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Double enzymatic staining of the ecmB-gal and ecmAO-gus fusion reporters revealed that some of the ALCs express the ecmB gene strongly and the ecmA gene very weakly (Jermyn et?al. 1996). These are called prestalk B (pstB) cells. PstB cells lie in the region where the slug contacts the substratum (Fig.?1A). During culmination, the pstB cells are sorted downward to reach the substratum (Fig.?1B), and some of them form the outer part of the basal disc while the others move upward to form the lower cup that provides subsidiary support for the spore mass. After prolonged slug migration, a portion of the pstB cells becomes enriched at the rear of the slug to form rear-guard cells. These cells terminally differentiate into vacuolated stalk [http://www.selleckchem.com/products/azd4547.html AZD4547 cost] cells once they are [http://en.wikipedia.org/wiki/INPP5D INPP5D] discarded into the slime sheath. Recently additional prestalk cell types, pstU cells and tip-organizer cells, have been defined. The pstU cells are intermingled within the upper cup cells, and the tip-organizer cells sit within the most anterior part of the pstA region (Fig.?1) and function as an organizer of slug behavior and integrity (Williams 2006; Yamada et?al. 2010). The prestalk cell-subtypes and expression patterns of the representative markers are summarized in Table?1. In contrast to the heterogeneity of the prestalk cells, prespore cells are a relatively homogenous population. They all have prespore vacuoles and they all differentiate into spores. There is, however, a proposed gradient of induction for cotC gene expression in the prespore region of the slug (Balint-Kurti et?al. 1998). Stalk differentiation can be divided into two steps; prestalk-cell differentiation followed by formation of mature, vacuolated stalk cells. The latter step can be induced in vitro with 8-Bromo-cAMP using cells taken from the prestalk region of slugs (Kwong et?al. 1988) (Maeda 1988; Inouye & Gross 1993) suggesting that DIF is not necessary for stalk maturation. Therefore, from a functional point of view, DIF can [http://www.selleckchem.com/products/Cisplatin.html Cisplatin] be regarded as a prestalk inducer, although it actually induces stalk cells in monolayer cells. In understanding DIF-dependent prestalk-cell differentiation, mutants that are defective in genes for DIF biosynthesis have provided many insights. The first characterized ��DIF-less�� mutant was created by inactivation of the methyl transferase gene, dmtA that catalyzes the addition of a methyl group in the terminal step in DIF biosynthesis (Thompson & Kay 2000). The mutant produces DIF at a very low level (