Cochlear whole mount in situ hybridization: identification of longitudinal and radial gradients.
The morphology of the organ of Corti has a radial asymmetry and also changes longitudinally from base to apex. Cellular localization of transcripts within the inner ear has relied primarily on the use of sectioned tissue with in situ hybridization. However, radial and longitudinal gradients of expression are not readily recognized using sectioned tissue owing to problems in visualization of signals with varying intensities. Herein, we describe the use of whole mount in situ hybridization for identification of cochlear longitudinal and radial expression gradients in the neurosensory epithelium, hair cells. Not only can these hair cell gradients be shown in adult tissues, but also the developmental up-regulation and down-regulation of genes and their associated spatio-temporal expression patterns can be demonstrated.
Judice
Tiffany N
TN
Center for Hereditary Communication Disorders, Boys Town National Research Hospital, Omaha, NE 68131, USA.
Nelson
Nathan C
NC
Beisel
Chase L
CL
Delimont
Duane C
DC
Fritzsch
Bernd
B
Beisel
Kirk W
KW
D000375:Aging / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D027321:Anion Transport Proteins; D016375:Antisense Elements (Genetics); D003051:Cochlea / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism*; D004210:Dissection / Q000295:instrumentation / Q000379:methods*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D006198:Hair Cells, Auditory / Q000166:cytology / Q000378:metabolism; D017403:In Situ Hybridization / Q000379:methods*; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D020409:Molecular Motor Proteins; D011506:Proteins / Q000235:genetics; D051381:Rats; D011978:Receptors, Nicotinic / Q000235:genetics; D000076988:Sulfate Transporters
D027321:Anion Transport Proteins; D016375:Antisense Elements (Genetics); D008565:Membrane Proteins; D020409:Molecular Motor Proteins; C118457:OTOF protein, human; C498720:Pres protein, mouse; D011506:Proteins; D011978:Receptors, Nicotinic; C409207:Slc26a5 protein, rat; D000076988:Sulfate Transporters; C443079:nAChR alpha9