Selective acquisition of individual cell types in the vestibular periphery for molecular biology studies.
OBJECTIVE
To develop a method for characterizing the transcriptome of individual cell types in the inner ear sensory epithelia.


METHODS
We employed the technique of laser capture microdissection to obtain enriched populations of hair cells and supporting cells. The respective mRNAs were extracted, reverse transcribed, and amplified using PCR.


RESULTS
We were able to isolate RNAs with good integrity from enriched cell populations obtained with laser capture microscopy and amplify specific mRNA targets.


CONCLUSIONS
We can now investigate the molecular differences between the different cell types in the inner ear sensory epithelia as identified by morphological criteria.


CONCLUSIONS
Analysis of gene expression profiles in the inner ear cell types has been hampered by the small size of this tissue and by the compact histoarchitecture of the sensory epithelia; however, the present technique offers new possibilities for the analysis of transcriptomes in the vestibular periphery using available high-throughput gene expression analysis methods.
Cristobal
Ricardo
R
Department of Otolaryngology and Communication Sciences, Medical College of Wisconsin, Milwaukee, WI 53226-3596, USA.
Wackym
P Ashley
PA
Cioffi
Joseph A
JA
Erbe
Christy B
CB
Popper
Paul
P
D000818:Animals; D004210:Dissection / Q000379:methods; D005240:Feasibility Studies; D006198:Hair Cells, Auditory / Q000473:pathology* / Q000502:physiology; D007760:Labyrinth Supporting Cells / Q000473:pathology* / Q000502:physiology; D053685:Laser Therapy / Q000379:methods*; D012333:RNA, Messenger; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger