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Fluorescence Activated Cell Sorting (FACS) and Gene Expression Analysis of Fos-expressing Neurons from Fresh and Frozen Rat Brain Tissue
Published on: August 27, 2016
Granule cells in the cochlear nucleus sensitive to sound activation detected by Fos protein expression
Y Yang1, R L Saint Marie, D L Oliver
1Department of Neuroscience, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030-3401, USA.
Abstract:
Granule cells are the smallest neuronal type in the cochlear nucleus (CN). Due to their small size, it is extremely difficult to record their sound-evoked activity with microelectrodes. Compared with large, non-granule cells, much less is known about their response properties to sound stimulation. Here, we use Fos, the nuclear regulatory protein, as a neuronal activity marker to determine the responsiveness of granule cells to sound in comparison to the larger neurons. The present study determined the threshold sensitivity and activation pattern of neurons in the three subdivisions of the CN with free-field sound stimulation in monaural, awake rats. Immunocytochemical localization of Fos was used as our metric for "sound activation." Neuronal types upregulating Fos expression in response to sound stimulation were further identified with Nissl counterstaining. Our results show that most CN cell types can upregulate Fos expression when sound activated and the number of Fos-expressing neurons is directly related to sound intensity. The threshold for Fos activation in granule cells is lower than that for non-granule cells. The number of Fos activated granule cells saturates at high sound intensity, while the number of Fos activated non-granule cells is a monotonic function. By comparing the patterns of sound-induced Fos expression in different CN cell types, it may be possible to predict features of sound-evoked activity in granule cells.
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