal condensation of mandibular condylar cartilage. Alcian blue analysis showed a significant decrease in the complex of condylar and angular cartilage formation in the lateral-anterior-miR-200a-mimic transfected samples compared to miR-200a-inhibitor-transfected and control samples. Conclusion miR-200a negatively regulates the formation of condylar cartilage in early mandibular development.

AB - Background MicroRNAs are non-coding small RNA molecules that regulate various cellular functions. In this study, we analyzed the function of microRNA-200a (miR-200a) during the development of primary and secondary cartilages in the mandible during the embryonic stage. Materials and methods Mandibular processes were excised from ICR mouse embryos. Meckel's cartilage and the complex of condylar and angular cartilages were examined as primary and secondary cartilages, respectively. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and in situ hybridization were performed to examine the expression of miR-200a during normal mandibular development. Transfection of a miR-200a-mimic and -inhibitor were performed to analyze the effect of miR-200a on mandibular cartilage formation. Prior to transfection, DiI labeling was performed to detect the transfection sites and examine the effect of miR-200a on Meckel's and condylar cartilage formation. Nine days after transfection, Alcian blue staining and quantification were performed to analyze the formation of the cartilage. Results The expression levels of miR-200a gradually increased from embryonic day 9–14 in mandibular processes. However, the expression levels of miR-200a in Meckel's cartilage were nearly identical from embryonic day 12–16. The positive hybridization signals were observed in Meckel's cartilage and mesenchymal condensation of mandibular condylar cartilage. Alcian blue analysis showed a significant decrease in the complex of condylar and angular cartilage formation in the lateral-anterior-miR-200a-mimic transfected samples compared to miR-200a-inhibitor-transfected and control samples. Conclusion miR-200a negatively regulates the formation of condylar cartilage in early mandibular development.

UR - http://www.scopus.com/inward/record.url?scp=85025161089&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85025161089&partnerID=8YFLogxK

U2 - 10.1016/j.odw.2017.06.001

DO - 10.1016/j.odw.2017.06.001

M3 - Article

AN - SCOPUS:85025161089

VL - 76

SP - 197

EP - 206

JO - Orthodontic Waves

JF - Orthodontic Waves

SN - 1344-0241

IS - 4

ER -