Gene expression changes during step-wise differentiation of embryonic stem cells along the inner ear hair cell pathway.
CONCLUSIONS
Our study outlines an alternative approach for the selection and investigation of genes involved in inner ear function.


OBJECTIVE
To gain understanding of the gene pathways involved in the development of the normal cochlea.


METHODS
Microarray technology currently offers the most efficient approach to investigate gene expression and identify pathways involved in cell differentiation. Epidermal growth factor (EGF) induces cultures derived from the organ of Corti to proliferate and produce new hair cells. Since pluripotent embryonic stem (ES) cells have the capacity to generate all tissues, we induced murine ES cells to differentiate towards ectodermal and neuroectodermal cell types and from there investigated their commitment towards the hair cell lineage in the presence of EGF. Cells were collected at three points along the differentiation pathway and their expression profiles were determined using the Soares NMIE mouse inner ear cDNA library printed in microarray format.


RESULTS
Three genes up-regulated after addition of EGF (serine (or cysteine) proteinase inhibitor, clade H, member 1 (Serpinh1), solute carrier family 2 (facilitated glucose transporter), member 10 (Slc2a10) and secreted acidic cysteine-rich glycoprotein (Sparc)) were selected for further analysis and characterization. Of the three genes, Serpinh1 and Slc2a10 have never been implicated in the hearing process.
De Silva
Michelle G
MG
Murdoch Childrens Research Institute, Royal Children's Hospital, Parkville, Vic 3052, Australia. michelle.desilva@mcri.edu.au
Hildebrand
Michael S
MS
Christopoulos
Helen
H
Newman
Michelle R
MR
Bell
Katrina
K
Ritchie
Matthew
M
Smyth
Gordon K
GK
Dahl
Hans-Henrik M
HH
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D002455:Cell Division / Q000235:genetics; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000502:physiology*; D051246:Glucose Transport Proteins, Facilitative / Q000235:genetics; D050958:HSP47 Heat-Shock Proteins / Q000235:genetics; D006199:Hair Cells, Auditory, Inner / Q000166:cytology*; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D009925:Organ of Corti / Q000166:cytology; D015676:Osteonectin / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000166:cytology*; D023822:Tissue Engineering; D015854:Up-Regulation / Q000235:genetics
D051246:Glucose Transport Proteins, Facilitative; D050958:HSP47 Heat-Shock Proteins; D015676:Osteonectin; C430746:SLC2A10 protein, human; C494661:Serpinh1 protein, mouse; D004815:Epidermal Growth Factor