title: RT-PCR analysis of Tecta, Coch, Eya4 and Strc in mouse cochlear explants.
abstract: Tecta, Coch, Eya4 and Strc are mouse orthologs of four human deafness-associated genes. Their expression is markedly restricted to specific cell types in cochleae. Cochleae were dissected on embryonic day 15 and cultured in vitro. Relative messenger RNA abundance of each gene was quantified by RT-PCR and compared in-vivo cochleae of equivalent embryonic age. After 48 h in culture, in-vivo and explant Strc expression levels were equivalent, Eya4 level reduced in explanted tissues, and expression of Tecta and Coch did not show the expected temporal rise. Expression of these genes was detectable even after 96 h. These results suggest that it is feasible to test the expression of inner ear specific genes in explanted cochleae.
authors.affiliation: Molecular Otolaryngology Research Laboratory, Department of Otolaryngology, The University of Iowa Hospitals and Clinics, Iowa City, IA 52242, USA.
authors.forename: Yukihide
authors.initials: Y
authors.lastname: Maeda
authors.forename: Kunihiro
authors.initials: K
authors.lastname: Fukushima
authors.forename: Masashi
authors.initials: M
authors.lastname: Kakiuchi
authors.forename: Yorihisa
authors.initials: Y
authors.lastname: Orita
authors.forename: Kazunori
authors.initials: K
authors.lastname: Nishizaki
authors.forename: Richard J H
authors.initials: RJ
authors.lastname: Smith
mesh_terms: D000818:Animals; D003051:Cochlea / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis / Q000235:genetics*; D005136:Eye Proteins / Q000096:biosynthesis / Q000235:genetics*; D005333:Fetus; D058851:GPI-Linked Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D009924:Organ Culture Techniques; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics*
chemical_list: C491729:Coch protein, mouse; D016326:Extracellular Matrix Proteins; D005136:Eye Proteins; D058851:GPI-Linked Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D011506:Proteins; C440007:Strc protein, mouse; C105350:Tecta protein, mouse; D015534:Trans-Activators; D012313:RNA