Establishment of mice expressing EGFP in the placode-derived inner ear sensory cell lineage and FACS-array analysis focused on the regional specificity of the otocyst.
In this study, we established a novel enhanced green fluorescent protein (EGFP) reporter mouse line that enables the visualization of the placode-derived inner ear sensory cell lineage. EGFP was initially expressed in the otic placode and throughout its differentiation process into the inner ear sensory patches. At embryonic day 10.5 (E10.5), EGFP was expressed in the ventral and dorsomedial region of the otocyst. These regions could mainly give rise to the cochlea, including the organ of Corti, and the saccule, including the macula and the endolymphatic duct. The region could also give rise to cells that will develop as either prosensory cells or statoacoustic ganglion neuroblasts. By using this line and fluorescence-activated cell sorting (FACS)-array technology, we developed a new gene expression profile of the regional specificity of the otocyst. EGFP-positive regions include the Otx1-positive region, which could be clearly distinguished from EGFP-negative regions. The signal log ratio of microarray data showed high efficiency in predicting the genes expressed mainly in the ventral and/or dorsomedial otocyst and the data could be mined to uncover many novel genes involved in inner ear morphogenesis and cell fate regulation. Additionally, these data suggest that some novel genes enriched in EGFP-positive regions may be potentially involved in human congenital sensorineural hearing loss. This reporter line could play important roles in the use of animal models for detailed analysis of the differentiation process into the sensory patches and the identification of regional-specific gene networks and novel gene functions in the developing inner ear.
Department of Physiological Chemistry and Metabolism, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
Chisato
C
Fujimoto
Hidenori
H
Ozeki
Yasunobu
Y
Uchijima
Keigo
K
Suzukawa
Akihisa
A
Mitani
Shigetomo
S
Fukuhara
Koichi
K
Nishiyama
Yukiko
Y
Kurihara
Kenji
K
Kondo
Hiroyuki
H
Aburatani
Kimitaka
K
Kaga
Tatsuya
T
Yamasoba
Hiroki
H
Kurihara
D000818:Animals; D019070:Cell Lineage / Q000235:genetics*; D007758:Ear, Inner / Q000166:cytology / Q000196:embryology*; D005434:Flow Cytometry / Q000379:methods; D005456:Fluorescent Dyes / Q000378:metabolism; D017930:Genes, Reporter / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D006198:Hair Cells, Auditory / Q000166:cytology / Q000378:metabolism*; D018069:Hair Cells, Vestibular / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D044022:Receptor, Endothelin A / Q000096:biosynthesis / Q000235:genetics*; D011984:Sensory Receptor Cells / Q000166:cytology / Q000378:metabolism*
D005456:Fluorescent Dyes; D044022:Receptor, Endothelin A; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins