A detailed procedure and dissection guide for the isolation of spiral ganglion cells of the guinea pig for electrophysiological experiments.
In the present study step-by-step instructions are provided for a preparative technique employed for the removal of the spiral ganglion from the inner ear of the guinea pig. Removal of the temporal bone is followed by opening of the bulla and excision of the modiolus. All major steps of the technique are illustrated with photographs. A procedure to obtain surviving, acutely separated spiral ganglion neurones is also described. By this procedure small tissue pieces are removed from the modiolus which contain the spiral ganglion neurones. The tissue fragments then undergo a mild enzyme treatment (collagenase and pronase). After the enzyme exposure, the tissue pieces are gently triturated, and the isolated cells are allowed to settle. Poly-D-lysine ensured the firm attachment of the spiral ganglion cells to the cover-slips. The application of this adhesive coating seemed to be desirable in functional studies when microelectrode techniques and/or rapid exchange of the extracellular solution were employed.
Szabó
Zsolt
Z
Department of Otolaryngology, BAZ County Infirmary, Szentpéteri kapu 72-76, Miskolc H-3529, Hungary.
Harasztosi
Csaba
C
Szûcs
Géza
G
Sziklai
István
I
Rusznák
Zoltán
Z
D000818:Animals; D002469:Cell Separation; D004210:Dissection / Q000379:methods*; D004594:Electrophysiology; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D005260:Female; D006168:Guinea Pigs; D007202:Indicators and Reagents; D008297:Male; D008566:Membranes / Q000502:physiology; D008853:Microscopy; D013136:Spiral Ganglion / Q000166:cytology*
D004791:Enzyme Inhibitors; D007202:Indicators and Reagents