Axons rely on guidance cues to reach remote targets during nervous system development. A well-studied model system for axon guidance is the retinotectal projection. The retina can be divided into halves; the nasal half, next to the nose, and the temporal half. A subset of retinal axons, those from the temporal half, is guided by repulsive cues expressed in a graded fashion in the optic tectum, part of the midbrain. Here we report the cloning and functional characterization of a membrane-associated glycoprotein, which we call RGM (repulsive guidance molecule). This molecule shares no sequence homology with known guidance cues, and its messenger RNA is distributed in a gradient with increasing concentration from the anterior to posterior pole of the embryonic tectum. Recombinant RGM at low nanomolar concentration induces collapse of temporal but not of nasal growth cones and guides temporal retinal axons in vitro, demonstrating its repulsive and axon-specific guiding activity.

The Eph family is thought to exert its function through the complementary expression of receptors and ligands. Here, we show that EphA receptors colocalize on retinal ganglion cell (RGC) axons with EphA ligands, which are expressed in a high-nasal-to-low-temporal pattern. In the stripe assay, only temporal axons are normally sensitive for repellent axon guidance cues of the caudal tectum. However, overexpression of ephrinA ligands on temporal axons abolishes this sensitivity, whereas treatment with PI-PLC both removes ephrinA ligands from retinal axons and induces a striped outgrowth of formerly insensitive nasal axons. In vivo, retinal overexpression of ephrinA2 leads to topographic targeting errors of temporal axons. These data suggest that differential ligand expression on retinal axons is a major determinant of topographic targeting in the retinotectal projection.

Two ligands for Eph‐related receptor tyrosine kinases, RAGS and ELF‐1, have been implicated in the control of development of the retinotectal projection. Both molecules are expressed in overlapping gradients in the tectum, the target area of retinal ganglion cell axons. In two in vitro assays ELF‐1 is shown to have a repellent axon guidance function for temporal, but apparently not for nasal axons. RAGS on the other hand is repellent for both types of axons, though to different degrees. Thus, RAGS and ELF‐1 share some and differ in other properties. The biological activities of these molecules correlate with the strength of interaction with their receptors expressed on RGC axons. The meaning of these findings for guidance of retinal axons in the tectum is discussed.

A gene encoding a member of the growing family of regulatory WD-repeat proteins has been identified in Hydra vulgaris. About 80% of its deduced amino acids are identical to RACK1, which has recently been identified in rat as a receptor for activated C kinase. The presence of a consensus sequence believed to be important for interaction with protein kinase C, prompted us to term the new sequence HvRACK1 (Hydra vulgaris RACK1). In situ hybridization revealed an abundant message restricted to 80% of the body column with the strongest signal in the upper body half. Terminally differentiated structures (foot, tentacles and apicalmost hypostomal cells) were completely free of expression. A corresponding prepattern was established in bisected animals at least 10 h before the regenerating structure began to form. In normal animals HvRACK1 transcripts were contained mainly in interstitial cells (i-cells), gland cells and digestive epithelial cells. Depletion of i-cells and their derivatives by hydroxyurea (HU) treatment induced expression of the gene in epithelio-muscular and -digestive cells with the overall expression level remaining stable as confirmed by northern blotting. Polyps consisting almost exclusively of epithelial cells expressed HvRACK1 differentially along the body axis with a maximum in the head (except tentacles and the apicalmost ectoderm of the hypostome) and fading out towards the foot. The enhanced expression level in epithelial cells of HU-treated animals indicates that these might take over regionspecific HvRACK1 functions usually inherent to interstitial cells and certain derivatives of this lineage.