Characterization of different neuron populations in mouse statoacoustic ganglion cultures

Abstract

Statoacoustic ganglion (SAG) cells were grown in primary culture and the appearance of different neuronal phenotypes was investigated. Analysis criteria were shape, size and staining for the immunocytochemical markers: neurofilament proteins (NF-200 kDa), neuron-specific enolase (NSE), calretinin, a calcium-binding protein and substance P, a neurotransmitter. Cultures were prepared from dissociated SAG cells of 13 gestation-day-old mouse embryos. Neurons were identified and counted after 7 days in vitro. At this stage, neurons were organized in small clusters forming an extensive network of neurites grown on a layer of fibroblasts and glia. Most neurons identified by NF or NSE immunoreactivity showed a typical adult-like bipolar profile. The diameters of the neurons were between 5.62 and 17.00 μm and displayed a normal distribution (mean: 10.6 μm). Two distinct subpopulations were identified by the expression of calretinin and substance P. Calretinin-immunoreactive neurons were large and very rare and had a mean diameter of 11.3 μm; the distribution of substance P was more extensive than that of calretinin and identified a population of small neurons with a mean diameter of 8.9 μm. The distributions of these two markers in SAG cultures were consistent with in vivo results. In conclusion, dissociated SAG cell cultures appear to be a suitable model for analyzing the development of the immunocytochemical and functional characteristics of the neurons of this inner ear ganglio.