The large number of different types of neurons in both vertebrate and invertebrate nervous systems raises the question of how the development of a particular neuronal cell type is specified. To pursue this question, Desai et al. (1988;Desai and Horvitz 1989) initiated a genetic analysis of the HSN serotonergic motor neurons in Caenorhabditis elegans. This analysis identified genes that are required for a number of distinct aspects of HSN development and placed these genes in a pathway of gene action. Some of these genes encode types of proteins that have been shown to control neuronal development in other organisms. Examples of such genes are egl-5, which encodes a homeo domain protein (Wang et al. 1993); unc-6, which encodes a member of the netrin family {Serafini et al. 1994); unc-33, which encodes a protein that is related to CRMP-62, a chick collapsin response mediator protein (Goshima et al. 1995); and unc-86, which encodes a POU homeo domain protein (Finney and Ruvkun 1990). This correlation between the types of proteins that control HSN development and the types of proteins that control neuronal development in other organisms suggests that study of HSN development will reveal general principles of how neuronal cell type is specified.~Corresponding author.The POU gene unc-86 is required for the normal pattern of HSN axonal outgrowth, normal HSN nuclear morphology, and HSN serotonin expression (Desai et al. 1988). POU homeo domain proteins have been shown to be required for neuronal development in both Drosophila (Bhat et al. 1995;Yeo et al. 1995) and mammals (for review, see Sharp and Morgan 1996). Mutation of the C. elegans gene sere-4 (sex muscle defective) causes the same spectrum of defects in HSN development as that caused by mutation of unc-86 (Desai et al. 1988). Because POU homeo domain proteins seem likely to play an evolutionarily conserved role in neuronal development, we are interested in determining how the SEM-4 and UNC-86 proteins interact in controlling HSN development. To understand how sem-4 functions in HSN development as well as in the development of other cell types, we have performed a genetic and molecular analysis of sere-4.