Molecular characterization and expression of maternally expressed gene 3 (Meg3/Gtl2) RNA in the mouse inner ear.
The pathways responsible for sound perception in the cochlea involve the coordinated and regulated expression of hundreds of genes. By using microarray analysis, we identified several transcripts enriched in the inner ear, including the maternally expressed gene 3 (Meg3/Gtl2), an imprinted noncoding RNA. Real-time PCR analysis demonstrated that Meg3/Gtl2 was highly expressed in the cochlea, brain, and eye. Molecular studies revealed the presence of several Meg3/Gtl2 RNA splice variants in the mouse cochlea, brain, and eye. In situ hybridizations showed intense Meg3/Gtl2 RNA staining in the nuclei of type I spiral ganglion cells and in cerebellum near the dorsal vestibular region of the cochlea. In embryonic mouse head sections, Meg3/Gtl2 RNA expression was observed in the otocyst, brain, eye, cartilage, connective tissue, and muscle. Meg3/Gtl2 RNA expression increased in the developing otocyst and localized to the spiral ganglion, stria vascularis, Reissner's membrane, and greater epithelial ridge (GER) in the cochlear duct. RT-PCR analysis performed on cell lines derived from the organ of Corti, representing neural, supporting, and hair cells, showed significantly elevated levels of Meg3/Gtl2 expression in differentiated neural cells. We propose that Meg3/Gtl2 RNA functions as a noncoding regulatory RNA in the inner ear and that it plays a role in pattern specification and differentiation of cells during otocyst development, as well as in the maintenance of a number of terminally differentiated cochlear cell types.
Manji
Shehnaaz S M
SS
Gene Identification and Expression, Murdoch Childrens Research Institute, Royal Children's Hospital, Parkville, Victoria, Australia. shehnaaz.manji@mcri.edu.au
Sørensen
Brita S
BS
Klockars
Tuomas
T
Lam
Timothy
T
Hutchison
Wendy
W
Dahl
Hans-Henrik M
HH
D000818:Animals; D015152:Blotting, Northern; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D007758:Ear, Inner / Q000254:growth & development* / Q000378:metabolism*; D004622:Embryo, Mammalian; D005123:Eye / Q000378:metabolism; D015870:Gene Expression / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D023281:Genomics / Q000379:methods; D006257:Head / Q000033:anatomy & histology / Q000196:embryology; D007150:Immunohistochemistry / Q000379:methods; D017403:In Situ Hybridization / Q000379:methods; D051379:Mice; D008813:Mice, Inbred ICR; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism; D009925:Organ of Corti / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012313:RNA / Q000378:metabolism; D062085:RNA, Long Noncoding; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
C422882:MEG3 non-coding RNA, mouse; D011506:Proteins; D062085:RNA, Long Noncoding; D012333:RNA, Messenger; C089785:myosin VI; D012313:RNA; D018995:Myosin Heavy Chains