Diversity of ganglion cells in the mouse retina: unsupervised morphological classification and its limits

The dendritic structures of retinal ganglion cells in the mouse retina were visualized by particle-mediated transfer of DiI, microinjection of Lucifer yellow, or visualization of green fluorescent protein expressed in a transgenic strain. The cells were imaged in three dimensions and the morphologies of a series of 219 cells were analyzed quantitatively. A total of 26 parameters were studied and automated cluster analysis was carried out using the k-means methods. An effective clustering, judged by silhouette analysis, was achieved using three parameters: level of stratification, extent of the dendritic field, and density of branching. An 11-cluster solution is illustrated. The cells within each cluster are visibly similar along morphological dimensions other than those used statistically to form the clusters. They could often be matched to ganglion cell types defined by previous studies. For reasons that are discussed, however, this classification must remain provisional. Some steps toward more definitive methods of unsupervised classification are pointed out.